Morphological and phylogenetic analyses reveal the taxonomic distinctiveness between Nymphaea pygmaea and N. tetragona (Nymphaeaceae)

Morphological and phylogenetic analyses reveal the taxonomic distinctiveness between Nymphaea pygmaea and N. tetragona (Nymphaeaceae) | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Morphological and phylogenetic analyses reveal the taxonomic distinctiveness between Nymphaea pygmaea and N. tetragona (Nymphaeaceae) Hanaka Naito, Syou Kato, Kohtaroh Shutoh, Takashi Shiga This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4414565/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In this study, we aimed to clarify the taxonomic identity between Nymphaea tetragona , which has a red to purple stigmatic disk and the widest distribution range among the genus, and Nymphaea pygmaea , which has a yellow stigmatic disk and is ordinarily treated as a synonym of N. tetragona , based on detailed morphological measurements and phylogenetic analyses. Through morphological analysis, we assessed 26 quantitative characters of 111 individuals from 15 populations. Seventeen characters were significantly different between the two species; however, these characters overlapped. Principal component analysis based on 13 morphological characters revealed different trends in the morphology of the two species. In DNA analysis, one nuclear ribosomal DNA region, internal transcribed spacer (ITS) region, and one chloroplast DNA region ( rps4-trnF ) were compared. The ITS region showed differences in 18–19 substitutions and two indels between N. pygmaea and N. tetragona . Additionally, the two species were closely related within the subgenus Nymphaea , and N. tetragona and Nymphaea leibergii formed a robust monophyletic group with N. pygmaea as a sister taxon. These results indicate that N. tetragona and N. pygmaea have differentiated morphologically and genetically, thus making these two species taxonomically distinct. aquatic plants internal transcribed spacer Japan Nymphaea leibergii rps4-trnF water lily Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Nymphaea L., commonly known as water lily, is a perennial floating-leaved plant distributed across all continents except Antarctica (Wiersema 1988). Among the six genera of Nymphaeaceae L., comprising approximately 60 species, Nymphaea is the largest genus, with 35–40 recognized species (Les 2017; Wiersema 1988). The genus is further divided into five subgenera: Anecphya (Casp.) Conard, Brachyceras (Casp.) Conard, Hydrocallis (Planch.) Conard, Lotos (DC.) Conard, and Nymphaea (Les 2017). The subgenus Nymphaea consists of seven species, namely Nymphaea alba L., Nymphaea candida J.Presl & C.Presl, Nymphaea leibergii Morong, Nymphaea loriana Wiersema, Hellquist & Borsch, Nymphaea mexicana Zucc., Nymphaea odorata Aiton, and Nymphaea tetragona Georgi (Borsch et al. 2014). These species exhibit cold tolerance and are distributed across subtropical to arctic zones (Slocum 2005). N. tetragona has the widest distribution within the genus (Wiersema 1988). Its key morphological characters include white petals and a tetragon flower bed receptacle (Georgi 1775). This species is known for its wide morphological variation (Borsch et al. 2014; Volkova and Shipunov 2007; Wiersema 1996), with several recognized infraspecific taxa, such as N. tetragona var. angusta Caspary, var. erythrostigmatica Koji Ito, and var. lata Caspary. Several taxa are recognized as synonyms, including N. crassifolia (Hand.-Mazz.) Nakai (Lee 1996), Nymphaea fennica Mela (Wiersema 1996), and Nymphaea pygmaea (Salisb.) W.T.Aiton (Caspary 1866; Fu et al. 2001; Lee 1996; Wiersema 1996). Among these, the taxonomic status of N. pygmaea is often debated in relation to N. tetragona , considering their morphological and flower color differences, distributional ranges, and phylogenetic relationships (Borsch et al. 2014; Dkhar 2010; Fu et al. 2001; Naito and Shiga 2024; Wiersema 1996). N. pygmaea was described by Richard Anthony Salisbury (1761–1829) as Castalia pygmaea Salisb. based on its small leaves (Salisbury 1805–1808). The plants considered as type materials were collected by William Kerr in China and introduced in 1805 from the ship Winchelsea, commanded by Capt. Campbell, under the auspices of the East India Company (Wiersema, 1996). Salisbury (1805–1808) noted that the transplanted plants had pygmy leaves, white petals, and slightly tetragon receptacles. The notable character of N. pygmaea is its yellow stigmatic disk and pollen sac (Salisbury 1805–1808), in contrast to N. tetragona in North America and Russia, which has red stigmatic disks (Borsch et al. 2014; Wiersema 1996). Subgenus Nymphaea comprises three sections: sect. Chamaenymphaea (Planch.) Wiersema, sect. Nymphaea , and sect. Xanthantha (Casp.) Wiersema. N. pygmaea serves as the type species for sect. Chamaenymphaea , including N. tetragona (Planchon 1853; Wiersema 1996). Caspary (1866) treated N. pygmaea as a synonym of N. tetragona without a detailed description. Subsequently, many taxonomists have followed this treatment (Lee 1996; Wiersema 1996). Fu et al. (2001) also treated N. pygmaea as a synonym of N. tetragona but noted differences in leaf thickness and distributional ranges. N. pygmaea has thicker leaves with impressed veins on the abaxial side, whereas N. tetragona has thinner leaves with raised veins on the abaxial side (Fu et al. 2001). Dakhr et al. (2010) performed a molecular phylogenetic analysis of Indian Nymphaea species, including N. tetragona . The study suggested that the Nymphaea specimen with a yellow-colored stigmatic disk is N. tetragona , and internal transcribed spacer (ITS) sequences revealed a close association with specimens from China rather than from Russia, although the location of the specimen in China remains unknown. N. pygmaea is distributed in China, Japan, and Vietnam, whereas N. tetragona occurs at latitudes above 42°N–50°N (Fu et al. 2001; Naito and Shiga 2024). Based on these results, Dakhr et al. (2010) suspected that the specimen could be N. pygmaea . Borsch et al. (2014) demonstrated that N. pygmaea and N. tetragona were phylogenetically distinct based on the molecular phylogenetic analysis of subgen. Nymphaea . This study relied solely on one cultivated N. pygmaea plant of unknown origin and performed no morphological examination. Thus, the previous studies may not be sufficient to clarify their taxonomic identities. In Japan, Ito (1968) classified plants with a yellow stigmatic disk as N. tetragona var. tetragona and described those with red to purple color as N. tetragona var. erythrostigmatica . These two varieties are widely recognized among Nymphaea species distributed in Japan (Kadono 1994, 2014; Shiga 2015). However, outside of Japan, particularly in Europe, North America, and Russia, only N. tetragona var. tetragona is recognized, without distinguishing infraspecific taxa based on stigmatic disk color (Volkova and Shipunov 2007; Wiersema 1996). Therefore, taxonomic classifications of N. tetragona , its infraspecific taxa, and its close relatives may be inconsistent, although detailed examination has not been conducted. In the present study, we aimed to clarify the taxonomic relationship between N. pygmaea and N. tetragona by comparing their quantitative morphological traits and conducting phylogenetic analyses based on nuclear ribosomal DNA (nrDNA) and chloroplast DNA (cpDNA) sequences. We used samples collected from various localities representing the known distribution areas of native Nymphaea species in Japan and Korea. Materials and methods Plant materials Nymphaea plants with yellow and red stigmatic disks were defined as N. pygmaea and N. tetragona , respectively (Fig. 1 ). Sampling was conducted from June to early October in 2019 and 2020 to cover the known distribution areas of native Nymphaea species in Japan and Korea (Kadono 1994, 2014; Lee 1996). N. pygmaea was collected from 34 populations spanning from Kyushu to Hokkaido, Japan, and one population in Korea. N. tetragona was obtained from six populations in Hokkaido, Japan (Fig. 2 and Table 1 ). Table 1 Population codes, coordinates, numbers of samples for morphological analyses (MA), ribo- and haplotypes, accession numbers of nrDNA (internal transcribed spacer [ITS] region) and cpDNA ( rps4–trnF region) sequences, and voucher information of 40 Nymphaea populations in Japan and Korea. One sample for each population was tested for DNA sequencing analysis. All voucher specimens were deposited to The Niigata University Herbarium (NGU). Pop. Code Sampling locality Coordinates Sequence type Accession no. MA Voucher Latitude Longitude ITS rps4-trnF ITS rps4-trnF Nymphaea pygmaea NP1 Ebino-shi, Miyazaki, Japan 32.05169 130.93358 nrN1 cpN1 LC775151 LC775191 10 NGU 11657 NP2 Sukumo-shi, Kochi, Japan 32.94522 132.78111 nrN1 cpN1 LC775152 LC775192 0 - NP3 Ureshino-shi, Saga, Japan 33.06744 129.97645 nrN2 cpN1 LC775153 LC775193 10 - NP4 Arita-cho, Saga, Japan 33.22014 129.87035 nrN2 cpN1 LC775154 LC775194 0 NGU 11655 NP5 Karatsu-shi, Saga, Japan 33.40537 130.10744 nrN1 cpN1 LC775155 LC775195 0 - NP6 Kama-shi, Fukuoka, Japan 33.54369 130.75550 nrN1 cpN1 LC775156 LC775196 0 - NP7 Chikujou-machi, Fukuoka, Japan 33.63237 131.04673 nrN1 cpN1 LC775157 LC775197 0 - NP8 Kitakyushu-shi, Fukuoka, Japan 33.78669 130.87847 nrN1 cpN1 LC775158 LC775198 0 - NP9 Imabari-shi, Ehime, Japan 33.99761 133.00780 nrN1 cpN1 LC775159 LC775199 0 NGU 11650 NP10 Nara-shi, Nara, Japan 34.60900 135.97352 nrN1 cpN1 LC775160 LC775200 0 - NP11 Ono-shi, Hyogo, Japan 34.87907 135.00076 nrN1 cpN1 LC775161 LC775201 10 NGU 8301 NP12 Iinan-cho, Shimane, Japan 35.01391 132.70516 nrN1 cpN2 LC775162 LC775202 0 NGU 8731 NP13 Taga-cho, Shiga, Japan 35.19983 136.30996 nrN1 cpN1 LC775163 LC775203 3 NGU 11317 NP14 Nasukarasuyama-shi, Tochigi, Japan 36.65837 140.11440 nrN1 cpN1 LC775164 LC775204 6 NGU 12378 NP15 Itoigawa-shi, Niigata, Japan 36.91476 137.90033 nrN1 cpN1 LC775165 LC775205 0 NGU 15565 NP16 Katashina-mura, Gunma, Japan 36.92194 139.21053 nrN1 cpN1 LC775166 LC775206 0 NGU 12948 NP17 Katashina-mura, Gunma, Japan 36.92575 139.22365 nrN1 cpN1 LC775167 LC775207 0 - NP18 Katashina-mura, Gunma, Japan 36.92834 139.22829 nrN1 cpN1 LC775168 LC775208 0 NGU 12954 NP19 Katashina-mura, Gunma, Japan 36.93177 139.23758 nrN1 cpN1 LC775169 LC775209 0 NGU 12952 NP20 Katashina-mura, Gunma, Japan 36.93417 139.22410 nrN1 cpN1 LC775170 LC775210 0 T. Shiga 11434 NP21 Katashina-mura, Gunma, Japan 36.94060 139.22847 nrN1 cpN1 LC775171 LC775211 0 NGU 12950 NP22 Tsunan-machi, Niigata, Japan 36.97462 138.59713 nrN1 cpN1 LC775172 LC775212 5 NGU 11675 NP23 Itoigawa-shi, Niigata, Japan 36.97462 138.59713 nrN1 cpN1 LC775173 LC775213 0 - NP24 Inawashiro-machi, Fukushima, Japan 37.51094 140.11169 nrN1 cpN1 LC775174 LC775214 0 NGU 11676 NP25 Goseong-gun, Gangwon-do, South Korea 38.25355 128.55626 nrN1 cpN1 LC775175 LC775215 6 NGU 8679 NP26 Murakami-shi, Yamagata, Japan 38.54518 140.36656 nrN1 cpN1 LC775176 LC775216 0 NGU 11672 NP27 Kami-machi, Miyagi, Japan 38.55432 140.68026 nrN1 cpN1 LC775177 LC775217 0 NGU 11659 NP28 Hanamaki-shi, Iwate, Japan 39.41820 141.18929 nrN1 cpN1 LC775178 LC775218 0 NGU 11681 NP29 Akita-shi, Akita, Japan 39.82303 140.08411 nrN1 cpN1 LC775179 LC775219 10 NGU 11155 NP30 Tsugaru-shi, Aomori, Japan 40.81180 140.27159 nrN1 cpN1 LC775180 LC775220 0 NGU 4952 NP31 Tsugaru-shi, Aomori, Japan 40.84641 140.30048 nrN1 cpN1 LC775181 LC775221 0 NGU 11670 NP32 Higashidori-mura, Aomori, Japan 41.29451 141.40436 nrN1 cpN1 LC775182 LC775222 8 NGU 11664 NP33 Atuma-cho, Hokkaido, Japan 42.67701 141.83335 nrN1 cpN1 LC775183 LC775223 0 NGU 11648 NP34 Ishikari-shi, Hokkaido, Japan 43.24029 141.34339 nrN1 cpN1 LC775184 LC775224 10 NGU 11678 N. tetragona NT1 Taiki-cho, Hokkaido, Japan 42.55410 143.47979 nrN3 cpN4 LC775185 LC775225 0 NGU 11700 NT2 Shibecha-cho, Hokkaido, Japan 43.14960 144.55105 nrN3 cpN4 LC775186 LC775226 6 NGU 11692 NT3 Nemuro-shi, Hokkaido, Japan 43.32029 145.61725 nrN3 cpN4 LC775187 LC775227 5 NGU 11712 NT4 Uryuu-cho, Hokkaido, Japan 43.69968 141.60428 nrN4 cpN3 LC775188 LC775228 2 NGU 11688 NT5 Kamikawa-cho, Hokkaido, Japan 43.93252 142.96418 nrN3 cpN3 LC775189 LC775229 10 NGU 11701 NT6 Sarufutsu-mura, Hokkaido, Japan 45.23455 142.21441 nrN3 cpN3 LC775190 LC775230 10 NGU 11694 Total 111 Morphological analyses For morphological analysis, leaves and flowers fixed in 70% ethanol (EtOH, v/v) from 3 to 10 individuals were sampled from 10 populations of N. pygmaea and five populations of N. tetragona , resulting in a total of 111 individuals from 15 populations being measured. We assessed 26 morphological characters, comprising 11 leaf characters and 15 flower characters, which have been previously considered important for the identification of subgenus Nymphaea (Salisbury 1805–1808; Wiersema 1996; Table 2 , Fig. S1 ). Characters L1–L6 were measured using ImageJ 1.53e (Schneider 2012) based on scanned images. Other characters were measured using a micrometer (L7 and L8), caliper (F1), and scale loupe (F2, F3, F5, F6, F8, F9, F10, F11, and F12). Table 2 Twenty-six morphological characters of Nymphaea pygmaea and Nymphaea tetragona investigated in this present study. Wilcoxon test (L1, L2, L3, L4, L5, L6, L11, F1, F3, F4, F7, F8, F9, and F13), Student’s t-test (L8, L9, L10, F2, F5, F6, F10, F11, F12, F14, and F15), and Welch’s t-test (L7) were subjected. Schematic images of the characters are shown in Fig. S2 Characters Nymphaea pygmaea (n = 78) N. tetragona (n = 33) p- value 2 PCA 3 Leaf L1: Leaf length (cm) 12.61 ± 4.05 11.48 ± 3.55 ns (5.54 − 23.61) (5.95 − 18.06) L2: Leaf width (cm) 10.55 ± 3.38 8.77 ± 2.96 ** (4.53 − 21.07) (4.26 − 14.78) L3: Leaf length from apex to base (cm) 6.83 ± 2.30 6.20 ± 2.04 *** + (2.92 − 12.61) (2.86 − 9.89) L4: L1-L3 (cm) 5.78 ± 1.80 5.29 ± 1.55 ns (2.47 − 11.01) (3.09 − 8.61) L5: Distance between lobe apexes (cm) 3.68 ± 1.42 2.17 ± 1.29 ns + (1.53 − 8.36) (0.69 − 5.03) L6: Overlapping length of lobes (cm) −0.40 ± 0.58 −0.07 ± 0.52 ** (− 2.32 − 1.00) (− 1.06 − 1.50) L7: Leaf thickness excluding leaf vein (mm) 0.337 ± 0.080 0.285 ± 0.057 *** + (0.148–0.560) (0.178 − 0.410) L8: Leaf thickness including leaf vein (mm) 0.421 ± 0.107 0.410 ± 0.117 ns + (0.158 − 0.700) (0.220 − 0.672) L9 1 : L2/L1 0.838 ± 0.044 0.757 ± 0.047 *** + (0.747 − 0.963) (0.648 − 0.853) L10: L5/L2 0.352 ± 0.088 0.239 ± 0.099 *** + (0.127 − 0.611) (0.073 − 0.514) L11: L3-L4 1.05 ± 0.73 0.91 ± 0.74 ns + (− 0.36 − 3.21) (− 0.41 − 2.79) Flower F1: Diagonal length of receptacle (cm) 11.49 ± 3.52 13.02 ± 3.43 * (4.80 − 24.55) (7.60 − 20.70) F2: Sepal length (mm) 32.6 ± 6.7 25.7 ± 5.2 *** + (16.8 − 55.5) (15.7 − 34.3) F3: Sepal width (mm) 15.2 ± 5.9 15.4 ± 4.0 *** + (4.3 − 40.1) (8.6 − 23.5) F4: Petal number 11.1 ± 2.6 16.4 ± 2.4 *** + (5 − 17) (12 − 21) F5: Petal length (mm) 29.4 ± 6.2 23.6 ± 4.7 *** (15.0 − 46.3) (15.0 − 32.6) F6: Petal width (mm) 10.9 ± 3.4 11.1 ± 2.9 ns (3.8 − 21.1) (6.8 − 16.7) F7: Stamen number 40.4 ± 9.3 61.6 ± 22.8 *** + (24 − 71) (32 − 116) F8: Stamen length (mm) 11.1 ± 2.0 8.9 ± 1.9 *** (7.2 − 17.6) (6.2 − 12.7) F9: Anther length (mm) 5.1 ± 1.2 3.6 ± 0.9 *** + (3.1 − 7.6) (1.8 − 5.4) F10: Maxmam filament width (mm) 2.2 ± 0.6 2.2 ± 0.4 ns (1.1 − 3.7) (1.5 − 2.9) F11: Length from maxmam filament width position to apex (mm) 7.4 ± 1.5 5.5 ± 1.2 *** (4.8 − 10.8) (3.6 − 7.5) F12: Carpellary appendage length (mm) 5.1 ± 5.6 4.8 ± 1.1 ns (1.5 − 4.2) (3.2 − 7.5) F13 1 : F3/F2 0.46 ± 0.11 0.60 ± 0.08 *** + (0.1 − 0.7) (0.4 − 0.8) F14: F9/F8 0.46 ± 0.05 0.40 ± 0.05 *** (0.34 − 0.59) (0.28 − 0.49) F15: F1×F1/2 72.1 ± 47.0 90.5 ± 47.9 ns (11.5 − 301.4) (28.9 − 214.2) 1 These characters underwent arcsine transformation 2 p-value are from statistical tests for each character (ns, p ≥ 0.05; *, < 0.05; **, < 0.01; ***, < 0.001) 3 “+” indicates that they were used for principal component analysis The following statistical analyses were conducted to compare morphological characters between the two Nymphaea groups: Wilcoxon test for L1, L2, L3, L4, L5, L6, L11, F1, F3, F4, F7, F8, F9, and F13; Student’s t-test for L8, L9, L10, F2, F5, F6, F10, F11, F12, F14, and F15; and Welch’s t-test for L7. Additionally, a preliminary principal component analysis (PCA) was conducted using 25 characters to select the morphological characters for the main PCA. When a character exhibited a high correlation (r > 0.8) with other characters, the one with the highest loading on the first three principal components was chosen. Characters with a low contribution (< 0.4) to the first three components were excluded. Consequently, 13 characters were retained for PCA (see Table 2 ). These statistical analyses were performed using JMP v.11.2.0 (SAS Institute Inc., Cary, North Carolina, USA). DNA analyses For DNA analyses, 40 individuals from 40 populations were used. Total genomic DNA was extracted from 5 × 5 mm sections of silica gel–dried leaves, taken from one individual in each population, using the cetyltrimethylammonium bromide method (Milligan 1992). One nrDNA region (ITS region: ITS1, 5.8S rRNA, and ITS2) and one cpDNA region ( rps4 - trnF region: rps4 - trnT spacer, trnT , trnT - trnL spacer, trnL 5ʹ-exon, trnL intron, trnL 3ʹ-exon, and trnL-trnF spacer) were amplified via PCR. Amplification primers for these regions were as follows: ITS5 and ITS4 (ITS region; White et al. 1990), trnL (USS) 5ʹ exonC and trnF (GAA) (Taberlet et al. 1991), rps4-5R (Sauquet et al. 2003), trnL110R (Borsch et al. 2003), and new primers (Nt_rps4-trnF_F1: CTTCTTCTTATAACTGGCAAGG, Nt_rps4-trnF_R1: ATTGAGAACACAAGACTTAGCT, and Nt_rps4-trnF_F2: CTTAACGCCTATGTAGTTACTA) designed using Primer3Plus (Untergasser et al. 2007) for the rps4 - trnF region (Table S1 ). Nt_rps4-trnF_F2 was used for direct sequencing. PCR was performed using a TaKaRa PCR Thermal Cycler TP650 Standard (TaKaRa Bio Inc., Tokyo, Japan) and an EmeraldAmp™ PCR Master Mix Kit (Takara Bio Inc., Tokyo, Japan). The PCR profile was as follows: one cycle at 94°C for 4 min (preliminary denaturation), followed by 35 cycles at 94°C for 1 min (denaturation), 53°C (ITS) and 52°C ( rps4-trnF ) for 1 min (annealing), 72°C for 2 min (extension), and a final step at 72°C for 7 min (final extension). PCR products were purified using the exoSAP method (Dugan et al. 2002). Purified PCR products were sequenced using a BigDye Terminator Cycle Sequencing Kit (Applied Biosystems, Foster, USA) with the same primers on an ABI Prism 3130 automated sequencer (Applied Biosystems, Foster, USA). Some samples were outsourced to Eurofin Genomics Co., Ltd. (Tokyo, Japan) DNA Sequence Service for sequencing. Chromatograms of nucleotide sequences were edited and assembled using Geneious software version 10.0.8 (Biomatters, Auckland, New Zealand). The resulting sequences were deposited in the DDBJ/GenBank/EMBL-Bank database (Table 1 ). To reveal the genetic relationships among N. pygmaea , N. tetragona , and closely related species, their haplotypes or genotypes were compared. In addition to the sequences determined in this study, 21 sequences of 12 Nymphaea taxa in the ITS and rps4-trnF regions were obtained from Borsch et al. (2014), Dakhr et al. (2010), and Woods et al. (2007) (Table S2). N. tetragona sequences with unknown stigmatic disk color were excluded from our analyses (e.g., accession nos.: AY707899, EU428056). Because the voucher specimen of N. tetragona (FJ597745) has a yellow stigmatic disk (Dakhr et al. 2010), it was treated as N. pygmaea . N. loriana was also excluded due to its hybrid origin. This species has two ITS region sequences, one matching N. leibergii and the other N. odorata (Borsch et al. 2014). The sequences of nrDNA (ITS region: 664–686 bp) and cpDNA ( rps4-trnF region: 1,678–1,679 bp) were aligned using MUSCLE ver. 3.5 (Edgar 2004). For phylogenetic analysis based on the ITS region, the sequences were trimmed at both ends, and the same range was used as accession no. “AY7718480 ( N. odorata )” (603–626 bp). Regarding the rps4-trnF region, a sequence range equivalent to the sequence trimmed at both ends (1–115 bp and 1,795–1,904 bp) of “LN_590041 ( N. tetragona )” was determined. Additionally, an insertion/deletion (indel) segment of simple sequence repeats, corresponding to 50,344–51,792 bp in “NC_006050 ( N. alba )”, was removed from this region. Following these procedures, a sequence range of 1,374–1,383 bp was obtained for phylogenetic analysis. To determine the phylogenetic position of N. pygmaea and N. tetragona within subgen. Nymphaea , Nymphaea amazonum Mart & Zucc., Nymphaea jamesoniana Planch., Nymphaea lingulata Wiersema, and Nymphaea novogranatensis Wiersema (subgen. Hydrocallis , Borsch et al. 2014) were selected as outgroup taxa. For analyzing the ITS region, the sequences of N. jamesoniana (FR717597) and N. lingulata (HG518070) were used. For analyzing the rps4-trnF region, the sequences of N. novogranatensis (AM422034) and N. amazonum (AM422026) were used. For phylogenetic analysis, samples representing a single haplotype were treated as a single operational taxonomic unit (OTU). For analyses of nrDNA and cpDNA, aligned data, excluding indels, consisted of 642 characters representing 12 OTUs from nine species in the ITS region and 1,401 characters representing 10 OTUs from nine species in the rps4-trnF region. These datasets were subjected to maximum likelihood (ML), neighbor-joining (NJ), and maximum parsimony (MP) analyses using MEGA 10.0.5 (Kumar et al., 2018). For ML analyses of nrDNA and cpDNA, K2 + G model (Kimura 2-parameter; Kimura, 1980) and T92 model (Tamura 3-parameter; Tamura, 1992) were employed, respectively. For NJ analyses of nrDNA and cpDNA, a T92 + G model was applied to the same alignment data. MP analyses of nrDNA and cpDNA were conducted using Subtree-Pruning-Regrafting (Nei and Kumar 2000). The robustness of each clade was tested using bootstrap analysis with 1,000 replications. Results Morphological analyses Six leaf characters (L2, L3, L6, L7, L9, and L10) and 11 flower characters (F1, F2, F3, F4, F5, F7, F8, F9, F11, F13, and F14) were significantly different between N. pygmaea and N. tetragona ( p < 0.05: Table 2 ). Regarding leaf shape (L9 = L2/L1), N. pygmaea exhibited wider floating leaves than N. tetragona ( N. pygmaea , L9 [L2/L1] = 0.838 ± 0.044 [0.747–0.963]; N. tetragona , 0.757 ± 0.047 [0.648–0.853]) (mean ± SD and [minimum–maximum values]) ( p < 0.001) (Table 2 ). In terms of flower characters, N. pygmaea displayed more elongated sepals than N. tetragona ( N. pygmaea , F13 [F3/F2] = 0.46 ± 0.11 [0.1–0.7]; N. tetragona , 0.60 ± 0.08 [0.4–0.8]) ( p < 0.001). N. pygmaea had fewer (F4 = 11.1 ± 2.6 [5–17]) and longer petals (F5 = 29.4 ± 6.2 mm [15.0–46.3]) than N. tetragona (F4 = 16.4 ± 2.4 [12–21]; F5 = 23.6 ± 4.7 mm [15.0–32.6]) ( p < 0.001). N. pygmaea also had fewer (F7 = 40.4 ± 9.3 [24–71]) and longer stamens (F8 = 11.1 ± 2.0 mm [7.2–17.6]) than N. tetragona (F7 = 61.6 ± 22.8 [32–116]; F8 = 8.90 ± 1.92 mm [6.20–1.92]) ( p < 0.001). Additionally, N. pygmaea had a higher ratio of anther length to total stamen length than N. tetragona ( N. pygmaea , F14 [F9/F8] = 0.46 ± 0.05 [0.34–0.59]; N. tetragona , 0.40 ± 0.05 [0.28–0.49]) ( p < 0.001). The PCA of 13 morphological characters (L3, L5, L7, L8, L9, L10, L11, F2, F3, F4, F7, F9, and F13) revealed well-defined morphological relationships between the two species (Fig. 3 ). Although Nymphaea plants were scattered continuously along principal component (PC) axis 1, the two species could be clearly distinguished on PC axis 2 (Fig. 3 a). The ranges of PCA values along axis 1 were − 4.584–7.195 (mean ± SD; 0.267 ± 2.321) for N. pygmaea and − 4.220–4.159 (− 0.630 ± 2.177) for N. tetragona . The ranges of PCA values along axis 2 were − 1.705–2.336 (0.971 ± 1.028) for N. pygmaea and − 3.841–0.352 (− 2.294 ± 0.810) for N. tetragona . Additionally, a scatter plot, averaging the PC scores for each population, showed a clear distinction between the two species on PC axis 2 (Fig. 3 b). The first three PCs accounted for 74.8% of the variance. PC axis 1 explained 40.9% of the total variance, attributed to the length from leaf tip to base (L3) and sepal width (F3) (Table S3). PC axis 2 explained 24.4% of the total variance, attributed to the number of petals (F4) and sepal shape (F13) (Table S3). DNA sequencing and phylogenetic analysis nrDNA analysis Among the ITS region sequences (684–686 bp) from 40 samples of Nymphaea plants, four ribotypes were distinguished (nrN1, nrN2, nrN3, and nrN4; Table 1 , Fig. 4 ). The ITS sequences of N. pygmaea were essentially identical (nrN1, n = 32), except for two samples from Saga Prefecture (Pref.) (NP3 and NP4; nrN2). Conversely, N. tetragona had nrN3 (n = 5), except for one sample from NT4 (nrN4). The sequence differences between N. pygmaea (nrN1 and nrN2) and N. tetragona (nrN3 and nrN4) were 18–19 substitutions and two indels (Table 3 ). The difference between nrN1 and nrN2 was one substitution, which was a heterozygous peak observed in the sequencing chromatogram in nrN2. nrN1 and nrN2 were similar to N. pygmaea sequences (nrN12, HG518076; nrN13, FJ597745), with zero to three substitutions and one indel between them. The difference between nrN3 and nrN4 was one substitution, which was a heterozygous peak in nrN4. nrN3 was identical to the ribotype of N. tetragona collected from Canada (HG518081). Table 3 Variable sites of four ribotypes of internal transcribed spacers (ITS 1 and 2) and 5.8S rRNA regions in Sect. Chamaenymphaea ( Nymphaea pygmaea , Nymphaea tetragona , and Nymphaea leibergii ). Numbers in parentheses after the sequence type indicate the number of samples 1 1 2 2 2 2 2 2 2 2 3 4 4 4 4 4 4 5 5 5 5 5 6 6 3 3 4 6 7 8 1 9 2 3 3 4 4 6 7 7 2 5 5 6 8 9 9 2 – 4 5 8 9 1 7 Species Ribotype 0 8 4 0 1 0 8 8 9 0 7 3 7 9 1 9 0 3 4 5 7 2 3 7 1 4 8 0 8 1 Nymphaea pygmaea nrN1 (32) T C A T T G A G - A A T - T - C C T A - C A C - C G G G T nrN2 (2) T C A T T A/G A G - A A T - T - C C T A - C A C - C G G G T nrN12 (1) T C A T T G A G - A A T - T - C C T A - C A C inserttion 2 C G G G T nrN13 (1) 1 T C A T C G A G - A A T - T - C T T A - C A C - C G G G T N. tetragona nrN3 (6) C C C G T G C A - G C C A C - C C C T C C C A - T C T T C nrN4 (1) C C C G T G C A - G C C A C - C/T C C T C C C A - T C T T C N. leibergii nrN6 (1) C T C T T G A G G G T C A C T C C T A C A C A - T C T T C 1 The sequence deposited for accession FJ597745 is shorter than the other ribotype sequences, lacking the terminal region from positions 1 to 13 observed in the other sequences 2 TTTTCGTCGGCGTTG The topologies of ML, NJ, and MP trees were identical (Fig. 4 , Supplementary Information 1). The phylogeny of subgenus Nymphaea comprised two clades. One clade included N. mexicana (nrN7 and nrN8) and N. odorata (nrN9, –nrN11), whereas the other consisted of five species (nrN1–nrN6, nrN12, and nrN13). Within the latter clade, two subclades were identified, comprising N. alba (nrN5) and N. candida (nrN5), as well as N. leibergii (nrN6), N. tetragona (nrN3 and nrN4), and N. pygmaea (nrN1, nrN2, nrN12, and nrN13). Regarding sect. Chamaenymphaea ( N. pygmaea , N. tetragona , and N. leibergii ), the section was monophyletic. In terms of inner relationships, N. tetragona and N. leibergii formed a monophyletic group, with N. pygmaea identified as their sister clade. All clades in these phylogenetic trees were supported by high bootstrap values (> 90%; Fig. 4 ). cpDNA analysis Among the rps4-trnF region from 40 Nymphaea samples, four haplotypes were distinguished (Tables 1 and S4). The rps4-trnF region of N. pygmaea was identical (cpN1, n = 34), except for one sample from Shimane Pref. (NP12; cpN2). N. tetragona exhibited two haplotypes (cpN3, n = 3; cpN4, n = 3). The differences among these four haplotypes were as follows: one substitution and two indels between cpN1 and cpN2, one substitution and one indel between cpN1 and cpN3 and between cpN2 and cpN4, two substitutions and one indel between cpN1 and cpN4, one indel between cpN2 and cpN3, and one substitution between cpN3 and cpN4 (Table S4). cpN1 was identical to N. pygmaea sequences (LN590038) deposited in DNA sequence databases. cpN3 and N. tetragona collected from Canada (HG518081) exhibited the same sequence. Among the cpDNA trees, the topologies of ML, NJ, and MP trees were nearly identical (Fig. S2, Supplementary Information 2). Similar to the ITS results, subgenus Nymphaea consisted of two clades: one included N. mexicana (cpN8 and cpN9) and N. odorata (cpN10–cpN12), whereas the other comprised five species, namely N. alba (cpN5), N. candida (cpN6), N. leibergii (cpN7), N. tetragona (cpN3 and cpN4), and N. pygmaea (cpN1 and cpN2). Although these cpDNA trees did not resolve the phylogenetic relationships among N. alba , N. candida , N. mexicana , and N. odorata , the monophyly among sect. Chamaenymphaea species ( N. leibergii , N. pygmaea , and N. tetragona ) was robust (Bootstrap values, 80–90%; Fig. S2). Discussion Taxonomic status of N. pygmaea and N. tetragona N. pygmaea and N. tetragona were morphologically and phylogenetically distinct (Figs. 3 and 4 and Tables 2 , 3 , and S4). In our nrDNA analysis, although we did not investigate reproductive isolation between the two taxa, we observed no individuals or populations with intermediate morphologies between the two species and no heterozygous peaks in the species-character sites of ITS region sequences. Moreover, the distinct distribution ranges of the two species suggest that they are reproductively well-isolated (Naito and Shiga 2024). Our results support the assertion that N. pygmaea should be treated as a distinct species and not a synonym of N. tetragona . However, one taxonomic issue remains unresolved: the type specimens of N. pygmaea have not been identified to date (Wiersema 1996). In our study, we were unable to examine the type specimens morphologically and genetically. To further clarify the taxonomic status of N. pygmaea and its variations, it is necessary to conduct comparisons using samples from southern China, which is considered the species’ type locality (Wiersema 1996). The morphological differences between the two species noted by Fu et al. (2001), namely stigmatic disk color and leaf thickness, are supported by the findings of the present study. Thus, the distribution of the two species in Eurasia and adjacent countries could be explained as suggested by Fu et al. (2010). In Japan, N. tetragona is unevenly distributed in the northern and eastern areas above 42°N in Hokkaido, with its distribution rarely overlapping with that of N. pygmaea , except in some areas. Only N. pygmaea is found south of Hokkaido (Naito and Shiga 2024). Consequently, a clear distributional boundary exists between the two species, indicating a high selective pressure that may have influenced each species’ distribution. This selection pressure could involve differences in ecological traits, particularly optimal growth temperature (Cox et al. 2016), and other factors such as pollinator and reproductive interference (Kuno 1992; Waser 1978). Comparative physiological, ecological, and population genetic studies of these two species can help elucidate the mechanisms underlying the formation of distribution patterns. Relationships with infraspecific taxa In the Korean peninsula, Nakai (1938) noted that N. tetragona is not distributed. He described the Korean Nymphaea as a variety, N. pygmaea var. minima Nakai (Nakai 1938), which he later reclassified as a species, Nymphaea minima (Nakai) Nakai (Nakai 1952). The key morphological characters of this taxon are primarily determined by leaf size (length = 1.8–5.5 cm, width = 1.6–4.3 cm) and flower size (sepal length = 1.7 cm, petal length = 1.5 cm), distinguishing it as smaller than N. pygmaea var. pygmaea (Nakai 1938). In the present study, small Nymphaea plants with yellow stigmatic disks were collected in Japan (leaf length = 2.9 cm, leaf width = 2.5 cm, sepal length = 1.4 cm, petal length = 1.3 cm [NGU 12124]) and Korea (leaf length = 4.8 cm, leaf width = 3.8 cm, sepal length = 2.0 cm, petal length = 1.5 cm [NGU 8679]); however, these populations also included larger individuals with a wide range of morphological variations, without genetic differences between Japanese and Korean N. pygmaea (Figs. 3 and 4 ). Therefore, it is reasonable to treat N. minima (= N. pygmaea var. minima Nakai) as a synonym of N. pygmaea . Caspary (1866) considered N. pygmaea to be synonymous with N. tetragona , and he described two infraspecific taxa, var. lata and var. angusta , without mentioning their stigmatic disk colors. var. lata was reported to be distributed in northern and eastern Asian regions (Southeastern Siberia, Kamchatka, and Manchuria), whereas var. angusta was found in China, Japan, and India (Caspary 1866). The distributional patterns of these two infraspecific taxa were similar to those of N. tetragona and N. pygmaea , respectively. According to Caspary (1866), the morphological differences between the two infraspecific taxa were in sepal and petal shapes (length: width = 1.16–2.7:1 [ lata ], 2.0–3.61:1 [ angusta ]) and leaf shape (leaf length: lobe length = 2.27:1 [ lata ], 1.9:1 [ angusta ]). However, the present study confirmed that sepal shapes (1.3–2.4:1 [ N. tetragona ]; 1.4–3.7:1 [ N. pygmaea ]) and leaf shapes (1.90–2.51:1 [ N. tetragona ]; 1.91–2.47:1 [ N. pygmaea ]) completely overlapped between the two species. Thus, var. lata and var. angusta did not correspond well to N. tetragona and N. pygmaea , respectively. Distinguishing the morphology between the two infraspecific taxa is difficult. Additionally, it remains unclear whether the two infraspecific taxa are synonymous with N. pygmaea and N. tetragona . In the present study, N. tetragona plants collected from both Canada and Japan exhibited identical DNA sequences. Our investigation of samples collected from the paratype locality of N. tetragona var. erythrostigmatica revealed DNA sequences and morphology identical to those of European N. tetragona var. tetragona , although N. tetragona var. erythrostigmatica plants are extinct in their holotype locality (Naito and Shiga 2021). Therefore, treating N. tetragona var. erythrostigmatica as synonymous with N. tetragona var. tetragona would be appropriate. Interspecific hybrids in subgen. Nymphaea Nymphaea species are known to readily crossbreed with each other, leading to many cultivated plants derived from artificial hybridization (Slocum 2005; Wiersema 1988). In subgenus Nymphaea , interspecific hybridization has been observed in areas where species distributions overlap, resulting in several reported hybrids: N. candida ( N. alba × N. tetragona ; Volkova et al. 2010), N. loriana ( N. leibergii × N. odorata ; Borsch et al. 2014), N. leibergii × N. tetragona , and N. mexicana × N. odorata (Borsch et al. 2014). In this study, hybrids between N. pygmaea and N. tetragona were not found. In Hokkaido Island, Japan, the two species grow allopatrically, except for one mire (Sasaki 2004, 2020). Naito and Shiga (2024) reported that the localities of collected specimens of N. pygmaea and N. tetragona were distinctly separated in Japan. Additionally, Fu et al. (2001) noted that N. tetragona was limited to boreal regions above 50°N latitude in North America and Europe. Even if the two species can cross, their hybrids may be rare due to their different distributional patterns. More detailed surveys regarding morphological variations and gene flows among the populations of these two species distributed at the boundaries of their distributional areas, as well as artificial crossing tests, are needed to confirm their isolation patterns and hybridization events. Taxonomic treatment Nymphaea pygmaea (Salisb.) W.T.Aiton, Hort. Kew., ed. 2, 3: 293 (1811)(Fig. 1 a–f). ≡ Castalia pygmaea Salisb., Parad. Lond. 1(2): 68 (1805–8). Type: unknown (see Wiersema 1996). = Nymphaea pygmaea var. minima Nakai, Bot. Mag. (Tokyo) 14(11): 746 (1938), syn. nov. ≡ Nymphaea minima (Nakai) Nakai, Bull. Nat. Sci. Mus., Tokyo 31: 25 (1952) ≡ Nymphaea tetragona var. minima (Nakai) W.Lee, Lineamenta Florae Koreae: 360 (1996). Holotype: South Korea; South Hwanghae Prov., 29 Jul. 1929, T. Nakai 12758 (TI!). = Nymphaea japono-koreana Nakai, J. Jap. Bot. 14: 751–752 (1938), nom. nud. =“ Nymphaea tetragona Georgi var. tetragona ” auct. non Georgi: Ohwi and Kitagawa, New Fl. Jap. 672–673 (1983); Kadono, Aquat. Pl. Jap. 109 (1994); Ito, Fl. Jap. 2a: 358 (2006); Choi H, Gen. Vas. Pl. Korea 161–162 (2007); Kadono, Fld. Guide Aquat. Pl. Jap. 50 (2014); Shiga, Wild Fl. Jap. 1: 48. pl. 37 (2015). Description : Herbs, perennial, aquatic. Rhizomes erect, cylindrical, unbranched, stolons absent (Fig. 1 f). Leaves dimorphic. Submerged leaves: leaf blade deltoid, sagittate, orbicular, 6–24 × 5–21 cm, membranous, margin entire, base cordate. Floating leaves: leaf blade obovate to orbicular, 5.5–24 × 4.5–21 cm, subcoriaceous, abaxially glabrous, base deeply cordate and basal lobes parallel to overlapping, thicker, 0.2–0.6 mm, impressed veins abaxially, margin entire (Fig. 1 a, e). Flowers floating, 3–6 cm in diameter (Fig. 1 c). Calyx insertion on receptacle tetragonal (Fig. 1 d). Sepals four, narrowly deltoid, 1.7–5.6 × 0.4–4.0 cm, obscurely veined, persistent (Fig. 1 b). Petals 5–17, white, broadly lanceolate, oblong, or narrowly obovate, 1.5–4.6 × 0.4–2.1 cm, transition to stamens gradual (Fig. 1 b). Stamens 24–71, 0.7–1.8 × 0.1–0.4 cm, yellow; filament of outer stamens wider than the anther, connective apically appendaged slightly (Fig. 1 b, c). Carpels completely united, walls between locules of the ovary single. Stigma rays 5–11, yellow (Fig. 1 c); carpellary appendages broadly lanceolate, 0.2–0.4 cm, yellow (Fig. 1 c). Fruit globose, 2.8–7.7 × 1.2–2.9 cm. Seeds ellipsoid to ovoid, 1.9–3.6 × 1.3–2.6 mm, smooth. Fl. Jun.–Oct. Regional name : Hitsuji-gusa (Japan: Ito 2016) and Kkoma-Suryeon (Korea: National Institute of Botanical Resources 2023) Distribution and habitat China, India, Japan, Korea, Nepal, and Vietnam. In ponds, lakes, marshes, moors, and paddy fields. Specimens examined : JAPAN. Hokkaido Pref.: Ebetsu-shi, 14 Jun. 2016, N. Nitta s.n. (SAPS); Iburi-no-kuni, Aug. 1893, Y. Tokubuchi s.n. (SAPS); 13 Jul. 1895, N. Date s.n. (SAPS); 21 Aug. 1899, J. Hanazawa s.n. (SAPS); 22 Aug. 1899, J. Hanazawa s.n. (SAPS); 17 Jun. 1975, A. Namimatsu s.n. (SAPS) (two specimens); 22 Jun. 1975, S. Nakayama s.n. (SAPS) (six specimens); Kuromatsunai-cho, 13 Jun. 1989, K. Ito s.n. (SAPS); Mukawa-cho, 17 Aug. 1964, Y. Takahashi s.n. (SAPS); 12 Jun. 1999, K. Oita 99-5930 (SAPS); 27 Aug. 2000, K. Oita 00-6048 (SAPS) (two specimens); 15 Jul. 2001, K. Oita 01-6609 (SAPS); Nanae-cho, 3 Aug. 1925, S. Miki s.n. (OSA230084); Oshamanbe-cho, 8 Aug. 2008, H. Takahashi s.n. (SAPS); Oshima-no-kuni, Aug. 1886, S. Nozawa s.n. (SAPS); 9 Jul. 1916, F. C. Grcatrex 189 (SAPS); Jun. 1917, S. Tanaka s.n. (SAPS); Sapporo-shi, 16 Aug. 1988, M. Hara 5758, 5990 (SAPS); 30 Aug. 1989, M. Hara 1256 (SAPS); Shiraoi-cho, 31 Jul. 1906, K. Saida 198 (TNS5549); 19 Aug. 1976, M. Hara 144 (SAPS); Tomakomai-shi, 29 Aug. 1969, M. Hara 132 (SAPS); 29 Aug. 1972, M. Hara 130 (SAPS); 28 Jun. 1974, S. Nakayama s.n. (SAPS); 27 Jun. 1987, J. Oginiawa s.n. (TNS988649); 17 Aug. 1989, Y. Kadono 6146 (OSA); 24 Aug. 1994, H. Takahashi 17548 (SAPS); 23 Aug. 1998, H. Takahashi 25789 (SAPS); Junsai-numa, 17 Aug. 1888, Y. Tokubuchi s.n. (SAPS). Aomori Pref.: Higashidori-mura, 28 Aug. 2019, H. Naito et al. 271, 272 (NGU11664, 11665); 19 Jun. 2022, J. Sasaki s.n. (NGU14396); 12 Aug. 2022 (S Kato s.n.), NGU14398; Tsugaru-shi, 26 Aug. 2019, H. Naito et al. 273 (NGU11668–11970). Iwate Pref.: Hanamaki-shi, 27 Aug. 2019, H. Naito et al. 267–269 (NGU11681–11684). Miyagi Pref.: Kami-machi, 28 Aug. 2019, H. Naito et al. 270 (NGU11659–11661). Akita Pref.: Akita-shi, 23 Aug. 2019, H. Naito et al. AM43, 44 (NGU11155, 11161, 11162, 11165). Yamagata Pref.: Murayama-shi, 28 Aug. 2019, S. Kato et al. TN176 (NGU10875). Fukushima Pref.: Inawashiro-machi, 23 Sep. 2019, S. Kato s.n. (NGU11676–11677). Tochigi Pref.: Nasukarasuyama-shi, 23 Sep. 2019, H. Naito et al. TN14, 15 (NGU12378–12380). Gunma Pref.: Katashina-mura, 10 Sep. 2019, H. Naito et al. 276, 277, 304, 306 (NGU12948–12955). Niigata Pref.: Tsunan-machi, 11 Aug. 2019, S. Kato et al. s.n. (NGU11675); Jouetsu-shi, 14 Sep. 2019, H. Naito et al. 222 (NGU11654, 11715). Gifu Pref.: Mizunami-shi, 7 Sep. 2020, H. Naito et al. 497, 498 (NGU11756, 11758). Aichi Pref.: Toyota-shi, 9 Sep. 2020, H. Naito et al. 601–604 (NGU11757, 11761, 11762). Mie Pref.: Tamaki-cho, 10 Sep. 2020, H. Naito et al. 662 (NGU11722). Shiga Pref.: Taga-cho, 7 Jul. 2019, T. Shiga 11317 (NGU10595, 10596). Hyogo Pref.: Ono-shi, 19 Sep. 2019, T. Shiga et al. 11229 (NGU8301, 8302). Shimane Pref.: Iinan-cho, 23 Jul. 2018, K. Tsubota & S. Midorikawa 232 (NGU 8731). Ehime Pref.: Imabari-shi, 22 Jul. 2019, Y. Fujiwara s.n. (NGU11650–11653). Saga Pref.: Arita-cho, 23 Jul. 2019, H. Naito et al. 228 (NGU11655, 11656). Miyazaki Pref.: Ebino-shi, 24 Jul. 2019, H. Naito et al. 227 (NGU11657–11658). KOREA. Gangwon-do: Goseong-gun, 8 Jun. 2019, T. Shiga et al. 10665, 10666 (NGU8679, 8680). Nymphaea tetragona Georgi, Bemerk. Reise Russ. Reich 1: 220 (1775)(Fig. 1 g–l). ≡ Castalia tetragona (Georgi) G. Lawson, Proc. & Trans. Roy. Soc. Canada 6, Sect. 4: 112 (1888). Lectotype: Angara, Siberia, Russia, Jul. 1772, J. Georgi (LE1009632!) (see Byalt and Krupkina 2017). = Nymphaea tetragona var. erythrostigmatica Koji Ito, J. Geobot. 16: 100 (1968), syn. nov.; Kadono, Aquat. Pl. Jap. 112 (1994); Kadono, Fld. Guide Aquat. Pl. Jap. 51 (2014); Shiga, Wild Fl. Jap. 1: 48 (2015). Holotype: Sarufutsu-mura, Kimomanuma, Japan, 21 Jul. 1967, Ko. Ito s.n. (SAPS72107!). = Nymphaea fennica Mela, Acta Soc. Fauna FI. Fenn. 14(3): 1 (1897). Syntype: Several syntypes at H (see Wiersema 1996). Description : Herbs, perennial, aquatic. Rhizomes erect, cylindrical, rarely branched; stolons absent (Fig. 1 l). Leaves dimorphic. Submerged leaves: leaf blade deltoid, sagittate, orbicular, 6–18 × 4–15 cm, membranous, margin entire, base cordate. Floating leaves: leaf blade cordate-ovate to ovate-elliptic, 6–18 × 4–15 cm, subcoriaceous, abaxially glabrous, base deeply cordate and basal lobes parallel to overlapping, thinner, 0.2–0.4 mm, raised veins abaxially, margin entire (Fig. 1 g, k). Flowers floating, 3–6 cm in diameter (Fig. 1 i). Calyx insertion on the receptacle prominently tetragonal (Fig. 1 j); sepals 4, broadly deltoid to narrowly ovate, 1.6–3.4 × 0.9–2.4 cm, veined, persistent (Fig. 1 h). Petals 12–21, white, rhombic to obovate, 1.5–3.3 × 0.7–1.7 cm, transition to stamens gradual (Fig. 1 h). Stamens 32–116, 0.6–1.3 × 0.2–0.3 cm, yellow–orange, abaxially usually tinged with red to purple in the inner stamens; filament of outer stamens wider than anther very prominent, connective apically appendaged slightly (Fig. 1 h, i). Carpels completely united, walls between locules of the ovary single. Stigma rays 4–12, dark red to purple (Fig. 1 i); carpellary appendages broadly lanceolate, 0.3–0.8 cm, yellow or dark red to purple, apically tinged with red to dark red (Fig. 1 i). Fruit globose, 2.6–3.7 × 1.0–2.2 cm. Seeds ellipsoid to ovoid, 2.5–2.9 × 1.5–1.9 mm, smooth. Fl. Jul.–Aug. Regional name : Pygmy water-lily (North America: Wiersema and Hellquist 1997), Small white water-lily (North America: Wiersema and Hellquist 1997), Nymphéa tetragonal (North America: Wiersema and Hellquist 1997), Shui lian (China: Fu et al. 2001), Ezo-Beni-Hitsuji-gusa (Japan: Ito 1968), and Suryeon (Korea: National Institute of Botanical Resources 2023) Distribution and habitat China, Japan, Russia; North America, Europe. In ponds, lakes, marshes, and moors. Specimens examined : JAPAN. Hokkaido Pref.: Akkeshi-cho, 14 Aug. 1925, S. Miki s.n. (OSA230085); Ikeda-cho, 20 Aug. 1934, H. Yokoyama s.n. (SAPS); Hamanaka-cho, 7 Aug. 2020, H. Naito et al. 398 (NGU11704); Kamikawa-cho, 14 Aug. 1947, M. Tatewaki 35570 (SAPS) (three specimens); 9 Jul. 1950, M. Kurashige 233 (SAPS); 1 Aug. 1967, K. Ito & A. Umezawa s.n. (SAPS) (two specimens); 2 Aug. 1967, K. Ito & A. Umezawa s.n. (SAPS); Kitami-shi, 10 Aug. 1925, collector unknown 231 (SAPS); 2 Aug. 1967, K. Ito & A. Umezawa s.n. (SAPS) (two specimens); 13 Aug. 2019, H. Naito et al. 214–216 (NGU11701–11703); 4 Jun. 2022, J. Sasaki s.n. (NGU14393, 14394); Kushiro-shi, 13 Aug. 1884, K. Miyabe s.n. (SAPS); 11 Jul. 1886, M. Nakamura B.149 (SAPS); 13 Aug. 1887, S Nogawa s.n. (SAPS); 10 Aug. 1951, M. Tatewaki s.n. (SAPS) (two specimens); 28 Jun. 1967, K. Seto 16511 (OSA); 25 Aug. 1975, Y. Kadono 1641 (OSA); 17 Jul. 1987, Y. Kadono 4689 (OSA); 7 Aug. 1991, Y. Kadono et al. 18 (OSA); 9 Aug. 1991, Y. Kadono et al. s.n. (OSA); Nemuro-shi, 28 Jul. 1928, M. Tatewaki & Y. Tokunaga s.n. (SAPS); 29 Aug. 2004, T. Shiga & S. Takebayashi 65 (OSA220192); 29 Aug. 2004, T. Shiga & S. Takebayashi 52 (OSA220147); collection date unknown, S. Yamane s.n. (SAPS); Sarufutsu-mura, 28 Jul. 1966, K. Ito s.n. (SAPS); 28 Jul. 1966, K. Ito s.n. (SAPS16635); 21 Jul. 1967, K. Ito s.n. (SAPS16636–16638); 21 Jul. 1967, K. Ito s.n. (SAPS);17 Jul. 1975, Y. Kadono 1977 (OSA); 21 Jul. 1987, Y. Kadono 4779 (OSA) (four specimens); 21 Jul. 1987, Y. Kadono 4784 (OSA); 5 Aug. 1998, H. Takahashi 25535 (SAPS15970); 26 Aug. 1989, J. Oginiawa 10643, 39380 (TNS960643, 989380); 15 Aug. 2019, H. Naito & T. Shiga 209–211, 301 (NGU11693–11696); 5 Aug. 2020, H. Naito et al. 281, 282, 558, 559 (NGU11711–11714, 11716); 5 Aug. 2020, H. Naito et al. 558, 559 (NGU11711–11714,11716); 6 Aug. 2020, H. Naito et al. 333, 356 (NGU11697, 11698); 11 Jul. 2022, J. Sasaki s.n. (NGU14395); Shibecha-cho, 9 Aug. 1969, N. Fujita 51 (TNS788665); 29 Aug. 1992, H. Takahashi & Y. Takashima 15143, 15162 (SAPS); 29 Jul. 2019, S. Kato s.n. (NGU11692); Taiki-cho, 28 Jul. 1979, E. Miki & M. Ito s.n. (TNS395745); 17 Jul. 1985, Y. Kadono & G. Wiegleb 859 (OSA) (two specimens); 22 Jun. 2019, K. Shutoh et al. 3681 (SAPS55004); 26 Jun. 2019, K Shutoh s.n. (NGU11700); 8 Aug. 2020, H. Naito et al. 429 (NGU11699); Takinoue-cho, 3 Aug. 1966, K. Ito s.n. (SAPS); Toyokoro-cho, 9 Aug. 2020, H Naito et al. 457, 560 (NGU11705–11708); Toyotomi-cho, 13 Jun. 1938, K. Miyabe & H. Yokoyama s.n. (SAPS); 2 Aug. 1968, K. Ito s.n. (SAPS); 16 Aug. 1980, K. Harusawa s.n. (OSA34528); Tsukigata-cho, 7 Aug. 2020, K. Katagiri K1, 2 (NGU11709, 11710); Uryu-cho, 8 Jul. 1998, H. Takahashi 25199 (SAPS16630); 23 Jul. 1998, H. Takahashi 25291, 25294, 25308 (SAPS16631, 16632, 16634); 12 Aug. 1998, H. Takahashi 25642 (SAPS); 6 Jun. 2004, M. Yamazaki s.n. (NGU7915); 6 Jun. 2004, M. Yamazaki s.n. (NGU); 12 Aug. 2009, M. Yamazaki s.n. (NGU13468) ; 23 Jul. 2019, K. Shutoh et al. 3658, 3659 (SAPS54921–54923); 11 Aug. 2020, H. Naito et al. 523–527, 532 (NUG11687–11691). Kurils: Furukamappu, 20 Aug. 1931, J. Ohwi 923 (TNS222126) (two specimens); Kunashiri, 26 Aug. 1923, M. Tatewaki 3411 (SAPS25573); 20 Aug. 1936, M. Tatewaki 25573 (SAPS); Tomari-mura, Aug. 1933, A. Tanaka s.n. (OSA230088). Sakhalin: Korsakov District, 29 Jul. 2001, H. Tatewaki 29516 (SAPS) (four specimens); Kusunnai, 21 Jun. 1906, T. Miyake s.n. (SAPS); Nyiwo, 15 Aug. 1922, Y. Kudo & M. Tatewaki 6584 (SAPS) (four specimens); Okhinsky District, 4 Aug. 2001, V.Y. Barkalov 10195 (SAPS); 4 Aug. 2001, T. Fukuda 1501 (SAPS); 11 Aug. 2001, T. Fukuda 1981 (SAPS); Shisuka-cho, 15 Aug. 1906, T. Miyake s.n. (SAPS); 29 Aug. 1929, collector unknown (TNS1066643); 1933, Y. Tanaka s.n. (OSA230087); 16 Aug. 1935, S. Sugawara 30155, 30157 (SAPS); collection date and collector unknown (SAPS) (two specimens); Shisuka-gun, 18 Jul. 1937, B. Yoshimura & M. Hara s.n. (TNS268313); Tominai-cho, 3 Aug. 1930, H. Otani & Y. Imai s.n. (SAPS); Ushiro-mura, 24 Jun. 1934, F. Fujita s.n. (OSA230086). Declarations Funding This study was partly funded by the Environment Research and Technology Development Fund (4-2001) of the Environmental Restoration and Conservation Agency and JST SPRING grant number JPMJSP2121. Author contributions All authors contributed to the study’s conception and design. Material preparation, data collection, and analysis were mainly performed by Hanaka Naito and Takashi Shiga. The first draft of the manuscript was written by Hanaka Naito, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Acknowledgments We thank the following people for their help with collecting plant samples: Dr. H. J. Choi, Mr. Y. Fujiwara, Mr. H. Kamiaka, Dr. Y. Kadono, Dr. K. Katagiri, Mr. Y. Komine, Dr. Y. Lee, Ms. E. Mashima, Mr. S. Midorikawa, Mr. J. Sasaki, Mr. T. Suzui, Dr. M. Suzuki, Mr. W. Takahashi, Mr. H. Tsurumaki, Mr. H. Yamada, Dr. T. Yamanouchi, and Ms. M. Yamazaki. Competing Interests: We declare that we have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. References Borsch T, Hilu KW, Quandt D, Wilde V, Neinhuis C, Barthlott W (2003) Noncoding plastid trnT-trnF sequences reveal a well resolved phylogeny of basal angiosperms. J Evol Biol 16:558–576. doi:10.1046/j.1420-9101.2003.00577.x Borsch T, Wiersema JH, Hellquist CB, Löhne C, Govers K (2014) Speciation in North American water lilies: Evidence for the hybrid origin of the newly discovered Canadian endemic Nymphaea loriana sp. nov. (Nymphaeaceae) in a past contact zone. Botany 92:867–882. doi:10.1139/cjb-2014-0060 Byalt VV, Krupkin LI (2017) Lectotypification of Nymphaea tetragona Georgi (Nymphaeaceae). Turczaninowia 20:182–186. doi: 10.14258/turczaninowia.20.1.14 Caspary R (1866) Nymphaeaceae. In: Miquel FAW (ed) Annales Musei Botanici Lugduno-Batavi, Leiden, Rijksherbarium, Netherlands, pp 241–256 Cox CB, Moore PD, Ladle RJ (2016) Biogeography: An ecological and evolutionary approach, 9th edn. Wiley, New York Dkhar J, Kumaria S, Rao SR, Tandon P (2010) Molecular phylogenetics and taxonomic reassessment of four Indian representatives of the genus Nymphaea . Aquat Bot 93:135–139. doi:10.1016/j.aquabot.2010.03.010 Dugan KA, Lawrence HS, Hares DR, Fisher CL, Budowle B (2002) An improved method for post-PCR purification for mtDNA sequence analysis. J Forensic Sci 47:811–818. doi:10.1520/JFS15459J Edgar RC (2004) MUSCLE: Multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797. doi:10.1093/nar/gkh340 Choi H (2007) Nymphaeaceae. In: Flora of Korea editorial committee (ed) The Genera of Vascular Plant of Korea. Academy Publishing Company, Seoul, pp 160–162 Fu D, Wiersema JH, Donald P (2001) Nymphaeaceae. In: Wu Z, Raven PH, Hong D (ed) Fl China, Missouri Botanical Garden, Missouri, pp 115–118 Georgi JG (1775) Bemerkungen einer Reise im Russischen Reich im Jahre 1772 vol. 1. Gedruckt bey der K Academie d Wissenschaften, St. Petersburg Ito K (1968) Observations on Northern Japanese plants (8). J Geobot 16:100–106 Kadono Y (1994) Aquatic plants of Japan. Bunichi Sogo Shuppan, Tokyo [in Japanese] Kadono Y (2014) A field guide to aquatic plants of Japan. Bunichi Sogo Shuppan, Tokyo [in Japanese] Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120. doi:10.1007/BF01731581 Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molec Biol Evol 35:1547–1549. doi:10.1093/molbev/msy096 Kuno E (1992) Competitive exclusion through reproductive interference. Res popul Eco 34:275–284. Doi: 10.1007/BF02514797 Lee WT (1996) Lineamenta florae Koreae. Academy Publishing, Seoul [in Korean] Les DH (2017) Aquatic dicotyledons of North America. CRC Press, Florida Milligan BG (1992) Plant DNA isolation. In: Hoelzel AR (ed) Molecular genetic analysis of populations: A practical approach. IRL Press, Oxford, pp 59–88 Naito H, Shiga T (2021) Disappearance of Nymphaea tetragona Georgi var. erythrostigmatica Koji Ito in the type locality, Lake Kimoma-numa, Japan. Bull Water Pl Soc Jap (111):41–44 Naito H, Shiga T (2024) Distribution of Nymphaea pygmaea and N. tetragona (Nymphaeaceae) in Hokkaido, Sakhalin island, and Kuril islands based on herbarium specimen records. J Asia-Pac Biodivers 17:81–86. doi:10.1016/j.japb.2023.10.010 Nakai T (1938) Notulae ad plantas Asiae orientalis (VI). J Jap Bot 14:734–751 Nakai T (1952) A synoptical sketch of Korean Flora. Bull Natl Sci Mus 31:1–153 National Institute of Botanical Resources (2023) National List of Korea. National Institute of Botanical Resources. https://www.kbr.go.kr/stat/ktsnfiledown/downpopup.do. Accessed 19 March 2024 Nei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, New York Ohwi J, Kitagawa M (1983) Nymphaeaceae. New flora of Japan. Shibundo, Tokyo, pp 669–673 [in Japanese] Planchon JE (1853) Etudes sur les Nymphaeacees. Ann Sci Nat Bot sér 3:17–63 Salisbury RA (1805–1808) The paradisus londinensis: Or coloured figures of plants cultivated in the vicinity of the metropolis. Shury, London Sasaki J (2004) Uyuu-numa no mizukusa [Aquatic plants of Uryuu-numa mire]. Hoppousansou (21):1–2 [in Japanese] Sasaki J (2022) Uyuu-numa shitsugen oyobi tozann-dou no ikannsoku-syokubutu risuto 2020 [List of Tracheophyte in the Uryuu-numa mire and mountain trail 2020]. Hoppousansou (39):79–91 [in Japanese] Sauquet H, Doyle JA, Scharaschkin T, Borsch T, Hilu KW, Chatrou LW, Le Thomas A (2003) Phylogenetic analysis of Magnoliales and Myristicaceae based on multiple data sets: Implications for character evolution. Bot J Linn Soc 142:125–186. doi:10.1046/j.1095-8339.2003.00171.x Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675. doi:10.1038/nmeth.2089 Shiga T (2015) Nymphaeaceae. In: Ohashi H, Kadota Y, Murata J, Yonekura K, Kihara H (ed) Wild Flowers of Japan Revised and Enlarged Edition 1. Heibonsha, Tokyo, pp 46–48 [in Japanese] Slocum PD (2005) Water lilies and lotuses. Timber Press, Seattle Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol Biol 17:1105–1109. doi:10.1007/BF00037152 Takakura K, Nishida S, Nishida T (2010) Reproductive interference and its implications for the biogeography and ecology of plants. Bunrui 10:151–162 [in Japanese] Tamura K (1992) Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G + C-content biases. Mol Biol Evol 9:678–687. doi:10.1093/oxfordjournals.molbev.a040752 Untergasser A, Nijveen H, Rao X, Bisseling T, Geurts R, Leunissen JA (2007) Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res 35 (Web server issue) (suppl_2):W71–W74. doi:10.1093/nar/gkm306 Volkova PA, Shipunov AB (2007) Morphological variation of Nymphaea (Nymphaeaceae) in European Russia. Nord J Bot 25:329–338. doi:10.1111/j.0107-055X.2007.00140.x Volkova PA, Trávníček P, Brochmann C (2010) Evolutionary dynamics across discontinuous freshwater systems: Rapid expansions and repeated allopolyploid origins in the Palearctic white water–lilies ( Nymphaea ). Taxon 59:483–494. doi:10.1002/tax.592013 Waser NM (1978) Interspecific pollen transfer and competition between co-occurring plant species. Oecologia 36:223–236.doi:10.1007/BF00349811 White TJ, Bruns T, Lee SJWT, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: A guide to methods and applications. Academic Press, New York, pp 315–322 Wiersema JH (1988) Reproductive biology of Nymphaea (Nymphaeaceae). Ann Mo Bot Gard 75:795–804. doi:10.2307/2399367 Wiersema JH (1996) Nymphaea tetragona and Nymphaea leibergii (Nymphaeaceae): Two species of diminutive water-lilies in North America. Brittonia 48:520–531. doi:10.2307/2807871 Wiersema JH, Hellquist CB (1997) Nymphaeaceae. In: Flora of North America Editorial Committee (eds) Flora North America North of Mexico vol. 3, Oxford University Press, Oxford, pp 66–77 Supplementary Files NymphaeapygmaeaSI2405132.pdf Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4414565","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":306152643,"identity":"1011d13f-4412-4046-be67-6c7eb44f72ac","order_by":0,"name":"Hanaka Naito","email":"","orcid":"","institution":"Niigata University: Niigata Daigaku","correspondingAuthor":false,"prefix":"","firstName":"Hanaka","middleName":"","lastName":"Naito","suffix":""},{"id":306152644,"identity":"7e378051-b557-4000-8670-d5639c15907b","order_by":1,"name":"Syou Kato","email":"","orcid":"","institution":"Niigata University: Niigata Daigaku","correspondingAuthor":false,"prefix":"","firstName":"Syou","middleName":"","lastName":"Kato","suffix":""},{"id":306152645,"identity":"6295e8dc-4b33-4c42-8028-1394d4ed6951","order_by":2,"name":"Kohtaroh Shutoh","email":"","orcid":"","institution":"Hokkaido University: Hokkaido Daigaku","correspondingAuthor":false,"prefix":"","firstName":"Kohtaroh","middleName":"","lastName":"Shutoh","suffix":""},{"id":306152646,"identity":"a5883fba-3d76-48c8-b107-7b3701bb74e8","order_by":3,"name":"Takashi Shiga","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEElEQVRIiWNgGAWjYDACCcYGBoYKBiDJ2HAASZyNgJYzcC0GxGgBYsY2kBYwMMCtEgbkZzc3v/g5z0a2Qbq58eCXmj/yDBIJjB9+MPDl4dJicOdgm2XvtjTjBpmDDYdljhkYNkgkMEv2MLAV49QikdhmwLvtcGKDRGLDYQk2A8b9NxIYpIF+SWzA5bAZiW2Gf+f8h2r5Z2APsuU3Pi0MNxKbH/M2HABrOfixzQDISGDDa4vBjcQ2ZpljycZtIFsY+4yTG3getln2GOD2i/yM9Mcf39TYyfZLABk/vsnZNrAnH77xo+IYzhADAjYJMAnEzDxgAVAcGRxLwKOF+QOMxfgDIVqDT8soGAWjYBSMLAAANTVbF47C6F4AAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0001-7292-4411","institution":"Niigata University: Niigata Daigaku","correspondingAuthor":true,"prefix":"","firstName":"Takashi","middleName":"","lastName":"Shiga","suffix":""}],"badges":[],"createdAt":"2024-05-13 16:51:48","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4414565/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4414565/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":58076550,"identity":"bf527fd1-9dbf-4350-a73b-7f890ffbb025","added_by":"auto","created_at":"2024-06-10 22:21:00","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":739748,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eNymphaea pygmaea\u003c/em\u003e (a–f) and \u003cem\u003eNymphaea tetragona\u003c/em\u003e (g–l). a, g, whole plants; b, h, sepals, petals, and stamens; c, i, flower; d, j, receptacle; e, k, veins of undersurface of leaf; f, l, vertical section of rhizome. Samples shown in e, h, and k were fixed with 70 % EtOH (v/v). Samples shown in f and l were from herbarium specimens NGU11660 and NGU11712, respectively\u003c/p\u003e","description":"","filename":"Fig1240513.png","url":"https://assets-eu.researchsquare.com/files/rs-4414565/v1/39b55576db1d9755e3a19241.png"},{"id":58076547,"identity":"19cba2ff-bac9-48a1-bba0-18f23d94e9f1","added_by":"auto","created_at":"2024-06-10 22:21:00","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":21527,"visible":true,"origin":"","legend":"\u003cp\u003eLocations of 40 populations of \u003cem\u003eNymphaea pygmaea\u003c/em\u003e and \u003cem\u003eNymphaea tetragona\u003c/em\u003e collected for morphological and genetic analyses\u003c/p\u003e","description":"","filename":"Fig2240514.png","url":"https://assets-eu.researchsquare.com/files/rs-4414565/v1/62de693566060d11913a182a.png"},{"id":58077157,"identity":"8076c311-fa8b-4fbd-acea-e6c3c550283d","added_by":"auto","created_at":"2024-06-10 22:29:00","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":44758,"visible":true,"origin":"","legend":"\u003cp\u003eResults of principal component analysis (PCA) of \u003cem\u003eNymphaea pygmaea\u003c/em\u003eand \u003cem\u003eNymphaea tetragona\u003c/em\u003e based on 11 morphological characters. Symbols indicate individual (a) or population (b) averages\u003c/p\u003e","description":"","filename":"Fig3240513.png","url":"https://assets-eu.researchsquare.com/files/rs-4414565/v1/4d4be537cc56f492f145fd36.png"},{"id":58076549,"identity":"97c88452-1929-4b50-84e2-03d87935bf8a","added_by":"auto","created_at":"2024-06-10 22:21:00","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":47430,"visible":true,"origin":"","legend":"\u003cp\u003eNeighbor-joining (NJ) tree based on 642 aligned characters of nuclear ribosomal DNA (internal transcribed spacer [ITS]1, 5.8S, and ITS2) of 15 ribotypes representing 10 \u003cem\u003eNymphaea\u003c/em\u003etaxa. Bootstrap values (\u0026gt;50 %) of each tree (maximum likelihood [ML], NJ, maximum parsimony [MP]) are shown above or under branches. Samples representing identical sequences in 642 characters of alignment matrix were treated as a single operational taxonomic unit. The classification of sections given by Wiersema (1996) are indicated on the right side of each clade. *: Including five samples from this study and one sequence from GenBank\u003c/p\u003e","description":"","filename":"Fig4240514.png","url":"https://assets-eu.researchsquare.com/files/rs-4414565/v1/f369c305ad88bde3d043e22b.png"},{"id":60947318,"identity":"a954e89e-79f3-453c-accf-0a16b58e5eb1","added_by":"auto","created_at":"2024-07-23 23:30:52","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2268166,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4414565/v1/aa9324cf-9c1a-4f9a-a5ad-542038e46790.pdf"},{"id":58076551,"identity":"cab78775-1885-4c40-b199-59844d606c32","added_by":"auto","created_at":"2024-06-10 22:21:00","extension":"pdf","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":616709,"visible":true,"origin":"","legend":"","description":"","filename":"NymphaeapygmaeaSI2405132.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4414565/v1/b643cb05da6c920d296f6681.pdf"}],"financialInterests":"","formattedTitle":"Morphological and phylogenetic analyses reveal the taxonomic distinctiveness between Nymphaea pygmaea and N. tetragona (Nymphaeaceae)","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eNymphaea\u003c/em\u003e L., commonly known as water lily, is a perennial floating-leaved plant distributed across all continents except Antarctica (Wiersema 1988). Among the six genera of Nymphaeaceae L., comprising approximately 60 species, \u003cem\u003eNymphaea\u003c/em\u003e is the largest genus, with 35\u0026ndash;40 recognized species (Les 2017; Wiersema 1988). The genus is further divided into five subgenera: \u003cem\u003eAnecphya\u003c/em\u003e (Casp.) Conard, \u003cem\u003eBrachyceras\u003c/em\u003e (Casp.) Conard, \u003cem\u003eHydrocallis\u003c/em\u003e (Planch.) Conard, \u003cem\u003eLotos\u003c/em\u003e (DC.) Conard, and \u003cem\u003eNymphaea\u003c/em\u003e (Les 2017). The subgenus \u003cem\u003eNymphaea\u003c/em\u003e consists of seven species, namely \u003cem\u003eNymphaea alba\u003c/em\u003e L., \u003cem\u003eNymphaea candida\u003c/em\u003e J.Presl \u0026amp; C.Presl, \u003cem\u003eNymphaea leibergii\u003c/em\u003e Morong, \u003cem\u003eNymphaea loriana\u003c/em\u003e Wiersema, Hellquist \u0026amp; Borsch, \u003cem\u003eNymphaea mexicana\u003c/em\u003e Zucc., \u003cem\u003eNymphaea odorata\u003c/em\u003e Aiton, and \u003cem\u003eNymphaea tetragona\u003c/em\u003e Georgi (Borsch et al. 2014). These species exhibit cold tolerance and are distributed across subtropical to arctic zones (Slocum 2005).\u003c/p\u003e \u003cp\u003e \u003cem\u003eN. tetragona\u003c/em\u003e has the widest distribution within the genus (Wiersema 1988). Its key morphological characters include white petals and a tetragon flower bed receptacle (Georgi 1775). This species is known for its wide morphological variation (Borsch et al. 2014; Volkova and Shipunov 2007; Wiersema 1996), with several recognized infraspecific taxa, such as \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003eangusta\u003c/em\u003e Caspary, var. \u003cem\u003eerythrostigmatica\u003c/em\u003e Koji Ito, and var. \u003cem\u003elata\u003c/em\u003e Caspary. Several taxa are recognized as synonyms, including \u003cem\u003eN. crassifolia\u003c/em\u003e (Hand.-Mazz.) Nakai (Lee 1996), \u003cem\u003eNymphaea fennica\u003c/em\u003e Mela (Wiersema 1996), and \u003cem\u003eNymphaea pygmaea\u003c/em\u003e (Salisb.) W.T.Aiton (Caspary 1866; Fu et al. 2001; Lee 1996; Wiersema 1996). Among these, the taxonomic status of \u003cem\u003eN. pygmaea\u003c/em\u003e is often debated in relation to \u003cem\u003eN. tetragona\u003c/em\u003e, considering their morphological and flower color differences, distributional ranges, and phylogenetic relationships (Borsch et al. 2014; Dkhar 2010; Fu et al. 2001; Naito and Shiga 2024; Wiersema 1996).\u003c/p\u003e \u003cp\u003e \u003cem\u003eN. pygmaea\u003c/em\u003e was described by Richard Anthony Salisbury (1761\u0026ndash;1829) as \u003cem\u003eCastalia pygmaea\u003c/em\u003e Salisb. based on its small leaves (Salisbury 1805\u0026ndash;1808). The plants considered as type materials were collected by William Kerr in China and introduced in 1805 from the ship Winchelsea, commanded by Capt. Campbell, under the auspices of the East India Company (Wiersema, 1996). Salisbury (1805\u0026ndash;1808) noted that the transplanted plants had pygmy leaves, white petals, and slightly tetragon receptacles. The notable character of \u003cem\u003eN. pygmaea\u003c/em\u003e is its yellow stigmatic disk and pollen sac (Salisbury 1805\u0026ndash;1808), in contrast to \u003cem\u003eN. tetragona\u003c/em\u003e in North America and Russia, which has red stigmatic disks (Borsch et al. 2014; Wiersema 1996). Subgenus \u003cem\u003eNymphaea\u003c/em\u003e comprises three sections: sect. \u003cem\u003eChamaenymphaea\u003c/em\u003e (Planch.) Wiersema, sect. \u003cem\u003eNymphaea\u003c/em\u003e, and sect. \u003cem\u003eXanthantha\u003c/em\u003e (Casp.) Wiersema. \u003cem\u003eN. pygmaea\u003c/em\u003e serves as the type species for sect. \u003cem\u003eChamaenymphaea\u003c/em\u003e, including \u003cem\u003eN. tetragona\u003c/em\u003e (Planchon 1853; Wiersema 1996). Caspary (1866) treated \u003cem\u003eN. pygmaea\u003c/em\u003e as a synonym of \u003cem\u003eN. tetragona\u003c/em\u003e without a detailed description. Subsequently, many taxonomists have followed this treatment (Lee 1996; Wiersema 1996). Fu et al. (2001) also treated \u003cem\u003eN. pygmaea\u003c/em\u003e as a synonym of \u003cem\u003eN. tetragona\u003c/em\u003e but noted differences in leaf thickness and distributional ranges. \u003cem\u003eN. pygmaea\u003c/em\u003e has thicker leaves with impressed veins on the abaxial side, whereas \u003cem\u003eN. tetragona\u003c/em\u003e has thinner leaves with raised veins on the abaxial side (Fu et al. 2001). Dakhr et al. (2010) performed a molecular phylogenetic analysis of Indian \u003cem\u003eNymphaea\u003c/em\u003e species, including \u003cem\u003eN. tetragona\u003c/em\u003e. The study suggested that the \u003cem\u003eNymphaea\u003c/em\u003e specimen with a yellow-colored stigmatic disk is \u003cem\u003eN. tetragona\u003c/em\u003e, and internal transcribed spacer (ITS) sequences revealed a close association with specimens from China rather than from Russia, although the location of the specimen in China remains unknown. \u003cem\u003eN. pygmaea\u003c/em\u003e is distributed in China, Japan, and Vietnam, whereas \u003cem\u003eN. tetragona\u003c/em\u003e occurs at latitudes above 42\u0026deg;N\u0026ndash;50\u0026deg;N (Fu et al. 2001; Naito and Shiga 2024). Based on these results, Dakhr et al. (2010) suspected that the specimen could be \u003cem\u003eN. pygmaea\u003c/em\u003e. Borsch et al. (2014) demonstrated that \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e were phylogenetically distinct based on the molecular phylogenetic analysis of subgen. \u003cem\u003eNymphaea\u003c/em\u003e. This study relied solely on one cultivated \u003cem\u003eN. pygmaea\u003c/em\u003e plant of unknown origin and performed no morphological examination. Thus, the previous studies may not be sufficient to clarify their taxonomic identities.\u003c/p\u003e \u003cp\u003eIn Japan, Ito (1968) classified plants with a yellow stigmatic disk as \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003etetragona\u003c/em\u003e and described those with red to purple color as \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003eerythrostigmatica\u003c/em\u003e. These two varieties are widely recognized among \u003cem\u003eNymphaea\u003c/em\u003e species distributed in Japan (Kadono 1994, 2014; Shiga 2015). However, outside of Japan, particularly in Europe, North America, and Russia, only \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003etetragona\u003c/em\u003e is recognized, without distinguishing infraspecific taxa based on stigmatic disk color (Volkova and Shipunov 2007; Wiersema 1996). Therefore, taxonomic classifications of \u003cem\u003eN. tetragona\u003c/em\u003e, its infraspecific taxa, and its close relatives may be inconsistent, although detailed examination has not been conducted.\u003c/p\u003e \u003cp\u003eIn the present study, we aimed to clarify the taxonomic relationship between \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e by comparing their quantitative morphological traits and conducting phylogenetic analyses based on nuclear ribosomal DNA (nrDNA) and chloroplast DNA (cpDNA) sequences. We used samples collected from various localities representing the known distribution areas of native \u003cem\u003eNymphaea\u003c/em\u003e species in Japan and Korea.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant materials\u003c/h2\u003e \u003cp\u003e \u003cem\u003eNymphaea\u003c/em\u003e plants with yellow and red stigmatic disks were defined as \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Sampling was conducted from June to early October in 2019 and 2020 to cover the known distribution areas of native \u003cem\u003eNymphaea\u003c/em\u003e species in Japan and Korea (Kadono 1994, 2014; Lee 1996). \u003cem\u003eN. pygmaea\u003c/em\u003e was collected from 34 populations spanning from Kyushu to Hokkaido, Japan, and one population in Korea. \u003cem\u003eN. tetragona\u003c/em\u003e was obtained from six populations in Hokkaido, Japan (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePopulation codes, coordinates, numbers of samples for morphological analyses (MA), ribo- and haplotypes, accession numbers of nrDNA (internal transcribed spacer [ITS] region) and cpDNA (\u003cem\u003erps4\u0026ndash;trnF\u003c/em\u003e region) sequences, and voucher information of 40 \u003cem\u003eNymphaea\u003c/em\u003e populations in Japan and Korea. One sample for each population was tested for DNA sequencing analysis. All voucher specimens were deposited to The Niigata University Herbarium (NGU).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePop. Code\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSampling locality\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eCoordinates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eSequence type\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eAccession no.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eVoucher\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLatitude\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLongitude\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eITS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003erps4-trnF\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eITS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cem\u003erps4-trnF\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNymphaea pygmaea\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEbino-shi, Miyazaki, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e32.05169\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e130.93358\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775151\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775191\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11657\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSukumo-shi, Kochi, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e32.94522\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e132.78111\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775152\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775192\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eUreshino-shi, Saga, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33.06744\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e129.97645\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775193\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eArita-cho, Saga, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33.22014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e129.87035\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775154\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775194\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11655\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKaratsu-shi, Saga, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33.40537\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e130.10744\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775155\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775195\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKama-shi, Fukuoka, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33.54369\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e130.75550\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775196\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eChikujou-machi, Fukuoka, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33.63237\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e131.04673\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775157\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775197\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKitakyushu-shi, Fukuoka, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33.78669\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e130.87847\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775158\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775198\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eImabari-shi, Ehime, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33.99761\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e133.00780\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775199\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11650\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNara-shi, Nara, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e34.60900\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e135.97352\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775160\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOno-shi, Hyogo, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e34.87907\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e135.00076\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775161\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 8301\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIinan-cho, Shimane, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e35.01391\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e132.70516\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775162\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775202\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 8731\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTaga-cho, Shiga, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e35.19983\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e136.30996\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775163\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775203\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11317\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNasukarasuyama-shi, Tochigi, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.65837\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140.11440\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775164\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775204\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 12378\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eItoigawa-shi, Niigata, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.91476\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e137.90033\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775165\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 15565\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKatashina-mura, Gunma, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.92194\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e139.21053\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775166\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775206\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 12948\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKatashina-mura, Gunma, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.92575\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e139.22365\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775207\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKatashina-mura, Gunma, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.92834\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e139.22829\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775168\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775208\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 12954\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKatashina-mura, Gunma, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.93177\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e139.23758\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775169\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775209\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 12952\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKatashina-mura, Gunma, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.93417\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e139.22410\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775170\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775210\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eT. Shiga 11434\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKatashina-mura, Gunma, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.94060\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e139.22847\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775171\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775211\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 12950\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTsunan-machi, Niigata, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.97462\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e138.59713\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775172\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775212\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11675\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eItoigawa-shi, Niigata, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e36.97462\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e138.59713\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775173\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775213\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eInawashiro-machi, Fukushima, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e37.51094\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140.11169\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775174\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775214\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11676\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGoseong-gun, Gangwon-do, South Korea\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e38.25355\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e128.55626\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775175\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 8679\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMurakami-shi, Yamagata, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e38.54518\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140.36656\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775176\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775216\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11672\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKami-machi, Miyagi, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e38.55432\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140.68026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775177\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775217\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11659\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHanamaki-shi, Iwate, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e39.41820\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e141.18929\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775178\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775218\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11681\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAkita-shi, Akita, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e39.82303\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140.08411\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775179\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775219\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11155\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTsugaru-shi, Aomori, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e40.81180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140.27159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775220\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 4952\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTsugaru-shi, Aomori, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e40.84641\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140.30048\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775181\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775221\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11670\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHigashidori-mura, Aomori, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e41.29451\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e141.40436\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775182\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775222\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11664\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAtuma-cho, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e42.67701\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e141.83335\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775223\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11648\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNP34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIshikari-shi, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e43.24029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e141.34339\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775184\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775224\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11678\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eN. tetragona\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNT1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTaiki-cho, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e42.55410\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e143.47979\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775185\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775225\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11700\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNT2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eShibecha-cho, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e43.14960\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e144.55105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775186\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775226\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11692\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNT3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNemuro-shi, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e43.32029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e145.61725\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775227\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11712\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNT4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eUryuu-cho, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e43.69968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e141.60428\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775188\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775228\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11688\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNT5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eKamikawa-cho, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e43.93252\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e142.96418\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775189\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775229\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11701\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNT6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSarufutsu-mura, Hokkaido, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e45.23455\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e142.21441\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003enrN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ecpN3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eLC775190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eLC775230\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eNGU 11694\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003e111\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eMorphological analyses\u003c/h2\u003e \u003cp\u003eFor morphological analysis, leaves and flowers fixed in 70% ethanol (EtOH, v/v) from 3 to 10 individuals were sampled from 10 populations of \u003cem\u003eN. pygmaea\u003c/em\u003e and five populations of \u003cem\u003eN. tetragona\u003c/em\u003e, resulting in a total of 111 individuals from 15 populations being measured. We assessed 26 morphological characters, comprising 11 leaf characters and 15 flower characters, which have been previously considered important for the identification of subgenus \u003cem\u003eNymphaea\u003c/em\u003e (Salisbury 1805\u0026ndash;1808; Wiersema 1996; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). Characters L1\u0026ndash;L6 were measured using ImageJ 1.53e (Schneider 2012) based on scanned images. Other characters were measured using a micrometer (L7 and L8), caliper (F1), and scale loupe (F2, F3, F5, F6, F8, F9, F10, F11, and F12).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTwenty-six morphological characters of \u003cem\u003eNymphaea pygmaea\u003c/em\u003e and \u003cem\u003eNymphaea tetragona\u003c/em\u003e investigated in this present study. Wilcoxon test (L1, L2, L3, L4, L5, L6, L11, F1, F3, F4, F7, F8, F9, and F13), Student\u0026rsquo;s t-test (L8, L9, L10, F2, F5, F6, F10, F11, F12, F14, and F15), and Welch\u0026rsquo;s t-test (L7) were subjected. Schematic images of the characters are shown in Fig. S2\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCharacters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eNymphaea pygmaea\u003c/em\u003e\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;78)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eN. tetragona\u003c/em\u003e\u003c/p\u003e \u003cp\u003e(n\u0026thinsp;=\u0026thinsp;33)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep- value\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePCA\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLeaf\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL1: Leaf length (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12.61\u0026thinsp;\u0026plusmn;\u0026thinsp;4.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.48\u0026thinsp;\u0026plusmn;\u0026thinsp;3.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(5.54\u0026thinsp;\u0026minus;\u0026thinsp;23.61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(5.95\u0026thinsp;\u0026minus;\u0026thinsp;18.06)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL2: Leaf width (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.55\u0026thinsp;\u0026plusmn;\u0026thinsp;3.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.77\u0026thinsp;\u0026plusmn;\u0026thinsp;2.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(4.53\u0026thinsp;\u0026minus;\u0026thinsp;21.07)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(4.26\u0026thinsp;\u0026minus;\u0026thinsp;14.78)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL3: Leaf length from apex to base (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.83\u0026thinsp;\u0026plusmn;\u0026thinsp;2.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.20\u0026thinsp;\u0026plusmn;\u0026thinsp;2.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(2.92\u0026thinsp;\u0026minus;\u0026thinsp;12.61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(2.86\u0026thinsp;\u0026minus;\u0026thinsp;9.89)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL4: L1-L3 (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.29\u0026thinsp;\u0026plusmn;\u0026thinsp;1.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(2.47\u0026thinsp;\u0026minus;\u0026thinsp;11.01)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(3.09\u0026thinsp;\u0026minus;\u0026thinsp;8.61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL5: Distance between lobe apexes (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.68\u0026thinsp;\u0026plusmn;\u0026thinsp;1.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.17\u0026thinsp;\u0026plusmn;\u0026thinsp;1.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(1.53\u0026thinsp;\u0026minus;\u0026thinsp;8.36)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(0.69\u0026thinsp;\u0026minus;\u0026thinsp;5.03)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL6: Overlapping length of lobes (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026minus;0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026minus;0.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e**\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(\u0026minus;\u0026thinsp;2.32\u0026thinsp;\u0026minus;\u0026thinsp;1.00)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(\u0026minus;\u0026thinsp;1.06\u0026thinsp;\u0026minus;\u0026thinsp;1.50)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL7: Leaf thickness excluding leaf vein (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.337\u0026thinsp;\u0026plusmn;\u0026thinsp;0.080\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.285\u0026thinsp;\u0026plusmn;\u0026thinsp;0.057\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0.148\u0026ndash;0.560)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(0.178\u0026thinsp;\u0026minus;\u0026thinsp;0.410)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL8: Leaf thickness including leaf vein (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.421\u0026thinsp;\u0026plusmn;\u0026thinsp;0.107\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.410\u0026thinsp;\u0026plusmn;\u0026thinsp;0.117\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0.158\u0026thinsp;\u0026minus;\u0026thinsp;0.700)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(0.220\u0026thinsp;\u0026minus;\u0026thinsp;0.672)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL9 \u003csup\u003e1\u003c/sup\u003e: L2/L1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.838\u0026thinsp;\u0026plusmn;\u0026thinsp;0.044\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.757\u0026thinsp;\u0026plusmn;\u0026thinsp;0.047\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0.747\u0026thinsp;\u0026minus;\u0026thinsp;0.963)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(0.648\u0026thinsp;\u0026minus;\u0026thinsp;0.853)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL10: L5/L2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.352\u0026thinsp;\u0026plusmn;\u0026thinsp;0.088\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.239\u0026thinsp;\u0026plusmn;\u0026thinsp;0.099\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0.127\u0026thinsp;\u0026minus;\u0026thinsp;0.611)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(0.073\u0026thinsp;\u0026minus;\u0026thinsp;0.514)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL11: L3-L4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(\u0026minus;\u0026thinsp;0.36\u0026thinsp;\u0026minus;\u0026thinsp;3.21)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(\u0026minus;\u0026thinsp;0.41\u0026thinsp;\u0026minus;\u0026thinsp;2.79)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eFlower\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF1: Diagonal length of receptacle (cm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.49\u0026thinsp;\u0026plusmn;\u0026thinsp;3.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e13.02\u0026thinsp;\u0026plusmn;\u0026thinsp;3.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(4.80\u0026thinsp;\u0026minus;\u0026thinsp;24.55)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(7.60\u0026thinsp;\u0026minus;\u0026thinsp;20.70)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF2: Sepal length (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e32.6\u0026thinsp;\u0026plusmn;\u0026thinsp;6.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25.7\u0026thinsp;\u0026plusmn;\u0026thinsp;5.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(16.8\u0026thinsp;\u0026minus;\u0026thinsp;55.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(15.7\u0026thinsp;\u0026minus;\u0026thinsp;34.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF3: Sepal width (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15.2\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;4.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(4.3\u0026thinsp;\u0026minus;\u0026thinsp;40.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(8.6\u0026thinsp;\u0026minus;\u0026thinsp;23.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF4: Petal number\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e16.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(5\u0026thinsp;\u0026minus;\u0026thinsp;17)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(12\u0026thinsp;\u0026minus;\u0026thinsp;21)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF5: Petal length (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29.4\u0026thinsp;\u0026plusmn;\u0026thinsp;6.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(15.0\u0026thinsp;\u0026minus;\u0026thinsp;46.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(15.0\u0026thinsp;\u0026minus;\u0026thinsp;32.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF6: Petal width (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e11.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(3.8\u0026thinsp;\u0026minus;\u0026thinsp;21.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(6.8\u0026thinsp;\u0026minus;\u0026thinsp;16.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF7: Stamen number\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e40.4\u0026thinsp;\u0026plusmn;\u0026thinsp;9.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e61.6\u0026thinsp;\u0026plusmn;\u0026thinsp;22.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(24\u0026thinsp;\u0026minus;\u0026thinsp;71)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(32\u0026thinsp;\u0026minus;\u0026thinsp;116)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF8: Stamen length (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e11.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(7.2\u0026thinsp;\u0026minus;\u0026thinsp;17.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(6.2\u0026thinsp;\u0026minus;\u0026thinsp;12.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF9: Anther length (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(3.1\u0026thinsp;\u0026minus;\u0026thinsp;7.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(1.8\u0026thinsp;\u0026minus;\u0026thinsp;5.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF10: Maxmam filament width (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(1.1\u0026thinsp;\u0026minus;\u0026thinsp;3.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(1.5\u0026thinsp;\u0026minus;\u0026thinsp;2.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF11: Length from maxmam filament width position to apex (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(4.8\u0026thinsp;\u0026minus;\u0026thinsp;10.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(3.6\u0026thinsp;\u0026minus;\u0026thinsp;7.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF12: Carpellary appendage length (mm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.1\u0026thinsp;\u0026plusmn;\u0026thinsp;5.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(1.5\u0026thinsp;\u0026minus;\u0026thinsp;4.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(3.2\u0026thinsp;\u0026minus;\u0026thinsp;7.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF13 \u003csup\u003e1\u003c/sup\u003e: F3/F2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0.1\u0026thinsp;\u0026minus;\u0026thinsp;0.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(0.4\u0026thinsp;\u0026minus;\u0026thinsp;0.8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF14: F9/F8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e***\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(0.34\u0026thinsp;\u0026minus;\u0026thinsp;0.59)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(0.28\u0026thinsp;\u0026minus;\u0026thinsp;0.49)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF15: F1\u0026times;F1/2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e72.1\u0026thinsp;\u0026plusmn;\u0026thinsp;47.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e90.5\u0026thinsp;\u0026plusmn;\u0026thinsp;47.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ens\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(11.5\u0026thinsp;\u0026minus;\u0026thinsp;301.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(28.9\u0026thinsp;\u0026minus;\u0026thinsp;214.2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e\u003csup\u003e1\u003c/sup\u003e These characters underwent arcsine transformation\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e\u003csup\u003e2\u003c/sup\u003e p-value are from statistical tests for each character (ns, p\u0026thinsp;\u0026ge;\u0026thinsp;0.05; *, \u0026lt; 0.05; **, \u0026lt; 0.01; ***, \u0026lt; 0.001)\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e\u003csup\u003e3\u003c/sup\u003e \u0026ldquo;+\u0026rdquo; indicates that they were used for principal component analysis\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe following statistical analyses were conducted to compare morphological characters between the two \u003cem\u003eNymphaea\u003c/em\u003e groups: Wilcoxon test for L1, L2, L3, L4, L5, L6, L11, F1, F3, F4, F7, F8, F9, and F13; Student\u0026rsquo;s t-test for L8, L9, L10, F2, F5, F6, F10, F11, F12, F14, and F15; and Welch\u0026rsquo;s t-test for L7. Additionally, a preliminary principal component analysis (PCA) was conducted using 25 characters to select the morphological characters for the main PCA. When a character exhibited a high correlation (r\u0026thinsp;\u0026gt;\u0026thinsp;0.8) with other characters, the one with the highest loading on the first three principal components was chosen. Characters with a low contribution (\u0026lt;\u0026thinsp;0.4) to the first three components were excluded. Consequently, 13 characters were retained for PCA (see Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). These statistical analyses were performed using JMP v.11.2.0 (SAS Institute Inc., Cary, North Carolina, USA).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eDNA analyses\u003c/h2\u003e \u003cp\u003eFor DNA analyses, 40 individuals from 40 populations were used. Total genomic DNA was extracted from 5 \u0026times; 5 mm sections of silica gel\u0026ndash;dried leaves, taken from one individual in each population, using the cetyltrimethylammonium bromide method (Milligan 1992). One nrDNA region (ITS region: ITS1, 5.8S rRNA, and ITS2) and one cpDNA region (\u003cem\u003erps4\u003c/em\u003e-\u003cem\u003etrnF\u003c/em\u003e region: \u003cem\u003erps4\u003c/em\u003e-\u003cem\u003etrnT\u003c/em\u003e spacer, \u003cem\u003etrnT\u003c/em\u003e, \u003cem\u003etrnT\u003c/em\u003e-\u003cem\u003etrnL\u003c/em\u003e spacer, \u003cem\u003etrnL\u003c/em\u003e 5ʹ-exon, \u003cem\u003etrnL\u003c/em\u003e intron, \u003cem\u003etrnL\u003c/em\u003e 3ʹ-exon, and \u003cem\u003etrnL-trnF\u003c/em\u003e spacer) were amplified via PCR. Amplification primers for these regions were as follows: ITS5 and ITS4 (ITS region; White et al. 1990), trnL (USS) 5ʹ exonC and trnF (GAA) (Taberlet et al. 1991), rps4-5R (Sauquet et al. 2003), trnL110R (Borsch et al. 2003), and new primers (Nt_rps4-trnF_F1: CTTCTTCTTATAACTGGCAAGG, Nt_rps4-trnF_R1: ATTGAGAACACAAGACTTAGCT, and Nt_rps4-trnF_F2: CTTAACGCCTATGTAGTTACTA) designed using Primer3Plus (Untergasser et al. 2007) for the \u003cem\u003erps4\u003c/em\u003e-\u003cem\u003etrnF\u003c/em\u003e region (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). Nt_rps4-trnF_F2 was used for direct sequencing.\u003c/p\u003e \u003cp\u003ePCR was performed using a TaKaRa PCR Thermal Cycler TP650 Standard (TaKaRa Bio Inc., Tokyo, Japan) and an EmeraldAmp\u0026trade; PCR Master Mix Kit (Takara Bio Inc., Tokyo, Japan). The PCR profile was as follows: one cycle at 94\u0026deg;C for 4 min (preliminary denaturation), followed by 35 cycles at 94\u0026deg;C for 1 min (denaturation), 53\u0026deg;C (ITS) and 52\u0026deg;C (\u003cem\u003erps4-trnF\u003c/em\u003e) for 1 min (annealing), 72\u0026deg;C for 2 min (extension), and a final step at 72\u0026deg;C for 7 min (final extension). PCR products were purified using the exoSAP method (Dugan et al. 2002). Purified PCR products were sequenced using a BigDye Terminator Cycle Sequencing Kit (Applied Biosystems, Foster, USA) with the same primers on an ABI Prism 3130 automated sequencer (Applied Biosystems, Foster, USA). Some samples were outsourced to Eurofin Genomics Co., Ltd. (Tokyo, Japan) DNA Sequence Service for sequencing. Chromatograms of nucleotide sequences were edited and assembled using Geneious software version 10.0.8 (Biomatters, Auckland, New Zealand). The resulting sequences were deposited in the DDBJ/GenBank/EMBL-Bank database (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTo reveal the genetic relationships among \u003cem\u003eN. pygmaea\u003c/em\u003e, \u003cem\u003eN. tetragona\u003c/em\u003e, and closely related species, their haplotypes or genotypes were compared. In addition to the sequences determined in this study, 21 sequences of 12 \u003cem\u003eNymphaea\u003c/em\u003e taxa in the ITS and \u003cem\u003erps4-trnF\u003c/em\u003e regions were obtained from Borsch et al. (2014), Dakhr et al. (2010), and Woods et al. (2007) (Table S2). \u003cem\u003eN. tetragona\u003c/em\u003e sequences with unknown stigmatic disk color were excluded from our analyses (e.g., accession nos.: AY707899, EU428056). Because the voucher specimen of \u003cem\u003eN. tetragona\u003c/em\u003e (FJ597745) has a yellow stigmatic disk (Dakhr et al. 2010), it was treated as \u003cem\u003eN. pygmaea\u003c/em\u003e. \u003cem\u003eN. loriana\u003c/em\u003e was also excluded due to its hybrid origin. This species has two ITS region sequences, one matching \u003cem\u003eN. leibergii\u003c/em\u003e and the other \u003cem\u003eN. odorata\u003c/em\u003e (Borsch et al. 2014). The sequences of nrDNA (ITS region: 664\u0026ndash;686 bp) and cpDNA (\u003cem\u003erps4-trnF\u003c/em\u003e region: 1,678\u0026ndash;1,679 bp) were aligned using MUSCLE ver. 3.5 (Edgar 2004). For phylogenetic analysis based on the ITS region, the sequences were trimmed at both ends, and the same range was used as accession no. \u0026ldquo;AY7718480 (\u003cem\u003eN. odorata\u003c/em\u003e)\u0026rdquo; (603\u0026ndash;626 bp). Regarding the \u003cem\u003erps4-trnF\u003c/em\u003e region, a sequence range equivalent to the sequence trimmed at both ends (1\u0026ndash;115 bp and 1,795\u0026ndash;1,904 bp) of \u0026ldquo;LN_590041 (\u003cem\u003eN. tetragona\u003c/em\u003e)\u0026rdquo; was determined. Additionally, an insertion/deletion (indel) segment of simple sequence repeats, corresponding to 50,344\u0026ndash;51,792 bp in \u0026ldquo;NC_006050 (\u003cem\u003eN. alba\u003c/em\u003e)\u0026rdquo;, was removed from this region. Following these procedures, a sequence range of 1,374\u0026ndash;1,383 bp was obtained for phylogenetic analysis.\u003c/p\u003e \u003cp\u003eTo determine the phylogenetic position of \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e within subgen. \u003cem\u003eNymphaea\u003c/em\u003e, \u003cem\u003eNymphaea amazonum\u003c/em\u003e Mart \u0026amp; Zucc., \u003cem\u003eNymphaea jamesoniana\u003c/em\u003e Planch., \u003cem\u003eNymphaea lingulata\u003c/em\u003e Wiersema, and \u003cem\u003eNymphaea novogranatensis\u003c/em\u003e Wiersema (subgen. \u003cem\u003eHydrocallis\u003c/em\u003e, Borsch et al. 2014) were selected as outgroup taxa. For analyzing the ITS region, the sequences of \u003cem\u003eN. jamesoniana\u003c/em\u003e (FR717597) and \u003cem\u003eN. lingulata\u003c/em\u003e (HG518070) were used. For analyzing the \u003cem\u003erps4-trnF\u003c/em\u003e region, the sequences of \u003cem\u003eN. novogranatensis\u003c/em\u003e (AM422034) and \u003cem\u003eN. amazonum\u003c/em\u003e (AM422026) were used.\u003c/p\u003e \u003cp\u003eFor phylogenetic analysis, samples representing a single haplotype were treated as a single operational taxonomic unit (OTU). For analyses of nrDNA and cpDNA, aligned data, excluding indels, consisted of 642 characters representing 12 OTUs from nine species in the ITS region and 1,401 characters representing 10 OTUs from nine species in the \u003cem\u003erps4-trnF\u003c/em\u003e region. These datasets were subjected to maximum likelihood (ML), neighbor-joining (NJ), and maximum parsimony (MP) analyses using MEGA 10.0.5 (Kumar et al., 2018). For ML analyses of nrDNA and cpDNA, K2\u0026thinsp;+\u0026thinsp;G model (Kimura 2-parameter; Kimura, 1980) and T92 model (Tamura 3-parameter; Tamura, 1992) were employed, respectively. For NJ analyses of nrDNA and cpDNA, a T92\u0026thinsp;+\u0026thinsp;G model was applied to the same alignment data. MP analyses of nrDNA and cpDNA were conducted using Subtree-Pruning-Regrafting (Nei and Kumar 2000). The robustness of each clade was tested using bootstrap analysis with 1,000 replications.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eMorphological analyses\u003c/h2\u003e \u003cp\u003eSix leaf characters (L2, L3, L6, L7, L9, and L10) and 11 flower characters (F1, F2, F3, F4, F5, F7, F8, F9, F11, F13, and F14) were significantly different between \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05: Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Regarding leaf shape (L9\u0026thinsp;=\u0026thinsp;L2/L1), \u003cem\u003eN. pygmaea\u003c/em\u003e exhibited wider floating leaves than \u003cem\u003eN. tetragona\u003c/em\u003e (\u003cem\u003eN. pygmaea\u003c/em\u003e, L9 [L2/L1]\u0026thinsp;=\u0026thinsp;0.838\u0026thinsp;\u0026plusmn;\u0026thinsp;0.044 [0.747\u0026ndash;0.963]; \u003cem\u003eN. tetragona\u003c/em\u003e, 0.757\u0026thinsp;\u0026plusmn;\u0026thinsp;0.047 [0.648\u0026ndash;0.853]) (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD and [minimum\u0026ndash;maximum values]) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In terms of flower characters, \u003cem\u003eN. pygmaea\u003c/em\u003e displayed more elongated sepals than \u003cem\u003eN. tetragona\u003c/em\u003e (\u003cem\u003eN. pygmaea\u003c/em\u003e, F13 [F3/F2]\u0026thinsp;=\u0026thinsp;0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11 [0.1\u0026ndash;0.7]; \u003cem\u003eN. tetragona\u003c/em\u003e, 0.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 [0.4\u0026ndash;0.8]) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). \u003cem\u003eN. pygmaea\u003c/em\u003e had fewer (F4\u0026thinsp;=\u0026thinsp;11.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6 [5\u0026ndash;17]) and longer petals (F5\u0026thinsp;=\u0026thinsp;29.4\u0026thinsp;\u0026plusmn;\u0026thinsp;6.2 mm [15.0\u0026ndash;46.3]) than \u003cem\u003eN. tetragona\u003c/em\u003e (F4\u0026thinsp;=\u0026thinsp;16.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4 [12\u0026ndash;21]; F5\u0026thinsp;=\u0026thinsp;23.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.7 mm [15.0\u0026ndash;32.6]) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). \u003cem\u003eN. pygmaea\u003c/em\u003e also had fewer (F7\u0026thinsp;=\u0026thinsp;40.4\u0026thinsp;\u0026plusmn;\u0026thinsp;9.3 [24\u0026ndash;71]) and longer stamens (F8\u0026thinsp;=\u0026thinsp;11.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0 mm [7.2\u0026ndash;17.6]) than \u003cem\u003eN. tetragona\u003c/em\u003e (F7\u0026thinsp;=\u0026thinsp;61.6\u0026thinsp;\u0026plusmn;\u0026thinsp;22.8 [32\u0026ndash;116]; F8\u0026thinsp;=\u0026thinsp;8.90\u0026thinsp;\u0026plusmn;\u0026thinsp;1.92 mm [6.20\u0026ndash;1.92]) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Additionally, \u003cem\u003eN. pygmaea\u003c/em\u003e had a higher ratio of anther length to total stamen length than \u003cem\u003eN. tetragona\u003c/em\u003e (\u003cem\u003eN. pygmaea\u003c/em\u003e, F14 [F9/F8]\u0026thinsp;=\u0026thinsp;0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05 [0.34\u0026ndash;0.59]; \u003cem\u003eN. tetragona\u003c/em\u003e, 0.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05 [0.28\u0026ndash;0.49]) (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003eThe PCA of 13 morphological characters (L3, L5, L7, L8, L9, L10, L11, F2, F3, F4, F7, F9, and F13) revealed well-defined morphological relationships between the two species (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Although \u003cem\u003eNymphaea\u003c/em\u003e plants were scattered continuously along principal component (PC) axis 1, the two species could be clearly distinguished on PC axis 2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea). The ranges of PCA values along axis 1 were \u0026minus;\u0026thinsp;4.584\u0026ndash;7.195 (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD; 0.267\u0026thinsp;\u0026plusmn;\u0026thinsp;2.321) for \u003cem\u003eN. pygmaea\u003c/em\u003e and \u0026minus;\u0026thinsp;4.220\u0026ndash;4.159 (\u0026minus;\u0026thinsp;0.630\u0026thinsp;\u0026plusmn;\u0026thinsp;2.177) for \u003cem\u003eN. tetragona\u003c/em\u003e. The ranges of PCA values along axis 2 were \u0026minus;\u0026thinsp;1.705\u0026ndash;2.336 (0.971\u0026thinsp;\u0026plusmn;\u0026thinsp;1.028) for \u003cem\u003eN. pygmaea\u003c/em\u003e and \u0026minus;\u0026thinsp;3.841\u0026ndash;0.352 (\u0026minus;\u0026thinsp;2.294\u0026thinsp;\u0026plusmn;\u0026thinsp;0.810) for \u003cem\u003eN. tetragona\u003c/em\u003e. Additionally, a scatter plot, averaging the PC scores for each population, showed a clear distinction between the two species on PC axis 2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe first three PCs accounted for 74.8% of the variance. PC axis 1 explained 40.9% of the total variance, attributed to the length from leaf tip to base (L3) and sepal width (F3) (Table S3). PC axis 2 explained 24.4% of the total variance, attributed to the number of petals (F4) and sepal shape (F13) (Table S3).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eDNA sequencing and phylogenetic analysis\u003c/h2\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003enrDNA analysis\u003c/h2\u003e \u003cp\u003eAmong the ITS region sequences (684\u0026ndash;686 bp) from 40 samples of \u003cem\u003eNymphaea\u003c/em\u003e plants, four ribotypes were distinguished (nrN1, nrN2, nrN3, and nrN4; Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The ITS sequences of \u003cem\u003eN. pygmaea\u003c/em\u003e were essentially identical (nrN1, n\u0026thinsp;=\u0026thinsp;32), except for two samples from Saga Prefecture (Pref.) (NP3 and NP4; nrN2). Conversely, \u003cem\u003eN. tetragona\u003c/em\u003e had nrN3 (n\u0026thinsp;=\u0026thinsp;5), except for one sample from NT4 (nrN4). The sequence differences between \u003cem\u003eN. pygmaea\u003c/em\u003e (nrN1 and nrN2) and \u003cem\u003eN. tetragona\u003c/em\u003e (nrN3 and nrN4) were 18\u0026ndash;19 substitutions and two indels (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The difference between nrN1 and nrN2 was one substitution, which was a heterozygous peak observed in the sequencing chromatogram in nrN2. nrN1 and nrN2 were similar to \u003cem\u003eN. pygmaea\u003c/em\u003e sequences (nrN12, HG518076; nrN13, FJ597745), with zero to three substitutions and one indel between them. The difference between nrN3 and nrN4 was one substitution, which was a heterozygous peak in nrN4. nrN3 was identical to the ribotype of \u003cem\u003eN. tetragona\u003c/em\u003e collected from Canada (HG518081).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eVariable sites of four ribotypes of internal transcribed spacers (ITS 1 and 2) and 5.8S rRNA regions in Sect. \u003cem\u003eChamaenymphaea\u003c/em\u003e (\u003cem\u003eNymphaea pygmaea\u003c/em\u003e, \u003cem\u003eNymphaea tetragona\u003c/em\u003e, and \u003cem\u003eNymphaea leibergii\u003c/em\u003e). Numbers in parentheses after the sequence type indicate the number of samples\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"33\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e 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colnum=\"30\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c31\" colnum=\"31\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c32\" colnum=\"32\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c33\" colnum=\"33\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c16\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c17\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c18\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c19\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c20\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c21\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c22\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c23\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c24\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c25\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c26\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c28\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c29\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c30\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c31\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c32\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c33\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c28\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRibotype\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c28\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eNymphaea pygmaea\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003enrN1 (32)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c28\" namest=\"c26\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003enrN2 (2)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eA/G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c28\" namest=\"c26\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003enrN12 (1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c28\" namest=\"c26\"\u003e \u003cp\u003einserttion\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003enrN13 (1)\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c28\" namest=\"c26\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eN. tetragona\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003enrN3 (6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c28\" namest=\"c26\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003enrN4 (1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003eC/T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c28\" namest=\"c26\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eN. leibergii\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003enrN6 (1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c28\" namest=\"c26\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c29\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c30\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c31\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c32\"\u003e \u003cp\u003eT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c33\"\u003e \u003cp\u003eC\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"33\"\u003e\u003csup\u003e1\u003c/sup\u003e The sequence deposited for accession FJ597745 is shorter than the other ribotype sequences, lacking the terminal region from positions 1 to 13 observed in the other sequences\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"33\"\u003e\u003csup\u003e2\u003c/sup\u003e TTTTCGTCGGCGTTG\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe topologies of ML, NJ, and MP trees were identical (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, Supplementary Information 1). The phylogeny of subgenus \u003cem\u003eNymphaea\u003c/em\u003e comprised two clades. One clade included \u003cem\u003eN. mexicana\u003c/em\u003e (nrN7 and nrN8) and \u003cem\u003eN. odorata\u003c/em\u003e (nrN9, \u0026ndash;nrN11), whereas the other consisted of five species (nrN1\u0026ndash;nrN6, nrN12, and nrN13). Within the latter clade, two subclades were identified, comprising \u003cem\u003eN. alba\u003c/em\u003e (nrN5) and \u003cem\u003eN. candida\u003c/em\u003e (nrN5), as well as \u003cem\u003eN. leibergii\u003c/em\u003e (nrN6), \u003cem\u003eN. tetragona\u003c/em\u003e (nrN3 and nrN4), and \u003cem\u003eN. pygmaea\u003c/em\u003e (nrN1, nrN2, nrN12, and nrN13). Regarding sect. \u003cem\u003eChamaenymphaea\u003c/em\u003e (\u003cem\u003eN. pygmaea\u003c/em\u003e, \u003cem\u003eN. tetragona\u003c/em\u003e, and \u003cem\u003eN. leibergii\u003c/em\u003e), the section was monophyletic. In terms of inner relationships, \u003cem\u003eN. tetragona\u003c/em\u003e and \u003cem\u003eN. leibergii\u003c/em\u003e formed a monophyletic group, with \u003cem\u003eN. pygmaea\u003c/em\u003e identified as their sister clade. All clades in these phylogenetic trees were supported by high bootstrap values (\u0026gt;\u0026thinsp;90%; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003ecpDNA analysis\u003c/h2\u003e \u003cp\u003eAmong the \u003cem\u003erps4-trnF\u003c/em\u003e region from 40 \u003cem\u003eNymphaea\u003c/em\u003e samples, four haplotypes were distinguished (Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and S4). The \u003cem\u003erps4-trnF\u003c/em\u003e region of \u003cem\u003eN. pygmaea\u003c/em\u003e was identical (cpN1, n\u0026thinsp;=\u0026thinsp;34), except for one sample from Shimane Pref. (NP12; cpN2). \u003cem\u003eN. tetragona\u003c/em\u003e exhibited two haplotypes (cpN3, n\u0026thinsp;=\u0026thinsp;3; cpN4, n\u0026thinsp;=\u0026thinsp;3). The differences among these four haplotypes were as follows: one substitution and two indels between cpN1 and cpN2, one substitution and one indel between cpN1 and cpN3 and between cpN2 and cpN4, two substitutions and one indel between cpN1 and cpN4, one indel between cpN2 and cpN3, and one substitution between cpN3 and cpN4 (Table S4). cpN1 was identical to \u003cem\u003eN. pygmaea\u003c/em\u003e sequences (LN590038) deposited in DNA sequence databases. cpN3 and \u003cem\u003eN. tetragona\u003c/em\u003e collected from Canada (HG518081) exhibited the same sequence.\u003c/p\u003e \u003cp\u003eAmong the cpDNA trees, the topologies of ML, NJ, and MP trees were nearly identical (Fig. S2, Supplementary Information 2). Similar to the ITS results, subgenus \u003cem\u003eNymphaea\u003c/em\u003e consisted of two clades: one included \u003cem\u003eN. mexicana\u003c/em\u003e (cpN8 and cpN9) and \u003cem\u003eN. odorata\u003c/em\u003e (cpN10\u0026ndash;cpN12), whereas the other comprised five species, namely \u003cem\u003eN. alba\u003c/em\u003e (cpN5), \u003cem\u003eN. candida\u003c/em\u003e (cpN6), \u003cem\u003eN. leibergii\u003c/em\u003e (cpN7), \u003cem\u003eN. tetragona\u003c/em\u003e (cpN3 and cpN4), and \u003cem\u003eN. pygmaea\u003c/em\u003e (cpN1 and cpN2). Although these cpDNA trees did not resolve the phylogenetic relationships among \u003cem\u003eN. alba\u003c/em\u003e, \u003cem\u003eN. candida\u003c/em\u003e, \u003cem\u003eN. mexicana\u003c/em\u003e, and \u003cem\u003eN. odorata\u003c/em\u003e, the monophyly among sect. \u003cem\u003eChamaenymphaea\u003c/em\u003e species (\u003cem\u003eN. leibergii\u003c/em\u003e, \u003cem\u003eN. pygmaea\u003c/em\u003e, and \u003cem\u003eN. tetragona\u003c/em\u003e) was robust (Bootstrap values, 80\u0026ndash;90%; Fig. S2).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003e \u003cb\u003eTaxonomic status of\u003c/b\u003e \u003cb\u003eN. pygmaea\u003c/b\u003e \u003cb\u003eand\u003c/b\u003e \u003cb\u003eN. tetragona\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eN. pygmaea and N. tetragona\u003c/em\u003e were morphologically and phylogenetically distinct (Figs.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e and Tables\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, and S4). In our nrDNA analysis, although we did not investigate reproductive isolation between the two taxa, we observed no individuals or populations with intermediate morphologies between the two species and no heterozygous peaks in the species-character sites of ITS region sequences. Moreover, the distinct distribution ranges of the two species suggest that they are reproductively well-isolated (Naito and Shiga 2024). Our results support the assertion that \u003cem\u003eN. pygmaea\u003c/em\u003e should be treated as a distinct species and not a synonym of \u003cem\u003eN. tetragona\u003c/em\u003e. However, one taxonomic issue remains unresolved: the type specimens of \u003cem\u003eN. pygmaea\u003c/em\u003e have not been identified to date (Wiersema 1996). In our study, we were unable to examine the type specimens morphologically and genetically. To further clarify the taxonomic status of \u003cem\u003eN. pygmaea\u003c/em\u003e and its variations, it is necessary to conduct comparisons using samples from southern China, which is considered the species\u0026rsquo; type locality (Wiersema 1996).\u003c/p\u003e \u003cp\u003eThe morphological differences between the two species noted by Fu et al. (2001), namely stigmatic disk color and leaf thickness, are supported by the findings of the present study. Thus, the distribution of the two species in Eurasia and adjacent countries could be explained as suggested by Fu et al. (2010). In Japan, \u003cem\u003eN. tetragona\u003c/em\u003e is unevenly distributed in the northern and eastern areas above 42\u0026deg;N in Hokkaido, with its distribution rarely overlapping with that of \u003cem\u003eN. pygmaea\u003c/em\u003e, except in some areas. Only \u003cem\u003eN. pygmaea\u003c/em\u003e is found south of Hokkaido (Naito and Shiga 2024). Consequently, a clear distributional boundary exists between the two species, indicating a high selective pressure that may have influenced each species\u0026rsquo; distribution. This selection pressure could involve differences in ecological traits, particularly optimal growth temperature (Cox et al. 2016), and other factors such as pollinator and reproductive interference (Kuno 1992; Waser 1978). Comparative physiological, ecological, and population genetic studies of these two species can help elucidate the mechanisms underlying the formation of distribution patterns.\u003c/p\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eRelationships with infraspecific taxa\u003c/h2\u003e \u003cp\u003eIn the Korean peninsula, Nakai (1938) noted that \u003cem\u003eN. tetragona\u003c/em\u003e is not distributed. He described the Korean \u003cem\u003eNymphaea\u003c/em\u003e as a variety, \u003cem\u003eN. pygmaea\u003c/em\u003e var. \u003cem\u003eminima\u003c/em\u003e Nakai (Nakai 1938), which he later reclassified as a species, \u003cem\u003eNymphaea minima\u003c/em\u003e (Nakai) Nakai (Nakai 1952). The key morphological characters of this taxon are primarily determined by leaf size (length\u0026thinsp;=\u0026thinsp;1.8\u0026ndash;5.5 cm, width\u0026thinsp;=\u0026thinsp;1.6\u0026ndash;4.3 cm) and flower size (sepal length\u0026thinsp;=\u0026thinsp;1.7 cm, petal length\u0026thinsp;=\u0026thinsp;1.5 cm), distinguishing it as smaller than \u003cem\u003eN. pygmaea\u003c/em\u003e var. \u003cem\u003epygmaea\u003c/em\u003e (Nakai 1938). In the present study, small \u003cem\u003eNymphaea\u003c/em\u003e plants with yellow stigmatic disks were collected in Japan (leaf length\u0026thinsp;=\u0026thinsp;2.9 cm, leaf width\u0026thinsp;=\u0026thinsp;2.5 cm, sepal length\u0026thinsp;=\u0026thinsp;1.4 cm, petal length\u0026thinsp;=\u0026thinsp;1.3 cm [NGU 12124]) and Korea (leaf length\u0026thinsp;=\u0026thinsp;4.8 cm, leaf width\u0026thinsp;=\u0026thinsp;3.8 cm, sepal length\u0026thinsp;=\u0026thinsp;2.0 cm, petal length\u0026thinsp;=\u0026thinsp;1.5 cm [NGU 8679]); however, these populations also included larger individuals with a wide range of morphological variations, without genetic differences between Japanese and Korean \u003cem\u003eN. pygmaea\u003c/em\u003e (Figs.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Therefore, it is reasonable to treat \u003cem\u003eN. minima\u003c/em\u003e (=\u0026thinsp;\u003cem\u003eN. pygmaea\u003c/em\u003e var. \u003cem\u003eminima\u003c/em\u003e Nakai) as a synonym of \u003cem\u003eN. pygmaea\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eCaspary (1866) considered \u003cem\u003eN. pygmaea\u003c/em\u003e to be synonymous with \u003cem\u003eN. tetragona\u003c/em\u003e, and he described two infraspecific taxa, var. \u003cem\u003elata\u003c/em\u003e and var. \u003cem\u003eangusta\u003c/em\u003e, without mentioning their stigmatic disk colors. var. \u003cem\u003elata\u003c/em\u003e was reported to be distributed in northern and eastern Asian regions (Southeastern Siberia, Kamchatka, and Manchuria), whereas var. \u003cem\u003eangusta\u003c/em\u003e was found in China, Japan, and India (Caspary 1866). The distributional patterns of these two infraspecific taxa were similar to those of \u003cem\u003eN. tetragona\u003c/em\u003e and \u003cem\u003eN. pygmaea\u003c/em\u003e, respectively. According to Caspary (1866), the morphological differences between the two infraspecific taxa were in sepal and petal shapes (length: width\u0026thinsp;=\u0026thinsp;1.16\u0026ndash;2.7:1 [\u003cem\u003elata\u003c/em\u003e], 2.0\u0026ndash;3.61:1 [\u003cem\u003eangusta\u003c/em\u003e]) and leaf shape (leaf length: lobe length\u0026thinsp;=\u0026thinsp;2.27:1 [\u003cem\u003elata\u003c/em\u003e], 1.9:1 [\u003cem\u003eangusta\u003c/em\u003e]). However, the present study confirmed that sepal shapes (1.3\u0026ndash;2.4:1 [\u003cem\u003eN. tetragona\u003c/em\u003e]; 1.4\u0026ndash;3.7:1 [\u003cem\u003eN. pygmaea\u003c/em\u003e]) and leaf shapes (1.90\u0026ndash;2.51:1 [\u003cem\u003eN. tetragona\u003c/em\u003e]; 1.91\u0026ndash;2.47:1 [\u003cem\u003eN. pygmaea\u003c/em\u003e]) completely overlapped between the two species. Thus, var. \u003cem\u003elata\u003c/em\u003e and var. \u003cem\u003eangusta\u003c/em\u003e did not correspond well to \u003cem\u003eN. tetragona\u003c/em\u003e and \u003cem\u003eN. pygmaea\u003c/em\u003e, respectively. Distinguishing the morphology between the two infraspecific taxa is difficult. Additionally, it remains unclear whether the two infraspecific taxa are synonymous with \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eIn the present study, \u003cem\u003eN. tetragona\u003c/em\u003e plants collected from both Canada and Japan exhibited identical DNA sequences. Our investigation of samples collected from the paratype locality of \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003eerythrostigmatica\u003c/em\u003e revealed DNA sequences and morphology identical to those of European \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003etetragona\u003c/em\u003e, although \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003eerythrostigmatica\u003c/em\u003e plants are extinct in their holotype locality (Naito and Shiga 2021). Therefore, treating \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003eerythrostigmatica\u003c/em\u003e as synonymous with \u003cem\u003eN. tetragona\u003c/em\u003e var. \u003cem\u003etetragona\u003c/em\u003e would be appropriate.\u003c/p\u003e \u003cp\u003e \u003cb\u003eInterspecific hybrids in subgen.\u003c/b\u003e \u003cb\u003eNymphaea\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eNymphaea\u003c/em\u003e species are known to readily crossbreed with each other, leading to many cultivated plants derived from artificial hybridization (Slocum 2005; Wiersema 1988). In subgenus \u003cem\u003eNymphaea\u003c/em\u003e, interspecific hybridization has been observed in areas where species distributions overlap, resulting in several reported hybrids: \u003cem\u003eN. candida\u003c/em\u003e (\u003cem\u003eN. alba\u003c/em\u003e \u0026times; \u003cem\u003eN. tetragona\u003c/em\u003e; Volkova et al. 2010), \u003cem\u003eN. loriana\u003c/em\u003e (\u003cem\u003eN. leibergii\u003c/em\u003e \u0026times; \u003cem\u003eN. odorata\u003c/em\u003e; Borsch et al. 2014), \u003cem\u003eN. leibergii\u003c/em\u003e \u0026times; \u003cem\u003eN. tetragona\u003c/em\u003e, and \u003cem\u003eN. mexicana\u003c/em\u003e \u0026times; \u003cem\u003eN. odorata\u003c/em\u003e (Borsch et al. 2014). In this study, hybrids between \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e were not found. In Hokkaido Island, Japan, the two species grow allopatrically, except for one mire (Sasaki 2004, 2020). Naito and Shiga (2024) reported that the localities of collected specimens of \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e were distinctly separated in Japan. Additionally, Fu et al. (2001) noted that \u003cem\u003eN. tetragona\u003c/em\u003e was limited to boreal regions above 50\u0026deg;N latitude in North America and Europe. Even if the two species can cross, their hybrids may be rare due to their different distributional patterns. More detailed surveys regarding morphological variations and gene flows among the populations of these two species distributed at the boundaries of their distributional areas, as well as artificial crossing tests, are needed to confirm their isolation patterns and hybridization events.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eTaxonomic treatment\u003c/h2\u003e \u003cp\u003e \u003cb\u003eNymphaea pygmaea\u003c/b\u003e (Salisb.) W.T.Aiton, Hort. Kew., ed. 2, 3: 293 (1811)(Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea\u0026ndash;f). \u0026equiv;\u003cem\u003eCastalia pygmaea\u003c/em\u003e Salisb., Parad. Lond. 1(2): 68 (1805\u0026ndash;8). Type: unknown (see Wiersema 1996).\u003c/p\u003e \u003cp\u003e=\u003cem\u003eNymphaea pygmaea\u003c/em\u003e var. \u003cem\u003eminima\u003c/em\u003e Nakai, Bot. Mag. (Tokyo) 14(11): 746 (1938), syn. nov. \u0026equiv;\u003cem\u003eNymphaea minima\u003c/em\u003e (Nakai) Nakai, Bull. Nat. Sci. Mus., Tokyo 31: 25 (1952)\u0026thinsp;\u0026equiv;\u0026thinsp;\u003cem\u003eNymphaea tetragona\u003c/em\u003e var. \u003cem\u003eminima\u003c/em\u003e (Nakai) W.Lee, Lineamenta Florae Koreae: 360 (1996). Holotype: South Korea; South Hwanghae Prov., 29 Jul. 1929, T. Nakai 12758 (TI!).\u003c/p\u003e \u003cp\u003e=\u003cem\u003eNymphaea japono-koreana\u003c/em\u003e Nakai, J. Jap. Bot. 14: 751\u0026ndash;752 (1938), nom. nud.\u003c/p\u003e \u003cp\u003e=\u0026ldquo;\u003cem\u003eNymphaea tetragona\u003c/em\u003e Georgi var. \u003cem\u003etetragona\u003c/em\u003e\u0026rdquo; auct. non Georgi: Ohwi and Kitagawa, New Fl. Jap. 672\u0026ndash;673 (1983); Kadono, Aquat. Pl. Jap. 109 (1994); Ito, Fl. Jap. 2a: 358 (2006); Choi H, Gen. Vas. Pl. Korea 161\u0026ndash;162 (2007); Kadono, Fld. Guide Aquat. Pl. Jap. 50 (2014); Shiga, Wild Fl. Jap. 1: 48. pl. 37 (2015).\u003c/p\u003e \u003cp\u003e \u003cb\u003eDescription\u003c/b\u003e: Herbs, perennial, aquatic. Rhizomes erect, cylindrical, unbranched, stolons absent (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ef). Leaves dimorphic. Submerged leaves: leaf blade deltoid, sagittate, orbicular, 6\u0026ndash;24 \u0026times; 5\u0026ndash;21 cm, membranous, margin entire, base cordate. Floating leaves: leaf blade obovate to orbicular, 5.5\u0026ndash;24 \u0026times; 4.5\u0026ndash;21 cm, subcoriaceous, abaxially glabrous, base deeply cordate and basal lobes parallel to overlapping, thicker, 0.2\u0026ndash;0.6 mm, impressed veins abaxially, margin entire (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, e). Flowers floating, 3\u0026ndash;6 cm in diameter (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec). Calyx insertion on receptacle tetragonal (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed). Sepals four, narrowly deltoid, 1.7\u0026ndash;5.6 \u0026times; 0.4\u0026ndash;4.0 cm, obscurely veined, persistent (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb). Petals 5\u0026ndash;17, white, broadly lanceolate, oblong, or narrowly obovate, 1.5\u0026ndash;4.6 \u0026times; 0.4\u0026ndash;2.1 cm, transition to stamens gradual (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb). Stamens 24\u0026ndash;71, 0.7\u0026ndash;1.8 \u0026times; 0.1\u0026ndash;0.4 cm, yellow; filament of outer stamens wider than the anther, connective apically appendaged slightly (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, c). Carpels completely united, walls between locules of the ovary single. Stigma rays 5\u0026ndash;11, yellow (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec); carpellary appendages broadly lanceolate, 0.2\u0026ndash;0.4 cm, yellow (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec). Fruit globose, 2.8\u0026ndash;7.7 \u0026times; 1.2\u0026ndash;2.9 cm. Seeds ellipsoid to ovoid, 1.9\u0026ndash;3.6 \u0026times; 1.3\u0026ndash;2.6 mm, smooth. Fl. Jun.\u0026ndash;Oct.\u003c/p\u003e \u003cp\u003e \u003cb\u003eRegional name\u003c/b\u003e: Hitsuji-gusa (Japan: Ito 2016) and Kkoma-Suryeon (Korea: National Institute of Botanical Resources 2023)\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eDistribution and habitat\u003c/strong\u003e \u003cp\u003eChina, India, Japan, Korea, Nepal, and Vietnam. In ponds, lakes, marshes, moors, and paddy fields.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eSpecimens examined\u003c/b\u003e: JAPAN. Hokkaido Pref.: Ebetsu-shi, 14 Jun. 2016, N. Nitta s.n. (SAPS); Iburi-no-kuni, Aug. 1893, Y. Tokubuchi s.n. (SAPS); 13 Jul. 1895, N. Date s.n. (SAPS); 21 Aug. 1899, J. Hanazawa s.n. (SAPS); 22 Aug. 1899, J. Hanazawa s.n. (SAPS); 17 Jun. 1975, A. Namimatsu s.n. (SAPS) (two specimens); 22 Jun. 1975, S. Nakayama s.n. (SAPS) (six specimens); Kuromatsunai-cho, 13 Jun. 1989, K. Ito s.n. (SAPS); Mukawa-cho, 17 Aug. 1964, Y. Takahashi s.n. (SAPS); 12 Jun. 1999, K. Oita 99-5930 (SAPS); 27 Aug. 2000, K. Oita 00-6048 (SAPS) (two specimens); 15 Jul. 2001, K. Oita 01-6609 (SAPS); Nanae-cho, 3 Aug. 1925, S. Miki s.n. (OSA230084); Oshamanbe-cho, 8 Aug. 2008, H. Takahashi s.n. (SAPS); Oshima-no-kuni, Aug. 1886, S. Nozawa s.n. (SAPS); 9 Jul. 1916, F. C. Grcatrex 189 (SAPS); Jun. 1917, S. Tanaka s.n. (SAPS); Sapporo-shi, 16 Aug. 1988, M. Hara 5758, 5990 (SAPS); 30 Aug. 1989, M. Hara 1256 (SAPS); Shiraoi-cho, 31 Jul. 1906, K. Saida 198 (TNS5549); 19 Aug. 1976, M. Hara 144 (SAPS); Tomakomai-shi, 29 Aug. 1969, M. Hara 132 (SAPS); 29 Aug. 1972, M. Hara 130 (SAPS); 28 Jun. 1974, S. Nakayama s.n. (SAPS); 27 Jun. 1987, J. Oginiawa s.n. (TNS988649); 17 Aug. 1989, Y. Kadono 6146 (OSA); 24 Aug. 1994, H. Takahashi 17548 (SAPS); 23 Aug. 1998, H. Takahashi 25789 (SAPS); Junsai-numa, 17 Aug. 1888, Y. Tokubuchi s.n. (SAPS). Aomori Pref.: Higashidori-mura, 28 Aug. 2019, H. Naito et al. 271, 272 (NGU11664, 11665); 19 Jun. 2022, J. Sasaki s.n. (NGU14396); 12 Aug. 2022 (S Kato s.n.), NGU14398; Tsugaru-shi, 26 Aug. 2019, H. Naito et al. 273 (NGU11668\u0026ndash;11970). Iwate Pref.: Hanamaki-shi, 27 Aug. 2019, H. Naito et al. 267\u0026ndash;269 (NGU11681\u0026ndash;11684). Miyagi Pref.: Kami-machi, 28 Aug. 2019, H. Naito et al. 270 (NGU11659\u0026ndash;11661). Akita Pref.: Akita-shi, 23 Aug. 2019, H. Naito et al. AM43, 44 (NGU11155, 11161, 11162, 11165). Yamagata Pref.: Murayama-shi, 28 Aug. 2019, S. Kato et al. TN176 (NGU10875). Fukushima Pref.: Inawashiro-machi, 23 Sep. 2019, S. Kato s.n. (NGU11676\u0026ndash;11677). Tochigi Pref.: Nasukarasuyama-shi, 23 Sep. 2019, H. Naito et al. TN14, 15 (NGU12378\u0026ndash;12380). Gunma Pref.: Katashina-mura, 10 Sep. 2019, H. Naito et al. 276, 277, 304, 306 (NGU12948\u0026ndash;12955). Niigata Pref.: Tsunan-machi, 11 Aug. 2019, S. Kato et al. s.n. (NGU11675); Jouetsu-shi, 14 Sep. 2019, H. Naito et al. 222 (NGU11654, 11715). Gifu Pref.: Mizunami-shi, 7 Sep. 2020, H. Naito et al. 497, 498 (NGU11756, 11758). Aichi Pref.: Toyota-shi, 9 Sep. 2020, H. Naito et al. 601\u0026ndash;604 (NGU11757, 11761, 11762). Mie Pref.: Tamaki-cho, 10 Sep. 2020, H. Naito et al. 662 (NGU11722). Shiga Pref.: Taga-cho, 7 Jul. 2019, T. Shiga 11317 (NGU10595, 10596). Hyogo Pref.: Ono-shi, 19 Sep. 2019, T. Shiga et al. 11229 (NGU8301, 8302). Shimane Pref.: Iinan-cho, 23 Jul. 2018, K. Tsubota \u0026amp; S. Midorikawa 232 (NGU 8731). Ehime Pref.: Imabari-shi, 22 Jul. 2019, Y. Fujiwara s.n. (NGU11650\u0026ndash;11653). Saga Pref.: Arita-cho, 23 Jul. 2019, H. Naito et al. 228 (NGU11655, 11656). Miyazaki Pref.: Ebino-shi, 24 Jul. 2019, H. Naito et al. 227 (NGU11657\u0026ndash;11658). KOREA. Gangwon-do: Goseong-gun, 8 Jun. 2019, T. Shiga et al. 10665, 10666 (NGU8679, 8680).\u003c/p\u003e \u003cp\u003e \u003cb\u003eNymphaea tetragona\u003c/b\u003e Georgi, Bemerk. Reise Russ. Reich 1: 220 (1775)(Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eg\u0026ndash;l). \u0026equiv;\u003cem\u003eCastalia tetragona\u003c/em\u003e (Georgi) G. Lawson, Proc. \u0026amp; Trans. Roy. Soc. Canada 6, Sect. 4: 112 (1888). Lectotype: Angara, Siberia, Russia, Jul. 1772, J. Georgi (LE1009632!) (see Byalt and Krupkina 2017).\u003c/p\u003e \u003cp\u003e=\u003cem\u003eNymphaea tetragona\u003c/em\u003e var. \u003cem\u003eerythrostigmatica\u003c/em\u003e Koji Ito, J. Geobot. 16: 100 (1968), syn. nov.; Kadono, Aquat. Pl. Jap. 112 (1994); Kadono, Fld. Guide Aquat. Pl. Jap. 51 (2014); Shiga, Wild Fl. Jap. 1: 48 (2015). Holotype: Sarufutsu-mura, Kimomanuma, Japan, 21 Jul. 1967, Ko. Ito s.n. (SAPS72107!).\u003c/p\u003e \u003cp\u003e=\u003cem\u003eNymphaea fennica\u003c/em\u003e Mela, Acta Soc. Fauna FI. Fenn. 14(3): 1 (1897). Syntype: Several syntypes at H (see Wiersema 1996).\u003c/p\u003e \u003cp\u003e \u003cb\u003eDescription\u003c/b\u003e: Herbs, perennial, aquatic. Rhizomes erect, cylindrical, rarely branched; stolons absent (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003el). Leaves dimorphic. Submerged leaves: leaf blade deltoid, sagittate, orbicular, 6\u0026ndash;18 \u0026times; 4\u0026ndash;15 cm, membranous, margin entire, base cordate. Floating leaves: leaf blade cordate-ovate to ovate-elliptic, 6\u0026ndash;18 \u0026times; 4\u0026ndash;15 cm, subcoriaceous, abaxially glabrous, base deeply cordate and basal lobes parallel to overlapping, thinner, 0.2\u0026ndash;0.4 mm, raised veins abaxially, margin entire (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eg, k). Flowers floating, 3\u0026ndash;6 cm in diameter (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ei). Calyx insertion on the receptacle prominently tetragonal (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ej); sepals 4, broadly deltoid to narrowly ovate, 1.6\u0026ndash;3.4 \u0026times; 0.9\u0026ndash;2.4 cm, veined, persistent (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eh). Petals 12\u0026ndash;21, white, rhombic to obovate, 1.5\u0026ndash;3.3 \u0026times; 0.7\u0026ndash;1.7 cm, transition to stamens gradual (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eh). Stamens 32\u0026ndash;116, 0.6\u0026ndash;1.3 \u0026times; 0.2\u0026ndash;0.3 cm, yellow\u0026ndash;orange, abaxially usually tinged with red to purple in the inner stamens; filament of outer stamens wider than anther very prominent, connective apically appendaged slightly (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eh, i). Carpels completely united, walls between locules of the ovary single. Stigma rays 4\u0026ndash;12, dark red to purple (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ei); carpellary appendages broadly lanceolate, 0.3\u0026ndash;0.8 cm, yellow or dark red to purple, apically tinged with red to dark red (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ei). Fruit globose, 2.6\u0026ndash;3.7 \u0026times; 1.0\u0026ndash;2.2 cm. Seeds ellipsoid to ovoid, 2.5\u0026ndash;2.9 \u0026times; 1.5\u0026ndash;1.9 mm, smooth. Fl. Jul.\u0026ndash;Aug.\u003c/p\u003e \u003cp\u003e \u003cb\u003eRegional name\u003c/b\u003e: Pygmy water-lily (North America: Wiersema and Hellquist 1997), Small white water-lily (North America: Wiersema and Hellquist 1997), Nymph\u0026eacute;a tetragonal (North America: Wiersema and Hellquist 1997), Shui lian (China: Fu et al. 2001), Ezo-Beni-Hitsuji-gusa (Japan: Ito 1968), and Suryeon (Korea: National Institute of Botanical Resources 2023)\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eDistribution and habitat\u003c/strong\u003e \u003cp\u003eChina, Japan, Russia; North America, Europe. In ponds, lakes, marshes, and moors.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eSpecimens examined\u003c/b\u003e: JAPAN. Hokkaido Pref.: Akkeshi-cho, 14 Aug. 1925, S. Miki s.n. (OSA230085); Ikeda-cho, 20 Aug. 1934, H. Yokoyama s.n. (SAPS); Hamanaka-cho, 7 Aug. 2020, H. Naito et al. 398 (NGU11704); Kamikawa-cho, 14 Aug. 1947, M. Tatewaki 35570 (SAPS) (three specimens); 9 Jul. 1950, M. Kurashige 233 (SAPS); 1 Aug. 1967, K. Ito \u0026amp; A. Umezawa s.n. (SAPS) (two specimens); 2 Aug. 1967, K. Ito \u0026amp; A. Umezawa s.n. (SAPS); Kitami-shi, 10 Aug. 1925, collector unknown 231 (SAPS); 2 Aug. 1967, K. Ito \u0026amp; A. Umezawa s.n. (SAPS) (two specimens); 13 Aug. 2019, H. Naito et al. 214\u0026ndash;216 (NGU11701\u0026ndash;11703); 4 Jun. 2022, J. Sasaki s.n. (NGU14393, 14394); Kushiro-shi, 13 Aug. 1884, K. Miyabe s.n. (SAPS); 11 Jul. 1886, M. Nakamura B.149 (SAPS); 13 Aug. 1887, S Nogawa s.n. (SAPS); 10 Aug. 1951, M. Tatewaki s.n. (SAPS) (two specimens); 28 Jun. 1967, K. Seto 16511 (OSA); 25 Aug. 1975, Y. Kadono 1641 (OSA); 17 Jul. 1987, Y. Kadono 4689 (OSA); 7 Aug. 1991, Y. Kadono et al. 18 (OSA); 9 Aug. 1991, Y. Kadono et al. s.n. (OSA); Nemuro-shi, 28 Jul. 1928, M. Tatewaki \u0026amp; Y. Tokunaga s.n. (SAPS); 29 Aug. 2004, T. Shiga \u0026amp; S. Takebayashi 65 (OSA220192); 29 Aug. 2004, T. Shiga \u0026amp; S. Takebayashi 52 (OSA220147); collection date unknown, S. Yamane s.n. (SAPS); Sarufutsu-mura, 28 Jul. 1966, K. Ito s.n. (SAPS); 28 Jul. 1966, K. Ito s.n. (SAPS16635); 21 Jul. 1967, K. Ito s.n. (SAPS16636\u0026ndash;16638); 21 Jul. 1967, K. Ito s.n. (SAPS);17 Jul. 1975, Y. Kadono 1977 (OSA); 21 Jul. 1987, Y. Kadono 4779 (OSA) (four specimens); 21 Jul. 1987, Y. Kadono 4784 (OSA); 5 Aug. 1998, H. Takahashi 25535 (SAPS15970); 26 Aug. 1989, J. Oginiawa 10643, 39380 (TNS960643, 989380); 15 Aug. 2019, H. Naito \u0026amp; T. Shiga 209\u0026ndash;211, 301 (NGU11693\u0026ndash;11696); 5 Aug. 2020, H. Naito et al. 281, 282, 558, 559 (NGU11711\u0026ndash;11714, 11716); 5 Aug. 2020, H. Naito et al. 558, 559 (NGU11711\u0026ndash;11714,11716); 6 Aug. 2020, H. Naito et al. 333, 356 (NGU11697, 11698); 11 Jul. 2022, J. Sasaki s.n. (NGU14395); Shibecha-cho, 9 Aug. 1969, N. Fujita 51 (TNS788665); 29 Aug. 1992, H. Takahashi \u0026amp; Y. Takashima 15143, 15162 (SAPS); 29 Jul. 2019, S. Kato s.n. (NGU11692); Taiki-cho, 28 Jul. 1979, E. Miki \u0026amp; M. Ito s.n. (TNS395745); 17 Jul. 1985, Y. Kadono \u0026amp; G. Wiegleb 859 (OSA) (two specimens); 22 Jun. 2019, K. Shutoh et al. 3681 (SAPS55004); 26 Jun. 2019, K Shutoh s.n. (NGU11700); 8 Aug. 2020, H. Naito et al. 429 (NGU11699); Takinoue-cho, 3 Aug. 1966, K. Ito s.n. (SAPS); Toyokoro-cho, 9 Aug. 2020, H Naito et al. 457, 560 (NGU11705\u0026ndash;11708); Toyotomi-cho, 13 Jun. 1938, K. Miyabe \u0026amp; H. Yokoyama s.n. (SAPS); 2 Aug. 1968, K. Ito s.n. (SAPS); 16 Aug. 1980, K. Harusawa s.n. (OSA34528); Tsukigata-cho, 7 Aug. 2020, K. Katagiri K1, 2 (NGU11709, 11710); Uryu-cho, 8 Jul. 1998, H. Takahashi 25199 (SAPS16630); 23 Jul. 1998, H. Takahashi 25291, 25294, 25308 (SAPS16631, 16632, 16634); 12 Aug. 1998, H. Takahashi 25642 (SAPS); 6 Jun. 2004, M. Yamazaki s.n. (NGU7915); 6 Jun. 2004, M. Yamazaki s.n. (NGU); 12 Aug. 2009, M. Yamazaki s.n. (NGU13468) ; 23 Jul. 2019, K. Shutoh et al. 3658, 3659 (SAPS54921\u0026ndash;54923); 11 Aug. 2020, H. Naito et al. 523\u0026ndash;527, 532 (NUG11687\u0026ndash;11691). Kurils: Furukamappu, 20 Aug. 1931, J. Ohwi 923 (TNS222126) (two specimens); Kunashiri, 26 Aug. 1923, M. Tatewaki 3411 (SAPS25573); 20 Aug. 1936, M. Tatewaki 25573 (SAPS); Tomari-mura, Aug. 1933, A. Tanaka s.n. (OSA230088). Sakhalin: Korsakov District, 29 Jul. 2001, H. Tatewaki 29516 (SAPS) (four specimens); Kusunnai, 21 Jun. 1906, T. Miyake s.n. (SAPS); Nyiwo, 15 Aug. 1922, Y. Kudo \u0026amp; M. Tatewaki 6584 (SAPS) (four specimens); Okhinsky District, 4 Aug. 2001, V.Y. Barkalov 10195 (SAPS); 4 Aug. 2001, T. Fukuda 1501 (SAPS); 11 Aug. 2001, T. Fukuda 1981 (SAPS); Shisuka-cho, 15 Aug. 1906, T. Miyake s.n. (SAPS); 29 Aug. 1929, collector unknown (TNS1066643); 1933, Y. Tanaka s.n. (OSA230087); 16 Aug. 1935, S. Sugawara 30155, 30157 (SAPS); collection date and collector unknown (SAPS) (two specimens); Shisuka-gun, 18 Jul. 1937, B. Yoshimura \u0026amp; M. Hara s.n. (TNS268313); Tominai-cho, 3 Aug. 1930, H. Otani \u0026amp; Y. Imai s.n. (SAPS); Ushiro-mura, 24 Jun. 1934, F. Fujita s.n. (OSA230086).\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis study was partly funded by the Environment Research and Technology Development Fund (4-2001) of the Environmental Restoration and Conservation Agency and JST SPRING grant number JPMJSP2121.\u003c/p\u003e\u003ch2\u003eAuthor contributions\u003c/h2\u003e \u003cp\u003eAll authors contributed to the study\u0026rsquo;s conception and design. Material preparation, data collection, and analysis were mainly performed by Hanaka Naito and Takashi Shiga. The first draft of the manuscript was written by Hanaka Naito, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eWe thank the following people for their help with collecting plant samples: Dr. H. J. Choi, Mr. Y. Fujiwara, Mr. H. Kamiaka, Dr. Y. Kadono, Dr. K. Katagiri, Mr. Y. Komine, Dr. Y. Lee, Ms. E. Mashima, Mr. S. Midorikawa, Mr. J. Sasaki, Mr. T. Suzui, Dr. M. Suzuki, Mr. W. Takahashi, Mr. H. Tsurumaki, Mr. H. Yamada, Dr. T. Yamanouchi, and Ms. M. Yamazaki.\u003c/p\u003e\u003cp\u003eCompeting Interests: We declare that we have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBorsch T, Hilu KW, Quandt D, Wilde V, Neinhuis C, Barthlott W (2003) Noncoding plastid \u003cem\u003etrnT-trnF\u003c/em\u003e sequences reveal a well resolved phylogeny of basal angiosperms. 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Bull Natl Sci Mus 31:1\u0026ndash;153\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNational Institute of Botanical Resources (2023) National List of Korea. National Institute of Botanical Resources. https://www.kbr.go.kr/stat/ktsnfiledown/downpopup.do. Accessed 19 March 2024\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, New York\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOhwi J, Kitagawa M (1983) Nymphaeaceae. New flora of Japan. Shibundo, Tokyo, pp 669\u0026ndash;673 [in Japanese]\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePlanchon JE (1853) Etudes sur les Nymphaeacees. Ann Sci Nat Bot s\u0026eacute;r 3:17\u0026ndash;63\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSalisbury RA (1805\u0026ndash;1808) The paradisus londinensis: Or coloured figures of plants cultivated in the vicinity of the metropolis. 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Nat Methods 9:671\u0026ndash;675. doi:10.1038/nmeth.2089\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShiga T (2015) Nymphaeaceae. In: Ohashi H, Kadota Y, Murata J, Yonekura K, Kihara H (ed) Wild Flowers of Japan Revised and Enlarged Edition 1. Heibonsha, Tokyo, pp 46\u0026ndash;48 [in Japanese]\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSlocum PD (2005) Water lilies and lotuses. Timber Press, Seattle\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTaberlet P, Gielly L, Pautou G, Bouvet J (1991) Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol Biol 17:1105\u0026ndash;1109. doi:10.1007/BF00037152\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTakakura K, Nishida S, Nishida T (2010) Reproductive interference and its implications for the biogeography and ecology of plants. 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Academic Press, New York, pp 315\u0026ndash;322\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWiersema JH (1988) Reproductive biology of \u003cem\u003eNymphaea\u003c/em\u003e (Nymphaeaceae). Ann Mo Bot Gard 75:795\u0026ndash;804. doi:10.2307/2399367\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWiersema JH (1996) \u003cem\u003eNymphaea tetragona\u003c/em\u003e and \u003cem\u003eNymphaea leibergii\u003c/em\u003e (Nymphaeaceae): Two species of diminutive water-lilies in North America. Brittonia 48:520\u0026ndash;531. doi:10.2307/2807871\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWiersema JH, Hellquist CB (1997) Nymphaeaceae. In: Flora of North America Editorial Committee (eds) Flora North America North of Mexico vol. 3, Oxford University Press, Oxford, pp 66\u0026ndash;77\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"aquatic plants, internal transcribed spacer, Japan, Nymphaea leibergii, rps4-trnF, water lily","lastPublishedDoi":"10.21203/rs.3.rs-4414565/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4414565/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn this study, we aimed to clarify the taxonomic identity between \u003cem\u003eNymphaea tetragona\u003c/em\u003e, which has a red to purple stigmatic disk and the widest distribution range among the genus, and \u003cem\u003eNymphaea pygmaea\u003c/em\u003e, which has a yellow stigmatic disk and is ordinarily treated as a synonym of \u003cem\u003eN. tetragona\u003c/em\u003e, based on detailed morphological measurements and phylogenetic analyses. Through morphological analysis, we assessed 26 quantitative characters of 111 individuals from 15 populations. Seventeen characters were significantly different between the two species; however, these characters overlapped. Principal component analysis based on 13 morphological characters revealed different trends in the morphology of the two species. In DNA analysis, one nuclear ribosomal DNA region, internal transcribed spacer (ITS) region, and one chloroplast DNA region (\u003cem\u003erps4-trnF\u003c/em\u003e) were compared. The ITS region showed differences in 18\u0026ndash;19 substitutions and two indels between \u003cem\u003eN. pygmaea\u003c/em\u003e and \u003cem\u003eN. tetragona\u003c/em\u003e. Additionally, the two species were closely related within the subgenus \u003cem\u003eNymphaea\u003c/em\u003e, and \u003cem\u003eN. tetragona\u003c/em\u003e and \u003cem\u003eNymphaea leibergii\u003c/em\u003e formed a robust monophyletic group with \u003cem\u003eN. pygmaea\u003c/em\u003e as a sister taxon. These results indicate that \u003cem\u003eN. tetragona\u003c/em\u003e and \u003cem\u003eN. pygmaea\u003c/em\u003e have differentiated morphologically and genetically, thus making these two species taxonomically distinct.\u003c/p\u003e","manuscriptTitle":"Morphological and phylogenetic analyses reveal the taxonomic distinctiveness between Nymphaea pygmaea and N. tetragona (Nymphaeaceae)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-10 22:20:55","doi":"10.21203/rs.3.rs-4414565/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0f2cc929-b0a5-4bd7-85e9-2c8e6510fb3b","owner":[],"postedDate":"June 10th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-23T23:22:44+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-10 22:20:55","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4414565","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4414565","identity":"rs-4414565","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}