Comparative karyotype analysis of eleven species of Lilium from China by FISH with rDNA oligo-probes

Comparative karyotype analysis of eleven species of Lilium from China by FISH with rDNA oligo-probes | 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 Article Comparative karyotype analysis of eleven species of Lilium from China by FISH with rDNA oligo-probes Ya-Di Yang, Xiang-Hui Jiang, Bang-Yue Zhang, Chao-Wen She This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8029111/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 17 Feb, 2026 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract The complexity of the classification of the genus Lilium requires more molecular cytogenetic researches to clarify the karyotype differentiation and chromosome evolution among species, verify the existing classification, and help resolve the remaining disputes in classification. In this study, the chromosomes of eleven species of Lilium , including China's main medicinal and edible lilies as well as seven species endemic to southwest and central China, were characterized using fluorescence in situ hybridization (FISH) with 5S and 35S rDNA oligonucleotide probes. Molecular cytogenetic karyotypes were quantitatively established using the dataset of chromosome measurements and FISH signals. Karyotype asymmetry indices were measured for elucidating their karyotype asymmetry and karyological relationships. Their karyotype structures could be differentiated by the scatter plot of M CA vs. CV CL . rDNA FISH showed that there were 1–2 5S rDNA loci and 2–6 35S rDNA loci in these species, and most of the 35S loci generated secondary constrictions. The combination of comparison of rDNA patterns and PCoA based on x , 2 n , TCL, M CA , CV CL and CV CI reveals the evolutionary relationships among these species. Our results cytogenetically confirm the rationality of the species classification of subset. Leucolirion 6a in the latest revised classification of Lilium . Biological sciences/Biological techniques Biological sciences/Genetics Biological sciences/Molecular biology Lilium karyotype karyotype asymmetry cytotaxonomy ribosomal RNA gene (rDNA) fluorescence in situ hybridization (FISH) Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction The genus Lilium L., which belongs to the tribe Lilieae in the family Liliaceae, consists approximately 125 species (now including the former genus Nomocharis ) originally found in temperate and alpine regions of the Northern Hemisphere, with centers of diversity in East Asia, southern Europe, the Caucasus, and eastern and western North America (Gao and Gao 2016 ; Givnish et al. 2020 ). Sixty-one Lilium species (including 6 species of the former genus Nomocharis , 37 endemic) are distributed in China, mainly in the southwestern, central and northeastern regions (Liang and Tamura 2000 ; Du et al. 2014 ; Gao and Gao 2016 ). Several studies have concluded that the Qinghai-Tibet Plateau (including the Himalayas and Hengduan Mountains) is a center of origin and diversification for Lilium (Patterson and Givnish 2002 ; Du et al. 2014 ; Givnish et al. 2020 ; Li et al. 2022 ). Lilies are of significant importance in horticulture. In China, four Lilium species have been widely used as traditional medicines and/or edible plants with long history and have potential for their diverse pharmacological activities (Zhou et al. 2021 ). The dried fleshy scaly leaves of L. lancifolium Thunb., L. brownii var. viridulum Baker and L. pumilum DC., known as Baihe (Lilii Bulbus), are traditional Chinese medicine (TCM) materials which have the effects of moistening the lung, dispelling fire, calming the mind, and killing insects and lice (Chinese Pharmacopoeia Commission 2020 ). The main edible lilies cultivated in China include Lanzhou lily (L. davidii var. willmottiae (E. H. Wilson) Raffill), Longya lily ( L. brownii var. viridulum ), and Juandan lily (cultivated varieties of L. lancifolium ). The classification of the genus Lilium has been complicated for long time. Since Linnaeus, many scholars have tried to classify Lilium . It was not until 1949 that Comber's classification of Lilium was widely accepted. Comber ( 1949 ) classified the naturally grown lilies into seven sections based on 15 phenotypic characters. However, under the influence of similar selection pressure, species with distant genetic relationships may converge in phenotype, while species with close genetic relationships may also diverge in phenotype in order to adapt to different environments or under the influence of survival competition. Therefore, sections of Lilium based largely on floral form may not always reflect actual evolutionary relationships (Givnish et al. 2020 ). It is necessary to continually revise the morphology-based classification of Lilium using the data of cytotaxonomy, molecular phylogenetics and hybridization breeding practice. In the latest revised classification of Lilium , this genus is still divided into seven sections with the names of some sections and the taxonomic locations of some species being adjusted compared to the Combers’ classification, but some classifications are still controversial (van Tuyl et al. 2018 ; Du et al. 2023). Karyotype analysis is an important method for studying the systematics and evolution in higher plants (e.g. Peruzzi et al. 2009 ; She et al. 2015 , 2023 ; Astuti et al. 2017 ; Wei et al. 2024 ). Conventional karyotype analysis is mainly based on chromosome number and morphological characteristics of chromosomes. However, in most cases, it is difficult to distinguish all chromosomes in a complement by morphological characteristics alone. To this end, researchers have successively developed Giemsa banding, fluorochrome banding and fluorescence in situ hybridization (FISH) of DNA sequences to generate chromosome markers for accurate karyotyping of plants. The most widely utilized DNA probes in plant FISH experiments are the ribosomal genes, 35S (18S-5.8S-26S) and 5S rDNAs, which are organized in tandem arrays with high copy numbers. Comparison of banding and/or FISH patterns (referring to the number and location of specific heterochromatic blocks or hybridization signals of specific DNA sequences) among species within a genus contributes to understanding of chromosome evolution and revealing evolutionary relationships among taxa (e.g. She et al. 2015 ; Han et al. 2018 ; Mitrenina et al. 2023 ; Dias et al. 2024 ; Wei et al. 2024 ; Sun et al. 2024 ). Evaluating karyotype asymmetry is one of the core aspects of karyotype analysis of plants. Karyotype asymmetry, as a measure of the structural heterogeneity of a chromosome complement, is a cornerstone of plant cytotaxonomy because it serves as a key indicator of evolutionary trends and karyological relationships (e.g. Stebbins 1971 ; Peruzzi et al. 2009 ; Astuti et al. 2017 ). In plant karyotype analysis, asymmetry has been assessed using a set of quantitative indices which fall into two categories for evaluating intrachromosomal asymmetry (the variation in centromere position within a complement) and interchromosomal asymmetry (the variation in chromosome length within a complement), respectively (for details and references see Paszko 2006 ; Peruzzi and Eroglu 2013 ). Recent critical reviews made by several authors confirm that mean centromeric asymmetry (M CA ) and coefffcient of variation of chromosome lengths (CV CL ) are the most appropriate indices for measuring the intrachromosomal asymmetry and interchromosomal asymmetry, respectively (Paszko 2006 ; Peruzzi and Eroglu 2013 ; Astuti et al. 2017 ). The best way for comparison of karyotype asymmetry among taxa is by means of bidimensional scatter plot of M CA vs. CV CL (Peruzzi and Eroglu 2013 ; Astuti et al. 2017 ; Kadluczka and Grzebelus 2021 ; She et al. 2023 ; Wei et al. 2024 ). For determination of karyological relationships among taxa, the most effective methodology is the principal coordinate analysis (PCoA) with six quantitative karyotypic parameters, x , 2 n , TCL (total chromosome length of the haploid complement), CV CI (coefficient of variation of centromeric index), M CA and CV CL (e.g. Peruzzi and Altınordu 2014 ; Kadluczka and Grzebelus 2021 ; She et al. 2023 ; Mitrenina et al. 2023 ; Wei et al. 2024 ; Sun et al. 2024 ). In terms of clarifying the evolutionary relationships among species, both banding and FISH patterns are often limited to comparison among species within the same genus, while PCoA, which is based on karyomorphological data, can not only be applied to comparison among species within a genus but also to comparison among species of different genera within the same family (Astuti et al. 2017 ; She et al. 2023 ). Comprehensive karyotypic features, including chromosome number, chromosome morphological features, karyotype asymmetry, FISH pattern of specific DNA sequences (e.g. rDNA), have proved of great use in elucidating evolutionary relationships and trends in chromosome evolution among taxa (e.g. Kadluczka and Grzebelus 2021 ; Mitrenina et al. 2023 ; She et al. 2023 ; Wei et al. 2024 ; Sun et al. 2024 ). Because of their large genome size, lilies have been long used as model plants by cytogeneticists for chromosome studies. Conventional karyotype analysis and C-banding analysis have revealed that the chromosome number of Lilium is very constant, ploidy variation is rare, and the differences between karyotypes of different species are small (Stewart 1947 ; Smyth et al. 1989 ; Noda 1991 ; Gao et al. 2011 , 2012 ; Liu et al. 2017 ). It is believed that small structural changes in chromosomes such as inversions and reciprocal translocations are the main driving forces for karyotype evolution in the genus Lilium (Noda 1991 ; Gao et al. 2011 ). The vast majority of species of Lilium are diploid (2n = 2x = 24), and only a few species have triploid, tetraploid or aneuploidy in addition to diploid (Stewart 1947 ; Noda 1986 , 1991 ; Gao et al. 2011 ; Lee et al. 2014 ; Nguyen et al. 2015 , 2016 ; Liu et al. 2017 ; Zhou et al. 2020 ). The haploid genome of species of Lilium is composed of two larger metacentric (m) or submetacentric (sm) chromosomes and ten smaller subtelocentric (st) or telocentric (t) chromosomes. The basic nomenclature of the karyotypes of species of Lilium is based on the order of chromosomes in sequences of decreasing length of short arms, with 1 representing the longest short arm and 12 the shortest (Stewart 1947 ). The number and location of secondary constrictions (SCs) varied greatly among species, but the karyotype asymmetry of the species that have been studied was all 3A and 3B types of Stebbins’ catergory (Gao et al. 2011 ; Wan et al. 2011 ; Liu et al. 2017 ), indicating that it is difficult to distinguish the karyotype structures of different species of Lilium by this asymmetry index. With the application of FISH technology in Lilium , it has greatly promoted the researches on cytotaxonomy, chromosome evolution, genetic diversity and hybrid identification of this genus (e.g. Lim et al. 2001 ; Marasek et al. 2004 ; Muratović et al. 2010 ; Sultana et al. 2010 ; Hwang et al. 2011 ; Wang et al. 2012 ; Lee et al. 2014 ; Wang et al. 2015 ; Nguyen et al. 2015 , 2016 ; Ahn et al. 2017 ; Liu et al. 2017 ; Cao et al. 2019 ; Zhou et al. 2020 ; Choi et al. 2021 ; Wu et al. 2021 ; Kang et al. 2024 ; Zhou et al. 2024 ; Zhu et al. 2025 ). According to our statistics, 53 species or varieties of Lilium (species originally belonging to Nomocharis are not included) have been studied for FISH mapping of both 5S and 35S rDNA (43/53) or 35S rDNA alone (10/53). The species (varieties) analyzed by FISH involved seven sections of the genus Lilium , including 6 in sect. Martagon , 3 in sect. Pseudolirium , 9 in sect. Lilium , 4 in sect. Archelirion , 18 in sect. Sinomartagon , 11 in sect. Leucolirion , and 2 in sect. Oxypetalum . In terms of geographical distribution, the species analyzed by FISH so far were mainly those distributed in Northeast Asia and Europe, while the species distributed in southwest and central China and North America were few studied by FISH. Most of the species of Lilium endemic to southwest and central China that have been studied by FISH had 35S rDNA localization only, and the results of rDNA mapping by different authors in some species were inconsistent (Wang et al. 2012 ; Liu et al. 2017 ; Zhou et al. 2020 ). Therefore, it is necessary to conduct dual-color FISH with 5S and 35S rDNA probes that include more species endemic to southwest and central China in order to molecular-cytogenetically characterize these species and analyze their evolutionary relationships. For the medicinal and edible lilies in China, although FISH localization of 5S and 35S rDNA has been reported for them, the results of different authors in some species differed or even were completely inconsistent (Wang et al. 2012 ; Lee et al. 2014 ; Nguyen et al. 2016 ; Liu et al. 2017 ; Zhou et al. 2020 ; Zhou et al. 2024 ), so further verification is needed to provide a reliable basis for molecular cytogenetic identification of these species. In present study, the chromosomes of eleven species of Lilium from China, including the main edible and medicinal lilies in China as well as seven species endemic to southwest and central China (including all the six species of subsect. Leucolirion 6a in the latest revised classification of Lilium ) were characterized by using FISH with 5S and 35S rDNA oligonucleotide probes. Their karyotypes were quantitatively constructed with chromosome measurement data and rDNA FISH signal information. The karyotype asymmetry indices that were recently critically reviewed and recognized were calculated for evaluating asymmetry of the karyotypes and karyological relationships among them. The data of karyotypic parameters and rDNA patterns were assessed to gain insights into the karyotype differentiation and the evolutionary relationships among these species, thereby providing cytotaxonomic support for the classification of Lilium . Materials and Methods Plant materials Twelve taxa belonging to eleven species of Lilium were collected different places of China and cultivated in Lily Germplasm Resources Engineering Center, Hunan University of Technology, Hunan Province, China (Table 1 ). The geographical distribution and source information of these materials are listed in Table S1 . The sections to which these species belong are listed in Table 2 . The plants were identified by Professor Si-Xiang Dong of Nuclear Agriculture and Chinese Medicinal Materials Institute of Hunan Academy of Agricultural Sciences. Table 1 The general karyotype features of the eleven species of Lilium Species Karyotype formula TCL ± SE (µm) C (µm) RRL CI ± SE CV CI M CA CV CL Stebinns’ types L. pumilum Redouté 2n = 24 = 4m(4SAT) + 14st(6SAT) + 6t 212.15 ± 21.06 17.68 6.20-12.17 19.73 ± 10.10 51.16 60.65 21.51 3A L. davidii var. willmottiae (E. H. Wilson) Raffill 2n = 24 = 2m(2SAT) + 2sm + 8st(2SAT) + 12t 222.96 ± 9.31 18.58 6.67–12.59 17.11 ± 11.15 65.17 66.46 20.31 3A L. lancifolium Thunb. 2n = 36 = 6m(6SAT) + 12st(2SAT) + 18t(5SAT) 221.75 ± 15.32 18.48 6.82–12.44 17.19 ± 11.73 68.22 65.43 21.84 3A L. lancifolium ‘Longshan’ 2n = 36 = 6m(6SAT) + 12st(3SAT) + 18t(6SAT) 212.44 ± 17.57 17.70 6.41–12.53 17.03 ± 11.46 67.28 66.21 22.20 3A L. jinfushanense L. J. Peng & B. N. Wang 2n = 24 = 2m(2SAT) + 2sm(2SAT) + 10st + 10t 220.38 ± 22.70 18.37 6.08–12.65 18.30 ± 11.05 60.38 63.77 22.70 3B L. brownii var. viridulum Baker 2n = 24 = 4m + 8st(4SAT) + 12t(2SAT) 215.68 ± 25.53 17.97 5.66–12.16 18.17 ± 11.74 64.58 63.91 23.43 3B L. leucanthum (Baker) Baker 2n = 24 = 4m(4SAT) + 8st(4SAT) + 12t 175.55 ± 15.66 14.63 5.80-12.14 17.88 ± 11.13 62.25 64.38 23.05 3B L. sargentiae E. H. Wilson 2n = 24 = 2m(2SAT) + 2sm(2SAT) + 8st(4SAT) + 12t 201.40 ± 3.88 16.78 6.63–12.52 17.54 ± 11.64 66.35 65.49 23.54 3B L. sulphureum Baker ex J. D. Hooker 2n = 24 = 4m(4SAT) + 10st(4SAT) + 10t 182.87 ± 7.66 15.24 6.06–12.25 18.21 ± 11.27 61.91 63.34 22.06 3B L. regale E. H. Wilson 2n = 24 = 2m(2SAT) + 2sm(2SAT) + 8st(6SAT) + 12t 214.65 ± 6.94 17.88 5.61–13.11 17.88 ± 11.57 64.69 64.83 25.38 3B L. henryi Baker 2n = 24 = 4m(2SAT) + 6st(2SAT) + 14t 197.73 ± 9.35 16.48 6.17–12.65 17.10 ± 10.84 63.39 65.33 22.09 3B L. rosthornii Diels 2n = 24 = 2m(2SAT) + 2sm + 8st(2SAT) + 12t 175.91 ± 7.26 14.66 6.06–12.10 17.35 ± 10.82 62.35 65.89 21.49 3A TCL, total length of the haploid complement (i.e. karyotype length); RRL, range of chromosome relative length; C, mean chromosome length; CI, mean centromeric index; CV CI , coefficient of variation of centromeric index; M CA , mean centromeric asymmetry; CV CL , coefficient of variation of chromosome lengths; Stebbin’s’ types, karyotype asymmetry category of Stebbins. Table 2 The sections to which the eleven species of Lilium belong and the number and location of their 5S and 45S rDNA loci Species Section (Comber 1949 /Du et al. 2014 /Du et al. 2023) 5S a 45S a L. pumilum Sinomartagon 5b / Sinomartagon clade II / Sinomartagon 5b One: 3L-INT (64.03%) Five: 1S-PRO (8.87%), 2S-INT (51.51%), 3L-INT (55.15%), 4-CEN, 5L-INT(60.61%) L. davidii var. willmottiae Sinomartagon 5a / Sinomartagon clade II / Sinomartagon 5a One: 3L-INT (66.45%) Four: 1S-PRO (8.82%), 4L-PRO (5.00%) b , 6L-INT(63.92%), 9S-PRO (21.74%) b L. lancifolium Sinomartagon 5a / Sinomartagon clade II / Sinomartagon 5a One: 3L-INT (71.52%) Six: 1S-PRO (7.93%), 2S-INT (58.86%), 6L-INT (58.34%) c , 7S-PRO (33.60%), 8L-PRO (20.19%) bd , 11L-INT(40.41%) c L. lancifolium ‘Longshan’ Sinomartagon 5a / Sinomartagon clade II / Sinomartagon 5a One: 3L-INT (71.25%) Five:1S-PRO(7.85%),2S-INT(62.88%), 6L-INT(62.7%), 7S-PRO (22.86%), 11L-INT (41.07%) L. jinfushanense Sinomartagon 5c / Sinomartagon clade III / Sinomartagon 5c One: 3L-PRO (9.72%) Two: 1S-PRO (11.50%), 2S-PRO (19.63%) L. brownii var. viridulum Archelirion / Leucolirion clade II / Leucolirion 6b One: 3L-INT (69.82%) Three: 4L-INT (46.05%), 5L-INT (66.87%), 7S-PRO (24%) L. leucanthum Leucolirion 6a / Leucolirion clade I / Leucolirion 6a One: 3L-PRO (6.66%) Four: 1S-PRO (7.54%), 2L-PRO (9.09%), 4S-PRO(20.00%), 5L-PRO (19.65%), L. sargentiae Leucolirion 6a / Leucolirion clade I / Leucolirion 6a One: 3L-PRO (6.17%) Four: 1S-PRO (10.24%), 2L-PRO (7.01%), 4S-PRO (15.79%), 5L-PRO (13.38%) L. sulphureum Leucolirion 6a / Leucolirion clade I / Leucolirion 6a One: 3L-PRO (8.13%) Four: 1S-PRO (9.35%), 2L-PRO (8.22%), 4S-PRO (13.51%), 5L-PRO (10.84%) L. regale Leucolirion 6a / Leucolirion clade I / Leucolirion 6a One: 3L-PRO (9.04%) Five: 1S-PRO (10.69%), 2L-PRO (7.14%), 4S-PRO (18.42%), 5L-PRO(8.98%), 6L-PRO (7.69%) L. henryi Sinomartagon 5a / Leucolirion clade I / Leucolirion 6a One: 3L-PRO + CEN Two: 1S-PRO(7.89%), 4L-INT(54.83%) L. rosthornii Sinomartagon 5a / Leucolirion clade I / Leucolirion 6a Two: 3L-PRO (15.27%), 6S-PRO (46.15%) Two: 1S-PRO (9.71%), 5L-INT (42.11%) a S and L represent short and long arms, respectively; CEN, PRO, INT, DIS and TER represent centromeric ( di = 0), proximal (0 < di < 25%), interstitial (25% ≤ di ≤ 75%), distal (75%< di < 100%) and terminal ( di = 100%) positions, respectively; figures ahead of the positions designate the homologous chromosome pair (triplet) involved; the percentages in square brackets are the percentage distance from centromere to rDNA locus. b indicates the 45S rDNA loci that did not generate secondary constrictions (SCs). SCs occurred at all other 45S loci. c Only two of the three members of the homologous triplet have hybridization signals. d Only one of the three members of the homologous triplet has hybridization signal. Chromosome preparation The bulbs used for cytogenetic experiments were cultivated in pots with mixed planting soil consisting of decomposed pine needle humus soil and sandy soil (v/v = 3:1). Water the bulbs with a solution of rooting powder diluted in water to promote the growth of new roots. When the new roots grew to 1–2 cm, the root tips of 2–3 mm were excised and pretreated with a mixture of 0.1% colchicine and 0.015% 8-hydroxyquinoline (1:1) at 28°C in the dark for 4.5 h, followed by fixation at 4°C in 3:1 (v/v) methanol/glacial acetic acid for more than 12 h. The chromosome slides were prepared using the enzymatic maceration and flame-drying (EMF) method (She et al. 2015 , 2023 ). The fixed root tips were thoroughly rinsed in deionized water and digested in a mixture of 1% cellulase RS and 1% pectolyase Y23 (Yakult Pharmaceutical Industry Co., Ltd.) in citric buffer (pH 4.5) at 37°C for 2 h. After the enzyme solution was replaced by double distilled water, 1–2 digested root tips were transferred to a glass slide and mashed by using fine-pointed forceps with the fixative solution. Then, the slides were flame-dried. The slides with well-spread metaphase chromosomes were selected under an Olympus BX51 phase contrast microscope and stored at -20°C until use. DNA probe and FISH For FISH experiments, the 5S and 35S rDNA oligonucleotide probes described by Han et al. ( 2018 ) were used. The 5S rDNA probes, 5S-1 and 5S-2, as well as the 35S rDNA probes, 35S-1, 35S-2 and 35S-3, were synthesized by Sangon Bioengineering Co., Ltd.. 5S-1 and 5S-2 were labeled with 6-carboxyl fluorescein (6-FAM) at the 5’-terminus and then mixed together to make the 5S rDNA probe solution. 35S-1, 35S-2, and 35S-3 were labeled with 6-carboxyl-tetramethyl rhodamine (TAMRA) at the 5’-terminus and mixed together to make the 35S rDNA probe solution. FISH with the two probes was performed as described by Wei et al. ( 2024 ). The slides were baked at 65℃ for 45 min, cooled, and then denatured in 70% deionized formamide at 85°C for 2.5 min. After denaturation, the slides were immediately placed in a series of 70%, 95%, and 100% ethanol at − 20°C for 5 min each and air dried. Then, hybridization solution was added onto the chromosome slide and then incubated in a moist box at 37°C overnight. The hybridization solution (each slide) was as follows: deionized formamide, 10 µL; 50% dextran sulphate, 4 µL; 20 × SSC, 2 µL; salmon sperm DNA, 2 µL (40 ng); 5S rDNA probe, 1 µL (40 ng); 35S rDNA probe, 1 µL (40 ng). The hybridized slides were washed twice each for 5 min with 2 × SSC at room temperature, and air dried in dark. Then, the chromosomes counterstained with 3 µg ml − 1 4,6–diamidino–2–phenylindole (DAPI) solution in Vectashield H-1000 (Vector Laboratories, Inc.) were visualized with an Olympus BX60 microscope equipped with a QImaging Retiga R6 CCD camera (Teledyne Photometrics) which was controlled using Ocular software (Teledyne Photometrics). Grey-scale images were digitally captured and merged by the Ocular software. Raw images were processed using Adobe Photoshop software (Adobe Systems Inc.). Karyotype analysis Karyotype analysis was performed using the methodology previously described by us (She et al. 2023 ; Wei et al. 2024 ). For each taxon, five metaphase cells were selected for measurement using Adobe Photoshop software. The length of long arm (L) and short arm (S) of each chromosome and the length between the center of FISH signal and centromere were measured. The following parameters were calculated for characterizing the karyotypes: (i) chromosome relative length (RL, % of haploid complement); (ii) arm ratio (AR = L/S); (iii) TCL (i.e. the karyotype length); (iv) mean chromosome length (C); (v) mean centromeric index (CI); (vi) four karyotype asymmetry indices including CV CI (Paszko 2006 ), CV CL (Paszko 2006 ), M CA (Peruzzi and Eroglu 2013 ) and Stebbins’ asymmetry category (Stebbins 1971 ); (vii) percent distance from centromere to DNA loci ( di = d × 100/ a ; d = distance of the center of FISH signal from the centromere, a = length of the corresponding chromosome arm). The chromosomes were classified as metacentric (m, AR = 1.00-1.70), submetacentric (sm, AR = 1.71-3.00), subtelocentric (st, AR = 3.01-7.00) and telocentric (t, AR > 7.01) types according to the nomenclature of Levan et al. ( 1964 ). Satellite chromosomes in which SCs generated were abbreviated as “SAT”. Chromosomes were arranged in order of decreasing length of the short arms according to Stewart ( 1947 ). Idiograms were drawn based on the dataset of chromosome measurements and rDNA FISH signals. To visualize karyotype asymmetry relationships among the analyzed taxa, a bidimensional scatter plot with parameters M CA vs. CV CL was plotted. To determine the karyological relationships among the 12 taxa, a principal coordinate analysis (PCoA) using Gower’s similarity coefficient were carried out based on six quantitative parameters, x , 2 n , TCL, CV CI , M CA and CV CL , as proposed by Peruzzi and Altınordu ( 2014 ). Results General karyotype features L. lancifolium and L. lancifolium ‘Longshan’ are triploid with somatic chromosome number 2n = 3x = 36, the other ten species are diploid with somatic chromosome number 2n = 2x = 24 (Table 1 ). No aneuploidy or B chromosomes were found in the taxa studied. All the eleven species have the same basic chromosome number x = 12. The TCL ranges from 175.55 µm ( L. leucanthum ) µm to 222.96 µm ( L. davidii var. willmottiae ) with a mean chromosome length from 14.63 µm to 18.58 µm. As for the range of relative length (RRL), the smallest RRL was observed in L. lancifolium , while the largest RRL was detected in L. regale . That is, L. lancifolium and L. regale exhibit the smallest and the largest variation in chromosome length, respectively. The CI of these complements varies between 17.03 ± 11.46 ( L. lancifolium ) and 19.73 ± 10.10 ( L. pumilum ). That is, L. lancifolium and L. pumilum are characterized by the smallest and the largest level of variation in the centromeric index, respectively. The haploid karyotypes of the 12 taxa show distinct bimodality, which are composed of two larger metacentric or submetacentric chromosomes and ten smaller subtelocentric or telocentric chromosomes (Table 1 , Table S2 ; Fig. 2 ). The karyotype formulas as well as the number and location of SCs are different among taxa (Table 1 ; Fig. 2 ; Fig. S1 ). All karyotypes fall into 3A or 3B type of Stebbins’ category, possessing moderate degree of asymmetry (Table 1 ). The ranges of CV CI , CV CL and M CA are as follow: CV CI = 51.16 ( L. pumilum ) − 68.22 ( L. lancifolium ), M CA = 60.65 ( L. pumilum ) − 66.46 ( L. davidii var. willmottiae ), CV CL = 20.31 ( L. davidii var. willmottiae ) − 25.38 ( L. regale ) (Table 1 ), revealing that, among the 12 taxa, L. pumilum and L. lancifolium have the least and the most heterogeneity in centromere position, respectively; L. pumilum and L. davidii var. willmottiae have the lowest and the highest intrachromosomal asymmetry, respectively; L. davidii var. willmottiae and L. regale have the lowest and highest level of interchromosomal asymmetry, respectively. The bidimensional scatter plot of M CA vs. CV CL shows that the karyotype structure of the twelve taxa can be completely distinguished by this couple of parameters because no overlap among taxa occurs (Fig. 3 ). The PCoA scatter plot shows that the 12 taxa are divided into two groups along the direction of PCoA1: L. pumilum , L. jinfushanense , L. brownii var. viridulum , L. regale , L. davidii var. willmottiae , L. lancifolium and L. lancifolium ‘Longshan’ in one group with L. pumilum occupying the most isolated position; while L. sulphureum , L. henryi , L. leucanthum , L. sargentiae , and L. rosthornii in another group (Fig. 4 ). Among the species clustered on the left, L. regale is the closest to L. henryi and L. sargentiae , which are clustered on the right (Fig. 4 ). FISH localization of 5S and 35S rDNA loci All taxa but L. rosthornii have a single locus of 5S rDNA, which in L. pumilum , L. davidii var. willmottiae , L. lancifolium , L. lancifolium ‘Longshan’ and L. brownii var. viridulum is situated in the interstitial regions of the long arms of chromosome 3 (Figs. 1 a, 1 b, 1 c, 1 d, 1 f, 2 a, 2 b, 2 c, 2 d, and 2 f; Table 2 ), while in L. jinfushanense , L. leucanthum , L. sargentiae , L. sulphureum , L. regale , L. henryi and L. rosthornii is located in the proximal regions of the long arms of chromosome 3 (Figs. 1 e, 1 g, 1 h, 1 i, 1 j, 1 k, 1 l, 2 e, 2 g, 2 h, 2 i, 2 j, 2 k, and 2 l; Table 2 ). In L. rosthornii , in addition to the 5S locus located on chromosome 3, another 5S locus located in the proximal regions of the short arms of chromosome 6 was detected (Figs. 1 l, and 2 l; Table 2 ). Particularly, the FISH signals of the 5S loci on chromosome 3 of both L. henryi and L. rosthornii are particularly strong, indicating a striking similarity between them. In L. pumilum and L. brownii var. viridulum , the intensity of 5S signal is different between homologous chromosomes, showing heterozygosity of the 5S locus (Figs. 1 a, f). The number of 35S rDNA loci ranges from two to six among the 12 taxa (Figs. 1 , and 2 ; Table 2 ). Most of the 35S loci (93.5%) generate SCs (Fig. S1 ; Table S2 ). In L. pumilum , five 35S loci are located in the proximal regions of the short arms of chromosome 1, the interstitial regions of the short arms of chromosome 2, the interstitial regions of the long arms of chromosome 3 and 5, the centromeric regions of chromosome 4, respectively (Figs. 1 a, and 2 a; Table 2 ). In L. davidii var. willmottiae , four 35S loci occur in the proximal regions of the short arms of chromosome 1 and 9, the proximal regions of the long arms of chromosome 4, the interstitial regions of the long arms of chromosome 6, respectively (Figs. 1 b, and 2 b; Table 2 ). Both L. lancifolium and L. lancifolium ‘Longshan’ have 35S loci on chromosome 1 and 2 at the same positions as chromosome 1 and 2 of L. pumilum , as well as other three 35S loci located in the interstitial regions of the long arms of chromosome 6 and 11, and the proximal regions of the short arms of chromosome 7, respectively (Figs. 1 c, 1 d, 2 c, and 2 d; Table 2 ). Compared with L. lancifolium ‘Longshan’, L. lancifolium has one more 35S locus on chromosome 8, and its 35S loci on chromosome 6, 8 and 11 appear only in one or two members of the triplets (Figs. 1 c, 1 d; Table 2 ). L. jinfushanense has only two 35S loci located in the proximal regions of the short arms of chromosome 1 and 2 (Figs. 1 e, and 2 e; Table 2 ). L. brownii var. viridulum possesses three 35S loci located in the interstitial regions of long arms of chromosome 4 and 5, and the proximal regions of the short arms of chromosome 7 (Figs. 1 f, and 2 f; Table 2 ). L. leucanthum , L. sargentiae , L. sulphureum and L. regale have four 35S loci at similar positions, i.e. the proximal regions of the short arms of chromosome 1 and 4, and the proximal regions of the long arms of chromosome 2 and 5 (Figs. 1 g, 1 h, 1 i, 1 j, 2 g, 2 h, 2 i, and 2 j; Table 2 ). L. regale has another 35S locus located in the proximal regions of the long arms of chromosome 6 (Figs. 1 j, and 2 j; Table 2 ). The percentage distances of the 35S locus on chromosome 5 vary among the four species (Figs. 2 g, 2 h, 2 i, and 2 j; Table 2 ). L. henryi and L. rosthornii have similar 35S patterns, with one locus in the proximal regions of the short arms of chromosome 1 and another locus in the interstitial regions of the long arms of a subtelocentric chromosome pair (Figs. 1 k, 1 l, 2 k, 2 l; Table 2 ). Discussion Karyotype variation and species differentiation A large number of previous cytogenetic studies have revealed that the karyotypes of Lilium are rather conserved among species. The karyotype conservatism of Lilium is mainly manifested in two aspects: first, except a few species that have triploid, tetraploid or aneuploidy in addition to diploid, the vast majority of species are all diploids with a chromosome number 2n = 24, and the basic chromosome number is x = 12; second, the haploid genome of all species is composed of two larger metacentric or submetacentric chromosomes and ten smaller subtelocentric or telocentric chromosomes with their karyotypes falling into 3A or 3B type as judged by Stebbins’ asymmetry index. Our study shows that except L. lancifolium and L. lancifolium ‘Longshan’ all other species are diploid. Extensive and in-depth cytogenetic studies have been conducted on L. lancifolium . This species is a cytologically conspicuous species with both diploids and triploids coexisting in nature. However, the diploid form has only been found in a few regions of South Korea and Japan (Noda 1986 , 1991 ; Nguyen et al. 2016 ). Our results indicate that the two populations of L. lancifolium studied herein are triploids, being consistent with the previously reported populations from China (Noda 1986 ; Nguyen et al. 2016 ; Liu et al. 2017 ; Zhou et al. 2020 ). Several previous studies have supported that the triploid L. lancifolium is an autopolyploid derived from the diploid L. lancifolium (Sultana et al. 2010 ; Nguyen et al. 2016 ). Our molecular-cytogenetic karyotyping shows that the chromosomal morphology and rDNA locus distribution in the same triplets of L. lancifolium are very similar, also providing evidence for its autopolyploid nature. When the chromosome number is the same, the karyomorphological differences between plant species or populations can be measured by a series of parameters (Paszko 2006 ; Peruzzi and Eroglu 2013 ). Nine parameters are given in this study, among which karyotype formula and karyotype asymmetry indices should be the main parameters for karyotypic comparison. In terms of the karyotype formula, L. lancifolium and L. lancifolium ‘Longshan’ are the same, and the karyotype formulas vary among the haploid genomes of the eleven species studied, including differences in the number of different chromosome types and in the number and location of SCs. Previous studies have suggested that the first two chromosome pairs are quite stable and likely evolutionary residues in Lilium that can serve as a specific trait of this genus (Gao et al. 2011 , 2012 ). Our results show that chromosome 1 of all species studied is m-type, while chromosome 2 is m- or sm-type (m-type in 6 species and sm-type in 5 species) (Table 1 ; Fig. 2 ). Among the 47 taxa of Lilium (39 species involved) from China studied by Gao et al. 2011 and Liu et al. 2017 , all taxa had chromosome 1 of m-type, 27 taxa had chromosome 2 of m-type, and 20 taxa had chromosome 2 of sm-type. These facts suggest that chromosome 1 is fairly stable during speciation in Lilium , while chromosome 2 is relatively prone to chromosomal rearrangement, resulting in changes in the length of its short and/or long arms. As for the ten smaller chromosomes, the haploid genomes consist of 7st + 3t, 5st + 5t, 4st + 6t and 3st + 7t chromosomes among the eleven species studied (Table 1 ; Fig. 2 ). In the karyotypes arranged in the order of decreasing length of the short arms, the RLs of the long arms of chromosomes 3 to 12 in the same numbered chromosomes of different species can vary significantly (Fig. 2 ). Even between closely related species such as L. leucanthum , L. sargentiae , L. sulphureum and L. regale , as well as L. henryi and L. rosthornii (detailed discussion on relationships see below), such differences also exist. For examples, the RLs of the long arms of chromosome 10 of L. sulphureum are significantly longer than those of L. leucanthum , L. sargentiae and L. regale ; the RLs of the long arms of chromosome 11 of L. sulphureum and L. regale are significantly shorter than those of L. leucanthum and L. sargentiae (Figs. 2 g, 2 h, 2 i, and 2 j; Table S2 ). For another example, there are significant differences in the RLs of the long arms of chromosomes 4, 5, 6, 9 and 12 between L. henryi and L. rosthornii (Figs. 2 k, and 2 l). The short arms of chromosomes 3 to 12 may also have their lengths altered due to rearrangements. For instance, chromosome 5 of L. rosthornii is similar to chromosome 4 of L. henryi because both have an interstitial 35S locus in the long arms (Fig. 2 k, and 2 l). The change in the order is likely caused by a chromosomal rearrangement, which has shortened the short arms of the corresponding chromosome pair in L. rosthornii . In a word, the differences in karyotype formula among different species and the variation in the lengths of the long arms of the same numbered chromosomes indicate that speciation in Lilium involves multiple chromosomal rearrangements (translocations, inversions, deletions, etc.). Karyotype asymmetry is an important parameter to measure the degree of genomic structural variation among species (Astuti et al. 2017 ). The use of statistically correct and reliable karyotype asymmetry indices as well as effective analysis methods can not only effectively distinguish the differences in karyotype structure among species and even among populations, but also reveal the karyological relationships among species (e.g. Peruzzi and Eroglu 2013 ; Peruzzi and Altınordu 2014 ; Kadluczka and Grzebelus 2021 ; She et al. 2023 ; Mitrenina et al. 2023 ; Wei et al. 2024 ). Our results show that all the 12 taxa of Lilium studied herein belong to type 3A or 3B of the Stebbins’ category, and this index cannot show their differences in karyotype structure, but the bidimensional scatter plots of M CA vs. CV CL can clearly distinguish all the 12 taxa, and there is no overlap even between the two populations of L. lancifolium . Our analysis demonstrates that calculating M CA and CV CL values and making dimensional scatter plot are an effective method to distinguish the karyotype structure of different species of Lilium , and also shows that the karyomorphology of Lilium is not the case that the karyotype differences between species are small and the major differences among species are mainly reflected in the number and position of SCs, as previously thought (Noda 1991 ; Gao et al. 2011 ). Variation of rDNA pattern and evolutionary relationships among species A large number of molecular cytogenetic studies have shown that the similarity and difference in rDNA pattern can directly reflect the closeness of relatedness between species in a plant genus (e.g. She et al. 2015 ; Mitrenina et al. 2023 ; Dias et al. 2024 ; Wei et al. 2024 ; Sun et al. 2024 ), and can also reflect the genetic differentiation between populations within a species (e.g. Nguyen et al. 2015 , 2016 ; Wei et al. 2024 ). Our rDNA FISH results show that there are more or less differences in rDNA patterns of the eleven species of Lilium studied. According to the previous reports on FISH mapping of rDNA in Lilium (Lim et al. 2001 ; Marasek et al. 2004 ; Muratović et al. 2010 ; Sultana et al. 2010 ; Lee et al. 2014 ; Nguyen et al. 2015 , 2016 ; Ahn et al. 2017 ; Zhou et al. 2020 ; Choi et al. 2021 ; Zhou et al. 2024 ; Kang et al. 2024 ; Zhu et al. 2025 ), to compare the rDNA patterns of different species of Lilium , we should first observe whether there are 35S loci on chromosome 1 and 2 and the differences in their locations, as well as the differences in the locations of the 5S locus on chromosome 3, and then should one check whether there are other 35S and 5S loci and the differences in their locations. In L. pumilum , the number and locations of the 35S loci detected by us were similar to those of Zhou et al. ( 2020 ). Lee et al. ( 2014 ) also detected five 35S loci and one 5S locus in L. pumilum , with four 35S loci and the 5S locus at the locations similar to ours. But instead of detecting 35S locus adjacent to the 5S locus, they detected one 35S locus on the short arms of chromosome 7, suggesting that there were differences in rDNA pattern among different populations of this species. In L. lancifolium and L. lancifolium ‘Longshan’, the number and locations of the 35S loci detected by us were the same as those of the triploid populations previously reported by most authors (Sultana et al. 2010 ; Hwang et al. 2011 ; Lee et al. 2014 ; Liu et al. 2017 ; Zhou et al. 2020 ), but somewhat different from those of the triploid populations reported by Nguyen et al. 2016 , in which one 35S locus was detected in the interstitial regions of the long arms of three or two or one members of triplet 4. The 35S locus located on the long arms of one member of triplet 8 was a new locus that hasn't been reported before in L. lancifolium . Previous reports have also found that the 35S locus on chromosomes 6 and 11 in some triploid populations appeared in only two members of the respective triplet (Nguyen et al. 2016 ). All triploid populations previously reported have two adjacent 5S loci located in the interstitial regions of long arms of chromosome 3 (Sultana et al. 2010 ; Hwang et al. 2011 ; Lee et al. 2014 ; Nguyen et al. 2016 ), but we only detected a single 5S locus. In a word, the rDNA loci are high variable among the triploid populations of L. lancifolium that rely on asexual reproduction only (Noda 1986 ). In L. davidii var. willmottiae , the number and locations of the 35S loci detected by us were similar to those of L. davidii var. willmottiae detected by Zhou et al. ( 2020 ), and those of L. davidii var. unicolor (Synonym L. davidii var. davidii ) detected by Wu et al. ( 2021 ). Wu et al. ( 2021 ) detected four 5S loci in L. davidii var. unicolor , that is, other three 5S loci were detected in addition to the 5S locus located in the interstitial regions of the long arms of chromosome 3. These facts indicate that the 35S loci of the two varieties of L. davidii are identical, but the number of 5S loci is quite different, although the 5S locus on chromosome 3 is conserved. The locations of the 35S loci on chromosomes 1 and 2 and the 5S loci on chromosome 3 of L. lancifolium and L. pumilum are similar. Moreover, the 35S locus on chromosome 6 of L. lancifolium is similar to that on chromosome 5 of L. pumilum (possibly due to chromosomal rearrangement that shortened the short arms and changed the numbering of the chromosome). These similarities indicate that L. lancifolium and L. pumilum have a close relationship. Both L. davidii var. willmottiae and L. lancifolium have 35S loci on chromosomes 1 and 6 and the positions are similar between them. Both L. davidii var. willmottia and L. pumilum have 35S loci on chromosomes 1 and 4 and the positions are similar between them. Moreover, the location of the 5S locus of L. davidii var. willmottiae is similar to that of L. pumilum and L. lancifolium. These similarities indicate that L. davidii is relatively closely related to L. lancifolium and L. pumilum . The relatedness among these three species revealed by our comparison of rDNA pattern is basically consistent with that previously revealed by molecular phylogenetic analyses (Du et al. 2014 ; Li et al. 2022 ; Zhou et al. 2023 ). In the PCoA scatter plot, L. pumilum, L. lancifolium and L. davidii var. willmottiae are clustered together in the same group along the direction of PCoA1, roughly confirming the relatedness of these species drawn from rDNA FISH and molecular phylogenetics. The 35S pattern of L. jinfushanense is different from that of any of the other species studied herein, but is the same as that of L. taliense reported by Zhou et al. 2020 . The location of the 5S locus on chromosome 3 of L. jinfushanense is different from that of the species belonging to Sinomartagon 5a and Sinomartagon 5b, but similar to that of the species belonging to Leucolirion 6a. The consistency of the 35S pattern between L. jinfushanense and L. taliense indicates their very close relationship. This is accordant with the result of molecular phylogenetic analysis based on ITS sequences and plastid genomes (Du et al. 2014 ; Li et al. 2022 ). It is worth in-depth study how the unique rDNA pattern of L. jinfushanense evolved and the evolutionary relationships with other species endemic to southwest China. The 35S pattern of L. brownii var. viridulum detected by us is the same as that of L. brownii var. viridulum and L. brownii reported by several authors (Liu et al. 2017 ; Wu et al. 2021 ; Zhou et al. 2020 ), but is completely inconsistent with that of L. brownii var. viridulum reported by Wu et al. 2012. As for the 5S locus of L. brownii var. viridulum , we only detected the 5S locus in the interstitial regions of the long arms of chromosome 3, but Wu et al. 2021 detected another five 5S loci. This inconsistency may be due to differences in material used. L. brownii was placed in sect. Archelirion by Comber ( 1949 ) and later adjusted to Leucolirion 6b based on molecular phylogenetic evidence (Du et al. 2014 ; Du et al. 2023). L. longiflorum and L. formosanum , two species also belonging to Leucolirion 6b, have a 35S pattern somewhat similar to that of L. brownii var. viridulum : they also have no 35S loci on chromosomes 1 and 2, and have a 35S locus in the proximal regions of short arms of chromosome 7 (Lim et al. 2001 ; Zhou et al. 2020 ). These facts indicate that the rDNA pattern of already studied species of Leucolirion 6b differ significantly from those of species of Leucolirion 6a studied herein. The locations of the 5S loci on chromosome 3 and the 35S loci on chromosomes 1, 2, 4 and 5 of L. leucanthum , L. sargentiae , L. sulphureum , and L. regale are the same or very similar, indicating their close relationships. PCoA also indicates that these species have close karyological relationships. Our study demonstrated from the perspective of cytogenetics that these four species are closely related to each other, in accordance with the conclusion inferred from molecular phylogenetic studies (Du et al. 2014 ; Li et al. 2022 ). It is worth noting that among these four species, the karyotype differentiation of L. regale is more significant. Not only does it have an additional 35s locus located on chromosome 6, but its karyotype parameters also alter greatly compared to the other three species, leading to its aggregation in another group in the PCoA plot. The rDNA localization of L. leucanthum has not been reported till now. Wang et al. 2012 reported the rDNA mapping of L. leucanthum var. centifolium , and showed no 35S loci on chromosome 1 and 2. Such 35S pattern of this variety is very different from that of L. leucanthum revealed by us. It is necessary to further determine the rDNA pattern of this variety by dual-color FISH. Previous studies have detected four 35S loci located on chromosomes 1 and 2 as well as two st-type chromosome pairs in L. sargentiae and L. sulphureum (Liu et al. 2017 ; Zhou et al. 2020 ; Zhu et al. 2025 ), and detected four (Zhou et al. 2024 ; Zhu et al. 2025 ), five (Cao et al. 2019 ; Zhou et al. 2020 ) and six (Liu et al. 2017 ) 35S loci (including the two loci on chromosomes 1 and 2) in L. regale , but these studies fail to accurately determine the chromosomal locations of some of these loci. Our comparative localization of 5S and 35S rDNA of L. henryi and L. rosthornii by dual-color FISH reveals the similarity of their rDNA patterns, further demonstrating the close relationship between them from the perspective of cytogenetics (Du et al. 2014 ; Zhou et al. 2020 ). Previous studies have reported the localization of both 5S and 35S (Marasek et al. 2004 ; Kang et al. 2024 ) and the localization of 35S alone (Du et al. 2014 ; Liu et al. 2017 ; Zhou et al. 2020 ) in L. henryi . Our result of rDNA mapping of this species is basically consistent with those of previous reports, except that the numbering of the st-type chromosome pair with 35S locus is different from theirs (Marasek et al. 2004 ; Du et al. 2014 ; Zhou et al. 2020 ; Kang et al. 2024 ). For L. rosthornii , previous studies only mapped 35S by FISH, revealing a similar pattern with ours (Du et al. 2014 ; Zhou et al. 2020 ). Earlier, L. rosthornii was assigned to Sinomartagon 5a (Comber 1949 ), and later transferred to Leucolirion 6a based on the conclusions of molecular phylogenetic analysis and 35S rDNA FISH (Du et al. 2014 ; van Tuyl et al. 2018 ; Zhou et al. 2020 ; Du et al. 2023). In the phylogenetic tree based on ITS sequences, L. henryi and L. rosthornii belong to Leucolirion clade I along with L. leucanthum , L. sargentiae , L. sulphureum and L. regale . The former two were clustered into one branch which showed a sister relationship with the branch where the latter four species were clustered (Du et al. 2014 ). In the latest revised classification of Lilium , these six species have been assigned to Leucolirion 6a (van Tuyl et al. 2018 ; Du et al. 2023). From the comparison of their rDNA patterns, the 5S locus of chromosome 3 and the 35S locus of chromosome 1 of the former two species are similar to those of the latter four species, but such similarities are not sufficient to indicate that there are close relationships between the species of the two branches. However, PCoA based on the six karyotypic parameters provides evidence. As shown in the PCoA scatter plot, L. henryi and L. rosthornii are clustered in the same group with L. leucanthum , L. sargentiae , and L. sulphureum along the direction of PCoA1. Conclusion Molecular cytogenetic karyotypes of eleven species of Lilium from China are established quantitatively using the dataset of chromosome measurements and FISH signals of 5S and 35S rDNA. Comparative karyotype analysis reveals variations in the karyotypic parameters and rDNA patterns among species. Their karyotype structures could be differentiated by the scatter plot of M CA vs. CV CL . The evolutionary relationships among them can be revealed by the combination of comparison of rDNA patterns as well as PCoA based on x , 2n, TCL, CV CI , M CA and CV CL . Our results confirm cytogenetically the rationality of the species classification of subset. Leucolirion 6a in the latest revised classification of Lilium. Declarations Conflict of interest The authors declare no competing interests. Funding The Natural Science Foundation of Hunan Province, China (2019JJ40231, 2024JJ7156). Author Contribution Conceptualization: CWS; Funding acquisition: CWS, BYZ; Investigation: YDY; Data curation: YDY, XHJ; Formal analysis: YDY, XHJ; Methodology: CWS, YDY, XHJ, BYZ; Project administration: CWS; Resources: BYZ, YDY; Supervision: CWS; Validation: YDY; Visualization: YDY; Writing - original draft: YDY; Writing - review & editing: CWS, XHJ, BYZ. Data Availability All data supporting the findings of this study are available in the article and its supplementary information files online. References Ahn, Y. J. et al. Investigation of karyotypic composition and evolution in Lilium species belonging to the section Martagon. Plant. Biotechnol. Rep. 11 , 407–416. https://doi.org/10.1007/s11816-017-0462-7 (2017). Astuti, G., Roma-Marzio, F. & Peruzzi, L. Traditional cytotaxonomic studies: can they still provide a solid basis in plant systematics? Fl Medit . 27 , 91–98. https://doi.org/10.7320/FlMedit27.091 (2017). Cao, Q. et al. Physical mapping of 45S rDNA loci in Lilium OT hybrids and interspecific hybrids with Lilium regale . Sci. Hortic. 252 , 48–54. https://doi.org/10.1016/j.scienta.2019.03.035 (2019). Chinese Pharmacopoeia Commission. Pharmacopoeia of the People’s Republic of China: 2020 version pp186–187 (Vol. I. China Medical Science, 2020). Choi, Y. H. et al. Using cytogenetic analysis to identify the genetic diversity in Lilium hansonii (Liliaceae), an endemic species of Ulleung Island, Korea. Hortic. Environ. Biotechnol. 62 , 795–804. https://doi.org/10.1007/s13580-021-00350-1 (2021). Comber, H. F. A new classification of the genus Lilium . Lily Yearb Roy Hort Soc. 13 , 86–105. https://doi.org/10.1007/s11032-010-9490-5 (1949). Dias, S. et al. do,, Cytomolecular diversity among Vigna Savi (Leguminosae) subgenera. Protoplasma 261:859–875. (2024). https://doi.org/10.1007/s00709-024-01944-z Du, F. Origin, classification and germplasm diversity in Lilium . J. China Agricultural Univ. 28 , 68–79. https://doi.org/10.11841/j.issn.1007-4333.2023.04.06 (2023). Du, Y. P. et al. Molecular phylogeny and genetic variation in the genus Lilium native to China based on the internal transcribed spacer sequences of nuclear ribosomal DNA. J. Plant. Res. 127 , 249–263. https://doi.org/10.1007/s10265-013-0600-4 (2014). Gao, Y. D. & Gao, X. F. Accommodating Nomocharis in Lilium (Liliaceae). Phytotaxa 277:205–210. (2016). https://doi.org/10.11646/phytotaxa.277.2.8 Gao, Y. D., Zhou, S. D. & He, X. J. Karyotype studies in thirty-two species of Lilium (Liliaceae) from China. Nord J. Bot. 29 , 746–761. https://doi.org/10.1111/j.1756-1051.2011.01069.x (2011). Gao, Y. D., Zhou, S. D., He, X. J. & Wan, J. Chromosome diversity and evolution in tribe Lilieae (Liliaceae) with emphasis on Chinese species. J. Plant. Res. 125 , 55–69. https://doi.org/10.1007/s10265-011-0422-1 (2012). Givnish, T. et al. Evolution, geographic spread and floral diversification of the genus Lilium with special reference to the lilies of North America. Lily Yearbook North. Am. Lily Soc. 74 , 26–44 (2020). Han, Y. L. et al. Comparative analysis of rDNA distribution in 29 species of Begonia sect. Coelocentrum Irmsch Phytotaxa . 381 , 141–152. https://doi.org/10.11646/PHYTOTAXA.381.1.18 (2018). Hwang, Y. J., Kim, H. H., Kim, J. B. & Lim, K. B. Karyotype analysis of Lilium tigrinum by FISH. Hort Environ. Biotechnol. 52 , 292–297. https://doi.org/10.1007/s13580-011-0225-2 (2011). Kadluczka, D. & Grzebelus, E. Using carrot centromeric repeats to study karyotype relationships in the genus Daucus (Apiaceae). BMC Genom. 22 , 508. https://doi.org/10.1186/s12864-021-07853-2 (2021). Kang, J. Y., Lim, K. B. & Ahn, Y. J. Cytogenetic analysis and visualization of genetic relationships in wild lilies. Genes Genom . 46 , 1253–1265. https://doi.org/10.1007/s13258-024-01568-w (2024). Lee, H., Younis, A., Hwang, Y. J., Kang, Y. I. & Lim, K. B. Molecular cytogenetic analysis and phylogenetic relationship of 5S and 45S ribosomal DNA in Sinomartagon Lilium species by fluorescence in situ hybridization (FISH). Hortic. Environ. Biotechnol. 55 , 514–523. https://doi.org/10.1007/s13580-014-0066-x (2014). Levan, A., Fredga, K. & Sandberg, A. Nomenclature for centromeric position in chromosomes. Hereditas 52 , 201–220. https://doi.org/10.1111/j.1601-5223.1964.tb01953.x (1964). Li, J. et al. Phylogeny, age, and evolution of tribe Lilieae (Liliaceae) based on whole plastid genomes. Front. Plant. Sci. 12 , 699226. https://doi.org/10.3389/fpls.2021.699226 (2022). Liang, S. Y. & Tamura, N. Lilium. In: (eds Wu, Z. Y. & Raven, P. H.) Flora of China. Science Press, Beijing, Miss Bot Gard, St. Louis, 24:135–149 (2000). Lim, K. B., Wennekes, J., de Jong, J. H., Jacobsen, E. & van Tuyl, J. M. Karyotype analysis of Lilium longiflorum and Lilium rubellum by chromosome banding and fluorescence in situ hybridisation. Genome 44 , 911–918. https://doi.org/10.1139/gen44-5-911 (2001). Liu, G. X. et al. Karyotype and fluorescence in situ hybridization analysis of 15 Lilium species from China. J. Am. Soc. Hortic. Sci. 142 , 298–305. https://doi.org/10.21273/JASHS04094-17 (2017). Marasek, A., Hasterok, R., Wiejacha, K. & Orlikowska, T. Determination by GISH and FISH of hybrid status in Lilium . Hereditas 140 , 1–7. https://doi.org/10.1111/j.1601-5223.2004.01721.x (2004). Mitrenina, E. Y. et al. Karyotypes and physical mapping of ribosomal DNA with Oligo-Probes in Eranthis sect. Eranthis (Ranunculaceae). Plants (Basel) . 13 , 47. https://doi.org/10.3390/plants13010047 (2023). Muratović, E., Robin, O., Bogunić, F., Šoljan, D. & Siljak-Yakovlev, S. Karyotype evolution and speciation of European lilies from Lilium sect. Liriotypus Taxon . 59 , 165–175. https://doi.org/10.2307/27757060 (2010). Nguyen, T. X. et al. Karyotype analysis of Korean Lilium maximowiczii Regal populations. J. Fac. Agr Kyushu Univ. 60 , 315–322. https://doi.org/10.5109/1526344 (2015). Nguyen, T. X., Lee, S. I., Rai, R., Kim, N. S. & Kim, J. H. Ribosomal DNA locus variation and REMAP analysis of the diploid and triploid complexes of Lilium lancifolium . Genome 59 , 551–564. https://doi.org/10.1139/gen-2016-0011 (2016). Noda, S. Cytogenetic behavior, chromosomal differentiations, and geographic distribution in Lilium lancifolium (Liliaceae). Plant. Spec. Biol. 1 , 69–78. https://doi.org/10.1111/j.1442-1984.1986.tb00016.x (1986). Noda, S. Chromosomal variation and evolution in the genus Lilium . In: Tsuchiya T, Gupta PK (eds), Chromosome engineering in plants: genetics, breeding, evolution, part B. Elsevier, pp 507–524. (1991). https://doi.org/10.1016/B978-0-444-88260-8.50032-X Paszko, B. A critical review and a new proposal of karyotype asymmetry indices. Plant. Syst. Evol. 258 , 39–48. https://doi.org/10.1007/s00606-005-0389-2 (2006). Patterson, T. B. & Givnish, T. J. Phylogeny, concerted convergence, and phylogenetic niche conservatism in the core Liliales: insights from rbcL and ndhF sequence data. Evolution 56 , 233–252. https://doi.org/10.1111/j.0014-3820.2002.tb01334.x (2002). Peruzzi, L. & Altınordu, F. A proposal for a multivariate quantitative approach to infer karyological relationships among taxa. Comp. Cytogenet. 8 , 337–349. https://doi.org/10.3897/CompCytogen.v8i4.8564 (2014). Peruzzi, L. & Eroglu, H. Karyotype asymmetry: again, how to measure and what to measure? Comp. Cytogenet. 7 , 1–9. https://doi.org/10.3897/CompCytogen.v7i1.4431 (2013). Peruzzi, L., Leitch, I. J. & Caparelli, K. F. Chromosome diversity and evolution in Liliaceae. Ann. Bot. 103 , 459–475. https://doi.org/10.1093/aob/mcn230 (2009). She, C. W., Jiang, X. H. & He, C. P. Comparative karyotype analysis of eight Cucurbitaceae crops using fluorochrome banding and 45S rDNA-FISH. Comp. Cytogenet. 17 , 31–58. https://doi.org/10.3897/compcytogen.v17.i1.99236 (2023). She, C. W. et al. Molecular cytogenetic characterisation and phylogenetic analysis of the seven cultivated Vigna species (Fabaceae). Plant. Biol. 17 , 268–280. https://doi.org/10.1111/plb.12174 (2015). Smyth, D. R., Kongsuwan, K. & Wisudharomn, S. A survey of C-band patterns in chromosomes of Lilium (Liliaceae). Plant. Syst. Evol. 163 , 53–69. https://doi.org/10.1007/BF00936153 (1989). Stebbins, G. L. Chromosomal evolution in higher plants (Addison-Wesley, 1971). pp220. Stewart, R. N. The morphology of somatic chromosomes in Lilium . Am. J. Bot. 34 , 9–26. https://doi.org/10.1002/j.1537-2197.1947.tb12953.x (1947). Sultana, S., Lee, S. H., Bang, J. W. & Choi, H. W. Physical mapping of rRNA gene loci and inter-specific relationships in wild Lilium distributed in Korea. J. Plant. Biol. 53 , 433–443. https://doi.org/10.1007/s12374-010-9133-8 (2010). Sun, J. et al. Comparative karyotype analysis provides cytogenetic evidence for the origin of sweetpotato. Chromosome Res. 32 , 14. https://doi.org/10.1007/s10577-024-09758-8 (2024). Van Tuyl, J. M. et al. Lilium. In: (ed Van Huylenbroeck, J.) Ornamental crops. Handbook of Plant Breeding, vol 11. Springer, Cham., 481–512. https://doi.org/10.1007/978-3-319-90698-0_20 (2018). Wan, J., Zhou, S., Gao, Y. & He, X. Karyotypes of twenty-five populations of thirteen species in Nomocharis and Lilium . Plant. Divers. Resour. 33 , 477–494. https://doi.org/10.3724/SP.J.1011.2011.00093 (2011). Wang, Q. et al. The application of fluorescence in situ hybridization in different ploidy levels cross-breeding of lily. PLoS One . 10 , e0126899. https://doi.org/10.1371/journal.pone.0126899 (2015). Wang, X., Xie, S., Zhang, Y. & Niu, L. Chromosome analysis and mapping of ribosomal genes by fluorescence in situ hybridization (FISH) in four endemic lily species ( Lilium ) in Qinling mountians, China. Pak J. Bot. 44 , 1319–1323 (2012). Wei, Y. F., Jiang, X. H., Song, R. & She, C. W. Molecular cytogenetic characterization of 9 populations of four species in the genus Polygonatum (Asparagaceae). Comp. Cytogenet. 18 , 73–95. https://doi.org/10.3897/compcytogen.18.122399 (2024). Wu, L. et al. Analysis of the cross-compatibility of Lilium brownii var. viridulum and L. davidii var. unicolor . Sci Hortic 284:110130. (2021). https://doi.org/10.1016/j.scienta.2021.110130 Zhou, Y. P. et al. Fluorescence in situ hybridization of 35S rDNA sites and karyotype of wild Lilium (Liliaceae) species from China: taxonomic and phylogenetic implications. Genet. Resour. Crop Evol. 67 , 1601–1617. https://doi.org/10.1007/s10722-020-00936-1 (2020). Zhou, J., An, R. & Huang, X. Genus Lilium : A review on traditional uses, phytochemistry and pharmacology. J. Ethnopharmacol. 270 , 113852. https://doi.org/10.1016/j.jep.2021.113852 (2021). Zhou, M. et al. Chromosomal analysis of progenies between Lilium intersectional hybrids and wild species using ND-FISH and GISH. Front. Plant. Sci. 15 , 1461798. https://doi.org/10.3389/fpls.2024.1461798 (2024). Zhou, N. et al. Historical biogeography and evolutionary diversification of Lilium (Liliaceae): New insights from plastome phylogenomics. Plant. Divers. 46 , 219–228. https://doi.org/10.1016/j.pld.2023.07.009 (2023). Zhu, Z., Sun, Y., Liu, S., Peng, M. & Zhou, S. Comparison of Lilium regale and L. sargentiae for lily breeding. Genet. Resour. Crop Evol. 72 , 6375–6386. https://doi.org/10.1007/s10722-025-02405-z (2025). Additional Declarations No competing interests reported. Supplementary Files Supplementarymaterial.docx Supplementarymaterialrevised.docx Cite Share Download PDF Status: Published Journal Publication published 17 Feb, 2026 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 23 Dec, 2025 Reviews received at journal 09 Dec, 2025 Reviewers agreed at journal 24 Nov, 2025 Reviews received at journal 24 Nov, 2025 Reviewers agreed at journal 19 Nov, 2025 Reviewers invited by journal 19 Nov, 2025 Editor assigned by journal 19 Nov, 2025 Editor invited by journal 18 Nov, 2025 Submission checks completed at journal 13 Nov, 2025 First submitted to journal 13 Nov, 2025 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8029111","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":550274522,"identity":"1ae63bcb-427a-46d4-916a-777480138725","order_by":0,"name":"Ya-Di Yang","email":"","orcid":"","institution":"Hunan University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Ya-Di","middleName":"","lastName":"Yang","suffix":""},{"id":550274523,"identity":"2fa09301-a94d-402e-9528-5c1c1d95f489","order_by":1,"name":"Xiang-Hui Jiang","email":"","orcid":"","institution":"Huaihua University","correspondingAuthor":false,"prefix":"","firstName":"Xiang-Hui","middleName":"","lastName":"Jiang","suffix":""},{"id":550274524,"identity":"6c114638-dfff-4bf9-a07f-b60b7eb8b185","order_by":2,"name":"Bang-Yue Zhang","email":"","orcid":"","institution":"Hunan University of Technology","correspondingAuthor":false,"prefix":"","firstName":"Bang-Yue","middleName":"","lastName":"Zhang","suffix":""},{"id":550274525,"identity":"9e78b5c2-9471-4654-bb3c-d2c0f5152d3e","order_by":3,"name":"Chao-Wen She","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3ElEQVRIiWNgGAWjYDACCQYGZgiL+QDDAwPStLAlMCSQqIXHgCGBGB3ys5sPfi6osUnsl+75+CGh4HDidgbmh49u4NFicOdYsvSMY2mJM+ec3SyRYHA4cWcDm7FxDj4tEjlmzDxsh3M33MjdANay4QAPmzQ+LfIz8r8x8/w7nLv/Rs7jH0RpYbiRw8bM2wa0RSKHjThbDG6kGUvz9qXVz7iRZmaRYJBuvOEwAb/Iz0h++Jnnm40x/4zkxzc+/LGW3XC8+eFjvA5DA83waCIa1JGofhSMglEwCkYCAACtmU/fb6cszgAAAABJRU5ErkJggg==","orcid":"","institution":"Hunan University of Technology","correspondingAuthor":true,"prefix":"","firstName":"Chao-Wen","middleName":"","lastName":"She","suffix":""}],"badges":[],"createdAt":"2025-11-04 13:08:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8029111/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8029111/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-026-37297-1","type":"published","date":"2026-02-17T15:56:53+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":97136070,"identity":"fd1e5e1f-2395-46e0-9f11-2c7b74e9eb32","added_by":"auto","created_at":"2025-12-01 09:55:20","extension":"tif","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":48684636,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.1.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/561ef44ee5110652dd0ecb7c.tif"},{"id":96943740,"identity":"c2f236b1-d3be-4fb4-b832-e5adf620b021","added_by":"auto","created_at":"2025-11-27 19:01:12","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":2070012,"visible":true,"origin":"","legend":"","description":"","filename":"ComparativekaryotypeanalysisofelevenspeciesofLiliumfromChinabyFISHwithrDNAoligoprobessecondrevision.docx","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/d0436f711d6a79453873d26e.docx"},{"id":97136039,"identity":"dff81e97-1d55-453c-a711-02086b1c8302","added_by":"auto","created_at":"2025-12-01 09:55:10","extension":"tif","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":55124828,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.2.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/773bbfdcc1dfce1e414fa7b7.tif"},{"id":96943741,"identity":"53b81c26-647e-4286-936e-5886f17a4f80","added_by":"auto","created_at":"2025-11-27 19:01:12","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":18928,"visible":true,"origin":"","legend":"","description":"","filename":"Table1.docx","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/5595a9a3feba914991ab7a82.docx"},{"id":96943743,"identity":"7d91f475-afc2-41dc-bbcd-a91133bfb552","added_by":"auto","created_at":"2025-11-27 19:01:12","extension":"docx","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":17618,"visible":true,"origin":"","legend":"","description":"","filename":"Table2.docx","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/d3688c27be33b9a1912bdfa7.docx"},{"id":96943773,"identity":"03bf0734-869b-42ba-91cd-5741be83abf0","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"tif","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":35860088,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.3.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/0b2871ba86823ec40355774b.tif"},{"id":96943759,"identity":"121fd69c-0898-45fe-9c36-263bc96ea00a","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"tif","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":33273124,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.4.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/18ec7f3d5617264becf7a072.tif"},{"id":97135851,"identity":"2b4ba621-9273-4721-b752-0e4f0b63b57d","added_by":"auto","created_at":"2025-12-01 09:54:02","extension":"json","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":5735,"visible":true,"origin":"","legend":"","description":"","filename":"122e4cb782844b559320a419372bde39.json","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/6182cb688161a90e4a98802d.json"},{"id":97136925,"identity":"cc79c1df-f84e-4bf7-a839-1df9e5494ea9","added_by":"auto","created_at":"2025-12-01 09:57:08","extension":"docx","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":6596238,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterialrevised.docx","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/5dcc4d89a2455d4ea36dcfa3.docx"},{"id":96943748,"identity":"c0bc44f3-ac68-4a9e-8640-288fb1a01370","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"docx","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1436339,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/1f7eedbf0c645a9fb90415d4.docx"},{"id":96943747,"identity":"8f163125-95a2-4aab-a3d1-d8382359b998","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"xml","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":212206,"visible":true,"origin":"","legend":"","description":"","filename":"122e4cb782844b559320a419372bde391enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/4840725f99e2ecd578b7a166.xml"},{"id":96943771,"identity":"f16a8a43-837d-469d-923d-9c63eda49225","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"tif","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":48684636,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.1.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/b953697854a1c51cd5a36915.tif"},{"id":96943768,"identity":"fe64374c-ef74-448f-929d-6491bd2e950b","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"tif","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":55124828,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.2.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/373f81e363ace1f7037a2df1.tif"},{"id":97136908,"identity":"34e2fe31-7dae-4ab3-9159-02b35e0fbfa0","added_by":"auto","created_at":"2025-12-01 09:57:06","extension":"tif","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":35860088,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.3.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/451e95eed13bd9b5a6ae3fe3.tif"},{"id":97136985,"identity":"8c9174be-7e33-46ec-81b9-64220cad6c36","added_by":"auto","created_at":"2025-12-01 09:57:15","extension":"tif","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":33273124,"visible":true,"origin":"","legend":"","description":"","filename":"Fig.4.tif","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/a0f04f1744b445d7883fd9c8.tif"},{"id":97136257,"identity":"d6c3758d-65d6-4603-8422-47d3b498a33a","added_by":"auto","created_at":"2025-12-01 09:56:14","extension":"jpeg","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1278084,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/19ed339428bce89a427acb32.jpeg"},{"id":97135812,"identity":"ab5f30ac-bce0-4778-bf6c-debed04cac53","added_by":"auto","created_at":"2025-12-01 09:53:50","extension":"jpeg","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":470714,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/addbf78eb153ecd7e9dd93ee.jpeg"},{"id":96943755,"identity":"2fe87b5e-9cfc-4f7c-83e5-71f1210b7612","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"jpeg","order_by":17,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":111592,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/207e612136e135c9a531dddc.jpeg"},{"id":96943756,"identity":"5aabba2d-260f-4452-a718-0ee74e539398","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"jpeg","order_by":18,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":129440,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/6f7c2fd71ab3847f5519f492.jpeg"},{"id":96943752,"identity":"60d8f379-643d-47d6-829a-4a5d4744e5a3","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"png","order_by":19,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":577941,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig.1.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/c45f07722f8e924edc14c490.png"},{"id":96943745,"identity":"a164c9c7-bd46-41cf-b8da-42addf4328ac","added_by":"auto","created_at":"2025-11-27 19:01:12","extension":"png","order_by":20,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":264800,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig.2.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/ad512d071800707a8e1d7ad2.png"},{"id":97135802,"identity":"3503b935-fcdd-4083-a001-cd223b3e6c47","added_by":"auto","created_at":"2025-12-01 09:53:44","extension":"png","order_by":21,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":253498,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig.3.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/f07ccabc4f08c04b1411f499.png"},{"id":96943762,"identity":"045f1bea-fa6c-468e-a270-e7a0791aedec","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"png","order_by":22,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":68043,"visible":true,"origin":"","legend":"","description":"","filename":"OnlineFig.4.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/e98804fa76686710c7e0ab7d.png"},{"id":96943763,"identity":"c09fb8ed-e450-454f-ae32-6dcd971aa10b","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"png","order_by":23,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":113279,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/7dfe008b67b1d4cdd834a7b8.png"},{"id":96943758,"identity":"8a5db9d8-11b6-4af2-adcf-42adff7c4e10","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"png","order_by":24,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":45196,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/90df93510c7d4847fe70f0ec.png"},{"id":97135988,"identity":"6a368683-8eb9-4e44-b0eb-2567c8f012cd","added_by":"auto","created_at":"2025-12-01 09:54:51","extension":"png","order_by":25,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":15915,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/b4cd3d58c7c8cc26e469079d.png"},{"id":96943765,"identity":"36ac695a-6974-4e8c-9e12-6df078350e0a","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"png","order_by":26,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":22663,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/00d7da802f6b4423a3d78616.png"},{"id":96943767,"identity":"cf3452f1-acbe-41ae-96ac-87495484f234","added_by":"auto","created_at":"2025-11-27 19:01:13","extension":"xml","order_by":27,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":211009,"visible":true,"origin":"","legend":"","description":"","filename":"122e4cb782844b559320a419372bde391structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/2f4a55b17c8f112c4b044eb5.xml"},{"id":97135838,"identity":"d0b04605-1a13-46b7-a483-d6c862844791","added_by":"auto","created_at":"2025-12-01 09:53:59","extension":"html","order_by":28,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":230891,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/a25720c4174f915fae640230.html"},{"id":97136962,"identity":"b1bb3f6b-5420-48f8-b947-6475b010bc51","added_by":"auto","created_at":"2025-12-01 09:57:13","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":33305485,"visible":true,"origin":"","legend":"\u003cp\u003eFISH to metaphase chromosomes of eleven species of \u003cem\u003eLilium\u003c/em\u003e, using 5S (green) and 35S (red) rDNA oligonucleotide probes. Chromosomes were counterstained with DAPI (blue). The serial numbers of chromosomes were designated by karyotyping. \u003cstrong\u003ea\u003c/strong\u003e \u003cem\u003eL. pumilum\u003c/em\u003e; \u003cstrong\u003eb\u003c/strong\u003e \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e; \u003cstrong\u003ec\u003c/strong\u003e \u003cem\u003eL. lancifolium\u003c/em\u003e; \u003cstrong\u003ed\u003c/strong\u003e \u003cem\u003eL. lancifolium\u003c/em\u003e ‘Longshan’; \u003cstrong\u003ee\u003c/strong\u003e \u003cem\u003eL. jinfushanense\u003c/em\u003e; \u003cstrong\u003ef\u003c/strong\u003e \u003cem\u003eL. brownii \u003c/em\u003evar. \u003cem\u003eviridulum\u003c/em\u003e; \u003cstrong\u003eg\u003c/strong\u003e \u003cem\u003eL. leucanthum\u003c/em\u003e; \u003cstrong\u003eh \u003c/strong\u003e\u003cem\u003eL. sargentiae\u003c/em\u003e; \u003cstrong\u003ei\u003c/strong\u003e \u003cem\u003eL. sulphureum\u003c/em\u003e; \u003cstrong\u003ej\u003c/strong\u003e \u003cem\u003eL. regale\u003c/em\u003e; \u003cstrong\u003ek\u003c/strong\u003e \u003cem\u003eL. henryi\u003c/em\u003e; \u003cstrong\u003el\u003c/strong\u003e \u003cem\u003eL. rosthornii\u003c/em\u003e. Scale bars = 10 µm.\u003c/p\u003e","description":"","filename":"Fig.1.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/e23f55dde7a43a051d23c25e.png"},{"id":97135709,"identity":"a271c350-a3e1-4231-9112-dc83054c56cd","added_by":"auto","created_at":"2025-12-01 09:53:00","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":14465662,"visible":true,"origin":"","legend":"\u003cp\u003eIdiograms of eleven species of \u003cem\u003eLilium \u003c/em\u003eshowing the chromosome measurements and the locations of 5S and 35S rDNA loci. \u003cstrong\u003ea\u003c/strong\u003e \u003cem\u003eL. pumilum\u003c/em\u003e; \u003cstrong\u003eb\u003c/strong\u003e \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e; \u003cstrong\u003ec\u003c/strong\u003e \u003cem\u003eL. lancifolium\u003c/em\u003e; \u003cstrong\u003ed\u003c/strong\u003e \u003cem\u003eL. lancifolium\u003c/em\u003e ‘Longshan’; \u003cstrong\u003ee\u003c/strong\u003e \u003cem\u003eL. jinfushanense\u003c/em\u003e; \u003cstrong\u003ef\u003c/strong\u003e \u003cem\u003eL. brownii \u003c/em\u003evar. \u003cem\u003eviridulum\u003c/em\u003e; \u003cstrong\u003eg\u003c/strong\u003e \u003cem\u003eL. leucanthum\u003c/em\u003e; \u003cstrong\u003eh \u003c/strong\u003e\u003cem\u003eL. sargentiae\u003c/em\u003e; \u003cstrong\u003ei\u003c/strong\u003e \u003cem\u003eL. sulphureum\u003c/em\u003e; \u003cstrong\u003ej\u003c/strong\u003e \u003cem\u003eL. regale\u003c/em\u003e; \u003cstrong\u003ek\u003c/strong\u003e \u003cem\u003eL. henryi\u003c/em\u003e; \u003cstrong\u003el\u003c/strong\u003e \u003cem\u003eL. rosthornii\u003c/em\u003e. The ordinate scale on the left indicates the relative length of chromosomes. The numbers at the bottom indicate the the serial numbers of chromosomes. The 35S rDNA loci with a gap in the middle indicates the formation of a secondary constriction.\u003c/p\u003e","description":"","filename":"Fig.2.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/4c01e2cb9249c6f6a6d376ed.png"},{"id":97135853,"identity":"d78e7b11-2861-4698-9640-6eec10b2bb73","added_by":"auto","created_at":"2025-12-01 09:54:02","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1722756,"visible":true,"origin":"","legend":"\u003cp\u003eBidimensional scatter plot of M\u003csub\u003eCA\u003c/sub\u003e vs. CV\u003csub\u003eCL\u003c/sub\u003e for eleven species of \u003cem\u003eLilium. \u003c/em\u003eL.pu. = \u003cem\u003eL. pumilum\u003c/em\u003e, L.da. = \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e, L.la. = \u003cem\u003eL. lancifolium\u003c/em\u003e, L.la.l = \u003cem\u003eL. lancifolium\u003c/em\u003e ‘Longshan’, L.ji. = \u003cem\u003eL. jinfushanense\u003c/em\u003e, L.br. = \u003cem\u003eL. brownii\u0026nbsp;\u003c/em\u003evar. \u003cem\u003eviridulum\u003c/em\u003e, L.le. = \u003cem\u003eL. leucanthum\u003c/em\u003e, L.sa. = \u003cem\u003eL. sargentiae\u003c/em\u003e, L.su. = \u003cem\u003eL. sulphureum, \u003c/em\u003eL.re.\u003cem\u003e =\u003c/em\u003e \u003cem\u003eL. regale\u003c/em\u003e, L.he. = \u003cem\u003eL. henryi\u003c/em\u003e, L.ro. = \u003cem\u003eL. rosthornii.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"Fig.3.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/4b39f349b80103508eedacf1.png"},{"id":96943738,"identity":"93b3121b-1a69-4a05-a120-2019e4d14925","added_by":"auto","created_at":"2025-11-27 19:01:12","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1003790,"visible":true,"origin":"","legend":"\u003cp\u003ePCoA for eleven species of \u003cem\u003eLilium\u003c/em\u003e based on \u003cem\u003ex\u003c/em\u003e, \u003cem\u003e2n\u003c/em\u003e, TCL, M\u003csub\u003eCA\u003c/sub\u003e, CV\u003csub\u003eCL\u003c/sub\u003e and CV\u003csub\u003eCI\u003c/sub\u003e. The taxa corresponding to the abbreviations are shown in the legend of Fig. 3. PCoA1 reflects the original data characteristics before the dimensionality reduction of 80.47%. PCoA2 reflects the character of the original data before the dimensionality reduction of 14.39%. The sum of the two percentages is 94.86%, indicating that the two-dimensional coordinate system can reflect the characteristics of 94.86% of the original data.\u003c/p\u003e","description":"","filename":"Fig.4.png","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/8fcced8873f5b1c0db7c0361.png"},{"id":103251013,"identity":"0d0a3836-8925-4cc7-a5b5-d7d744462e22","added_by":"auto","created_at":"2026-02-23 16:00:39","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":38578263,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/a876aa1b-136d-4d3e-af61-b1ac38e49309.pdf"},{"id":97136524,"identity":"e41cde07-77bd-434e-b341-9505e2ec76ca","added_by":"auto","created_at":"2025-12-01 09:56:42","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":1436339,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/d727491995c9ca03ddd7535e.docx"},{"id":97136629,"identity":"6f9799c6-158f-4f57-b346-fb0ba3f83d5c","added_by":"auto","created_at":"2025-12-01 09:56:49","extension":"docx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":6596238,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterialrevised.docx","url":"https://assets-eu.researchsquare.com/files/rs-8029111/v1/6b20b3b934b00214c21af925.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comparative karyotype analysis of eleven species of Lilium from China by FISH with rDNA oligo-probes","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe genus \u003cem\u003eLilium\u003c/em\u003e L., which belongs to the tribe Lilieae in the family Liliaceae, consists approximately 125 species (now including the former genus \u003cem\u003eNomocharis\u003c/em\u003e) originally found in temperate and alpine regions of the Northern Hemisphere, with centers of diversity in East Asia, southern Europe, the Caucasus, and eastern and western North America (Gao and Gao \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Givnish et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Sixty-one \u003cem\u003eLilium\u003c/em\u003e species (including 6 species of the former genus \u003cem\u003eNomocharis\u003c/em\u003e, 37 endemic) are distributed in China, mainly in the southwestern, central and northeastern regions (Liang and Tamura \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Gao and Gao \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Several studies have concluded that the Qinghai-Tibet Plateau (including the Himalayas and Hengduan Mountains) is a center of origin and diversification for \u003cem\u003eLilium\u003c/em\u003e (Patterson and Givnish \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Givnish et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Lilies are of significant importance in horticulture. In China, four \u003cem\u003eLilium\u003c/em\u003e species have been widely used as traditional medicines and/or edible plants with long history and have potential for their diverse pharmacological activities (Zhou et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The dried fleshy scaly leaves of \u003cem\u003eL. lancifolium\u003c/em\u003e Thunb., \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e Baker and \u003cem\u003eL. pumilum\u003c/em\u003e DC., known as Baihe (Lilii Bulbus), are traditional Chinese medicine (TCM) materials which have the effects of moistening the lung, dispelling fire, calming the mind, and killing insects and lice (Chinese Pharmacopoeia Commission \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The main edible lilies cultivated in China include Lanzhou lily \u003cem\u003e(L. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e (E. H. Wilson) Raffill), Longya lily (\u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e), and Juandan lily (cultivated varieties of \u003cem\u003eL. lancifolium\u003c/em\u003e).\u003c/p\u003e\u003cp\u003eThe classification of the genus \u003cem\u003eLilium\u003c/em\u003e has been complicated for long time. Since Linnaeus, many scholars have tried to classify \u003cem\u003eLilium\u003c/em\u003e. It was not until 1949 that Comber's classification of \u003cem\u003eLilium\u003c/em\u003e was widely accepted. Comber (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1949\u003c/span\u003e) classified the naturally grown lilies into seven sections based on 15 phenotypic characters. However, under the influence of similar selection pressure, species with distant genetic relationships may converge in phenotype, while species with close genetic relationships may also diverge in phenotype in order to adapt to different environments or under the influence of survival competition. Therefore, sections of \u003cem\u003eLilium\u003c/em\u003e based largely on floral form may not always reflect actual evolutionary relationships (Givnish et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). It is necessary to continually revise the morphology-based classification of \u003cem\u003eLilium\u003c/em\u003e using the data of cytotaxonomy, molecular phylogenetics and hybridization breeding practice. In the latest revised classification of \u003cem\u003eLilium\u003c/em\u003e, this genus is still divided into seven sections with the names of some sections and the taxonomic locations of some species being adjusted compared to the Combers\u0026rsquo; classification, but some classifications are still controversial (van Tuyl et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Du et al. 2023).\u003c/p\u003e\u003cp\u003eKaryotype analysis is an important method for studying the systematics and evolution in higher plants (e.g. Peruzzi et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; She et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2015\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Astuti et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Conventional karyotype analysis is mainly based on chromosome number and morphological characteristics of chromosomes. However, in most cases, it is difficult to distinguish all chromosomes in a complement by morphological characteristics alone. To this end, researchers have successively developed Giemsa banding, fluorochrome banding and fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization (FISH) of DNA sequences to generate chromosome markers for accurate karyotyping of plants. The most widely utilized DNA probes in plant FISH experiments are the ribosomal genes, 35S (18S-5.8S-26S) and 5S rDNAs, which are organized in tandem arrays with high copy numbers. Comparison of banding and/or FISH patterns (referring to the number and location of specific heterochromatic blocks or hybridization signals of specific DNA sequences) among species within a genus contributes to understanding of chromosome evolution and revealing evolutionary relationships among taxa (e.g. She et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Han et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Mitrenina et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Dias et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Sun et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Evaluating karyotype asymmetry is one of the core aspects of karyotype analysis of plants. Karyotype asymmetry, as a measure of the structural heterogeneity of a chromosome complement, is a cornerstone of plant cytotaxonomy because it serves as a key indicator of evolutionary trends and karyological relationships (e.g. Stebbins \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e1971\u003c/span\u003e; Peruzzi et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Astuti et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). In plant karyotype analysis, asymmetry has been assessed using a set of quantitative indices which fall into two categories for evaluating intrachromosomal asymmetry (the variation in centromere position within a complement) and interchromosomal asymmetry (the variation in chromosome length within a complement), respectively (for details and references see Paszko \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Peruzzi and Eroglu \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Recent critical reviews made by several authors confirm that mean centromeric asymmetry (M\u003csub\u003eCA\u003c/sub\u003e) and coefffcient of variation of chromosome lengths (CV\u003csub\u003eCL\u003c/sub\u003e) are the most appropriate indices for measuring the intrachromosomal asymmetry and interchromosomal asymmetry, respectively (Paszko \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Peruzzi and Eroglu \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Astuti et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The best way for comparison of karyotype asymmetry among taxa is by means of bidimensional scatter plot of M\u003csub\u003eCA\u003c/sub\u003e vs. CV\u003csub\u003eCL\u003c/sub\u003e (Peruzzi and Eroglu \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Astuti et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Kadluczka and Grzebelus \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; She et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). For determination of karyological relationships among taxa, the most effective methodology is the principal coordinate analysis (PCoA) with six quantitative karyotypic parameters, \u003cem\u003ex\u003c/em\u003e, 2\u003cem\u003en\u003c/em\u003e, TCL (total chromosome length of the haploid complement), CV\u003csub\u003eCI\u003c/sub\u003e (coefficient of variation of centromeric index), M\u003csub\u003eCA\u003c/sub\u003e and CV\u003csub\u003eCL\u003c/sub\u003e (e.g. Peruzzi and Altınordu \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Kadluczka and Grzebelus \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; She et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Mitrenina et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Sun et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). In terms of clarifying the evolutionary relationships among species, both banding and FISH patterns are often limited to comparison among species within the same genus, while PCoA, which is based on karyomorphological data, can not only be applied to comparison among species within a genus but also to comparison among species of different genera within the same family (Astuti et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; She et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Comprehensive karyotypic features, including chromosome number, chromosome morphological features, karyotype asymmetry, FISH pattern of specific DNA sequences (e.g. rDNA), have proved of great use in elucidating evolutionary relationships and trends in chromosome evolution among taxa (e.g. Kadluczka and Grzebelus \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Mitrenina et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; She et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Sun et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBecause of their large genome size, lilies have been long used as model plants by cytogeneticists for chromosome studies. Conventional karyotype analysis and C-banding analysis have revealed that the chromosome number of \u003cem\u003eLilium\u003c/em\u003e is very constant, ploidy variation is rare, and the differences between karyotypes of different species are small (Stewart \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1947\u003c/span\u003e; Smyth et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; Noda \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). It is believed that small structural changes in chromosomes such as inversions and reciprocal translocations are the main driving forces for karyotype evolution in the genus \u003cem\u003eLilium\u003c/em\u003e (Noda \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The vast majority of species of \u003cem\u003eLilium\u003c/em\u003e are diploid (2n\u0026thinsp;=\u0026thinsp;2x\u0026thinsp;=\u0026thinsp;24), and only a few species have triploid, tetraploid or aneuploidy in addition to diploid (Stewart \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1947\u003c/span\u003e; Noda \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1986\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Lee et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The haploid genome of species of \u003cem\u003eLilium\u003c/em\u003e is composed of two larger metacentric (m) or submetacentric (sm) chromosomes and ten smaller subtelocentric (st) or telocentric (t) chromosomes. The basic nomenclature of the karyotypes of species of \u003cem\u003eLilium\u003c/em\u003e is based on the order of chromosomes in sequences of decreasing length of short arms, with 1 representing the longest short arm and 12 the shortest (Stewart \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1947\u003c/span\u003e). The number and location of secondary constrictions (SCs) varied greatly among species, but the karyotype asymmetry of the species that have been studied was all 3A and 3B types of Stebbins\u0026rsquo; catergory (Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Wan et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), indicating that it is difficult to distinguish the karyotype structures of different species of \u003cem\u003eLilium\u003c/em\u003e by this asymmetry index. With the application of FISH technology in \u003cem\u003eLilium\u003c/em\u003e, it has greatly promoted the researches on cytotaxonomy, chromosome evolution, genetic diversity and hybrid identification of this genus (e.g. Lim et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Marasek et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Muratović et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Sultana et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Hwang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Wang et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Lee et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Wang et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Ahn et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Cao et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Choi et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Wu et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Kang et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Zhu et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). According to our statistics, 53 species or varieties of \u003cem\u003eLilium\u003c/em\u003e (species originally belonging to \u003cem\u003eNomocharis\u003c/em\u003e are not included) have been studied for FISH mapping of both 5S and 35S rDNA (43/53) or 35S rDNA alone (10/53). The species (varieties) analyzed by FISH involved seven sections of the genus \u003cem\u003eLilium\u003c/em\u003e, including 6 in sect. \u003cem\u003eMartagon\u003c/em\u003e, 3 in sect. \u003cem\u003ePseudolirium\u003c/em\u003e, 9 in sect. \u003cem\u003eLilium\u003c/em\u003e, 4 in sect. \u003cem\u003eArchelirion\u003c/em\u003e, 18 in sect. \u003cem\u003eSinomartagon\u003c/em\u003e, 11 in sect. \u003cem\u003eLeucolirion\u003c/em\u003e, and 2 in sect. \u003cem\u003eOxypetalum\u003c/em\u003e. In terms of geographical distribution, the species analyzed by FISH so far were mainly those distributed in Northeast Asia and Europe, while the species distributed in southwest and central China and North America were few studied by FISH. Most of the species of \u003cem\u003eLilium\u003c/em\u003e endemic to southwest and central China that have been studied by FISH had 35S rDNA localization only, and the results of rDNA mapping by different authors in some species were inconsistent (Wang et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Therefore, it is necessary to conduct dual-color FISH with 5S and 35S rDNA probes that include more species endemic to southwest and central China in order to molecular-cytogenetically characterize these species and analyze their evolutionary relationships. For the medicinal and edible lilies in China, although FISH localization of 5S and 35S rDNA has been reported for them, the results of different authors in some species differed or even were completely inconsistent (Wang et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Lee et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), so further verification is needed to provide a reliable basis for molecular cytogenetic identification of these species.\u003c/p\u003e\u003cp\u003eIn present study, the chromosomes of eleven species of \u003cem\u003eLilium\u003c/em\u003e from China, including the main edible and medicinal lilies in China as well as seven species endemic to southwest and central China (including all the six species of subsect. \u003cem\u003eLeucolirion\u003c/em\u003e 6a in the latest revised classification of \u003cem\u003eLilium\u003c/em\u003e) were characterized by using FISH with 5S and 35S rDNA oligonucleotide probes. Their karyotypes were quantitatively constructed with chromosome measurement data and rDNA FISH signal information. The karyotype asymmetry indices that were recently critically reviewed and recognized were calculated for evaluating asymmetry of the karyotypes and karyological relationships among them. The data of karyotypic parameters and rDNA patterns were assessed to gain insights into the karyotype differentiation and the evolutionary relationships among these species, thereby providing cytotaxonomic support for the classification of \u003cem\u003eLilium\u003c/em\u003e.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003ePlant materials\u003c/h2\u003e\u003cp\u003eTwelve taxa belonging to eleven species of \u003cem\u003eLilium\u003c/em\u003e were collected different places of China and cultivated in Lily Germplasm Resources Engineering Center, Hunan University of Technology, Hunan Province, China (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The geographical distribution and source information of these materials are listed in Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e. The sections to which these species belong are listed in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The plants were identified by Professor Si-Xiang Dong of Nuclear Agriculture and Chinese Medicinal Materials Institute of Hunan Academy of Agricultural Sciences.\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\u003eThe general karyotype features of the eleven species of \u003cem\u003eLilium\u003c/em\u003e\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"10\"\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=\"char\" char=\"\u0026plusmn;\" 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=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSpecies\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eKaryotype formula\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTCL\u0026thinsp;\u0026plusmn;\u0026thinsp;SE (\u0026micro;m)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC (\u0026micro;m)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eRRL\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eCI\u0026thinsp;\u0026plusmn;\u0026thinsp;SE\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eCV\u003csub\u003eCI\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eM\u003csub\u003eCA\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eCV\u003csub\u003eCL\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eStebinns\u0026rsquo; types\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. pumilum\u003c/em\u003e Redout\u0026eacute;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;4m(4SAT)\u0026thinsp;+\u0026thinsp;14st(6SAT)\u0026thinsp;+\u0026thinsp;6t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e212.15\u0026thinsp;\u0026plusmn;\u0026thinsp;21.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e17.68\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.20-12.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e19.73\u0026thinsp;\u0026plusmn;\u0026thinsp;10.10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e51.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e60.65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e21.51\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3A\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e (E. H. Wilson) Raffill\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;2m(2SAT)\u0026thinsp;+\u0026thinsp;2sm\u0026thinsp;+\u0026thinsp;8st(2SAT)\u0026thinsp;+\u0026thinsp;12t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e222.96\u0026thinsp;\u0026plusmn;\u0026thinsp;9.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e18.58\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.67\u0026ndash;12.59\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.11\u0026thinsp;\u0026plusmn;\u0026thinsp;11.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e65.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e66.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e20.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3A\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. lancifolium\u003c/em\u003e Thunb.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;36\u0026thinsp;=\u0026thinsp;6m(6SAT)\u0026thinsp;+\u0026thinsp;12st(2SAT)\u0026thinsp;+\u0026thinsp;18t(5SAT)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e221.75\u0026thinsp;\u0026plusmn;\u0026thinsp;15.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e18.48\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.82\u0026ndash;12.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.19\u0026thinsp;\u0026plusmn;\u0026thinsp;11.73\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e68.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e65.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e21.84\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3A\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;36\u0026thinsp;=\u0026thinsp;6m(6SAT)\u0026thinsp;+\u0026thinsp;12st(3SAT)\u0026thinsp;+\u0026thinsp;18t(6SAT)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e212.44\u0026thinsp;\u0026plusmn;\u0026thinsp;17.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e17.70\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.41\u0026ndash;12.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.03\u0026thinsp;\u0026plusmn;\u0026thinsp;11.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e67.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e66.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e22.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3A\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. jinfushanense\u003c/em\u003e L. J. Peng \u0026amp; B. N. Wang\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;2m(2SAT)\u0026thinsp;+\u0026thinsp;2sm(2SAT)\u0026thinsp;+\u0026thinsp;10st\u0026thinsp;+\u0026thinsp;10t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e220.38\u0026thinsp;\u0026plusmn;\u0026thinsp;22.70\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e18.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.08\u0026ndash;12.65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e18.30\u0026thinsp;\u0026plusmn;\u0026thinsp;11.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e60.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e63.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e22.70\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3B\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e Baker\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;4m\u0026thinsp;+\u0026thinsp;8st(4SAT)\u0026thinsp;+\u0026thinsp;12t(2SAT)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e215.68\u0026thinsp;\u0026plusmn;\u0026thinsp;25.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e17.97\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.66\u0026ndash;12.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e18.17\u0026thinsp;\u0026plusmn;\u0026thinsp;11.74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e64.58\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e63.91\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e23.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3B\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. leucanthum\u003c/em\u003e (Baker) Baker\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;4m(4SAT)\u0026thinsp;+\u0026thinsp;8st(4SAT)\u0026thinsp;+\u0026thinsp;12t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e175.55\u0026thinsp;\u0026plusmn;\u0026thinsp;15.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e14.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.80-12.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.88\u0026thinsp;\u0026plusmn;\u0026thinsp;11.13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e62.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e64.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e23.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3B\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. sargentiae\u003c/em\u003e E. H. Wilson\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;2m(2SAT)\u0026thinsp;+\u0026thinsp;2sm(2SAT)\u0026thinsp;+\u0026thinsp;8st(4SAT)\u0026thinsp;+\u0026thinsp;12t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e201.40\u0026thinsp;\u0026plusmn;\u0026thinsp;3.88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e16.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.63\u0026ndash;12.52\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.54\u0026thinsp;\u0026plusmn;\u0026thinsp;11.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e66.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e65.49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e23.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3B\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. sulphureum\u003c/em\u003e Baker ex J. D. Hooker\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;4m(4SAT)\u0026thinsp;+\u0026thinsp;10st(4SAT)\u0026thinsp;+\u0026thinsp;10t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e182.87\u0026thinsp;\u0026plusmn;\u0026thinsp;7.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.06\u0026ndash;12.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e18.21\u0026thinsp;\u0026plusmn;\u0026thinsp;11.27\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e61.91\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e63.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e22.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3B\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. regale\u003c/em\u003e E. H. Wilson\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;2m(2SAT)\u0026thinsp;+\u0026thinsp;2sm(2SAT)\u0026thinsp;+\u0026thinsp;8st(6SAT)\u0026thinsp;+\u0026thinsp;12t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e214.65\u0026thinsp;\u0026plusmn;\u0026thinsp;6.94\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e17.88\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.61\u0026ndash;13.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.88\u0026thinsp;\u0026plusmn;\u0026thinsp;11.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e64.69\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e64.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e25.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3B\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. henryi\u003c/em\u003e Baker\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;4m(2SAT)\u0026thinsp;+\u0026thinsp;6st(2SAT)\u0026thinsp;+\u0026thinsp;14t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e197.73\u0026thinsp;\u0026plusmn;\u0026thinsp;9.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e16.48\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.17\u0026ndash;12.65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.10\u0026thinsp;\u0026plusmn;\u0026thinsp;10.84\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e63.39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e65.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e22.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3B\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. rosthornii\u003c/em\u003e Diels\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2n\u0026thinsp;=\u0026thinsp;24\u0026thinsp;=\u0026thinsp;2m(2SAT)\u0026thinsp;+\u0026thinsp;2sm\u0026thinsp;+\u0026thinsp;8st(2SAT)\u0026thinsp;+\u0026thinsp;12t\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e175.91\u0026thinsp;\u0026plusmn;\u0026thinsp;7.26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e14.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.06\u0026ndash;12.10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e17.35\u0026thinsp;\u0026plusmn;\u0026thinsp;10.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e62.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e65.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e21.49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3A\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"10\"\u003eTCL, total length of the haploid complement (i.e. karyotype length); RRL, range of chromosome relative length; C, mean chromosome length; CI, mean centromeric index; CV\u003csub\u003eCI\u003c/sub\u003e, coefficient of variation of centromeric index; M\u003csub\u003eCA\u003c/sub\u003e, mean centromeric asymmetry; CV\u003csub\u003eCL\u003c/sub\u003e, coefficient of variation of chromosome lengths; Stebbin\u0026rsquo;s\u0026rsquo; types, karyotype asymmetry category of Stebbins.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\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\u003eThe sections to which the eleven species of \u003cem\u003eLilium\u003c/em\u003e belong and the number and location of their 5S and 45S rDNA loci\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\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\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSpecies\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSection\u003c/p\u003e\u003cp\u003e(Comber \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1949\u003c/span\u003e/Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e/Du et al. 2023)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5S\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e45S\u003csup\u003ea\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\u003cem\u003eL. pumilum\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSinomartagon 5b / Sinomartagon clade II / Sinomartagon 5b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-INT (64.03%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFive: 1S-PRO (8.87%), 2S-INT (51.51%), 3L-INT (55.15%), 4-CEN, 5L-INT(60.61%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSinomartagon 5a / Sinomartagon clade II / Sinomartagon 5a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-INT (66.45%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFour: 1S-PRO (8.82%), 4L-PRO (5.00%)\u003csup\u003eb\u003c/sup\u003e, 6L-INT(63.92%), 9S-PRO (21.74%)\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. lancifolium\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSinomartagon 5a / Sinomartagon clade II / Sinomartagon 5a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-INT (71.52%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eSix: 1S-PRO (7.93%), 2S-INT (58.86%), 6L-INT (58.34%)\u003csup\u003ec\u003c/sup\u003e, 7S-PRO (33.60%), 8L-PRO (20.19%)\u003csup\u003ebd\u003c/sup\u003e, 11L-INT(40.41%)\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSinomartagon 5a / Sinomartagon clade II / Sinomartagon 5a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-INT (71.25%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFive:1S-PRO(7.85%),2S-INT(62.88%), 6L-INT(62.7%), 7S-PRO (22.86%), 11L-INT (41.07%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. jinfushanense\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSinomartagon 5c / Sinomartagon clade III / Sinomartagon 5c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-PRO (9.72%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTwo: 1S-PRO (11.50%), 2S-PRO (19.63%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. brownii\u003c/em\u003e\u0026nbsp;var. \u003cem\u003eviridulum\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eArchelirion / Leucolirion clade II / Leucolirion 6b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-INT (69.82%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eThree: 4L-INT (46.05%), 5L-INT (66.87%), 7S-PRO (24%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. leucanthum\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeucolirion 6a / Leucolirion clade I / Leucolirion 6a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-PRO (6.66%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFour: 1S-PRO (7.54%), 2L-PRO (9.09%), 4S-PRO(20.00%), 5L-PRO (19.65%),\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. sargentiae\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeucolirion 6a / Leucolirion clade I / Leucolirion 6a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-PRO (6.17%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFour: 1S-PRO (10.24%), 2L-PRO (7.01%), 4S-PRO (15.79%), 5L-PRO (13.38%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. sulphureum\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeucolirion 6a / Leucolirion clade I / Leucolirion 6a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-PRO (8.13%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFour: 1S-PRO (9.35%), 2L-PRO (8.22%),\u003c/p\u003e\u003cp\u003e4S-PRO (13.51%), 5L-PRO (10.84%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. regale\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeucolirion 6a / Leucolirion clade I / Leucolirion 6a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-PRO (9.04%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eFive: 1S-PRO (10.69%), 2L-PRO (7.14%), 4S-PRO (18.42%), 5L-PRO(8.98%), 6L-PRO (7.69%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. henryi\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSinomartagon 5a / Leucolirion clade I / Leucolirion 6a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eOne: 3L-PRO\u0026thinsp;+\u0026thinsp;CEN\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTwo: 1S-PRO(7.89%), 4L-INT(54.83%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eL. rosthornii\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSinomartagon 5a / Leucolirion clade I / Leucolirion 6a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eTwo: 3L-PRO (15.27%),\u003c/p\u003e\u003cp\u003e6S-PRO (46.15%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTwo: 1S-PRO (9.71%), 5L-INT (42.11%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003e\u003csup\u003ea\u003c/sup\u003e S and L represent short and long arms, respectively; CEN, PRO, INT, DIS and TER represent centromeric (\u003cem\u003edi\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0), proximal (0\u0026thinsp;\u0026lt;\u0026thinsp;\u003cem\u003edi\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;25%), interstitial (25%\u0026thinsp;\u0026le;\u0026thinsp;\u003cem\u003edi\u003c/em\u003e\u0026thinsp;\u0026le;\u0026thinsp;75%), distal (75%\u0026lt;\u003cem\u003edi\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;100%) and terminal (\u003cem\u003edi\u003c/em\u003e\u0026thinsp;=\u0026thinsp;100%) positions, respectively; figures ahead of the positions designate the homologous chromosome pair (triplet) involved; the percentages in square brackets are the percentage distance from centromere to rDNA locus. \u003csup\u003eb\u003c/sup\u003e indicates the 45S rDNA loci that did not generate secondary constrictions (SCs). SCs occurred at all other 45S loci. \u003csup\u003ec\u003c/sup\u003e Only two of the three members of the homologous triplet have hybridization signals. \u003csup\u003ed\u003c/sup\u003e Only one of the three members of the homologous triplet has hybridization signal.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eChromosome preparation\u003c/h3\u003e\n\u003cp\u003eThe bulbs used for cytogenetic experiments were cultivated in pots with mixed planting soil consisting of decomposed pine needle humus soil and sandy soil (v/v\u0026thinsp;=\u0026thinsp;3:1). Water the bulbs with a solution of rooting powder diluted in water to promote the growth of new roots. When the new roots grew to 1\u0026ndash;2 cm, the root tips of 2\u0026ndash;3 mm were excised and pretreated with a mixture of 0.1% colchicine and 0.015% 8-hydroxyquinoline (1:1) at 28\u0026deg;C in the dark for 4.5 h, followed by fixation at 4\u0026deg;C in 3:1 (v/v) methanol/glacial acetic acid for more than 12 h. The chromosome slides were prepared using the enzymatic maceration and flame-drying (EMF) method (She et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2015\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The fixed root tips were thoroughly rinsed in deionized water and digested in a mixture of 1% cellulase RS and 1% pectolyase Y23 (Yakult Pharmaceutical Industry Co., Ltd.) in citric buffer (pH 4.5) at 37\u0026deg;C for 2 h. After the enzyme solution was replaced by double distilled water, 1\u0026ndash;2 digested root tips were transferred to a glass slide and mashed by using fine-pointed forceps with the fixative solution. Then, the slides were flame-dried. The slides with well-spread metaphase chromosomes were selected under an Olympus BX51 phase contrast microscope and stored at -20\u0026deg;C until use.\u003c/p\u003e\n\u003ch3\u003eDNA probe and FISH\u003c/h3\u003e\n\u003cp\u003eFor FISH experiments, the 5S and 35S rDNA oligonucleotide probes described by Han et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) were used. The 5S rDNA probes, 5S-1 and 5S-2, as well as the 35S rDNA probes, 35S-1, 35S-2 and 35S-3, were synthesized by Sangon Bioengineering Co., Ltd.. 5S-1 and 5S-2 were labeled with 6-carboxyl fluorescein (6-FAM) at the 5\u0026rsquo;-terminus and then mixed together to make the 5S rDNA probe solution. 35S-1, 35S-2, and 35S-3 were labeled with 6-carboxyl-tetramethyl rhodamine (TAMRA) at the 5\u0026rsquo;-terminus and mixed together to make the 35S rDNA probe solution.\u003c/p\u003e\u003cp\u003eFISH with the two probes was performed as described by Wei et al. (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The slides were baked at 65℃ for 45 min, cooled, and then denatured in 70% deionized formamide at 85\u0026deg;C for 2.5 min. After denaturation, the slides were immediately placed in a series of 70%, 95%, and 100% ethanol at \u0026minus;\u0026thinsp;20\u0026deg;C for 5 min each and air dried. Then, hybridization solution was added onto the chromosome slide and then incubated in a moist box at 37\u0026deg;C overnight. The hybridization solution (each slide) was as follows: deionized formamide, 10 \u0026micro;L; 50% dextran sulphate, 4 \u0026micro;L; 20 \u0026times; SSC, 2 \u0026micro;L; salmon sperm DNA, 2 \u0026micro;L (40 ng); 5S rDNA probe, 1 \u0026micro;L (40 ng); 35S rDNA probe, 1 \u0026micro;L (40 ng). The hybridized slides were washed twice each for 5 min with 2 \u0026times; SSC at room temperature, and air dried in dark. Then, the chromosomes counterstained with 3 \u0026micro;g ml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e 4,6\u0026ndash;diamidino\u0026ndash;2\u0026ndash;phenylindole (DAPI) solution in Vectashield H-1000 (Vector Laboratories, Inc.) were visualized with an Olympus BX60 microscope equipped with a QImaging Retiga R6 CCD camera (Teledyne Photometrics) which was controlled using Ocular software (Teledyne Photometrics). Grey-scale images were digitally captured and merged by the Ocular software. Raw images were processed using Adobe Photoshop software (Adobe Systems Inc.).\u003c/p\u003e\n\u003ch3\u003eKaryotype analysis\u003c/h3\u003e\n\u003cp\u003eKaryotype analysis was performed using the methodology previously described by us (She et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). For each taxon, five metaphase cells were selected for measurement using Adobe Photoshop software. The length of long arm (L) and short arm (S) of each chromosome and the length between the center of FISH signal and centromere were measured. The following parameters were calculated for characterizing the karyotypes: (i) chromosome relative length (RL, % of haploid complement); (ii) arm ratio (AR\u0026thinsp;=\u0026thinsp;L/S); (iii) TCL (i.e. the karyotype length); (iv) mean chromosome length (C); (v) mean centromeric index (CI); (vi) four karyotype asymmetry indices including CV\u003csub\u003eCI\u003c/sub\u003e (Paszko \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e ), CV\u003csub\u003eCL\u003c/sub\u003e (Paszko \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), M\u003csub\u003eCA\u003c/sub\u003e (Peruzzi and Eroglu \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) and Stebbins\u0026rsquo; asymmetry category (Stebbins \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e1971\u003c/span\u003e); (vii) percent distance from centromere to DNA loci (\u003cem\u003edi\u003c/em\u003e\u0026thinsp;=\u0026thinsp;\u003cem\u003ed\u003c/em\u003e \u0026times; 100/\u003cem\u003ea\u003c/em\u003e; \u003cem\u003ed\u003c/em\u003e\u0026thinsp;=\u0026thinsp;distance of the center of FISH signal from the centromere, \u003cem\u003ea\u003c/em\u003e\u0026thinsp;=\u0026thinsp;length of the corresponding chromosome arm). The chromosomes were classified as metacentric (m, AR\u0026thinsp;=\u0026thinsp;1.00-1.70), submetacentric (sm, AR\u0026thinsp;=\u0026thinsp;1.71-3.00), subtelocentric (st, AR\u0026thinsp;=\u0026thinsp;3.01-7.00) and telocentric (t, AR\u0026thinsp;\u0026gt;\u0026thinsp;7.01) types according to the nomenclature of Levan et al. (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1964\u003c/span\u003e). Satellite chromosomes in which SCs generated were abbreviated as \u0026ldquo;SAT\u0026rdquo;. Chromosomes were arranged in order of decreasing length of the short arms according to Stewart (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1947\u003c/span\u003e). Idiograms were drawn based on the dataset of chromosome measurements and rDNA FISH signals.\u003c/p\u003e\u003cp\u003eTo visualize karyotype asymmetry relationships among the analyzed taxa, a bidimensional scatter plot with parameters M\u003csub\u003eCA\u003c/sub\u003e vs. CV\u003csub\u003eCL\u003c/sub\u003e was plotted. To determine the karyological relationships among the 12 taxa, a principal coordinate analysis (PCoA) using Gower\u0026rsquo;s similarity coefficient were carried out based on six quantitative parameters, \u003cem\u003ex\u003c/em\u003e, 2\u003cem\u003en\u003c/em\u003e, TCL, CV\u003csub\u003eCI\u003c/sub\u003e, M\u003csub\u003eCA\u003c/sub\u003e and CV\u003csub\u003eCL\u003c/sub\u003e, as proposed by Peruzzi and Altınordu (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eGeneral karyotype features\u003c/h2\u003e\u003cp\u003e\u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo; are triploid with somatic chromosome number 2n\u0026thinsp;=\u0026thinsp;3x\u0026thinsp;=\u0026thinsp;36, the other ten species are diploid with somatic chromosome number 2n\u0026thinsp;=\u0026thinsp;2x\u0026thinsp;=\u0026thinsp;24 (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). No aneuploidy or B chromosomes were found in the taxa studied. All the eleven species have the same basic chromosome number x\u0026thinsp;=\u0026thinsp;12. The TCL ranges from 175.55 \u0026micro;m (\u003cem\u003eL. leucanthum\u003c/em\u003e) \u0026micro;m to 222.96 \u0026micro;m (\u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e) with a mean chromosome length from 14.63 \u0026micro;m to 18.58 \u0026micro;m. As for the range of relative length (RRL), the smallest RRL was observed in \u003cem\u003eL. lancifolium\u003c/em\u003e, while the largest RRL was detected in \u003cem\u003eL. regale\u003c/em\u003e. That is, \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. regale\u003c/em\u003e exhibit the smallest and the largest variation in chromosome length, respectively. The CI of these complements varies between 17.03\u0026thinsp;\u0026plusmn;\u0026thinsp;11.46 (\u003cem\u003eL. lancifolium\u003c/em\u003e) and 19.73\u0026thinsp;\u0026plusmn;\u0026thinsp;10.10 (\u003cem\u003eL. pumilum\u003c/em\u003e). That is, \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. pumilum\u003c/em\u003e are characterized by the smallest and the largest level of variation in the centromeric index, respectively.\u003c/p\u003e\u003cp\u003eThe haploid karyotypes of the 12 taxa show distinct bimodality, which are composed of two larger metacentric or submetacentric chromosomes and ten smaller subtelocentric or telocentric chromosomes (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The karyotype formulas as well as the number and location of SCs are different among taxa (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e; Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). All karyotypes fall into 3A or 3B type of Stebbins\u0026rsquo; category, possessing moderate degree of asymmetry (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The ranges of CV\u003csub\u003eCI\u003c/sub\u003e, CV\u003csub\u003eCL\u003c/sub\u003e and M\u003csub\u003eCA\u003c/sub\u003e are as follow: CV\u003csub\u003eCI\u003c/sub\u003e = 51.16 (\u003cem\u003eL. pumilum\u003c/em\u003e) \u0026minus;\u0026thinsp;68.22 (\u003cem\u003eL. lancifolium\u003c/em\u003e), M\u003csub\u003eCA\u003c/sub\u003e = 60.65 (\u003cem\u003eL. pumilum\u003c/em\u003e) \u0026minus;\u0026thinsp;66.46 (\u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e), CV\u003csub\u003eCL\u003c/sub\u003e = 20.31 (\u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e) \u0026minus;\u0026thinsp;25.38 (\u003cem\u003eL. regale\u003c/em\u003e) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), revealing that, among the 12 taxa, \u003cem\u003eL. pumilum\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e have the least and the most heterogeneity in centromere position, respectively; \u003cem\u003eL. pumilum\u003c/em\u003e and \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e have the lowest and the highest intrachromosomal asymmetry, respectively; \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e and \u003cem\u003eL. regale\u003c/em\u003e have the lowest and highest level of interchromosomal asymmetry, respectively. The bidimensional scatter plot of M\u003csub\u003eCA\u003c/sub\u003e vs. CV\u003csub\u003eCL\u003c/sub\u003e shows that the karyotype structure of the twelve taxa can be completely distinguished by this couple of parameters because no overlap among taxa occurs (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The PCoA scatter plot shows that the 12 taxa are divided into two groups along the direction of PCoA1: \u003cem\u003eL. pumilum\u003c/em\u003e, \u003cem\u003eL. jinfushanense\u003c/em\u003e, \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e, \u003cem\u003eL. regale\u003c/em\u003e, \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e, \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo; in one group with \u003cem\u003eL. pumilum\u003c/em\u003e occupying the most isolated position; while \u003cem\u003eL. sulphureum\u003c/em\u003e, \u003cem\u003eL. henryi\u003c/em\u003e, \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e, and \u003cem\u003eL. rosthornii\u003c/em\u003e in another group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Among the species clustered on the left, \u003cem\u003eL. regale\u003c/em\u003e is the closest to \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. sargentiae\u003c/em\u003e, which are clustered on the right (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eFISH localization of 5S and 35S rDNA loci\u003c/h3\u003e\n\u003cp\u003eAll taxa but \u003cem\u003eL. rosthornii\u003c/em\u003e have a single locus of 5S rDNA, which in \u003cem\u003eL. pumilum\u003c/em\u003e, \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e, \u003cem\u003eL. lancifolium\u003c/em\u003e, \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo; and \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e is situated in the interstitial regions of the long arms of chromosome 3 (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ec, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ed, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ef, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ea, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eb, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ec, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ed, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ef; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), while in \u003cem\u003eL. jinfushanense\u003c/em\u003e, \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e, \u003cem\u003eL. sulphureum\u003c/em\u003e, \u003cem\u003eL. regale\u003c/em\u003e, \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e is located in the proximal regions of the long arms of chromosome 3 (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ee, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003eg, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003eh, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ei, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ej, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ek, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003el, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ee, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eg, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eh, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ei, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ej, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ek, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003el; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In \u003cem\u003eL. rosthornii\u003c/em\u003e, in addition to the 5S locus located on chromosome 3, another 5S locus located in the proximal regions of the short arms of chromosome 6 was detected (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003el, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003el; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Particularly, the FISH signals of the 5S loci on chromosome 3 of both \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e are particularly strong, indicating a striking similarity between them. In \u003cem\u003eL. pumilum\u003c/em\u003e and \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e, the intensity of 5S signal is different between homologous chromosomes, showing heterozygosity of the 5S locus (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, f).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe number of 35S rDNA loci ranges from two to six among the 12 taxa (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003e, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Most of the 35S loci (93.5%) generate SCs (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e; Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). In \u003cem\u003eL. pumilum\u003c/em\u003e, five 35S loci are located in the proximal regions of the short arms of chromosome 1, the interstitial regions of the short arms of chromosome 2, the interstitial regions of the long arms of chromosome 3 and 5, the centromeric regions of chromosome 4, respectively (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ea; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e, four 35S loci occur in the proximal regions of the short arms of chromosome 1 and 9, the proximal regions of the long arms of chromosome 4, the interstitial regions of the long arms of chromosome 6, respectively (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eb; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Both \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo; have 35S loci on chromosome 1 and 2 at the same positions as chromosome 1 and 2 of \u003cem\u003eL. pumilum\u003c/em\u003e, as well as other three 35S loci located in the interstitial regions of the long arms of chromosome 6 and 11, and the proximal regions of the short arms of chromosome 7, respectively (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ec, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ed, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ec, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ed; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Compared with \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo;, \u003cem\u003eL. lancifolium\u003c/em\u003e has one more 35S locus on chromosome 8, and its 35S loci on chromosome 6, 8 and 11 appear only in one or two members of the triplets (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ec, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ed; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). \u003cem\u003eL. jinfushanense\u003c/em\u003e has only two 35S loci located in the proximal regions of the short arms of chromosome 1 and 2 (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ee, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ee; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e possesses three 35S loci located in the interstitial regions of long arms of chromosome 4 and 5, and the proximal regions of the short arms of chromosome 7 (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ef, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ef; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e, \u003cem\u003eL. sulphureum\u003c/em\u003e and \u003cem\u003eL. regale\u003c/em\u003e have four 35S loci at similar positions, i.e. the proximal regions of the short arms of chromosome 1 and 4, and the proximal regions of the long arms of chromosome 2 and 5 (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003eg, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003eh, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ei, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ej, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eg, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eh, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ei, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ej; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). \u003cem\u003eL. regale\u003c/em\u003e has another 35S locus located in the proximal regions of the long arms of chromosome 6 (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ej, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ej; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The percentage distances of the 35S locus on chromosome 5 vary among the four species (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eg, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eh, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ei, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ej; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e have similar 35S patterns, with one locus in the proximal regions of the short arms of chromosome 1 and another locus in the interstitial regions of the long arms of a subtelocentric chromosome pair (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003ek, \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003el, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ek, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003el; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eKaryotype variation and species differentiation\u003c/h2\u003e\u003cp\u003eA large number of previous cytogenetic studies have revealed that the karyotypes of \u003cem\u003eLilium\u003c/em\u003e are rather conserved among species. The karyotype conservatism of \u003cem\u003eLilium\u003c/em\u003e is mainly manifested in two aspects: first, except a few species that have triploid, tetraploid or aneuploidy in addition to diploid, the vast majority of species are all diploids with a chromosome number 2n\u0026thinsp;=\u0026thinsp;24, and the basic chromosome number is x\u0026thinsp;=\u0026thinsp;12; second, the haploid genome of all species is composed of two larger metacentric or submetacentric chromosomes and ten smaller subtelocentric or telocentric chromosomes with their karyotypes falling into 3A or 3B type as judged by Stebbins\u0026rsquo; asymmetry index.\u003c/p\u003e\u003cp\u003eOur study shows that except \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo; all other species are diploid. Extensive and in-depth cytogenetic studies have been conducted on \u003cem\u003eL. lancifolium\u003c/em\u003e. This species is a cytologically conspicuous species with both diploids and triploids coexisting in nature. However, the diploid form has only been found in a few regions of South Korea and Japan (Noda \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1986\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Our results indicate that the two populations of \u003cem\u003eL. lancifolium\u003c/em\u003e studied herein are triploids, being consistent with the previously reported populations from China (Noda \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1986\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Several previous studies have supported that the triploid \u003cem\u003eL. lancifolium\u003c/em\u003e is an autopolyploid derived from the diploid \u003cem\u003eL. lancifolium\u003c/em\u003e (Sultana et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Our molecular-cytogenetic karyotyping shows that the chromosomal morphology and rDNA locus distribution in the same triplets of \u003cem\u003eL. lancifolium\u003c/em\u003e are very similar, also providing evidence for its autopolyploid nature.\u003c/p\u003e\u003cp\u003eWhen the chromosome number is the same, the karyomorphological differences between plant species or populations can be measured by a series of parameters (Paszko \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Peruzzi and Eroglu \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Nine parameters are given in this study, among which karyotype formula and karyotype asymmetry indices should be the main parameters for karyotypic comparison. In terms of the karyotype formula, \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo; are the same, and the karyotype formulas vary among the haploid genomes of the eleven species studied, including differences in the number of different chromosome types and in the number and location of SCs.\u003c/p\u003e\u003cp\u003ePrevious studies have suggested that the first two chromosome pairs are quite stable and likely evolutionary residues in \u003cem\u003eLilium\u003c/em\u003e that can serve as a specific trait of this genus (Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Our results show that chromosome 1 of all species studied is m-type, while chromosome 2 is m- or sm-type (m-type in 6 species and sm-type in 5 species) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Among the 47 taxa of \u003cem\u003eLilium\u003c/em\u003e (39 species involved) from China studied by Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e and Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e, all taxa had chromosome 1 of m-type, 27 taxa had chromosome 2 of m-type, and 20 taxa had chromosome 2 of sm-type. These facts suggest that chromosome 1 is fairly stable during speciation in \u003cem\u003eLilium\u003c/em\u003e, while chromosome 2 is relatively prone to chromosomal rearrangement, resulting in changes in the length of its short and/or long arms. As for the ten smaller chromosomes, the haploid genomes consist of 7st\u0026thinsp;+\u0026thinsp;3t, 5st\u0026thinsp;+\u0026thinsp;5t, 4st\u0026thinsp;+\u0026thinsp;6t and 3st\u0026thinsp;+\u0026thinsp;7t chromosomes among the eleven species studied (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In the karyotypes arranged in the order of decreasing length of the short arms, the RLs of the long arms of chromosomes 3 to 12 in the same numbered chromosomes of different species can vary significantly (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Even between closely related species such as \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e, \u003cem\u003eL. sulphureum\u003c/em\u003e and \u003cem\u003eL. regale\u003c/em\u003e, as well as \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e (detailed discussion on relationships see below), such differences also exist. For examples, the RLs of the long arms of chromosome 10 of \u003cem\u003eL. sulphureum\u003c/em\u003e are significantly longer than those of \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e and \u003cem\u003eL. regale\u003c/em\u003e; the RLs of the long arms of chromosome 11 of \u003cem\u003eL. sulphureum\u003c/em\u003e and \u003cem\u003eL. regale\u003c/em\u003e are significantly shorter than those of \u003cem\u003eL. leucanthum\u003c/em\u003e and \u003cem\u003eL. sargentiae\u003c/em\u003e (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eg, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eh, \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ei, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ej; Table \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). For another example, there are significant differences in the RLs of the long arms of chromosomes 4, 5, 6, 9 and 12 between \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ek, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003el). The short arms of chromosomes 3 to 12 may also have their lengths altered due to rearrangements. For instance, chromosome 5 of \u003cem\u003eL. rosthornii\u003c/em\u003e is similar to chromosome 4 of \u003cem\u003eL. henryi\u003c/em\u003e because both have an interstitial 35S locus in the long arms (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ek, and \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003el). The change in the order is likely caused by a chromosomal rearrangement, which has shortened the short arms of the corresponding chromosome pair in \u003cem\u003eL. rosthornii\u003c/em\u003e. In a word, the differences in karyotype formula among different species and the variation in the lengths of the long arms of the same numbered chromosomes indicate that speciation in \u003cem\u003eLilium\u003c/em\u003e involves multiple chromosomal rearrangements (translocations, inversions, deletions, etc.).\u003c/p\u003e\u003cp\u003eKaryotype asymmetry is an important parameter to measure the degree of genomic structural variation among species (Astuti et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The use of statistically correct and reliable karyotype asymmetry indices as well as effective analysis methods can not only effectively distinguish the differences in karyotype structure among species and even among populations, but also reveal the karyological relationships among species (e.g. Peruzzi and Eroglu \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Peruzzi and Altınordu \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Kadluczka and Grzebelus \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; She et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Mitrenina et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Our results show that all the 12 taxa of \u003cem\u003eLilium\u003c/em\u003e studied herein belong to type 3A or 3B of the Stebbins\u0026rsquo; category, and this index cannot show their differences in karyotype structure, but the bidimensional scatter plots of M\u003csub\u003eCA\u003c/sub\u003e vs. CV\u003csub\u003eCL\u003c/sub\u003e can clearly distinguish all the 12 taxa, and there is no overlap even between the two populations of \u003cem\u003eL. lancifolium\u003c/em\u003e. Our analysis demonstrates that calculating M\u003csub\u003eCA\u003c/sub\u003e and CV\u003csub\u003eCL\u003c/sub\u003e values and making dimensional scatter plot are an effective method to distinguish the karyotype structure of different species of \u003cem\u003eLilium\u003c/em\u003e, and also shows that the karyomorphology of \u003cem\u003eLilium\u003c/em\u003e is not the case that the karyotype differences between species are small and the major differences among species are mainly reflected in the number and position of SCs, as previously thought (Noda \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Gao et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eVariation of rDNA pattern and evolutionary relationships among species\u003c/h2\u003e\u003cp\u003eA large number of molecular cytogenetic studies have shown that the similarity and difference in rDNA pattern can directly reflect the closeness of relatedness between species in a plant genus (e.g. She et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Mitrenina et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Dias et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Sun et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), and can also reflect the genetic differentiation between populations within a species (e.g. Nguyen et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Wei et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Our rDNA FISH results show that there are more or less differences in rDNA patterns of the eleven species of \u003cem\u003eLilium\u003c/em\u003e studied. According to the previous reports on FISH mapping of rDNA in \u003cem\u003eLilium\u003c/em\u003e (Lim et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Marasek et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Muratović et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Sultana et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Lee et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2015\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Ahn et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Choi et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Kang et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Zhu et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), to compare the rDNA patterns of different species of \u003cem\u003eLilium\u003c/em\u003e, we should first observe whether there are 35S loci on chromosome 1 and 2 and the differences in their locations, as well as the differences in the locations of the 5S locus on chromosome 3, and then should one check whether there are other 35S and 5S loci and the differences in their locations.\u003c/p\u003e\u003cp\u003eIn \u003cem\u003eL. pumilum\u003c/em\u003e, the number and locations of the 35S loci detected by us were similar to those of Zhou et al. (\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Lee et al. (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) also detected five 35S loci and one 5S locus in \u003cem\u003eL. pumilum\u003c/em\u003e, with four 35S loci and the 5S locus at the locations similar to ours. But instead of detecting 35S locus adjacent to the 5S locus, they detected one 35S locus on the short arms of chromosome 7, suggesting that there were differences in rDNA pattern among different populations of this species.\u003c/p\u003e\u003cp\u003eIn \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e \u0026lsquo;Longshan\u0026rsquo;, the number and locations of the 35S loci detected by us were the same as those of the triploid populations previously reported by most authors (Sultana et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Hwang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Lee et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), but somewhat different from those of the triploid populations reported by Nguyen et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e, in which one 35S locus was detected in the interstitial regions of the long arms of three or two or one members of triplet 4. The 35S locus located on the long arms of one member of triplet 8 was a new locus that hasn't been reported before in \u003cem\u003eL. lancifolium\u003c/em\u003e. Previous reports have also found that the 35S locus on chromosomes 6 and 11 in some triploid populations appeared in only two members of the respective triplet (Nguyen et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). All triploid populations previously reported have two adjacent 5S loci located in the interstitial regions of long arms of chromosome 3 (Sultana et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Hwang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Lee et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Nguyen et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), but we only detected a single 5S locus. In a word, the rDNA loci are high variable among the triploid populations of \u003cem\u003eL. lancifolium\u003c/em\u003e that rely on asexual reproduction only (Noda \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1986\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e, the number and locations of the 35S loci detected by us were similar to those of \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e detected by Zhou et al. (\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and those of \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003eunicolor\u003c/em\u003e (Synonym \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003edavidii\u003c/em\u003e) detected by Wu et al. (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Wu et al. (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) detected four 5S loci in \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003eunicolor\u003c/em\u003e, that is, other three 5S loci were detected in addition to the 5S locus located in the interstitial regions of the long arms of chromosome 3. These facts indicate that the 35S loci of the two varieties of \u003cem\u003eL. davidii\u003c/em\u003e are identical, but the number of 5S loci is quite different, although the 5S locus on chromosome 3 is conserved.\u003c/p\u003e\u003cp\u003eThe locations of the 35S loci on chromosomes 1 and 2 and the 5S loci on chromosome 3 of \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. pumilum\u003c/em\u003e are similar. Moreover, the 35S locus on chromosome 6 of \u003cem\u003eL. lancifolium\u003c/em\u003e is similar to that on chromosome 5 of \u003cem\u003eL. pumilum\u003c/em\u003e (possibly due to chromosomal rearrangement that shortened the short arms and changed the numbering of the chromosome). These similarities indicate that \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. pumilum\u003c/em\u003e have a close relationship. Both \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e and \u003cem\u003eL. lancifolium\u003c/em\u003e have 35S loci on chromosomes 1 and 6 and the positions are similar between them. Both \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottia\u003c/em\u003e and \u003cem\u003eL. pumilum\u003c/em\u003e have 35S loci on chromosomes 1 and 4 and the positions are similar between them. Moreover, the location of the 5S locus of \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e is similar to that of \u003cem\u003eL. pumilum\u003c/em\u003e and \u003cem\u003eL. lancifolium.\u003c/em\u003e These similarities indicate that \u003cem\u003eL. davidii\u003c/em\u003e is relatively closely related to \u003cem\u003eL. lancifolium\u003c/em\u003e and \u003cem\u003eL. pumilum\u003c/em\u003e. The relatedness among these three species revealed by our comparison of rDNA pattern is basically consistent with that previously revealed by molecular phylogenetic analyses (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In the PCoA scatter plot, \u003cem\u003eL. pumilum, L. lancifolium\u003c/em\u003e and \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003ewillmottiae\u003c/em\u003e are clustered together in the same group along the direction of PCoA1, roughly confirming the relatedness of these species drawn from rDNA FISH and molecular phylogenetics.\u003c/p\u003e\u003cp\u003eThe 35S pattern of \u003cem\u003eL. jinfushanense\u003c/em\u003e is different from that of any of the other species studied herein, but is the same as that of \u003cem\u003eL. taliense\u003c/em\u003e reported by Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e. The location of the 5S locus on chromosome 3 of \u003cem\u003eL. jinfushanense\u003c/em\u003e is different from that of the species belonging to \u003cem\u003eSinomartagon\u003c/em\u003e 5a and \u003cem\u003eSinomartagon\u003c/em\u003e 5b, but similar to that of the species belonging to \u003cem\u003eLeucolirion\u003c/em\u003e 6a. The consistency of the 35S pattern between \u003cem\u003eL. jinfushanense\u003c/em\u003e and \u003cem\u003eL. taliense\u003c/em\u003e indicates their very close relationship. This is accordant with the result of molecular phylogenetic analysis based on ITS sequences and plastid genomes (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). It is worth in-depth study how the unique rDNA pattern of \u003cem\u003eL. jinfushanense\u003c/em\u003e evolved and the evolutionary relationships with other species endemic to southwest China.\u003c/p\u003e\u003cp\u003eThe 35S pattern of \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e detected by us is the same as that of \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e and \u003cem\u003eL. brownii\u003c/em\u003e reported by several authors (Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Wu et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), but is completely inconsistent with that of \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e reported by Wu et al. 2012. As for the 5S locus of L. \u003cem\u003ebrownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e, we only detected the 5S locus in the interstitial regions of the long arms of chromosome 3, but Wu et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2021\u003c/span\u003e detected another five 5S loci. This inconsistency may be due to differences in material used. \u003cem\u003eL. brownii\u003c/em\u003e was placed in sect. \u003cem\u003eArchelirion\u003c/em\u003e by Comber (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1949\u003c/span\u003e) and later adjusted to \u003cem\u003eLeucolirion\u003c/em\u003e 6b based on molecular phylogenetic evidence (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Du et al. 2023). \u003cem\u003eL. longiflorum\u003c/em\u003e and \u003cem\u003eL. formosanum\u003c/em\u003e, two species also belonging to \u003cem\u003eLeucolirion\u003c/em\u003e 6b, have a 35S pattern somewhat similar to that of \u003cem\u003eL. brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e: they also have no 35S loci on chromosomes 1 and 2, and have a 35S locus in the proximal regions of short arms of chromosome 7 (Lim et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These facts indicate that the rDNA pattern of already studied species of \u003cem\u003eLeucolirion\u003c/em\u003e 6b differ significantly from those of species of \u003cem\u003eLeucolirion\u003c/em\u003e 6a studied herein.\u003c/p\u003e\u003cp\u003eThe locations of the 5S loci on chromosome 3 and the 35S loci on chromosomes 1, 2, 4 and 5 of \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e, \u003cem\u003eL. sulphureum\u003c/em\u003e, and \u003cem\u003eL. regale\u003c/em\u003e are the same or very similar, indicating their close relationships. PCoA also indicates that these species have close karyological relationships. Our study demonstrated from the perspective of cytogenetics that these four species are closely related to each other, in accordance with the conclusion inferred from molecular phylogenetic studies (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). It is worth noting that among these four species, the karyotype differentiation of \u003cem\u003eL. regale\u003c/em\u003e is more significant. Not only does it have an additional 35s locus located on chromosome 6, but its karyotype parameters also alter greatly compared to the other three species, leading to its aggregation in another group in the PCoA plot. The rDNA localization of \u003cem\u003eL. leucanthum\u003c/em\u003e has not been reported till now. Wang et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e reported the rDNA mapping of \u003cem\u003eL. leucanthum\u003c/em\u003e var. \u003cem\u003ecentifolium\u003c/em\u003e, and showed no 35S loci on chromosome 1 and 2. Such 35S pattern of this variety is very different from that of \u003cem\u003eL. leucanthum\u003c/em\u003e revealed by us. It is necessary to further determine the rDNA pattern of this variety by dual-color FISH. Previous studies have detected four 35S loci located on chromosomes 1 and 2 as well as two st-type chromosome pairs in \u003cem\u003eL. sargentiae\u003c/em\u003e and \u003cem\u003eL. sulphureum\u003c/em\u003e (Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Zhu et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), and detected four (Zhou et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Zhu et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), five (Cao et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) and six (Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) 35S loci (including the two loci on chromosomes 1 and 2) in \u003cem\u003eL. regale\u003c/em\u003e, but these studies fail to accurately determine the chromosomal locations of some of these loci.\u003c/p\u003e\u003cp\u003eOur comparative localization of 5S and 35S rDNA of \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e by dual-color FISH reveals the similarity of their rDNA patterns, further demonstrating the close relationship between them from the perspective of cytogenetics (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Previous studies have reported the localization of both 5S and 35S (Marasek et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Kang et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and the localization of 35S alone (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) in \u003cem\u003eL. henryi\u003c/em\u003e. Our result of rDNA mapping of this species is basically consistent with those of previous reports, except that the numbering of the st-type chromosome pair with 35S locus is different from theirs (Marasek et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Kang et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). For \u003cem\u003eL. rosthornii\u003c/em\u003e, previous studies only mapped 35S by FISH, revealing a similar pattern with ours (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Earlier, \u003cem\u003eL. rosthornii\u003c/em\u003e was assigned to \u003cem\u003eSinomartagon\u003c/em\u003e 5a (Comber \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1949\u003c/span\u003e), and later transferred to \u003cem\u003eLeucolirion\u003c/em\u003e 6a based on the conclusions of molecular phylogenetic analysis and 35S rDNA FISH (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; van Tuyl et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Du et al. 2023). In the phylogenetic tree based on ITS sequences, \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e belong to \u003cem\u003eLeucolirion\u003c/em\u003e clade I along with \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e, \u003cem\u003eL. sulphureum\u003c/em\u003e and \u003cem\u003eL. regale\u003c/em\u003e. The former two were clustered into one branch which showed a sister relationship with the branch where the latter four species were clustered (Du et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). In the latest revised classification of \u003cem\u003eLilium\u003c/em\u003e, these six species have been assigned to \u003cem\u003eLeucolirion\u003c/em\u003e 6a (van Tuyl et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Du et al. 2023). From the comparison of their rDNA patterns, the 5S locus of chromosome 3 and the 35S locus of chromosome 1 of the former two species are similar to those of the latter four species, but such similarities are not sufficient to indicate that there are close relationships between the species of the two branches. However, PCoA based on the six karyotypic parameters provides evidence. As shown in the PCoA scatter plot, \u003cem\u003eL. henryi\u003c/em\u003e and \u003cem\u003eL. rosthornii\u003c/em\u003e are clustered in the same group with \u003cem\u003eL. leucanthum\u003c/em\u003e, \u003cem\u003eL. sargentiae\u003c/em\u003e, and \u003cem\u003eL. sulphureum\u003c/em\u003e along the direction of PCoA1.\u003c/p\u003e\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eMolecular cytogenetic karyotypes of eleven species of \u003cem\u003eLilium\u003c/em\u003e from China are established quantitatively using the dataset of chromosome measurements and FISH signals of 5S and 35S rDNA. Comparative karyotype analysis reveals variations in the karyotypic parameters and rDNA patterns among species. Their karyotype structures could be differentiated by the scatter plot of M\u003csub\u003eCA\u003c/sub\u003e vs. CV\u003csub\u003eCL\u003c/sub\u003e. The evolutionary relationships among them can be revealed by the combination of comparison of rDNA patterns as well as PCoA based on \u003cem\u003ex\u003c/em\u003e, 2n, TCL, CV\u003csub\u003eCI\u003c/sub\u003e, M\u003csub\u003eCA\u003c/sub\u003e and CV\u003csub\u003eCL\u003c/sub\u003e. Our results confirm cytogenetically the rationality of the species classification of subset. \u003cem\u003eLeucolirion\u003c/em\u003e 6a in the latest revised classification of \u003cem\u003eLilium.\u003c/em\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eThe Natural Science Foundation of Hunan Province, China (2019JJ40231, 2024JJ7156).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eConceptualization: CWS; Funding acquisition: CWS, BYZ; Investigation: YDY; Data curation: YDY, XHJ; Formal analysis: YDY, XHJ; Methodology: CWS, YDY, XHJ, BYZ; Project administration: CWS; Resources: BYZ, YDY; Supervision: CWS; Validation: YDY; Visualization: YDY; Writing - original draft: YDY; Writing - review \u0026amp; editing: CWS, XHJ, BYZ.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eAll data supporting the findings of this study are available in the article and its supplementary information files online.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAhn, Y. J. et al. Investigation of karyotypic composition and evolution in \u003cem\u003eLilium\u003c/em\u003e species belonging to the section Martagon. \u003cem\u003ePlant. Biotechnol. Rep.\u003c/em\u003e \u003cb\u003e11\u003c/b\u003e, 407\u0026ndash;416. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11816-017-0462-7\u003c/span\u003e\u003cspan address=\"10.1007/s11816-017-0462-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAstuti, G., Roma-Marzio, F. \u0026amp; Peruzzi, L. Traditional cytotaxonomic studies: can they still provide a solid basis in plant systematics? \u003cem\u003eFl Medit\u003c/em\u003e. \u003cb\u003e27\u003c/b\u003e, 91\u0026ndash;98. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.7320/FlMedit27.091\u003c/span\u003e\u003cspan address=\"10.7320/FlMedit27.091\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCao, Q. et al. Physical mapping of 45S rDNA loci in \u003cem\u003eLilium\u003c/em\u003e OT hybrids and interspecific hybrids with \u003cem\u003eLilium regale\u003c/em\u003e. \u003cem\u003eSci. Hortic.\u003c/em\u003e \u003cb\u003e252\u003c/b\u003e, 48\u0026ndash;54. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scienta.2019.03.035\u003c/span\u003e\u003cspan address=\"10.1016/j.scienta.2019.03.035\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChinese Pharmacopoeia Commission. \u003cem\u003ePharmacopoeia of the People\u0026rsquo;s Republic of China: 2020 version\u003c/em\u003e pp186\u0026ndash;187 (Vol. I. China Medical Science, 2020).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChoi, Y. H. et al. Using cytogenetic analysis to identify the genetic diversity in \u003cem\u003eLilium hansonii\u003c/em\u003e(Liliaceae), an endemic species of Ulleung Island, Korea. \u003cem\u003eHortic. Environ. Biotechnol.\u003c/em\u003e \u003cb\u003e62\u003c/b\u003e, 795\u0026ndash;804. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s13580-021-00350-1\u003c/span\u003e\u003cspan address=\"10.1007/s13580-021-00350-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eComber, H. F. A new classification of the genus \u003cem\u003eLilium\u003c/em\u003e. \u003cem\u003eLily Yearb Roy Hort Soc.\u003c/em\u003e \u003cb\u003e13\u003c/b\u003e, 86\u0026ndash;105. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11032-010-9490-5\u003c/span\u003e\u003cspan address=\"10.1007/s11032-010-9490-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1949).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDias, S. et al. do,, Cytomolecular diversity among \u003cem\u003eVigna\u003c/em\u003e Savi (Leguminosae) subgenera. Protoplasma 261:859\u0026ndash;875. (2024). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00709-024-01944-z\u003c/span\u003e\u003cspan address=\"10.1007/s00709-024-01944-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDu, F. Origin, classification and germplasm diversity in \u003cem\u003eLilium\u003c/em\u003e. \u003cem\u003eJ. China Agricultural Univ.\u003c/em\u003e \u003cb\u003e28\u003c/b\u003e, 68\u0026ndash;79. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.11841/j.issn.1007-4333.2023.04.06\u003c/span\u003e\u003cspan address=\"10.11841/j.issn.1007-4333.2023.04.06\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDu, Y. P. et al. Molecular phylogeny and genetic variation in the genus \u003cem\u003eLilium\u003c/em\u003e native to China based on the internal transcribed spacer sequences of nuclear ribosomal DNA. \u003cem\u003eJ. Plant. Res.\u003c/em\u003e \u003cb\u003e127\u003c/b\u003e, 249\u0026ndash;263. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10265-013-0600-4\u003c/span\u003e\u003cspan address=\"10.1007/s10265-013-0600-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGao, Y. D. \u0026amp; Gao, X. F. Accommodating \u003cem\u003eNomocharis\u003c/em\u003e in \u003cem\u003eLilium\u003c/em\u003e (Liliaceae). Phytotaxa 277:205\u0026ndash;210. (2016). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.11646/phytotaxa.277.2.8\u003c/span\u003e\u003cspan address=\"10.11646/phytotaxa.277.2.8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGao, Y. D., Zhou, S. D. \u0026amp; He, X. J. Karyotype studies in thirty-two species of \u003cem\u003eLilium\u003c/em\u003e (Liliaceae) from China. \u003cem\u003eNord J. Bot.\u003c/em\u003e \u003cb\u003e29\u003c/b\u003e, 746\u0026ndash;761. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1756-1051.2011.01069.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1756-1051.2011.01069.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2011).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGao, Y. D., Zhou, S. D., He, X. J. \u0026amp; Wan, J. Chromosome diversity and evolution in tribe Lilieae (Liliaceae) with emphasis on Chinese species. \u003cem\u003eJ. Plant. Res.\u003c/em\u003e \u003cb\u003e125\u003c/b\u003e, 55\u0026ndash;69. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10265-011-0422-1\u003c/span\u003e\u003cspan address=\"10.1007/s10265-011-0422-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGivnish, T. et al. Evolution, geographic spread and floral diversification of the genus \u003cem\u003eLilium\u003c/em\u003e with special reference to the lilies of North America. \u003cem\u003eLily Yearbook North. Am. Lily Soc.\u003c/em\u003e \u003cb\u003e74\u003c/b\u003e, 26\u0026ndash;44 (2020).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHan, Y. L. et al. Comparative analysis of rDNA distribution in 29 species of \u003cem\u003eBegonia\u003c/em\u003e sect. \u003cem\u003eCoelocentrum Irmsch Phytotaxa\u003c/em\u003e. \u003cb\u003e381\u003c/b\u003e, 141\u0026ndash;152. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.11646/PHYTOTAXA.381.1.18\u003c/span\u003e\u003cspan address=\"10.11646/PHYTOTAXA.381.1.18\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHwang, Y. J., Kim, H. H., Kim, J. B. \u0026amp; Lim, K. B. Karyotype analysis of \u003cem\u003eLilium tigrinum\u003c/em\u003e by FISH. \u003cem\u003eHort Environ. Biotechnol.\u003c/em\u003e \u003cb\u003e52\u003c/b\u003e, 292\u0026ndash;297. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s13580-011-0225-2\u003c/span\u003e\u003cspan address=\"10.1007/s13580-011-0225-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2011).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKadluczka, D. \u0026amp; Grzebelus, E. Using carrot centromeric repeats to study karyotype relationships in the genus \u003cem\u003eDaucus\u003c/em\u003e (Apiaceae). \u003cem\u003eBMC Genom.\u003c/em\u003e \u003cb\u003e22\u003c/b\u003e, 508. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12864-021-07853-2\u003c/span\u003e\u003cspan address=\"10.1186/s12864-021-07853-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKang, J. Y., Lim, K. B. \u0026amp; Ahn, Y. J. Cytogenetic analysis and visualization of genetic relationships in wild lilies. \u003cem\u003eGenes Genom\u003c/em\u003e. \u003cb\u003e46\u003c/b\u003e, 1253\u0026ndash;1265. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s13258-024-01568-w\u003c/span\u003e\u003cspan address=\"10.1007/s13258-024-01568-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLee, H., Younis, A., Hwang, Y. J., Kang, Y. I. \u0026amp; Lim, K. B. Molecular cytogenetic analysis and phylogenetic relationship of 5S and 45S ribosomal DNA in Sinomartagon \u003cem\u003eLilium\u003c/em\u003e species by fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization (FISH). \u003cem\u003eHortic. Environ. Biotechnol.\u003c/em\u003e \u003cb\u003e55\u003c/b\u003e, 514\u0026ndash;523. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s13580-014-0066-x\u003c/span\u003e\u003cspan address=\"10.1007/s13580-014-0066-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLevan, A., Fredga, K. \u0026amp; Sandberg, A. Nomenclature for centromeric position in chromosomes. \u003cem\u003eHereditas\u003c/em\u003e \u003cb\u003e52\u003c/b\u003e, 201\u0026ndash;220. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1601-5223.1964.tb01953.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1601-5223.1964.tb01953.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1964).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi, J. et al. Phylogeny, age, and evolution of tribe Lilieae (Liliaceae) based on whole plastid genomes. \u003cem\u003eFront. Plant. Sci.\u003c/em\u003e \u003cb\u003e12\u003c/b\u003e, 699226. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2021.699226\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2021.699226\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLiang, S. Y. \u0026amp; Tamura, N. Lilium. In: (eds Wu, Z. Y. \u0026amp; Raven, P. H.) Flora of China. Science Press, Beijing, Miss Bot Gard, St. Louis, 24:135\u0026ndash;149 (2000).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLim, K. B., Wennekes, J., de Jong, J. H., Jacobsen, E. \u0026amp; van Tuyl, J. M. Karyotype analysis of \u003cem\u003eLilium longiflorum\u003c/em\u003e and \u003cem\u003eLilium rubellum\u003c/em\u003e by chromosome banding and fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridisation. \u003cem\u003eGenome\u003c/em\u003e \u003cb\u003e44\u003c/b\u003e, 911\u0026ndash;918. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1139/gen44-5-911\u003c/span\u003e\u003cspan address=\"10.1139/gen44-5-911\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2001).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLiu, G. X. et al. Karyotype and fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization analysis of 15 \u003cem\u003eLilium\u003c/em\u003e species from China. \u003cem\u003eJ. Am. Soc. Hortic. Sci.\u003c/em\u003e \u003cb\u003e142\u003c/b\u003e, 298\u0026ndash;305. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.21273/JASHS04094-17\u003c/span\u003e\u003cspan address=\"10.21273/JASHS04094-17\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMarasek, A., Hasterok, R., Wiejacha, K. \u0026amp; Orlikowska, T. Determination by GISH and FISH of hybrid status in \u003cem\u003eLilium\u003c/em\u003e. \u003cem\u003eHereditas\u003c/em\u003e \u003cb\u003e140\u003c/b\u003e, 1\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1601-5223.2004.01721.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1601-5223.2004.01721.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2004).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMitrenina, E. Y. et al. Karyotypes and physical mapping of ribosomal DNA with Oligo-Probes in \u003cem\u003eEranthis\u003c/em\u003e sect. Eranthis (Ranunculaceae). \u003cem\u003ePlants (Basel)\u003c/em\u003e. \u003cb\u003e13\u003c/b\u003e, 47. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/plants13010047\u003c/span\u003e\u003cspan address=\"10.3390/plants13010047\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMuratović, E., Robin, O., Bogunić, F., Šoljan, D. \u0026amp; Siljak-Yakovlev, S. Karyotype evolution and speciation of European lilies from \u003cem\u003eLilium\u003c/em\u003e sect. \u003cem\u003eLiriotypus Taxon\u003c/em\u003e. \u003cb\u003e59\u003c/b\u003e, 165\u0026ndash;175. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2307/27757060\u003c/span\u003e\u003cspan address=\"10.2307/27757060\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNguyen, T. X. et al. Karyotype analysis of Korean \u003cem\u003eLilium maximowiczii\u003c/em\u003e Regal populations. \u003cem\u003eJ. Fac. Agr Kyushu Univ.\u003c/em\u003e \u003cb\u003e60\u003c/b\u003e, 315\u0026ndash;322. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5109/1526344\u003c/span\u003e\u003cspan address=\"10.5109/1526344\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNguyen, T. X., Lee, S. I., Rai, R., Kim, N. S. \u0026amp; Kim, J. H. Ribosomal DNA locus variation and REMAP analysis of the diploid and triploid complexes of \u003cem\u003eLilium lancifolium\u003c/em\u003e. \u003cem\u003eGenome\u003c/em\u003e \u003cb\u003e59\u003c/b\u003e, 551\u0026ndash;564. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1139/gen-2016-0011\u003c/span\u003e\u003cspan address=\"10.1139/gen-2016-0011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNoda, S. Cytogenetic behavior, chromosomal differentiations, and geographic distribution in \u003cem\u003eLilium lancifolium\u003c/em\u003e (Liliaceae). \u003cem\u003ePlant. Spec. Biol.\u003c/em\u003e \u003cb\u003e1\u003c/b\u003e, 69\u0026ndash;78. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1442-1984.1986.tb00016.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1442-1984.1986.tb00016.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1986).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNoda, S. Chromosomal variation and evolution in the genus \u003cem\u003eLilium\u003c/em\u003e. In: Tsuchiya T, Gupta PK (eds), Chromosome engineering in plants: genetics, breeding, evolution, part B. Elsevier, pp 507\u0026ndash;524. (1991). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/B978-0-444-88260-8.50032-X\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-444-88260-8.50032-X\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePaszko, B. A critical review and a new proposal of karyotype asymmetry indices. \u003cem\u003ePlant. Syst. Evol.\u003c/em\u003e \u003cb\u003e258\u003c/b\u003e, 39\u0026ndash;48. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00606-005-0389-2\u003c/span\u003e\u003cspan address=\"10.1007/s00606-005-0389-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2006).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePatterson, T. B. \u0026amp; Givnish, T. J. Phylogeny, concerted convergence, and phylogenetic niche conservatism in the core Liliales: insights from \u003cem\u003erbcL\u003c/em\u003e and \u003cem\u003endhF\u003c/em\u003e sequence data. \u003cem\u003eEvolution\u003c/em\u003e \u003cb\u003e56\u003c/b\u003e, 233\u0026ndash;252. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.0014-3820.2002.tb01334.x\u003c/span\u003e\u003cspan address=\"10.1111/j.0014-3820.2002.tb01334.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2002).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePeruzzi, L. \u0026amp; Altınordu, F. A proposal for a multivariate quantitative approach to infer karyological relationships among taxa. \u003cem\u003eComp. Cytogenet.\u003c/em\u003e \u003cb\u003e8\u003c/b\u003e, 337\u0026ndash;349. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3897/CompCytogen.v8i4.8564\u003c/span\u003e\u003cspan address=\"10.3897/CompCytogen.v8i4.8564\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePeruzzi, L. \u0026amp; Eroglu, H. Karyotype asymmetry: again, how to measure and what to measure? \u003cem\u003eComp. Cytogenet.\u003c/em\u003e \u003cb\u003e7\u003c/b\u003e, 1\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3897/CompCytogen.v7i1.4431\u003c/span\u003e\u003cspan address=\"10.3897/CompCytogen.v7i1.4431\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePeruzzi, L., Leitch, I. J. \u0026amp; Caparelli, K. F. Chromosome diversity and evolution in Liliaceae. \u003cem\u003eAnn. Bot.\u003c/em\u003e \u003cb\u003e103\u003c/b\u003e, 459\u0026ndash;475. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/aob/mcn230\u003c/span\u003e\u003cspan address=\"10.1093/aob/mcn230\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2009).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eShe, C. W., Jiang, X. H. \u0026amp; He, C. P. Comparative karyotype analysis of eight Cucurbitaceae crops using fluorochrome banding and 45S rDNA-FISH. \u003cem\u003eComp. Cytogenet.\u003c/em\u003e \u003cb\u003e17\u003c/b\u003e, 31\u0026ndash;58. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3897/compcytogen.v17.i1.99236\u003c/span\u003e\u003cspan address=\"10.3897/compcytogen.v17.i1.99236\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eShe, C. W. et al. Molecular cytogenetic characterisation and phylogenetic analysis of the seven cultivated \u003cem\u003eVigna\u003c/em\u003e species (Fabaceae). \u003cem\u003ePlant. Biol.\u003c/em\u003e \u003cb\u003e17\u003c/b\u003e, 268\u0026ndash;280. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/plb.12174\u003c/span\u003e\u003cspan address=\"10.1111/plb.12174\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSmyth, D. R., Kongsuwan, K. \u0026amp; Wisudharomn, S. A survey of C-band patterns in chromosomes of \u003cem\u003eLilium\u003c/em\u003e (Liliaceae). \u003cem\u003ePlant. Syst. Evol.\u003c/em\u003e \u003cb\u003e163\u003c/b\u003e, 53\u0026ndash;69. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00936153\u003c/span\u003e\u003cspan address=\"10.1007/BF00936153\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1989).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStebbins, G. L. \u003cem\u003eChromosomal evolution in higher plants\u003c/em\u003e (Addison-Wesley, 1971). pp220.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStewart, R. N. The morphology of somatic chromosomes in \u003cem\u003eLilium\u003c/em\u003e. \u003cem\u003eAm. J. Bot.\u003c/em\u003e \u003cb\u003e34\u003c/b\u003e, 9\u0026ndash;26. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/j.1537-2197.1947.tb12953.x\u003c/span\u003e\u003cspan address=\"10.1002/j.1537-2197.1947.tb12953.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1947).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSultana, S., Lee, S. H., Bang, J. W. \u0026amp; Choi, H. W. Physical mapping of rRNA gene loci and inter-specific relationships in wild \u003cem\u003eLilium\u003c/em\u003e distributed in Korea. \u003cem\u003eJ. Plant. Biol.\u003c/em\u003e \u003cb\u003e53\u003c/b\u003e, 433\u0026ndash;443. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s12374-010-9133-8\u003c/span\u003e\u003cspan address=\"10.1007/s12374-010-9133-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSun, J. et al. Comparative karyotype analysis provides cytogenetic evidence for the origin of sweetpotato. \u003cem\u003eChromosome Res.\u003c/em\u003e \u003cb\u003e32\u003c/b\u003e, 14. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10577-024-09758-8\u003c/span\u003e\u003cspan address=\"10.1007/s10577-024-09758-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVan Tuyl, J. M. et al. Lilium. In: (ed Van Huylenbroeck, J.) Ornamental crops. Handbook of Plant Breeding, vol 11. Springer, Cham., 481\u0026ndash;512. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-3-319-90698-0_20\u003c/span\u003e\u003cspan address=\"10.1007/978-3-319-90698-0_20\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWan, J., Zhou, S., Gao, Y. \u0026amp; He, X. Karyotypes of twenty-five populations of thirteen species in \u003cem\u003eNomocharis\u003c/em\u003e and \u003cem\u003eLilium\u003c/em\u003e. \u003cem\u003ePlant. Divers. Resour.\u003c/em\u003e \u003cb\u003e33\u003c/b\u003e, 477\u0026ndash;494. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3724/SP.J.1011.2011.00093\u003c/span\u003e\u003cspan address=\"10.3724/SP.J.1011.2011.00093\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2011).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang, Q. et al. The application of fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization in different ploidy levels cross-breeding of lily. \u003cem\u003ePLoS One\u003c/em\u003e. \u003cb\u003e10\u003c/b\u003e, e0126899. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0126899\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0126899\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang, X., Xie, S., Zhang, Y. \u0026amp; Niu, L. Chromosome analysis and mapping of ribosomal genes by fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization (FISH) in four endemic lily species (\u003cem\u003eLilium\u003c/em\u003e) in Qinling mountians, China. \u003cem\u003ePak J. Bot.\u003c/em\u003e \u003cb\u003e44\u003c/b\u003e, 1319\u0026ndash;1323 (2012).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWei, Y. F., Jiang, X. H., Song, R. \u0026amp; She, C. W. Molecular cytogenetic characterization of 9 populations of four species in the genus \u003cem\u003ePolygonatum\u003c/em\u003e (Asparagaceae). \u003cem\u003eComp. Cytogenet.\u003c/em\u003e \u003cb\u003e18\u003c/b\u003e, 73\u0026ndash;95. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3897/compcytogen.18.122399\u003c/span\u003e\u003cspan address=\"10.3897/compcytogen.18.122399\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWu, L. et al. Analysis of the cross-compatibility of \u003cem\u003eLilium brownii\u003c/em\u003e var. \u003cem\u003eviridulum\u003c/em\u003e and \u003cem\u003eL. davidii\u003c/em\u003e var. \u003cem\u003eunicolor\u003c/em\u003e. Sci Hortic 284:110130. (2021). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scienta.2021.110130\u003c/span\u003e\u003cspan address=\"10.1016/j.scienta.2021.110130\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou, Y. P. et al. Fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization of 35S rDNA sites and karyotype of wild \u003cem\u003eLilium\u003c/em\u003e (Liliaceae) species from China: taxonomic and phylogenetic implications. \u003cem\u003eGenet. Resour. Crop Evol.\u003c/em\u003e \u003cb\u003e67\u003c/b\u003e, 1601\u0026ndash;1617. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10722-020-00936-1\u003c/span\u003e\u003cspan address=\"10.1007/s10722-020-00936-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou, J., An, R. \u0026amp; Huang, X. Genus \u003cem\u003eLilium\u003c/em\u003e: A review on traditional uses, phytochemistry and pharmacology. \u003cem\u003eJ. Ethnopharmacol.\u003c/em\u003e \u003cb\u003e270\u003c/b\u003e, 113852. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jep.2021.113852\u003c/span\u003e\u003cspan address=\"10.1016/j.jep.2021.113852\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou, M. et al. Chromosomal analysis of progenies between \u003cem\u003eLilium\u003c/em\u003e intersectional hybrids and wild species using ND-FISH and GISH. \u003cem\u003eFront. Plant. Sci.\u003c/em\u003e \u003cb\u003e15\u003c/b\u003e, 1461798. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2024.1461798\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2024.1461798\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhou, N. et al. Historical biogeography and evolutionary diversification of \u003cem\u003eLilium\u003c/em\u003e (Liliaceae): New insights from plastome phylogenomics. \u003cem\u003ePlant. Divers.\u003c/em\u003e \u003cb\u003e46\u003c/b\u003e, 219\u0026ndash;228. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.pld.2023.07.009\u003c/span\u003e\u003cspan address=\"10.1016/j.pld.2023.07.009\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eZhu, Z., Sun, Y., Liu, S., Peng, M. \u0026amp; Zhou, S. Comparison of \u003cem\u003eLilium regale\u003c/em\u003e and \u003cem\u003eL. sargentiae\u003c/em\u003e for lily breeding. \u003cem\u003eGenet. Resour. Crop Evol.\u003c/em\u003e \u003cb\u003e72\u003c/b\u003e, 6375\u0026ndash;6386. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10722-025-02405-z\u003c/span\u003e\u003cspan address=\"10.1007/s10722-025-02405-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2025).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Lilium, karyotype, karyotype asymmetry, cytotaxonomy, ribosomal RNA gene (rDNA), fluorescence in situ hybridization (FISH)","lastPublishedDoi":"10.21203/rs.3.rs-8029111/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8029111/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe complexity of the classification of the genus \u003cem\u003eLilium\u003c/em\u003e requires more molecular cytogenetic researches to clarify the karyotype differentiation and chromosome evolution among species, verify the existing classification, and help resolve the remaining disputes in classification. In this study, the chromosomes of eleven species of \u003cem\u003eLilium\u003c/em\u003e, including China's main medicinal and edible lilies as well as seven species endemic to southwest and central China, were characterized using fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization (FISH) with 5S and 35S rDNA oligonucleotide probes. Molecular cytogenetic karyotypes were quantitatively established using the dataset of chromosome measurements and FISH signals. Karyotype asymmetry indices were measured for elucidating their karyotype asymmetry and karyological relationships. Their karyotype structures could be differentiated by the scatter plot of M\u003csub\u003eCA\u003c/sub\u003e vs. CV\u003csub\u003eCL\u003c/sub\u003e. rDNA FISH showed that there were 1\u0026ndash;2 5S rDNA loci and 2\u0026ndash;6 35S rDNA loci in these species, and most of the 35S loci generated secondary constrictions. The combination of comparison of rDNA patterns and PCoA based on \u003cem\u003ex\u003c/em\u003e, 2\u003cem\u003en\u003c/em\u003e, TCL, M\u003csub\u003eCA\u003c/sub\u003e, CV\u003csub\u003eCL\u003c/sub\u003e and CV\u003csub\u003eCI\u003c/sub\u003e reveals the evolutionary relationships among these species. Our results cytogenetically confirm the rationality of the species classification of subset. \u003cem\u003eLeucolirion\u003c/em\u003e 6a in the latest revised classification of \u003cem\u003eLilium\u003c/em\u003e.\u003c/p\u003e","manuscriptTitle":"Comparative karyotype analysis of eleven species of Lilium from China by FISH with rDNA oligo-probes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-27 19:01:07","doi":"10.21203/rs.3.rs-8029111/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-12-23T05:56:26+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-10T03:46:38+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"339171481587324336225764202660573117043","date":"2025-11-25T03:21:27+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-25T02:13:29+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"79898746474085136258341441351591197830","date":"2025-11-19T12:46:23+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-19T12:02:26+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-19T11:57:01+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-11-18T05:03:11+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-13T15:16:37+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-11-13T15:10:59+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b037ca11-710d-41fe-8aad-8cdd48f0df9a","owner":[],"postedDate":"November 27th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":58547946,"name":"Biological sciences/Biological techniques"},{"id":58547947,"name":"Biological sciences/Genetics"},{"id":58547948,"name":"Biological sciences/Molecular biology"}],"tags":[],"updatedAt":"2026-02-23T15:59:57+00:00","versionOfRecord":{"articleIdentity":"rs-8029111","link":"https://doi.org/10.1038/s41598-026-37297-1","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-02-17 15:56:53","publishedOnDateReadable":"February 17th, 2026"},"versionCreatedAt":"2025-11-27 19:01:07","video":"","vorDoi":"10.1038/s41598-026-37297-1","vorDoiUrl":"https://doi.org/10.1038/s41598-026-37297-1","workflowStages":[]},"version":"v1","identity":"rs-8029111","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8029111","identity":"rs-8029111","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}