Production and Partial Purification of Pectinases Produced by Three Novel Cladosporium Species Isolated from Egypt

Production and Partial Purification of Pectinases Produced by Three Novel Cladosporium Species Isolated from Egypt | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Production and Partial Purification of Pectinases Produced by Three Novel Cladosporium Species Isolated from Egypt Ahmed Mohamed Moharram, Abdel-Naser Ahmed Zohri, Abd El-Latif Hesham, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1291889/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Based on phenotypic characters and sequencing of the internal transcribed spacers (ITS), 28S Large Subunit (LSU) and partial actin (ACT), three novel species of Cladosporium were described in this study as C. parasphaerospermum , C. chlamydosporum , and C. compactisporum . Activities of pectinases produced by the three strains were 5700, 3720, and 3480 U mg − 1 enzymes, at the optimum pH (6.0, 7.0, and 8.0) and temperature (35, 30, and 35 ºC), respectively. The ions potassium and manganese had a strong activating impact on the activity of C. compactisporum pectinase (124.4 and 172%, respectively). When tested under optimal conditions, EDTA and the other metal ions inhibited the activity of the pectinases in varied degrees. The strongest inhibitory effects of C. parasphaerospermum , C. chlamydosporum , and C. compactisporum pectinases were with Ni + 2 , Zn + 2 , and EDTA, respectively. Cladosporium species have a long history of biotechnological uses, and here we add another: pectinase makers. We offer three unique suppliers of low-temperature active fungal pectinases for biotechnological applications such as the management of Pectinacious wastes in the juice industry. Biodiversity Fungi Pectinase Phylogeny Taxonomy Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Cladosporium is one of the largest and most complex genera of hyphomycetes, which currently includes more than 728 names. Until recently, all types of unrelated dematiaceous hyphomycetes characterized by amero-to-phragmosporous conidia formed in acropetal chains had been referred to Cladosporium 1 . Species of Cladosporium are well adapted to be spread easily in large numbers over long distances, therefore they are cosmopolitan and widely present in all various types of plants and other debris, mostly isolated from air, soil, seeds, grains, food, paint, textiles and other organic matter 1–9 .Other species of this genus are plant pathogenic causing leaf spots and other lesions 10 , or they occur as hyperparasites on other fungi 11 . Cladosporium species are also known to be common endophytes 12–14 as well as phylloplane fungi 15–18 . Some Cladosporium species including C . cladosporioides , C . chlorocephalum and C . uredinicola were recorded as entomopathogens of Aphids and whiteflies 19,20 . The Genus Cladosporium is considered a rich source of diverse and bioactive natural compounds 21 . Some species were reported to produce anticancer compounds such as L-asparaginase 22 , paclitaxel 23 , as well as enzymes like cellulases 24 , and pectinases 25–27 . Pectinases are enzymes that target pectin and depolymerize it by hydrolysis, transelimination, and deesterification processes, which hydrolyze the ester connection between pectin's carboxyl and methyl groups. These enzymes degrade pectin, a complex polysaccharide present in the cell walls of higher plants that serves as a cementing substance for the cellulose network. Pectinases account for 10% of all industrial enzymes manufactured globally, and their market is growing by the day 28–30 . As a result, the current study attempted to identify some unique Cladosporium species isolated from air and fruits in some Egyptian governorates, as well as to produce and partially purify pectinases in submerged fermentation. Results Molecular studies The ITS of Cladosporium strain AUMC 10865 was 99.64% (554/556) similar to C . cladosporioides (KJ767065), C. parahalotolerans (MK262909) and C. halotolerans (MK258720). ACT sequence was 96.21% (127/132) similar to C. halotolerans (MF084398 and MF084399) and C. omanense (MH716046). LSU sequence was 98.96% (1140/1152) similar to C. delicatulum (JQ732984 and JQ732983). ITS of Cladosporium strain AUMC 11340 was 99.27% (549/553) similar to C. floccosum (MK460809). ACT sequence was 99.13% (228/230) to C. herbarum (EF679510). Comparing the ACT sequence to the type strains, it was 98.26% (226/230) similar to C. macrocarpum (EF679529), 97.83% (225/230) to C. herbarum (EF679516), and 95.15% (216/227) to C. versiforme (KT600613). LSU was 98.74% (1184/1199) similar to C. herbarum (MH047193), and C. allicinum (GU214408). ITS sequence of Cladosporium strain AUMC 11366 was 98.9% (548/554) similar to C. cladosporioides (MT367253, MF077224 and MF319920). By comparing to ITS sequences of the type species, it was 98.55% (542/550) similar to C. verrucocladosporioides (MH863939), 98.53% (536/544) to C. vicinum (MF473311) and 98.88% (530/536) to C. kenpeggii (KY646222). ACT of Cladosporium strain AUMC 11366 was 100% similar to C. cladosporioides (KY886457, MZ734607 and HM148501). By comparing with ACT sequences of the type materials, it was 97.38% similar to C. proteacearum (MZ344213), 98.15% to C. devikae (MZ344212), and 95.65% similar to C. cladosporioides (HM148490). LSU was 98.77% (1206/1221) similar to C. delicatulum (JQ732985) and C. uredinicola (EU019264). This analysis included 55 sequences. The full parsimony dataset consisted of 808 characters with 437 constant characters (no gaps, no N), 114 variable characters which were parsimony-uninformative (26.1% of constant characters), and 33 characters were counted as parsimony informative (7.6% of constant). Tamura 3-parameter (T92 + G) was the best nucleotide substitution model. The most parsimonious tree, resulted from the maximum parsimony analysis with the highest log likelihood (-5359.90), a tree length of 802 steps, consistency index of 0.520478, retention index of 0.807270, and a composite index of 0.524071 for all sites and parsimony-informative sites, is shown in Fig. 1 . Nexus file of the sequence alignments for all data sets (ITS + ACT) were uploaded to TreeBASE http://purl.org/phylo/treebase/phylows/study/TB2:S23783 (study no. 29214). Taxonomy Cladosporium parasphaerospermum sp. nov ., Moharram AM, Zohri AA, Maher MA and Al-Bedak OA MycoBank: MB839546 Etymology Name refers to globose to subglobose conidia near to that of C. sphaerospermum . Holotype : Egypt, Beni Suef, Air, Maher MA, AUMC 10865. Ex-type culture: EMCCN: 2062 Macroscopic and microscopic characteristics Colonies on PDA reaching 18–20 mm diameter after 7 d at 28 ºC, raised at the center, radially sulcate, radially furrowed under the colony, olive, olive green at the center (2F6). Margin curled, paler than the colony center (3F8). Sporulation profuse. Exudates absent. Colonies on SNA reaching 17–19 mm in diameter after 7 d at 28 ºC, flat, slightly raised at the center, circular, olive, olive green (3F4-6). Margin entire. Sporulation abundant. Exudates absent. On OA colonies attaining 16–19 mm in diameter after 7 d at 28 ºC, circular, flat, somewhat lanuginose, dark olive, dark olive green (1F4-6). Margin entire. Sporulation abundant. Exudates absent. Mycelium abundantly formed, branched, 3–5 µm wide, septate, pale brown to brown, smooth. Conidiophores macronematous, micronematous, abundantly formed, arising terminally or laterally, more or less straight to flexuous, cylindrical, pale brown to brown, smooth, septate, commonly (–35) 75–100 × 3–5 µm (av. 87.5 × 4) µm (n = 50), not constricted at septa. Ramoconidia integrated, terminal, intercalary, cylindrical, smooth, thick-walled, 0–1 septate, 8–18 × 3–5 µm (av. 13 × 4) µm (n = 50), with 1–3 loci per cell. Loci usually confined to small lateral shoulders, protuberant, conspicuous, short cylindrical, 1–2 µm wide, up to 1–2 µm high. Conidia brown to dark brown, smooth, thick-walled, globose, subglobose, lemon-shaped, 0–septate, 4–6 × 3–5 µm (av. 5 × 4) µm (n = 50). Chlamydospores not formed (Fig. 2 ). Cladosporium chlamydosporigenum sp. nov ., Moharram AM, Zohri AA, Maher MA and Al-Bedak OA MycoBank: MB839547 Etymology refers to the formation of chlamydospores in culture. Holotype : Egypt, Sohag, Grapevine fruits, Maher MA, AUMC 11340. Ex-type culture: EMCCN: 2332. Macroscopic and microscopic characteristics Colonies on PDA reaching 15–17 mm diameter after 7 d at 28 ºC, raised at the center, wrinkled, irregular, olive, olive green (3F6-7). Margin undulate, narrow, paler than the center (3E5-6). Sporulation abundant. Exudates absent. On SNA colonies attaining 9–11 mm in diameter after 7 d at 28 ºC, flat, filamentous, olive, olive green (3E2). Margin filiform, narrow, (3E6-7). µm (av. 250 × 5) µm (n = 50), not constricted at septa. Ramoconidia integrated, terminal, intercalary, cylindrical, 11–22 × 6–8 µm (av. 16.5 × 7) µm (n = 50), verruculose to finely roughened, 0–2 septa with 1–3 loci per cell, Loci usually confined to small lateral shoulders, protuberant, conspicuous, short cylindrical, 1–2 µm wide, up to 1 µm high. Conidial chains unbranched or branched, conidia pale brown, straight, subglobose, obovoid to ellipsoid, verruculose to finely roughened, 0–septate, 4–11 × 5–7 µm (av. 7.5 × 6) µm (n = 50). Chlamydospores produced in hyphae, intercalary, aggregated, brown to dark brown, thick-walled, globose, subglobose, 15–25 × µm (Fig. 3 ). Cladosporium compactisporum sp. nov ., Moharram AM, Zohri AA, Maher MA and Al-Bedak OA MycoBank: MB839548 Etymology refers to the compact conidial chains. Holotype : Egypt, Qena, Air, Maher MA, AUMC 11366. Ex-type culture: EMCCN: 2358. Macroscopic and microscopic characteristics. Colonies on PDA attaining 25–28 mm after 7 d at 28 ºC, raised, umbonate, circular, olive to olive green (3E3-3F4). Margin entire, narrow, about 3.0 mm in width, paler than the colony center (3E1-3). Sporulation profuse. Exudates absent. Colonies on SNA attaining 17–20 mm diameter after 7 d at 28 ºC, raised, umbonate, olive to olive green (3F6-7). Margin entire, about 3.0 mm in width, paler than the colony center (3E2-3). Sporulation abundant. Exudates absent. Colonies on OA attaining 19–23 mm in diameter after 7 d at 28 ºC, raised, umbonate, lanuginose, olive grey (2E1-2). Margin undulate, narrow, dark olive grey (2F2). Sporulation abundant. Exudates lacking. Mycelium abundantly formed, branched, 3–5 µm wide, septate, swollen, pale brown to brown, smooth. Conidiophores macronematous and micronematous, abundantly formed, arising terminally or laterally, more or less straight to flexuous, nodulose, geniculate at the upper part, cylindrical, pale brown to brown, smooth, septate, branched, 100–300 µm × 3.0–6.0 µm (av. 200 × 4.5) µm (n = 50). Ramoconidia integrated, terminal, intercalary, cylindrical, 7–22 × 3–4 µm (av. 14.5 × 3.5) µm (n = 50), smooth, 0–1 septa with 1–3 loci per cell. Loci usually confined to small lateral shoulders, protuberant, conspicuous, short cylindrical, 1 µm wide, up to 1–2 µm high. Conidia formed in compact and branched chains, pale brown, subglobose, obovoid to ellipsoid, smooth, 0-septate, 4–6 × 3–4 µm (av. 5 × 3.5) µm (n = 50). Chlamydospores not formed (Fig. 4 ). Yield and Activity of Pectinases at pH and temperature The three fungi generated pectinases with a rather high output in submerged fermentation. Per liter of fermentation media, C. parasphaerospermum produced 3.0 g pectinase powder, C. chlamydosporum produced 3.65 g, and C. compactisporum produced 2.85 g. The pectinases appeared to be active; the greatest activity for the microbial pectinases was 1840, 893, and 930 U ml − 1 min − 1 at pH 6.0, 7.0, and 8.0, respectively. C. parasphaerospermum , C. chlamydosporum , and C. compactisporum had specific activity of 4221 ± 138 Ug − 1 , 1698 ± 63 Ug − 1 , and 2120 ± 15 Ug − 1 pectinase, respectively (Fig. 5 ). The optimum temperature was 35, 30, and 35 ºC, at pH 6.0, 7.0, and 8.0 for the three microbial pectinases produced by C. parasphaerospermum , C. chlamydosporum , and C. compactisporum , respectively. The specific activity values increased to 5758 ± 159 Ug − 1 , 3716 ± 3.8 Ug − 1 , and 3482 ± 50 Ug − 1 pectinase, respectively (Fig. 6 ). Effect of some ions and EDTA on pectinases activity The ions potassium and manganese exhibited a high activating influence (124.4% and 172%, respectively) on the activity of Cladosporium compactisporum pectinase. When evaluated under the optimal circumstances, EDTA and the metal ions showed varying inhibitory effects on the activity of the pectinases generated. In the case of C. parasphaerospermum , C. chlamydosporum and C. compactisporum pectinases, the highest inhibitory effects were with Ni, Zn, and EDTA, respectively (Table 1). Discussion Identification of novel species is at the heart of biodiversity research, and in recent years biodiversity efforts have been favouring DNA based method to morphology-based ones 31–35 . Understanding of the microbial composition aids awareness of host-microbe interactions and their environmental function, revealing a complex and delicate balance that can be easily upset 36–38 . Due to the complexity of fungal genomes and the lack of verified databases documenting appropriate biodiversity, such metagenomics studies in fungi have been limited. Identification of novel species is crucial in biodiversity research, which has lately adopted DNA-based methodologies. In this work, Cladosporium parasphaerospermum , C. chlamydosporigenum , and C. compactisporum were identified as new Cladosporium species based on phenotypic features of their cultures and sequencing of the ITS, ACT, and LSU. Morphologically, C. parasphaerospermum AUMC 10865 has conidiophores that are 75–100 µm shorter than C. cladosporioides (350 µm). It may also be recognized from C. cladosporioides 1 , C. halotolerans 39 , and C. parahalotolerans 39 by its smaller ramoconidia (8–18 µm), which measure 15–50 µm, 15–37 µm, and 24–37 µm, respectively. It may also be recognized from C. omanense 40 by its slower developing colonies on PDA (18–20 mm diam/7 days) compared to C. omanense 's 83 mm/14 days. Cladosporium omanense hyphae are in a polysaccharide-like substance and produce brief cross connections between hyphae, which C. parasphaerospermum lacks. Cladosporium chlamydosporigenum AUMC 11340 has smaller conidia (4–11 µm) than C. floccosum 39 and C. versiforme 41 , which have terminal (6–15 µm and 4–18µm) or intercalary (10–21 µm and 6–21 µm) conidia, respectively. It also has smaller ramoconidia (11–22 µm) with 0–2 septa than C. versiforme , whereas C. versiforme generates 45 µm ramoconidia with up to 5 septa. C. herbarum 1 has several nodulose swellings in quick succession, but C. allicinum 1 has head-like swellings with additional intercalary swellings, which C. chlamydosporigenum lacks. C. chlamydosporigenum may also be distinguished from C. floccosum , C. herbarum , C. allicinum , and C. macrocarpum by the presence of chlamydospores. Cladosporium compactisporum AUMC 11366 contains smaller ramoconidia (7–22 µm) with 1–3 loci compared to C. cladosporioides (15–50 µm) with up to 4 loci packed at the apex, and C. vicinum 39 which measures 20–60 (-70) µm with up to 7 loci packed at the apex plus 1–2 loci. Cladosporium compactisporum differs from C. verrucocladosporioides 1 and C. delicatulum 39 by having nodulose and smooth conidiophores, as well as smaller ramoconidia (7–22 µm) compared to non-nodulose conidiophores and larger ramoconidia of C. verrucocladosporioides . Cladosporium uredinicola 1 possesses branching, aggregated hyphae that create thick hyphal nets, but C. compactisporum does not. The three new strains, Cladosporium parasphaerospermum , C. chlamydosporum , and C. compactisporum were used in this study to produce pectinases with high specific activities at low temperatures (10 o C) in SmF. Because of their biodegradability, non-toxicity, high selectivity, and high yields, microbial enzymes are superior to chemical synthesis (Bruno et al. 2019). The global enzyme market was valued at $ 9.9 billion in 2019 and is expected to grow at a 7.1% annual rate from 2020 to 2027 42 . Pectinase has lengthy been used in commercial food processing to degrade pectin and aid in various processing steps such as liquefaction, clarification, and juice extraction 43 . Pectinases are among the most widely used enzymes, accounting for 40% of all food enzymes 44,45 . Several Cladosporium species have been shown to produce active pectinases as well 9,25,26,46−48 . Due to minor changes in methodology, it is difficult to compare the values of enzyme activity between different researches. As a result, comparisons should be made with caution. The three novel strains could create active pectinases at pH 6.0, 7.0, and 8.0, respectively, while the optimum temperatures for C. parasphaerospermum , C. chlamydosporum , and C. compactisporum were 35, 30, and 35 o C, respectively. The majority of commercial enzymes, including pectinase, are now mesophilic or thermophilic. In the food sector, and particularly in the fruit processing sector, there has been an increasing desire to replace high-temperature procedures with low-temperature processes. Specific economic and environmental benefits, such as energy savings, retention of biologically inert and aromatic fragrance components, contamination mitigation, and eradication of any residual enzyme activity, which is deactivation of enzyme when temperature is raised, are driving this shift in trend 42,43,49−51 . Conclusion In the current study, three novel Cladosporium species were introduced and described as C. parasphaerospermum , C. chlamydosporum , and C. compactisporum . The three novel species appeared to produce mesophilic pectinases that had high activity at pH 6.0 and 35 ºC, pH 7.0 and 30 ºC, and pH 8.0 and 35 ºC, respectively, of which C. parasphaerospermum was the most active. We may now add the generation of active pectinases to the vast list of Cladosporium species that have been documented. We also describe three novel species that can be exploited as active microbial pectinase producers in biotechnological solutions. Although further research is needed, these unique species might be employed to degrade troublesome and resistant pertinacious wastes as well as clear fruit juices. Materials And Methods Isolation and Maintenance of Cladosporium Strains Three Cladosporium isolates involved in the current study, of which two were isolated from air of Beni Suef and Qena cities and one from grapevine fruits in Sohag city, Egypt. Exposure method 52 was employed for isolation of Cladosporium from air and direct plating technique 53 for isolation from grapevine fruits. Czapek’s Dox agar was used as an isolation medium. The isolation medium composed of (g/L): Sucrose, 30, Na 2 NO 3 , 2, K 2 HPO 4 , 1, KCl, 0.5, MgSO 4 .7H 2 O, 0.5, FeSO 4 , 0.01, ZnSO 4 , 0.01, CuSO 4 , 0.005, Rose Bengal, 0.05, chloramphenicol, 0.25, agar, 15 and the final pH 7.3. The interesting isolates obtained were preserved as frozen and lyophilized cultures in the culture collection of Assiut University Mycological Centre (AUMC) and in the Egyptian Microbial Culture Collection Network (EMCCN) as AUMC 10865 = EMCCN 2062 (air, Beni Suef), AUMC 11340 = EMCCN 2332 (grapevine fruits, Sohag) and AUMC 11366 = EMCCN 2358 (air, Qena). The novel species were registered at the MycoBank with their description with accession numbers of MB839546, MB839547 and MB839548, respectively. Morphological studies of the Cladosporium strains Spore suspension (prepared in a 30 % glycerol, 0.2 % agar and 0.05 % Tween 80 solution) of the three Cladosporium was inoculated in a three-point pattern using inoculum size of 1.0 μl/spot on potato dextrose agar (PDA, 54 46 54 54 54 54 54 33 54 [54] (Smith & Onions, 1994) (54) [54] [54] [33] [33] (Smith and Onions 1994) (Smith and Onions 1994) [33], synthetic nutrient agar (SNA) and Oat meal agar (OA) as described by 54,55 . Morphological characteristics and growth rates on SNA were examined in lacto-phenol cotton blue. Colony colors were identified according to 56 . Molecular identification of the Cladosporium strains DNA extraction For DNA isolation, a small portion from fungal growth of 7-day-old colonies of Cladosporium isolates AUMC 10865, AUMC 11340 and AUMC 11366 grown on PDA at 25 °C were collected and transferred individually to 2 ml-Eppendorf tube. The DNA extraction was performed following CTAB method described in Moubasher, et al. 57 . PCR and sequencing of ITS, ACT and LSU The universal primers ITS1 and ITS4 58 were used for amplification of the internal transcribed spacer (ITS) region, LROR and LR7 primers 59 for amplification of the large subunit (LSU) and ACT512F and ACT783R for amplification of ACT gene 60 . PCR was done following Al-Bedak and Moubasher 61 . Alignments and phylogenetic analyses Sequences of Cladosporium species (ITS, ACT, LSU) in this study were compared to sequences of the type and ex-type species in GenBank (Table 2). The ITS and ACT sequences of the strains in this study and those downloaded from GenBank were concatenated using Sequence Matrix version 1.8 62 . The produced concatenated sequences were aligned together using MAFFT (version 6.861b) with the default options 63 , and Cercospora beticola CBS 116456 was used as the outgroup. Alignment gaps and parsimony uninformative characters were treated by BMGE 64 . Maximum-likelihood (ML) and Maximum parsimony (MP) phylogenetic analyses were performed using PhyML 3.0 65 . The robustness of the most parsimonious trees was evaluated by 1000 replications 66 . The best optimal model of nucleotide substitution for the ML analyses was determined using Smart Model Selection (SMS) version 1.8.1 67 . The phylogenetic tree was drawn and visualized using MEGA X 10.2.6 68 . The resulting tree was edited using Microsoft Power Point (2016) and saved as TIF file 9 . Production of Pectinases in Submerged Fermentation (SmF) Previously, the three Cladosporium species AUMC (10865, 11340 and 11366) were discovered to be capable of producing large quantities of pectinases in SmF at 10 ºC reaching 8.0 U/mg, 1.78 U/mg and 7.52 U/mg, respectively 27 . For pectinases production by the three strains, Cladosporium species were employed in Erlenmeyer flasks (500 ml) to produce pectinases in SmF at 10 o C using sucrose-free Czapek's (initial pH 7.0) supplemented with 1.0 % pectin (as only carbon source). Cladosporium species were inoculated with 1.5 x 10 8 spore/ml spore suspensions obtained from 7-day-old cultures. The incubation period lasted 7 days at 10 o C and 150 rpm. Following the incubation time, cell-free supernatants were recovered by centrifuging at 10,000 rpm for 10 min. At 4 o C, total protein was isolated using 70 % saturation of ammonium sulphate. A freeze dryer (VirTis, model #6KBTES-55, NY, USA) was used to separate and lyophilize the precipitated total protein. Lyophilized protein was dissolved in citrate buffer (pH 5.0) and dialyzed twice for 2 hours at room temperature (cutoffs: 12-14 KD) against the same buffer, eliminating the buffer each time, before being refrigerated overnight at 4 ºC to remove small molecules. The dialyzed protein was then lyophilized, weighed, and used in enzyme characterization experiments as a partly pure fungal pectinases enzymes. Impact of pH, Temperature and Some Ions and Inhibitors on Pectinases Activities A 0.01 g enzyme powder and 0.01 g citrus peel pectin (each dissolved in 1.0 ml of 50 mmol buffer solution) were included in this test in a water bath. The impact of pH (3.0–10.0) and temperature (30–60 ºC) on pectinases activity were investigated. After the reaction time (20 min), the reaction was terminated by introducing 2.0 ml of 3,5-dinitrosalicylic acid (DNS) according to Miller 69 and Jayani, et al. 70 , and the pectinase activity was determined as the amount of the enzyme that releases 1 µmol ml -1 min -1 galacturonic acid under standard assay conditions. The buffers used were citrate buffer (pH 3.0–6.0), phosphate buffer (pH 7.0–8.0), and borate buffer (pH 9.0–10.0). Also, some ions (Na + , K + , Ca +2 , Co +2 , Ni +2 , Cu +2 , Fe +2 , Mg +2 , Mn +2 , and Zn +2 ) were evaluated by introducing them at 5 mmol/ml concentrations as NaCl, KCl, CaCl 2 , CoCl 2 , NiSO 4 , CuSO 4 , FeSO 4 , MgSO 4 , MnSO 4 , and ZnSO 4 . 5 mmol/ml ethylenediaminetetraacetic acid was used to evaluate an enzyme inhibitor (EDTA). Under standard conditions, the activity of the microbial pectinases in the absence of metal ions or EDTA was evaluated to define 100 % activity. All experiments were conducted in three repetitions. Accessibility of data: Pure cultures of the type materials of the novel species are deposited in the culture collection of Assiut University Mycological Centre as AUMC 10865, AUMC 11340 and AUMC 11366, and in the Egyptian Microbial Culture Collection Network as EMCCN 2062, EMCCN 2332 and EMCCN 2358. Nexus file of the sequence alignments for all data sets were uploaded to TreeBASE http://purl.org/phylo/treebase/phylows/study/TB2:S23783 (study no. 29214). Declarations Author Contributions: A.M.M.: Supervision of the study, writing, revising, A-N.A.Z.: Supervision of the study, writing, revising, M.A.M.: Fungal isolation, enzymatic production, A.E.H.: Editing, revising, O.AA.-B.: Molecular work, data analysis, writing, revising. All authors contributed to data analysis, drafting, or revising the manuscript. All authors have read and agreed to the published version of the manuscript. Data Availability Statement: “The datasets generated and/or analysed during the current study are available in the GenBank repository, [https://www.ncbi.nlm.nih.gov/genbank/]”. Conflicts of Interest: The authors declare that there are no potential conflict of interest regarding the publication of this paper. Data Availability: All datasets generated or analyzed during this study are included in the manuscript. References Bensch, K., Braun, U., Groenewald, J. Z. & Crous, P. W. The genus cladosporium . Studies in mycology 72 , 1–401, https://doi.org/10.3114/sim0003 (2012). Ellis, M. More dematiaceous hyphomycetes (p. 507). Kew, Surrey, England: Commonwealth Mycological Institute (1976). Yehia, R. S., Osman, G. H., Assaggaf, H., Salem, R. & Mohamed, M. S. Isolation of potential antimicrobial metabolites from endophytic fungus Cladosporium cladosporioides from endemic plant Zygophyllum mandavillei . South African Journal of Botany 134 , 296–302, https://doi.org/10.1016/j.sajb.2020.02.033 (2020). Moubasher, A. Soil fungi in Qatar and other Arab countries. The Centre for Scientific and Applied Research, University of Qatar (1993). Ismail, M., Moubasher, A., Mohamed, R. & Al-Bedak, O. Extremophilic fungi and chemical analysis of hypersaline, alkaline lakes of Wadi-El-Natrun, Egypt. International Journal of Technical Research and Science 1 , 345–363 (2017). Farr, D. F., Bills, G. F., Chamuris, G. P. & Rossman, A. Y. Fungi on plants and plant products in the United States. APS press (1989). Haas, C. N. Microorganisms in home and indoor work environments: diversity, health impacts, investigation and control edited by Brian Flannigan, Robert A. Samson, and J. David Miller. Risk Analysis 31 , 1327–1327, DOI: 10.1111/j.1539-6924.2011.01665.x (2011). Flannigan, B., Samson, R. A. & Miller, J. D. Microorganisms in home and indoor work environments: diversity, health impacts, investigation and control. CRC Press (2016). Al-Bedak, O. A., Teama, E.A.E., Ali, E.A., Said, M.T., Shalaby, E.M. & Moharram, Z.A.M. Impact of fumigation with phosphine on viability of wheat grains stored for six months at two levels of moisture content, in addition to description of four new records of associated fungi and assessment of their potential for enzymatic production. Journal of Basic & Applied Mycology (Egypt) 11 , 77–97 (2020). Schubert, K. & Braun, U. Taxonomic revision of the genus Cladosporium s. lat. 2. Cladosporium species occurring on hosts of the families Bignoniaceae and Orchidaceae. Sydowia 56 , 296–317 (2004). Heuchert, B., Braun, U. & Schubert, K. Morphotaxonomic revision of fungicolous Cladosporium species (hyphomycetes). Schlechtendalia 13 , 1–78 (2005). Riesen, T. & Sieber, T. Endophytic fungi in winter wheat ( Titicum aestivum L.). (Zurich (Switzerland), Swiss Federal Institute of Technology , 190 p. (1985). Brown, K. Preliminary studies on endophytic fungal communities of Musa acuminata species complex in Hong Kong and Australia. Fung. Divers. 1 , 27–51 (1998). El-Morsy, E. Fungi isolated from the endorhizosphere of halophytic plants from the Red Sea Coast of Egypt. Fungal Diversity 5 , 43–54 (2000). Islam, M. & Hasin, F. Studies on the phylloplane mycoflora of Amaranthus viridis . L. National Academy Science letters 23 , 121–123 (2000). Inácio, J. et al. Estimation and diversity of phylloplane mycobiota on selected plants in a mediterranean–type ecosystem in Portugal. Microbial Ecology 44 , 344–353 (2002). Stohr, S. & Dighton, J. Effects of species diversity on establishment and coexistence: A phylloplane fungal community model system. Microbial Ecology 48 , 431–438 (2004). Levetin, E. & Dorsey, K. Contribution of leaf surface fungi to the air spora. Aerobiologia 22 , 3–12 (2006). Abdel-Baky, N., Arafat, N. S. & Abdel-Salam, A. Three Cladosporium spp. as promising biological control candidates for controlling whiteflies ( Bemisia spp.) in Egypt. Pak. J. Biol. Sci 1 , 188–195 (1998). Abdel-Baky, N. & Abdel-Salam, A. Natural incidence of Cladosporium spp. as a bio-control agent against whiteflies and aphids in Egypt. (2003). Salvatore, M. M., Andolfi, A. & Nicoletti, R. The Genus Cladosporium : A Rich Source of Diverse and Bioactive Natural Compounds. Molecules 26 , 3959 (2021). Hamed, M., Osman, A. A. & Ateş, M. Statistical optimization of L-asparaginase production by Cladosporium tenuissimum . Egyptian Pharmaceutical Journal 20 , 51 (2021). El-Sabbagh, S. M., Eissa, O. A. E. & Sallam, M. H. E. Taxol production by an endophytic fungus Cladosporium cladosporioides isolated from Catheranthus roseus . Vázquez-Montoya, E. L., Castro-Ochoa, L. D., Maldonado-Mendoza, I. E., Luna-Suárez, S. & Castro-Martínez, C. Moringa straw as cellulase production inducer and cellulolytic fungi source. Revista Argentina de microbiologia 52 , 4–12 (2020). Bastos, S. C. et al. Pectinases from a new strain of Cladosporium cladosporioides (Fres.) De Vries isolated from coffee bean. World J Agric Sci 9 , 167–172 (2013). Al-Bedak, O. A., Abdel-Sater, M. A., Abdel-Latif, A. M. & Abdel-Wahab, D. A. Endophytic mycobiota and enzymatic capacity in wild and cultivated plants in New Valley, Egypt: A comparative analysis. Journal of Basic & Applied Mycology (Egypt) 12 , 27–41 (2021). Moharram, A., Zohri, A., Hesham, A., Maher, M. & Al-Bedak, O. Production of Cocktail Enzymes by Three Cladosporium Isolates and Bioconversion of Orange Peel Wastes into Valuable Enzymes. J Pure Appl Microbiol 15 , 2336–2346 (2021). Amin, F., Bhatti, H. N. & Bilal, M. Recent advances in the production strategies of microbial pectinases—A review. International journal of biological macromolecules 122 , 1017–1026 (2019). Saranraj, P. & Naidu, M. Microbial pectinases: a review. Global J Trad Med Syst 3 , 1–9 (2014). Salikin, N. H. & Makhtar, M. M. Z. in Waste Management, Processing and Valorisation 175–206 (Springer, 2022). Hebert, P. D., Cywinska, A., Ball, S. L. & Dewaard, J. R. Biological identifications through DNA barcodes. Proceedings of the Royal Society of London. Series B: Biological Sciences 270 , 313–321 (2003). De Queiroz, K. Species concepts and species delimitation. Systematic biology 56 , 879–886 (2007). Verkley, G., Quaedvlieg, W., Shin, H.-D. & Crous, P. A new approach to species delimitation in Septoria. Studies in Mycology 75 , 213–305 (2013). Woudenberg, J., Groenewald, J., Binder, M. & Crous, P. Alternaria redefined. Studies in mycology 75 , 171–212 (2013). Liu, X.-Z. et al. Phylogeny of tremellomycetous yeasts and related dimorphic and filamentous basidiomycetes reconstructed from multiple gene sequence analyses. Studies in mycology 81 , 1–26 (2015). Fuhrman, J. A. Microbial community structure and its functional implications. Nature 459 , 193–199 (2009). Garza, D. R., Van Verk, M. C., Huynen, M. A. & Dutilh, B. E. Bottom-up ecology of the human microbiome: from metagenomes to metabolomes. BioRxiv , 060673 (2016). Levy, M., Blacher, E. & Elinav, E. Microbiome, metabolites and host immunity. Current opinion in microbiology 35 , 8–15 (2017). Bensch, K. et al. Cladosporium species in indoor environments. Studies in mycology 89 , 177–301 (2018). Halo, B., Maharachchikumbura, S. S., Al-Yahyai, R. A. & Al-Sadi, A. M. Cladosporium omanense , a new endophytic species from Zygophyllum coccineum in Oman. Phytotaxa 388 , 145–154 (2019). Bensch, K. et al. Common but different: The expanding realm of Cladosporium . Studies in Mycology 82 , 23–74 (2015). Kumar, A., Mukhia, S. & Kumar, R. Industrial applications of cold-adapted enzymes: challenges, innovations and future perspective. 3 Biotech 11 , 1–18 (2021). Adapa, V., Ramya, L., Pulicherla, K. & Rao, K. S. Cold active pectinases: advancing the food industry to the next generation. Applied biochemistry and biotechnology 172 , 2324–2337 (2014). Pedrolli, D. B., Monteiro, A. C., Gomes, E. & Carmona, E. C. Pectin and pectinases: production, characterization and industrial application of microbial pectinolytic enzymes. Open Biotechnology Journal, 9–18 (2009). Semenova, M. et al. Use of a preparation from fungal pectin lyase in the food industry. Applied Biochemistry and Microbiology 42 , 598–602 (2006). Poveda, G., Gil-Durán, C., Vaca, I., Levicán, G. & Chávez, R. Cold-active pectinolytic activity produced by filamentous fungi associated with Antarctic marine sponges. Biological research 51 (2018). Ming, X., Huai, Q., Huamei, W., Zhuyan, L. & Jian, Z. On kinetics of batch fermentation for pectinase from mutant of Cladosporium herbarum (pers) link ex gray. Guizhou science, 02 (1997). Bastos, S. C. et al. in Microorganisms In Industry And Environment: From Scientific and Industrial Research to Consumer Products 538–542 (World Scientific, 2011). Van Truong, L., Tuyen, H., Helmke, E., Binh, L. T. & Schweder, T. Cloning of two pectate lyase genes from the marine Antarctic bacterium Pseudoalteromonas haloplanktis strain ANT/505 and characterization of the enzymes. Extremophiles 5 , 35–44 (2001). Pulicherla, K., Ghosh, M., Kumar, P. & Rao, K. Psychrozymes–the next generation industrial enzymes. J Marine Sci Res Development 1 (2011). Mangiagalli, M., Brocca, S., Orlando, M. & Lotti, M. The “cold revolution”. Present and future applications of cold-active enzymes and ice-binding proteins. New biotechnology 55 , 5–11 (2020). Flannigan, B. Air sampling for fungi in indoor environments. Journal of Aerosol Science 28 , 381–392 (1997). Berry, E. D. & Wells, J. E. A direct plating method for estimating populations of Escherichia coli O157 in bovine manure and manure-based materials. Journal of food protection 71 , 2233–2238 (2008). Smith, D. & Onions, A. H. The preservation and maintenance of living fungi. (CAB international, 1994). Gerlach, W., Nirenberg, H., Eckart, I., Rummland, I. & Schwarz, R. The genus Fusarium: a pictorial atlas. Vol. 209 (Kommissionsverlag P. Parey Berlin, 1982). Kornerup, A. & Wanscher, J. (1978). Moubasher, A., Ismail, M., Al-Bedak, O. & Mohamed, R. Ramophialophora chlamydospora , a new species from an alkaline lake of Wadi-El-Natron, Egypt. Asian Journal of Mycology 2 , 110–117 (2019). White, T. J., Bruns, T., Lee, S. & Taylor, J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications 18 , 315–322 (1990). Rehner, S. A. & Samuels, G. J. Taxonomy and phylogeny of Gliocladium analysed from nuclear large subunit ribosomal DNA sequences. Mycological Research 98 , 625–634 (1994). Carbone, I. & Kohn, L. M. A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 91 , 553–556 (1999). Al-Bedak, O. & Moubasher, A. Aspergillus gaarensis , a new addition to section Circumdati from soil of Lake El-Gaar in Wadi-El-Natron, Egypt. Studies in Fungi 5 , 59–65 (2020). Vaidya, G., Lohman, D. J. & Meier, R. SequenceMatrix: concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics 27 , 171–180 (2011). Katoh, K. & Standley, D. M. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular biology and evolution 30 , 772–780 (2013). Criscuolo, A. & Gribaldo, S. BMGE (Block Mapping and Gathering with Entropy): a new software for selection of phylogenetic informative regions from multiple sequence alignments. BMC evolutionary biology 10 , 210 (2010). Guindon, S. et al. New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Systematic biology 59 , 307–321 (2010). Felsenstein, J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39 , 783–791 (1985). Lefort, V., Longueville, J.-E. & Gascuel, O. SMS: smart model selection in PhyML. Molecular biology and evolution 34 , 2422–2424 (2017). Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular biology and evolution 35 , 1547–1549 (2018). Miller, G. L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical chemistry 31 , 426–428 (1959). Jayani, R. S., Saxena, S. & Gupta, R. Microbial pectinolytic enzymes: a review. Process Biochemistry 40 , 2931–2944 (2005). Tables Table 1. Effect of metal ions and EDTA (5 mmol mL - 1 ) on pectinase activity produced by C. parasphaerospermum , C. chlamydosporum , and C. compactisporum (mean ± SD, n = 3). The results are expressed as the activity in the tested inhibitory conditions compared to the pectinase activity in the control without inhibitors (in bold). Metal ions and inhibitors C. parasphaerospermum C. chlamydosporum C. compactisporum Specific activity U g -1 Specific activity U g -1 Specific activity U g -1 Control 5758±159 3716±3.8 3482±50 Na + 4027±22 3600±40 2025±18 K + 3272±24 3086±26 4287±85 Fe +2 3068±71 759±55 1865±15 Cu +2 3406±49 2851±38 1336±14 Ca +2 3875±52 2734±15 3379±67 Mg +2 2231±45 1682±15 3215±33 Zn +2 4185±30 913±15 2182±26 Ni +2 2106±19 2954±31 3432±30 Co +2 3473±106 1553±31 2390±36 Mn +2 2955±26 1553±29 5992±51 EDTA 2383±15 2165±100 1253±22 Table 2. List of specimens and GenBank accession numbers of sequences used in this study. Bold accession numbers were generated from this study. Cladosporium species Strain Locality ITS ACT LSU C. parasphaerospermum AUMC 10865 Egypt MN826828 OL514008 MW205008 C. chlamydosporigenum AUMC 11340 Egypt MN826919 OL514009 MW205067 C. compactisporum AUMC 11366 Egypt MN826822 OL514010 MW205086 C. aerium DTO:323-B4 China MF472897 MF473747 - DTO:323-G7 China MF472899 MF473749 - DTO:323-G6 China MF472898 MF473748 - C. allicinum CBS 121624 Belgium MH863126 EF679502 MH874678 DTO 111-A5 Denmark KP701924 KP702047 - DTO 249-G3 Netherlands KP701975 KP702097 - C. antarcticum CBS 690.92 Antarctica NR_121332 EF679484 - C. cladosporioides CBS 112388 Germany NR_119839 HM148490 - CPC 15167 Slovenia HM148052 HM148539 - GZYQ-08-01 China MK852271 MK852272 - C. colombiae CBS 274.80B Colombia MH861262 FJ936166 - C. cycadicola CBS 137970 Australia NR_156279 KJ869227 NG_058881 C. delicatulum DTO 145-C4 Germany KP701940 KP702062 - CPC 14372 Denmark HM148089 HM148578 - CPC 14363 Denmark HM148088 HM148577 - C. domesticum CPC 22307 USA NR_156348 MF473805 - DTO 308-B1 USA MF472966 MF473816 - CPC:22413 USA MF472961 MF473811 - C. exasperatum CBS 125986 Australia MH863865 HM148579 MH875326 C. floccosum DTO 323-H6 China MF472979 MF473829 - CPC 22968 USA MF472978 MF473828 - CPC 22399 USA MF472977 MF473827 - C. halotolerans CBS 119416 USA NR_119605 EF101397 - DTO 257-F4 Netherlands KP701989 KP702111 - C. herbaroides CBS 121626 Israel NR_119655 EF679509 - C. herbarum CBS 289.49 Switzerland MH856530 EF679558 MH868058 CPC 12183 Netherlands EF679368 EF679521 - C. iridis CBS 107.20 Netherlands MH854682 EF679522 MH866199 CBS 138.40 Netherlands NR_111271 EF679523 - C. kenpeggii CPC 19248 Australia KY646222 KY646225 - C. macrocarpum CPC 12759 USA EF679380 EF679534 - CPC 14305 Denmark MF473141 MF473990 - C. ossifragi CBS 842.91 Norway MH862342 EF679535 - C. parahalotolerans DTO 307-H4 Mexico MF473161 MF474009 - DTO 324-B7 China MF473169 MF474017 - C. perangustum CBS 167.54 Netherlands HM148124 HM148613 - CBS 126365 USA MH863940 HM148612 MH875401 C. sphaerospermum CBS 193.54 Slovenia NR_111222 EU570269 - DTO 255-H7 Netherlands KP701988 KP702110 - C. spinulosum CBS 119907 Slovenia NR_119660 EF679542 - C. subcinereum UTHSC DI-13-257 USA NR_148193 LN834617 - C. subinflatum CBS 121630 Slovenia MH863129 EF679543 MH874681 C. tenuissimum CBS 125995 USA MH864840 HM148687 MH876286 C. tuberosum UTHSC DI-13-217 Spain LN834417 LN834601 MH878168 C. uredinicola CPC 5390 USA AY251071 HM148712 EU019264 C. variabile CBS 121635 USA MH863131 EF679557 MH874683 C. verrucocladosporioides CBS 126363 South Korea MH863939 HM148717 - C. versiforme CBS 140491 Netherlands NR_152297 KT600613 - C. vicinum CPC 22316 USA MF473311 MF474161 - CPC 15457 New Zealand HM148060 HM148547 - C. wyomingense CPC 22310 USA MF473315 MF474165 - Cercospora beticola CBS 116456 USA NR_121315 AY840458 NG_068999 Abbreviations used: AUMC = Assiut University Mycological Centre, Assiut University, Assiut, Egypt, CBS = Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, CPC = Culture Collection of Pedro Crous, housed at CBS, Utrecht, The Netherlands, DTO = Culture Collection of Jos Houbraken, UTHSC = The University of Tennessee Health Science Center, Monroe Avenue, Memphis, Tennessee, USA. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-1291889","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":84358114,"identity":"ae6a5edb-581a-4f42-9c09-6250987223da","order_by":0,"name":"Ahmed Mohamed Moharram","email":"","orcid":"","institution":"Assiut University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ahmed","middleName":"Mohamed","lastName":"Moharram","suffix":""},{"id":84358116,"identity":"f1f3c61b-036b-4ff6-b1c1-7d5e400e47b0","order_by":1,"name":"Abdel-Naser Ahmed Zohri","email":"","orcid":"","institution":"Assiut University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Abdel-Naser","middleName":"Ahmed","lastName":"Zohri","suffix":""},{"id":84358117,"identity":"c55897c8-7d6c-4f64-9900-f0f46159a0dd","order_by":2,"name":"Abd El-Latif Hesham","email":"data:image/png;base64,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","orcid":"","institution":"Beni-Suef University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Abd","middleName":"El-Latif","lastName":"Hesham","suffix":""},{"id":84358118,"identity":"b014703f-157d-48a2-8ec6-cfe2ebe5632a","order_by":3,"name":"Mohamed Al-Amin Maher","email":"","orcid":"","institution":"Assiut University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"Al-Amin","lastName":"Maher","suffix":""},{"id":84358119,"identity":"42f6bdbe-ea0f-4d93-9a23-4cb3a665e5ed","order_by":4,"name":"Osama Abdel-Hafeez Al-Bedak","email":"","orcid":"","institution":"Assiut University Mycological Centre, Assiut University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Osama","middleName":"Abdel-Hafeez","lastName":"Al-Bedak","suffix":""}],"badges":[],"createdAt":"2022-01-24 13:14:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1291889/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1291889/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":18373186,"identity":"b1eb233f-fdee-4d46-81ad-def12e60d494","added_by":"auto","created_at":"2022-02-18 17:12:32","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":428886,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum likelihood tree based on concatenated ITS and ACT sequences of \u003cem\u003eCladosporium parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporigenum \u003c/em\u003eand \u003cem\u003eC. compactisporum \u003c/em\u003ein this study (in bold blue color) with the most similar sequences belonging to \u003cem\u003eCladosporium \u003c/em\u003ein GenBank. Bootstrap support values (1000 replications) for ML/MP ≥50% are indicated near the respective nodes. The tree was rooted to \u003cem\u003eCercospora beticola \u003c/em\u003eCBS 116456 as outgroup (in bold red color).\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-1291889/v1/66f9ad7d492d8c7fad2aba1d.png"},{"id":18373346,"identity":"7d865153-c84b-4743-b06a-b52ad613240d","added_by":"auto","created_at":"2022-02-18 17:15:32","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2351195,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eCladosporium parasphaerospermum \u003c/em\u003e(AUMC 10865). A-C.7-day-old colonies on PDA, SNA and OA at 25 ºC. D-F. Macronematous conidiophores and conidial chains. G. Globose to subglobose conidia. Scale bar = 20 µm (D), 10 µm (E-G).\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-1291889/v1/03a9ab9a95e68edf0f1dfbac.png"},{"id":18373183,"identity":"9a295476-4540-4cb2-86f4-8637518f8397","added_by":"auto","created_at":"2022-02-18 17:12:32","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":3809867,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eCladosporium chlamydosporigenum \u003c/em\u003e(AUMC 11340). A-C.7-day-old colonies on PDA, SNA and OA at 25 ºC. D. Mycelium with abundant chlamydospores. E-H. Macronematous conidiophores and conidial chains. I. Aggregated chlamydospores. Scale bar = 20 µm.\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-1291889/v1/786283ab29d1643a44939cd3.png"},{"id":18373184,"identity":"6212fae9-4bd1-4222-955c-b721e8885db7","added_by":"auto","created_at":"2022-02-18 17:12:32","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":2529581,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eCladosporium compactisporum \u003c/em\u003e(AUMC 11366). A-C. 7-day-old colonies on PDA, SNA and OA at 25 ºC. D. Geniculate conidiophore. E. Swollen conidiophore. F. Compact chains of conidia. Scale bar = 20 µm.\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-1291889/v1/093c609bc396e57cfbae231e.png"},{"id":18373181,"identity":"f067e64c-774d-46ac-9cef-4a113ce2ba86","added_by":"auto","created_at":"2022-02-18 17:12:32","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":57355,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of pH on the activity of pectinases produced by \u003cem\u003eCladosporium \u003c/em\u003estrains.\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-1291889/v1/cdf1c079b59331823af5a84b.png"},{"id":18373345,"identity":"3bd31d26-02ce-401f-bd47-1323c3642cd1","added_by":"auto","created_at":"2022-02-18 17:15:32","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":65695,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of temperature on the activity of pectinases produced by \u003cem\u003eCladosporium \u003c/em\u003estrains\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-1291889/v1/28f4e7551bac816f2477f7f1.png"},{"id":18373347,"identity":"d4622b38-6d35-4002-82b7-ed3b10f9f978","added_by":"auto","created_at":"2022-02-18 17:15:35","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":676439,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1291889/v1/e6a72958-e437-485f-8ec9-2fbe272edeee.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Production and Partial Purification of Pectinases Produced by Three Novel Cladosporium Species Isolated from Egypt","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eCladosporium\u003c/em\u003e is one of the largest and most complex genera of hyphomycetes, which currently includes more than 728 names. Until recently, all types of unrelated dematiaceous hyphomycetes characterized by amero-to-phragmosporous conidia formed in acropetal chains had been referred to \u003cem\u003eCladosporium\u003c/em\u003e\u003csup\u003e1\u003c/sup\u003e. Species of \u003cem\u003eCladosporium\u003c/em\u003e are well adapted to be spread easily in large numbers over long distances, therefore they are cosmopolitan and widely present in all various types of plants and other debris, mostly isolated from air, soil, seeds, grains, food, paint, textiles and other organic matter\u003csup\u003e1\u0026ndash;9\u003c/sup\u003e.Other species of this genus are plant pathogenic causing leaf spots and other lesions\u003csup\u003e10\u003c/sup\u003e, or they occur as hyperparasites on other fungi\u003csup\u003e11\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003e \u003cem\u003eCladosporium\u003c/em\u003e species are also known to be common endophytes\u003csup\u003e12\u0026ndash;14\u003c/sup\u003e as well as phylloplane fungi\u003csup\u003e15\u0026ndash;18\u003c/sup\u003e. Some \u003cem\u003eCladosporium\u003c/em\u003e species including \u003cem\u003eC\u003c/em\u003e. \u003cem\u003ecladosporioides\u003c/em\u003e, \u003cem\u003eC\u003c/em\u003e. \u003cem\u003echlorocephalum\u003c/em\u003e and \u003cem\u003eC\u003c/em\u003e. \u003cem\u003euredinicola\u003c/em\u003e were recorded as entomopathogens of Aphids and whiteflies\u003csup\u003e19,20\u003c/sup\u003e. The Genus \u003cem\u003eCladosporium\u003c/em\u003e is considered a rich source of diverse and bioactive natural compounds\u003csup\u003e21\u003c/sup\u003e. Some species were reported to produce anticancer compounds such as L-asparaginase\u003csup\u003e22\u003c/sup\u003e, paclitaxel\u003csup\u003e23\u003c/sup\u003e, as well as enzymes like cellulases\u003csup\u003e24\u003c/sup\u003e, and pectinases\u003csup\u003e25\u0026ndash;27\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003ePectinases are enzymes that target pectin and depolymerize it by hydrolysis, transelimination, and deesterification processes, which hydrolyze the ester connection between pectin's carboxyl and methyl groups. These enzymes degrade pectin, a complex polysaccharide present in the cell walls of higher plants that serves as a cementing substance for the cellulose network. Pectinases account for 10% of all industrial enzymes manufactured globally, and their market is growing by the day\u003csup\u003e28\u0026ndash;30\u003c/sup\u003e. As a result, the current study attempted to identify some unique \u003cem\u003eCladosporium\u003c/em\u003e species isolated from air and fruits in some Egyptian governorates, as well as to produce and partially purify pectinases in submerged fermentation.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMolecular studies\u003c/h2\u003e \u003cp\u003eThe ITS of \u003cem\u003eCladosporium\u003c/em\u003e strain AUMC 10865 was 99.64% (554/556) similar to \u003cem\u003eC\u003c/em\u003e. \u003cem\u003ecladosporioides\u003c/em\u003e (KJ767065), \u003cem\u003eC. parahalotolerans\u003c/em\u003e (MK262909) and \u003cem\u003eC. halotolerans\u003c/em\u003e (MK258720). ACT sequence was 96.21% (127/132) similar to \u003cem\u003eC. halotolerans\u003c/em\u003e (MF084398 and MF084399) and \u003cem\u003eC. omanense\u003c/em\u003e (MH716046). LSU sequence was 98.96% (1140/1152) similar to \u003cem\u003eC. delicatulum\u003c/em\u003e (JQ732984 and JQ732983). ITS of \u003cem\u003eCladosporium\u003c/em\u003e strain AUMC 11340 was 99.27% (549/553) similar to \u003cem\u003eC. floccosum\u003c/em\u003e (MK460809). ACT sequence was 99.13% (228/230) to \u003cem\u003eC. herbarum\u003c/em\u003e (EF679510). Comparing the ACT sequence to the type strains, it was 98.26% (226/230) similar to \u003cem\u003eC. macrocarpum\u003c/em\u003e (EF679529), 97.83% (225/230) to \u003cem\u003eC. herbarum\u003c/em\u003e (EF679516), and 95.15% (216/227) to \u003cem\u003eC. versiforme\u003c/em\u003e (KT600613). LSU was 98.74% (1184/1199) similar to \u003cem\u003eC. herbarum\u003c/em\u003e (MH047193), and \u003cem\u003eC. allicinum\u003c/em\u003e (GU214408).\u003c/p\u003e \u003cp\u003eITS sequence of \u003cem\u003eCladosporium\u003c/em\u003e strain AUMC 11366 was 98.9% (548/554) similar to \u003cem\u003eC. cladosporioides\u003c/em\u003e (MT367253, MF077224 and MF319920). By comparing to ITS sequences of the type species, it was 98.55% (542/550) similar to \u003cem\u003eC. verrucocladosporioides\u003c/em\u003e (MH863939), 98.53% (536/544) to \u003cem\u003eC. vicinum\u003c/em\u003e (MF473311) and 98.88% (530/536) to \u003cem\u003eC. kenpeggii\u003c/em\u003e (KY646222). ACT of \u003cem\u003eCladosporium\u003c/em\u003e strain AUMC 11366 was 100% similar to \u003cem\u003eC. cladosporioides\u003c/em\u003e (KY886457, MZ734607 and HM148501). By comparing with ACT sequences of the type materials, it was 97.38% similar to \u003cem\u003eC. proteacearum\u003c/em\u003e (MZ344213), 98.15% to \u003cem\u003eC. devikae\u003c/em\u003e (MZ344212), and 95.65% similar to \u003cem\u003eC. cladosporioides\u003c/em\u003e (HM148490). LSU was 98.77% (1206/1221) similar to \u003cem\u003eC. delicatulum\u003c/em\u003e (JQ732985) and \u003cem\u003eC. uredinicola\u003c/em\u003e (EU019264).\u003c/p\u003e \u003cp\u003eThis analysis included 55 sequences. The full parsimony dataset consisted of 808 characters with 437 constant characters (no gaps, no N), 114 variable characters which were parsimony-uninformative (26.1% of constant characters), and 33 characters were counted as parsimony informative (7.6% of constant). Tamura 3-parameter (T92\u0026thinsp;+\u0026thinsp;G) was the best nucleotide substitution model. The most parsimonious tree, resulted from the maximum parsimony analysis with the highest log likelihood (-5359.90), a tree length of 802 steps, consistency index of 0.520478, retention index of 0.807270, and a composite index of 0.524071 for all sites and parsimony-informative sites, is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Nexus file of the sequence alignments for all data sets (ITS\u0026thinsp;+\u0026thinsp;ACT) were uploaded to TreeBASE \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://purl.org/phylo/treebase/phylows/study/TB2:S23783\u003c/span\u003e\u003c/span\u003e (study no. 29214).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eTaxonomy\u003c/h2\u003e \u003cp\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eCladosporium parasphaerospermum sp. nov\u003c/span\u003e., Moharram AM, Zohri AA, Maher MA and Al-Bedak OA\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eMycoBank: MB839546\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eEtymology\u003c/strong\u003e \u003cp\u003eName refers to globose to subglobose conidia near to that of \u003cem\u003eC. sphaerospermum\u003c/em\u003e.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eHolotype\u003c/b\u003e: Egypt, Beni Suef, Air, Maher MA, AUMC 10865. Ex-type culture: EMCCN: 2062\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eMacroscopic and microscopic characteristics\u003c/strong\u003e \u003cp\u003eColonies on PDA reaching 18\u0026ndash;20 mm diameter after 7 d at 28 \u0026ordm;C, raised at the center, radially sulcate, radially furrowed under the colony, olive, olive green at the center (2F6). Margin curled, paler than the colony center (3F8). Sporulation profuse. Exudates absent. Colonies on SNA reaching 17\u0026ndash;19 mm in diameter after 7 d at 28 \u0026ordm;C, flat, slightly raised at the center, circular, olive, olive green (3F4-6). Margin entire. Sporulation abundant. Exudates absent. On OA colonies attaining 16\u0026ndash;19 mm in diameter after 7 d at 28 \u0026ordm;C, circular, flat, somewhat lanuginose, dark olive, dark olive green (1F4-6). Margin entire. Sporulation abundant. Exudates absent. Mycelium abundantly formed, branched, 3\u0026ndash;5 \u0026micro;m wide, septate, pale brown to brown, smooth. Conidiophores macronematous, micronematous, abundantly formed, arising terminally or laterally, more or less straight to flexuous, cylindrical, pale brown to brown, smooth, septate, commonly (\u0026ndash;35) 75\u0026ndash;100 \u0026times; 3\u0026ndash;5 \u0026micro;m (av. 87.5 \u0026times; 4) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50), not constricted at septa. Ramoconidia integrated, terminal, intercalary, cylindrical, smooth, thick-walled, 0\u0026ndash;1 septate, 8\u0026ndash;18 \u0026times; 3\u0026ndash;5 \u0026micro;m (av. 13 \u0026times; 4) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50), with 1\u0026ndash;3 loci per cell. Loci usually confined to small lateral shoulders, protuberant, conspicuous, short cylindrical, 1\u0026ndash;2 \u0026micro;m wide, up to 1\u0026ndash;2 \u0026micro;m high. Conidia brown to dark brown, smooth, thick-walled, globose, subglobose, lemon-shaped, 0\u0026ndash;septate, 4\u0026ndash;6 \u0026times; 3\u0026ndash;5 \u0026micro;m (av. 5 \u0026times; 4) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50). Chlamydospores not formed (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eCladosporium chlamydosporigenum sp. nov\u003c/span\u003e., Moharram AM, Zohri AA, Maher MA and Al-Bedak OA\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eMycoBank: MB839547\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eEtymology\u003c/strong\u003e \u003cp\u003erefers to the formation of chlamydospores in culture.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eHolotype\u003c/b\u003e: Egypt, Sohag, Grapevine fruits, Maher MA, AUMC 11340. Ex-type culture: EMCCN: 2332.\u003c/p\u003e \u003cp\u003e \u003cstrong\u003eMacroscopic and microscopic characteristics\u003c/strong\u003e \u003cp\u003eColonies on PDA reaching 15\u0026ndash;17 mm diameter after 7 d at 28 \u0026ordm;C, raised at the center, wrinkled, irregular, olive, olive green (3F6-7). Margin undulate, narrow, paler than the center (3E5-6). Sporulation abundant. Exudates absent. On SNA colonies attaining 9\u0026ndash;11 mm in diameter after 7 d at 28 \u0026ordm;C, flat, filamentous, olive, olive green (3E2). Margin filiform, narrow, (3E6-7). \u0026micro;m (av. 250 \u0026times; 5) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50), not constricted at septa. Ramoconidia integrated, terminal, intercalary, cylindrical, 11\u0026ndash;22 \u0026times; 6\u0026ndash;8 \u0026micro;m (av. 16.5 \u0026times; 7) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50), verruculose to finely roughened, 0\u0026ndash;2 septa with 1\u0026ndash;3 loci per cell, Loci usually confined to small lateral shoulders, protuberant, conspicuous, short cylindrical, 1\u0026ndash;2 \u0026micro;m wide, up to 1 \u0026micro;m high. Conidial chains unbranched or branched, conidia pale brown, straight, subglobose, obovoid to ellipsoid, verruculose to finely roughened, 0\u0026ndash;septate, 4\u0026ndash;11 \u0026times; 5\u0026ndash;7 \u0026micro;m (av. 7.5 \u0026times; 6) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50). Chlamydospores produced in hyphae, intercalary, aggregated, brown to dark brown, thick-walled, globose, subglobose, 15\u0026ndash;25\u0026thinsp;\u0026times;\u0026thinsp;\u0026micro;m (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cspan type=\"BoldItalic\" class=\"BoldItalic\" name=\"Emphasis\"\u003eCladosporium compactisporum sp. nov\u003c/span\u003e., Moharram AM, Zohri AA, Maher MA and Al-Bedak OA\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eMycoBank: MB839548\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eEtymology\u003c/strong\u003e \u003cp\u003erefers to the compact conidial chains.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eHolotype\u003c/b\u003e: Egypt, Qena, Air, Maher MA, AUMC 11366. Ex-type culture: EMCCN: 2358.\u003c/p\u003e \u003cp\u003e \u003cb\u003eMacroscopic and microscopic characteristics.\u003c/b\u003e Colonies on PDA attaining 25\u0026ndash;28 mm after 7 d at 28 \u0026ordm;C, raised, umbonate, circular, olive to olive green (3E3-3F4). Margin entire, narrow, about 3.0 mm in width, paler than the colony center (3E1-3). Sporulation profuse. Exudates absent. Colonies on SNA attaining 17\u0026ndash;20 mm diameter after 7 d at 28 \u0026ordm;C, raised, umbonate, olive to olive green (3F6-7). Margin entire, about 3.0 mm in width, paler than the colony center (3E2-3). Sporulation abundant. Exudates absent. Colonies on OA attaining 19\u0026ndash;23 mm in diameter after 7 d at 28 \u0026ordm;C, raised, umbonate, lanuginose, olive grey (2E1-2). Margin undulate, narrow, dark olive grey (2F2). Sporulation abundant. Exudates lacking. Mycelium abundantly formed, branched, 3\u0026ndash;5 \u0026micro;m wide, septate, swollen, pale brown to brown, smooth. Conidiophores macronematous and micronematous, abundantly formed, arising terminally or laterally, more or less straight to flexuous, nodulose, geniculate at the upper part, cylindrical, pale brown to brown, smooth, septate, branched, 100\u0026ndash;300 \u0026micro;m \u0026times; 3.0\u0026ndash;6.0 \u0026micro;m (av. 200 \u0026times; 4.5) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50). Ramoconidia integrated, terminal, intercalary, cylindrical, 7\u0026ndash;22 \u0026times; 3\u0026ndash;4 \u0026micro;m (av. 14.5 \u0026times; 3.5) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50), smooth, 0\u0026ndash;1 septa with 1\u0026ndash;3 loci per cell. Loci usually confined to small lateral shoulders, protuberant, conspicuous, short cylindrical, 1 \u0026micro;m wide, up to 1\u0026ndash;2 \u0026micro;m high. Conidia formed in compact and branched chains, pale brown, subglobose, obovoid to ellipsoid, smooth, 0-septate, 4\u0026ndash;6 \u0026times; 3\u0026ndash;4 \u0026micro;m (av. 5 \u0026times; 3.5) \u0026micro;m (n\u0026thinsp;=\u0026thinsp;50). Chlamydospores not formed (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eYield and Activity of Pectinases at pH and temperature\u003c/h2\u003e \u003cp\u003eThe three fungi generated pectinases with a rather high output in submerged fermentation. Per liter of fermentation media, \u003cem\u003eC. parasphaerospermum\u003c/em\u003e produced 3.0 g pectinase powder, \u003cem\u003eC. chlamydosporum\u003c/em\u003e produced 3.65 g, and \u003cem\u003eC. compactisporum\u003c/em\u003e produced 2.85 g. The pectinases appeared to be active; the greatest activity for the microbial pectinases was 1840, 893, and 930 U ml\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e at pH 6.0, 7.0, and 8.0, respectively. \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e had specific activity of 4221\u0026thinsp;\u0026plusmn;\u0026thinsp;138 Ug\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 1698\u0026thinsp;\u0026plusmn;\u0026thinsp;63 Ug\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, and 2120\u0026thinsp;\u0026plusmn;\u0026thinsp;15 Ug\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e pectinase, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). The optimum temperature was 35, 30, and 35 \u0026ordm;C, at pH 6.0, 7.0, and 8.0 for the three microbial pectinases produced by \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e, respectively. The specific activity values increased to 5758\u0026thinsp;\u0026plusmn;\u0026thinsp;159 Ug\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 3716\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8 Ug\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, and 3482\u0026thinsp;\u0026plusmn;\u0026thinsp;50 Ug\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e pectinase, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eEffect of some ions and EDTA on pectinases activity\u003c/h2\u003e \u003cp\u003eThe ions potassium and manganese exhibited a high activating influence (124.4% and 172%, respectively) on the activity of \u003cem\u003eCladosporium compactisporum\u003c/em\u003e pectinase. When evaluated under the optimal circumstances, EDTA and the metal ions showed varying inhibitory effects on the activity of the pectinases generated. In the case of \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e and \u003cem\u003eC. compactisporum\u003c/em\u003e pectinases, the highest inhibitory effects were with Ni, Zn, and EDTA, respectively (Table\u0026nbsp;1).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIdentification of novel species is at the heart of biodiversity research, and in recent years biodiversity efforts have been favouring DNA based method to morphology-based ones \u003csup\u003e31\u0026ndash;35\u003c/sup\u003e. Understanding of the microbial composition aids awareness of host-microbe interactions and their environmental function, revealing a complex and delicate balance that can be easily upset \u003csup\u003e36\u0026ndash;38\u003c/sup\u003e. Due to the complexity of fungal genomes and the lack of verified databases documenting appropriate biodiversity, such metagenomics studies in fungi have been limited. Identification of novel species is crucial in biodiversity research, which has lately adopted DNA-based methodologies. In this work, \u003cem\u003eCladosporium parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporigenum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e were identified as new \u003cem\u003eCladosporium\u003c/em\u003e species based on phenotypic features of their cultures and sequencing of the ITS, ACT, and LSU.\u003c/p\u003e \u003cp\u003eMorphologically, \u003cem\u003eC. parasphaerospermum\u003c/em\u003e AUMC 10865 has conidiophores that are 75\u0026ndash;100 \u0026micro;m shorter than \u003cem\u003eC. cladosporioides\u003c/em\u003e (350 \u0026micro;m). It may also be recognized from \u003cem\u003eC. cladosporioides\u003c/em\u003e \u003csup\u003e1\u003c/sup\u003e, \u003cem\u003eC. halotolerans\u003c/em\u003e \u003csup\u003e39\u003c/sup\u003e, and \u003cem\u003eC. parahalotolerans\u003c/em\u003e \u003csup\u003e39\u003c/sup\u003e by its smaller ramoconidia (8\u0026ndash;18 \u0026micro;m), which measure 15\u0026ndash;50 \u0026micro;m, 15\u0026ndash;37 \u0026micro;m, and 24\u0026ndash;37 \u0026micro;m, respectively. It may also be recognized from \u003cem\u003eC. omanense\u003c/em\u003e \u003csup\u003e40\u003c/sup\u003e by its slower developing colonies on PDA (18\u0026ndash;20 mm diam/7 days) compared to \u003cem\u003eC. omanense\u003c/em\u003e's 83 mm/14 days. \u003cem\u003eCladosporium omanense\u003c/em\u003e hyphae are in a polysaccharide-like substance and produce brief cross connections between hyphae, which \u003cem\u003eC. parasphaerospermum\u003c/em\u003e lacks. \u003cem\u003eCladosporium chlamydosporigenum\u003c/em\u003e AUMC 11340 has smaller conidia (4\u0026ndash;11 \u0026micro;m) than \u003cem\u003eC. floccosum\u003c/em\u003e \u003csup\u003e39\u003c/sup\u003e and \u003cem\u003eC. versiforme\u003c/em\u003e \u003csup\u003e41\u003c/sup\u003e, which have terminal (6\u0026ndash;15 \u0026micro;m and 4\u0026ndash;18\u0026micro;m) or intercalary (10\u0026ndash;21 \u0026micro;m and 6\u0026ndash;21 \u0026micro;m) conidia, respectively. It also has smaller ramoconidia (11\u0026ndash;22 \u0026micro;m) with 0\u0026ndash;2 septa than \u003cem\u003eC. versiforme\u003c/em\u003e, whereas \u003cem\u003eC. versiforme\u003c/em\u003e generates 45 \u0026micro;m ramoconidia with up to 5 septa. \u003cem\u003eC. herbarum\u003c/em\u003e \u003csup\u003e1\u003c/sup\u003e has several nodulose swellings in quick succession, but \u003cem\u003eC. allicinum\u003c/em\u003e \u003csup\u003e1\u003c/sup\u003e has head-like swellings with additional intercalary swellings, which \u003cem\u003eC. chlamydosporigenum\u003c/em\u003e lacks. \u003cem\u003eC. chlamydosporigenum\u003c/em\u003e may also be distinguished from \u003cem\u003eC. floccosum\u003c/em\u003e, \u003cem\u003eC. herbarum\u003c/em\u003e, \u003cem\u003eC. allicinum\u003c/em\u003e, and \u003cem\u003eC. macrocarpum\u003c/em\u003e by the presence of chlamydospores.\u003c/p\u003e \u003cp\u003e \u003cem\u003eCladosporium compactisporum\u003c/em\u003e AUMC 11366 contains smaller ramoconidia (7\u0026ndash;22 \u0026micro;m) with 1\u0026ndash;3 loci compared to \u003cem\u003eC. cladosporioides\u003c/em\u003e (15\u0026ndash;50 \u0026micro;m) with up to 4 loci packed at the apex, and \u003cem\u003eC. vicinum\u003c/em\u003e \u003csup\u003e39\u003c/sup\u003e which measures 20\u0026ndash;60 (-70) \u0026micro;m with up to 7 loci packed at the apex plus 1\u0026ndash;2 loci. \u003cem\u003eCladosporium compactisporum\u003c/em\u003e differs from \u003cem\u003eC. verrucocladosporioides\u003c/em\u003e \u003csup\u003e1\u003c/sup\u003e and \u003cem\u003eC. delicatulum\u003c/em\u003e \u003csup\u003e39\u003c/sup\u003e by having nodulose and smooth conidiophores, as well as smaller ramoconidia (7\u0026ndash;22 \u0026micro;m) compared to non-nodulose conidiophores and larger ramoconidia of \u003cem\u003eC. verrucocladosporioides\u003c/em\u003e. \u003cem\u003eCladosporium uredinicola\u003c/em\u003e \u003csup\u003e1\u003c/sup\u003e possesses branching, aggregated hyphae that create thick hyphal nets, but \u003cem\u003eC. compactisporum\u003c/em\u003e does not. The three new strains, \u003cem\u003eCladosporium parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e were used in this study to produce pectinases with high specific activities at low temperatures (10 \u003csup\u003eo\u003c/sup\u003eC) in SmF. Because of their biodegradability, non-toxicity, high selectivity, and high yields, microbial enzymes are superior to chemical synthesis (Bruno et al. 2019). The global enzyme market was valued at \u003cspan\u003e$\u003c/span\u003e9.9\u0026nbsp;billion in 2019 and is expected to grow at a 7.1% annual rate from 2020 to 2027 \u003csup\u003e42\u003c/sup\u003e. Pectinase has lengthy been used in commercial food processing to degrade pectin and aid in various processing steps such as liquefaction, clarification, and juice extraction \u003csup\u003e43\u003c/sup\u003e. Pectinases are among the most widely used enzymes, accounting for 40% of all food enzymes \u003csup\u003e44,45\u003c/sup\u003e. Several \u003cem\u003eCladosporium\u003c/em\u003e species have been shown to produce active pectinases as well \u003csup\u003e9,25,26,46\u0026minus;48\u003c/sup\u003e. Due to minor changes in methodology, it is difficult to compare the values of enzyme activity between different researches. As a result, comparisons should be made with caution.\u003c/p\u003e \u003cp\u003eThe three novel strains could create active pectinases at pH 6.0, 7.0, and 8.0, respectively, while the optimum temperatures for \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e were 35, 30, and 35 \u003csup\u003eo\u003c/sup\u003eC, respectively. The majority of commercial enzymes, including pectinase, are now mesophilic or thermophilic. In the food sector, and particularly in the fruit processing sector, there has been an increasing desire to replace high-temperature procedures with low-temperature processes. Specific economic and environmental benefits, such as energy savings, retention of biologically inert and aromatic fragrance components, contamination mitigation, and eradication of any residual enzyme activity, which is deactivation of enzyme when temperature is raised, are driving this shift in trend \u003csup\u003e42,43,49\u0026minus;51\u003c/sup\u003e.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn the current study, three novel \u003cem\u003eCladosporium\u003c/em\u003e species were introduced and described as \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e. The three novel species appeared to produce mesophilic pectinases that had high activity at pH 6.0 and 35 \u0026ordm;C, pH 7.0 and 30 \u0026ordm;C, and pH 8.0 and 35 \u0026ordm;C, respectively, of which \u003cem\u003eC. parasphaerospermum\u003c/em\u003e was the most active. We may now add the generation of active pectinases to the vast list of \u003cem\u003eCladosporium\u003c/em\u003e species that have been documented. We also describe three novel species that can be exploited as active microbial pectinase producers in biotechnological solutions. Although further research is needed, these unique species might be employed to degrade troublesome and resistant pertinacious wastes as well as clear fruit juices.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e\u003cstrong\u003eIsolation and Maintenance of \u003cem\u003eCladosporium\u003c/em\u003e Strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThree \u003cem\u003eCladosporium\u003c/em\u003e isolates involved in the current study, of which two were isolated from air of Beni Suef and Qena cities and one from grapevine fruits in Sohag city, Egypt. Exposure method \u003csup\u003e52\u003c/sup\u003e was employed for isolation of \u003cem\u003eCladosporium\u0026nbsp;\u003c/em\u003efrom air and direct plating technique \u003csup\u003e53\u003c/sup\u003e for isolation from grapevine fruits. Czapek\u0026rsquo;s Dox agar was used as an isolation medium. The isolation medium composed of (g/L): Sucrose, 30, Na\u003csub\u003e2\u003c/sub\u003eNO\u003csub\u003e3\u003c/sub\u003e, 2, K\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e, 1, KCl, 0.5, MgSO\u003csub\u003e4\u003c/sub\u003e.7H\u003csub\u003e2\u003c/sub\u003eO, 0.5, FeSO\u003csub\u003e4\u003c/sub\u003e, 0.01, ZnSO\u003csub\u003e4\u003c/sub\u003e, 0.01, CuSO\u003csub\u003e4\u003c/sub\u003e, 0.005, Rose Bengal, 0.05, chloramphenicol, 0.25, agar, 15 and the final pH 7.3. The interesting isolates obtained were preserved as frozen and lyophilized cultures in the culture collection of Assiut University Mycological Centre (AUMC) and in the Egyptian Microbial Culture Collection Network (EMCCN) as AUMC 10865 = EMCCN 2062 (air, Beni Suef), AUMC 11340 = EMCCN 2332 (grapevine fruits, Sohag) and AUMC 11366 = EMCCN 2358 (air, Qena). The novel species were registered at the MycoBank with their description with accession numbers of MB839546, MB839547\u0026nbsp;and MB839548, respectively.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMorphological studies of the \u003cem\u003eCladosporium\u003c/em\u003e strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSpore suspension (prepared in a 30 % glycerol, 0.2 % agar and 0.05 % Tween 80 solution) of the three \u003cem\u003eCladosporium\u0026nbsp;\u003c/em\u003ewas inoculated in a three-point pattern using inoculum size of 1.0 \u0026mu;l/spot on potato dextrose agar \u0026nbsp;(PDA,\u003csup\u003e\u0026nbsp;54 46 54 54 54 54 54 33 54\u003c/sup\u003e [54] (Smith \u0026amp; Onions, 1994) \u0026nbsp;(54) [54] [54] [33] [33] \u0026nbsp;(Smith and Onions 1994) (Smith and Onions 1994) [33], synthetic nutrient agar (SNA) and Oat meal agar (OA) as described by \u003csup\u003e54,55\u003c/sup\u003e. \u0026nbsp;Morphological characteristics and growth rates on SNA were examined in lacto-phenol cotton blue. Colony colors were identified according to \u003csup\u003e56\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMolecular identification of the \u003cem\u003eCladosporium\u003c/em\u003e strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDNA extraction\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor DNA isolation, a small portion from fungal growth of 7-day-old colonies of \u003cem\u003eCladosporium\u003c/em\u003e isolates AUMC 10865, AUMC 11340 and AUMC 11366 grown on PDA at 25 \u0026deg;C were collected and transferred individually to 2 ml-Eppendorf tube. The DNA extraction was performed following CTAB method described in Moubasher, et al. \u003csup\u003e57\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePCR and sequencing of ITS, ACT and LSU\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe universal primers ITS1 and ITS4 \u003csup\u003e58\u003c/sup\u003e were used for amplification of the internal transcribed spacer (ITS) region, LROR and LR7 primers \u003csup\u003e59\u003c/sup\u003e for amplification of the large subunit (LSU) and ACT512F and ACT783R for amplification of ACT gene \u003csup\u003e60\u003c/sup\u003e. PCR was done following Al-Bedak and Moubasher \u003csup\u003e61\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAlignments and phylogenetic analyses\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSequences of \u003cem\u003eCladosporium\u003c/em\u003e species (ITS, ACT, LSU) in this study were compared to sequences of the type and ex-type species in GenBank (Table 2). The ITS and ACT sequences of the strains in this study and those downloaded from GenBank were concatenated using Sequence Matrix version 1.8 \u003csup\u003e62\u003c/sup\u003e. The produced concatenated sequences were aligned together using MAFFT (version 6.861b) with the default options \u003csup\u003e63\u003c/sup\u003e, and \u003cem\u003eCercospora beticola\u0026nbsp;\u003c/em\u003eCBS 116456 was used as the outgroup. Alignment gaps and parsimony uninformative characters were treated by BMGE \u003csup\u003e64\u003c/sup\u003e. Maximum-likelihood (ML) and Maximum parsimony (MP) phylogenetic analyses were performed using PhyML 3.0 \u003csup\u003e65\u003c/sup\u003e. The robustness of the most parsimonious trees was evaluated by 1000 replications \u003csup\u003e66\u003c/sup\u003e. The best optimal model of nucleotide substitution for the ML analyses was determined using Smart Model Selection (SMS) version 1.8.1 \u003csup\u003e67\u003c/sup\u003e. The phylogenetic tree was drawn and visualized using MEGA X 10.2.6 \u003csup\u003e68\u003c/sup\u003e. The resulting tree was edited using Microsoft Power Point (2016) and saved as TIF file \u003csup\u003e9\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eProduction of Pectinases in Submerged Fermentation (SmF)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePreviously, the three Cladosporium species AUMC (10865, 11340 and 11366) were discovered to be capable of producing large quantities of pectinases in SmF at 10 \u0026ordm;C reaching 8.0 U/mg, 1.78 U/mg and 7.52 U/mg, respectively \u003csup\u003e27\u003c/sup\u003e. For pectinases production by the three strains, Cladosporium species were employed in Erlenmeyer flasks (500 ml) to produce pectinases in SmF at 10 \u003csup\u003eo\u003c/sup\u003eC using sucrose-free Czapek\u0026apos;s (initial pH 7.0) supplemented with 1.0 % pectin (as only carbon source).\u0026nbsp;Cladosporium\u0026nbsp;species were inoculated with 1.5 x 10\u003csup\u003e8\u003c/sup\u003e spore/ml spore suspensions obtained from 7-day-old cultures. The incubation period lasted 7 days at 10\u003csup\u003eo\u003c/sup\u003eC and 150 rpm. Following the incubation time, cell-free supernatants were recovered by centrifuging at 10,000 rpm for 10 min. At 4 \u003csup\u003eo\u003c/sup\u003eC, total protein was isolated using 70 % saturation of ammonium sulphate.\u0026nbsp;A freeze dryer (VirTis, model #6KBTES-55, NY, USA) was used to separate and lyophilize the precipitated total protein. Lyophilized protein was dissolved in citrate buffer (pH 5.0) and dialyzed twice for 2 hours at room temperature (cutoffs: 12-14 KD) against the same buffer, eliminating the buffer each time, before being refrigerated overnight at 4 \u0026ordm;C to remove small molecules. The dialyzed protein was then lyophilized, weighed, and used in enzyme characterization experiments as a partly pure fungal pectinases enzymes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eImpact of pH, Temperature and Some Ions and Inhibitors on Pectinases Activities\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA 0.01 g enzyme powder and 0.01 g citrus peel pectin (each dissolved in 1.0 ml of 50 mmol buffer solution) were included in this test in a water bath. The impact of pH (3.0\u0026ndash;10.0) and temperature (30\u0026ndash;60 \u0026ordm;C) on pectinases activity were investigated. After the reaction time (20 min), the reaction was terminated by introducing 2.0 ml of 3,5-dinitrosalicylic acid (DNS) according to Miller \u003csup\u003e69\u003c/sup\u003e and Jayani, et al. \u003csup\u003e70\u003c/sup\u003e, and the pectinase activity was determined as the amount of the enzyme that releases 1 \u0026micro;mol ml\u003csup\u003e-1\u003c/sup\u003e min\u003csup\u003e-1\u003c/sup\u003e galacturonic acid under standard assay conditions. The buffers used were citrate buffer (pH 3.0\u0026ndash;6.0), phosphate buffer (pH 7.0\u0026ndash;8.0), and borate buffer (pH 9.0\u0026ndash;10.0). Also, some ions (Na\u003csup\u003e+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e+2\u003c/sup\u003e, Co\u003csup\u003e+2\u003c/sup\u003e, Ni\u003csup\u003e+2\u003c/sup\u003e, Cu\u003csup\u003e+2\u003c/sup\u003e, Fe\u003csup\u003e+2\u003c/sup\u003e, Mg\u003csup\u003e+2\u003c/sup\u003e, Mn\u003csup\u003e+2\u003c/sup\u003e, and Zn\u003csup\u003e+2\u003c/sup\u003e) were evaluated by introducing them at 5 mmol/ml concentrations as NaCl, KCl, CaCl\u003csub\u003e2\u003c/sub\u003e, CoCl\u003csub\u003e2\u003c/sub\u003e, NiSO\u003csub\u003e4\u003c/sub\u003e, CuSO\u003csub\u003e4\u003c/sub\u003e, FeSO\u003csub\u003e4\u003c/sub\u003e, MgSO\u003csub\u003e4\u003c/sub\u003e, MnSO\u003csub\u003e4\u003c/sub\u003e, and ZnSO\u003csub\u003e4\u003c/sub\u003e. 5 mmol/ml ethylenediaminetetraacetic acid was used to evaluate an enzyme inhibitor (EDTA). Under standard conditions, the activity of the microbial pectinases in the absence of metal ions or EDTA was evaluated to define 100 % activity. All experiments were conducted in three repetitions. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAccessibility of data:\u0026nbsp;\u003c/strong\u003ePure cultures of the type materials of the novel species are deposited in the culture collection of Assiut University Mycological Centre as AUMC 10865, AUMC 11340 and AUMC 11366, and in the Egyptian Microbial Culture Collection Network as EMCCN 2062, EMCCN 2332 and EMCCN 2358. Nexus file of the sequence alignments for all\u003cbr\u003e\u0026nbsp;data sets were uploaded to TreeBASE http://purl.org/phylo/treebase/phylows/study/TB2:S23783 \u0026nbsp;(study no. 29214).\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u0026nbsp;\u003c/strong\u003eA.M.M.: Supervision of the study, writing, revising, A-N.A.Z.: Supervision of the study, writing, revising, M.A.M.: Fungal isolation, enzymatic production, A.E.H.: Editing, revising, O.AA.-B.: Molecular work, data analysis, writing, revising. All authors contributed to data analysis, drafting, or revising the manuscript. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u0026nbsp;\u003c/strong\u003e\u0026ldquo;The datasets generated and/or analysed during the current study are available in the GenBank repository, [https://www.ncbi.nlm.nih.gov/genbank/]\u0026rdquo;.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest:\u0026nbsp;\u003c/strong\u003eThe authors declare that there are no potential conflict of interest regarding the publication of this paper.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability:\u0026nbsp;\u003c/strong\u003eAll datasets generated or analyzed during this study are included in the manuscript.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBensch, K., Braun, U., Groenewald, J. Z. \u0026amp; Crous, P. W. The genus \u003cem\u003ecladosporium\u003c/em\u003e. Studies in mycology \u003cb\u003e72\u003c/b\u003e, 1\u0026ndash;401, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3114/sim0003\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEllis, M. More dematiaceous hyphomycetes (p.\u0026nbsp;507). \u003cem\u003eKew, Surrey, England: Commonwealth Mycological Institute\u003c/em\u003e (1976).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYehia, R. S., Osman, G. H., Assaggaf, H., Salem, R. \u0026amp; Mohamed, M. S. Isolation of potential antimicrobial metabolites from endophytic fungus \u003cem\u003eCladosporium cladosporioides\u003c/em\u003e from endemic plant \u003cem\u003eZygophyllum mandavillei\u003c/em\u003e. South African Journal of Botany \u003cb\u003e134\u003c/b\u003e, 296\u0026ndash;302, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.sajb.2020.02.033\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoubasher, A. Soil fungi in Qatar and other Arab countries. \u003cem\u003eThe Centre for Scientific and Applied Research, University of Qatar\u003c/em\u003e (1993).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIsmail, M., Moubasher, A., Mohamed, R. \u0026amp; Al-Bedak, O. Extremophilic fungi and chemical analysis of hypersaline, alkaline lakes of Wadi-El-Natrun, Egypt. International Journal of Technical Research and Science \u003cb\u003e1\u003c/b\u003e, 345\u0026ndash;363 (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFarr, D. F., Bills, G. F., Chamuris, G. P. \u0026amp; Rossman, A. Y. Fungi on plants and plant products in the United States. \u003cem\u003eAPS press\u003c/em\u003e (1989).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHaas, C. N. Microorganisms in home and indoor work environments: diversity, health impacts, investigation and control edited by Brian Flannigan, Robert A. Samson, and J. David Miller. \u003cem\u003eRisk Analysis\u003c/em\u003e \u003cb\u003e31\u003c/b\u003e, 1327\u0026ndash;1327, DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1539-6924.2011.01665.x\u003c/span\u003e\u003c/span\u003e (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFlannigan, B., Samson, R. A. \u0026amp; Miller, J. D. Microorganisms in home and indoor work environments: diversity, health impacts, investigation and control. CRC Press (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAl-Bedak, O. A., Teama, E.A.E., Ali, E.A., Said, M.T., Shalaby, E.M. \u0026amp; Moharram, Z.A.M. Impact of fumigation with phosphine on viability of wheat grains stored for six months at two levels of moisture content, in addition to description of four new records of associated fungi and assessment of their potential for enzymatic production. Journal of Basic \u0026amp; Applied Mycology (Egypt) \u003cb\u003e11\u003c/b\u003e, 77\u0026ndash;97 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchubert, K. \u0026amp; Braun, U. Taxonomic revision of the genus \u003cem\u003eCladosporium\u003c/em\u003e s. lat. 2. \u003cem\u003eCladosporium\u003c/em\u003e species occurring on hosts of the families Bignoniaceae and Orchidaceae. Sydowia \u003cb\u003e56\u003c/b\u003e, 296\u0026ndash;317 (2004).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHeuchert, B., Braun, U. \u0026amp; Schubert, K. Morphotaxonomic revision of fungicolous \u003cem\u003eCladosporium\u003c/em\u003e species (hyphomycetes). Schlechtendalia \u003cb\u003e13\u003c/b\u003e, 1\u0026ndash;78 (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRiesen, T. \u0026amp; Sieber, T. Endophytic fungi in winter wheat (\u003cem\u003eTiticum aestivum\u003c/em\u003e L.). (Zurich (Switzerland), \u003cem\u003eSwiss Federal Institute of Technology\u003c/em\u003e, 190 p. (1985).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBrown, K. Preliminary studies on endophytic fungal communities of \u003cem\u003eMusa acuminata\u003c/em\u003e species complex in Hong Kong and Australia. Fung. Divers. \u003cb\u003e1\u003c/b\u003e, 27\u0026ndash;51 (1998).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Morsy, E. Fungi isolated from the endorhizosphere of halophytic plants from the Red Sea Coast of Egypt. Fungal Diversity \u003cb\u003e5\u003c/b\u003e, 43\u0026ndash;54 (2000).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIslam, M. \u0026amp; Hasin, F. Studies on the phylloplane mycoflora of \u003cem\u003eAmaranthus viridis\u003c/em\u003e. L. National Academy Science letters \u003cb\u003e23\u003c/b\u003e, 121\u0026ndash;123 (2000).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIn\u0026aacute;cio, J. \u003cem\u003eet al.\u003c/em\u003e Estimation and diversity of phylloplane mycobiota on selected plants in a mediterranean\u0026ndash;type ecosystem in Portugal. Microbial Ecology \u003cb\u003e44\u003c/b\u003e, 344\u0026ndash;353 (2002).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStohr, S. \u0026amp; Dighton, J. Effects of species diversity on establishment and coexistence: A phylloplane fungal community model system. Microbial Ecology \u003cb\u003e48\u003c/b\u003e, 431\u0026ndash;438 (2004).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLevetin, E. \u0026amp; Dorsey, K. Contribution of leaf surface fungi to the air spora. Aerobiologia \u003cb\u003e22\u003c/b\u003e, 3\u0026ndash;12 (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbdel-Baky, N., Arafat, N. S. \u0026amp; Abdel-Salam, A. Three \u003cem\u003eCladosporium\u003c/em\u003e spp. as promising biological control candidates for controlling whiteflies (\u003cem\u003eBemisia\u003c/em\u003e spp.) in Egypt. Pak. J. Biol. Sci \u003cb\u003e1\u003c/b\u003e, 188\u0026ndash;195 (1998).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbdel-Baky, N. \u0026amp; Abdel-Salam, A. Natural incidence of \u003cem\u003eCladosporium\u003c/em\u003e spp. as a bio-control agent against whiteflies and aphids in Egypt. (2003).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSalvatore, M. M., Andolfi, A. \u0026amp; Nicoletti, R. The Genus \u003cem\u003eCladosporium\u003c/em\u003e: A Rich Source of Diverse and Bioactive Natural Compounds. Molecules \u003cb\u003e26\u003c/b\u003e, 3959 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHamed, M., Osman, A. A. \u0026amp; Ateş, M. Statistical optimization of L-asparaginase production by \u003cem\u003eCladosporium tenuissimum\u003c/em\u003e. Egyptian Pharmaceutical Journal \u003cb\u003e20\u003c/b\u003e, 51 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Sabbagh, S. M., Eissa, O. A. E. \u0026amp; Sallam, M. H. E. Taxol production by an endophytic fungus \u003cem\u003eCladosporium cladosporioides\u003c/em\u003e isolated from \u003cem\u003eCatheranthus roseus\u003c/em\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eV\u0026aacute;zquez-Montoya, E. L., Castro-Ochoa, L. D., Maldonado-Mendoza, I. E., Luna-Su\u0026aacute;rez, S. \u0026amp; Castro-Mart\u0026iacute;nez, C. Moringa straw as cellulase production inducer and cellulolytic fungi source. Revista Argentina de microbiologia \u003cb\u003e52\u003c/b\u003e, 4\u0026ndash;12 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBastos, S. C. \u003cem\u003eet al.\u003c/em\u003e Pectinases from a new strain of \u003cem\u003eCladosporium cladosporioides\u003c/em\u003e (Fres.) De Vries isolated from coffee bean. World J Agric Sci \u003cb\u003e9\u003c/b\u003e, 167\u0026ndash;172 (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAl-Bedak, O. A., Abdel-Sater, M. A., Abdel-Latif, A. M. \u0026amp; Abdel-Wahab, D. A. Endophytic mycobiota and enzymatic capacity in wild and cultivated plants in New Valley, Egypt: A comparative analysis. Journal of Basic \u0026amp; Applied Mycology (Egypt) \u003cb\u003e12\u003c/b\u003e, 27\u0026ndash;41 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoharram, A., Zohri, A., Hesham, A., Maher, M. \u0026amp; Al-Bedak, O. Production of Cocktail Enzymes by Three \u003cem\u003eCladosporium\u003c/em\u003e Isolates and Bioconversion of Orange Peel Wastes into Valuable Enzymes. J Pure Appl Microbiol \u003cb\u003e15\u003c/b\u003e, 2336\u0026ndash;2346 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmin, F., Bhatti, H. N. \u0026amp; Bilal, M. Recent advances in the production strategies of microbial pectinases\u0026mdash;A review. International journal of biological macromolecules \u003cb\u003e122\u003c/b\u003e, 1017\u0026ndash;1026 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaranraj, P. \u0026amp; Naidu, M. Microbial pectinases: a review. Global J Trad Med Syst \u003cb\u003e3\u003c/b\u003e, 1\u0026ndash;9 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSalikin, N. H. \u0026amp; Makhtar, M. M. Z. in \u003cem\u003eWaste Management, Processing and Valorisation\u003c/em\u003e 175\u0026ndash;206 (Springer, 2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHebert, P. D., Cywinska, A., Ball, S. L. \u0026amp; Dewaard, J. R. Biological identifications through DNA barcodes. \u003cem\u003eProceedings of the Royal Society of London. Series B: Biological Sciences\u003c/em\u003e \u003cb\u003e270\u003c/b\u003e, 313\u0026ndash;321 (2003).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Queiroz, K. Species concepts and species delimitation. Systematic biology \u003cb\u003e56\u003c/b\u003e, 879\u0026ndash;886 (2007).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVerkley, G., Quaedvlieg, W., Shin, H.-D. \u0026amp; Crous, P. A new approach to species delimitation in Septoria. Studies in Mycology \u003cb\u003e75\u003c/b\u003e, 213\u0026ndash;305 (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWoudenberg, J., Groenewald, J., Binder, M. \u0026amp; Crous, P. \u003cem\u003eAlternaria\u003c/em\u003e redefined. \u003cem\u003eStudies in mycology\u003c/em\u003e \u003cb\u003e75\u003c/b\u003e, 171\u0026ndash;212 (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu, X.-Z. \u003cem\u003eet al.\u003c/em\u003e Phylogeny of tremellomycetous yeasts and related dimorphic and filamentous basidiomycetes reconstructed from multiple gene sequence analyses. Studies in mycology \u003cb\u003e81\u003c/b\u003e, 1\u0026ndash;26 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFuhrman, J. A. Microbial community structure and its functional implications. Nature \u003cb\u003e459\u003c/b\u003e, 193\u0026ndash;199 (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarza, D. R., Van Verk, M. C., Huynen, M. A. \u0026amp; Dutilh, B. E. Bottom-up ecology of the human microbiome: from metagenomes to metabolomes. \u003cem\u003eBioRxiv\u003c/em\u003e, 060673 (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLevy, M., Blacher, E. \u0026amp; Elinav, E. Microbiome, metabolites and host immunity. Current opinion in microbiology \u003cb\u003e35\u003c/b\u003e, 8\u0026ndash;15 (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBensch, K. \u003cem\u003eet al.\u003c/em\u003e Cladosporium species in indoor environments. Studies in mycology \u003cb\u003e89\u003c/b\u003e, 177\u0026ndash;301 (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHalo, B., Maharachchikumbura, S. S., Al-Yahyai, R. A. \u0026amp; Al-Sadi, A. M. \u003cem\u003eCladosporium omanense\u003c/em\u003e, a new endophytic species from \u003cem\u003eZygophyllum coccineum\u003c/em\u003e in Oman. Phytotaxa \u003cb\u003e388\u003c/b\u003e, 145\u0026ndash;154 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBensch, K. \u003cem\u003eet al.\u003c/em\u003e Common but different: The expanding realm of \u003cem\u003eCladosporium\u003c/em\u003e. Studies in Mycology \u003cb\u003e82\u003c/b\u003e, 23\u0026ndash;74 (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, A., Mukhia, S. \u0026amp; Kumar, R. Industrial applications of cold-adapted enzymes: challenges, innovations and future perspective. 3 Biotech \u003cb\u003e11\u003c/b\u003e, 1\u0026ndash;18 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAdapa, V., Ramya, L., Pulicherla, K. \u0026amp; Rao, K. S. Cold active pectinases: advancing the food industry to the next generation. Applied biochemistry and biotechnology \u003cb\u003e172\u003c/b\u003e, 2324\u0026ndash;2337 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePedrolli, D. B., Monteiro, A. C., Gomes, E. \u0026amp; Carmona, E. C. Pectin and pectinases: production, characterization and industrial application of microbial pectinolytic enzymes. Open Biotechnology Journal, 9\u0026ndash;18 (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSemenova, M. \u003cem\u003eet al.\u003c/em\u003e Use of a preparation from fungal pectin lyase in the food industry. Applied Biochemistry and Microbiology \u003cb\u003e42\u003c/b\u003e, 598\u0026ndash;602 (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePoveda, G., Gil-Dur\u0026aacute;n, C., Vaca, I., Levic\u0026aacute;n, G. \u0026amp; Ch\u0026aacute;vez, R. Cold-active pectinolytic activity produced by filamentous fungi associated with Antarctic marine sponges. Biological research \u003cb\u003e51\u003c/b\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMing, X., Huai, Q., Huamei, W., Zhuyan, L. \u0026amp; Jian, Z. On kinetics of batch fermentation for pectinase from mutant of \u003cem\u003eCladosporium herbarum\u003c/em\u003e (pers) link ex gray. Guizhou science, 02 (1997).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBastos, S. C. \u003cem\u003eet al.\u003c/em\u003e in Microorganisms In Industry And Environment: From Scientific and Industrial Research to Consumer Products 538\u0026ndash;542 (World Scientific, 2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVan Truong, L., Tuyen, H., Helmke, E., Binh, L. T. \u0026amp; Schweder, T. Cloning of two pectate lyase genes from the marine Antarctic bacterium \u003cem\u003ePseudoalteromonas haloplanktis\u003c/em\u003e strain ANT/505 and characterization of the enzymes. Extremophiles \u003cb\u003e5\u003c/b\u003e, 35\u0026ndash;44 (2001).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePulicherla, K., Ghosh, M., Kumar, P. \u0026amp; Rao, K. Psychrozymes\u0026ndash;the next generation industrial enzymes. J Marine Sci Res Development \u003cb\u003e1\u003c/b\u003e (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMangiagalli, M., Brocca, S., Orlando, M. \u0026amp; Lotti, M. The \u0026ldquo;cold revolution\u0026rdquo;. Present and future applications of cold-active enzymes and ice-binding proteins. New biotechnology \u003cb\u003e55\u003c/b\u003e, 5\u0026ndash;11 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFlannigan, B. Air sampling for fungi in indoor environments. Journal of Aerosol Science \u003cb\u003e28\u003c/b\u003e, 381\u0026ndash;392 (1997).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBerry, E. D. \u0026amp; Wells, J. E. A direct plating method for estimating populations of Escherichia coli O157 in bovine manure and manure-based materials. Journal of food protection \u003cb\u003e71\u003c/b\u003e, 2233\u0026ndash;2238 (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSmith, D. \u0026amp; Onions, A. H. The preservation and maintenance of living fungi. (CAB international, 1994).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGerlach, W., Nirenberg, H., Eckart, I., Rummland, I. \u0026amp; Schwarz, R. The genus Fusarium: a pictorial atlas. Vol.\u0026nbsp;209 (Kommissionsverlag P. Parey Berlin, 1982).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKornerup, A. \u0026amp; Wanscher, J. (1978).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoubasher, A., Ismail, M., Al-Bedak, O. \u0026amp; Mohamed, R. \u003cem\u003eRamophialophora chlamydospora\u003c/em\u003e, a new species from an alkaline lake of Wadi-El-Natron, Egypt. Asian Journal of Mycology \u003cb\u003e2\u003c/b\u003e, 110\u0026ndash;117 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWhite, T. J., Bruns, T., Lee, S. \u0026amp; Taylor, J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR protocols: a guide to methods and applications \u003cb\u003e18\u003c/b\u003e, 315\u0026ndash;322 (1990).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRehner, S. A. \u0026amp; Samuels, G. J. Taxonomy and phylogeny of \u003cem\u003eGliocladium\u003c/em\u003e analysed from nuclear large subunit ribosomal DNA sequences. Mycological Research \u003cb\u003e98\u003c/b\u003e, 625\u0026ndash;634 (1994).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarbone, I. \u0026amp; Kohn, L. M. A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia \u003cb\u003e91\u003c/b\u003e, 553\u0026ndash;556 (1999).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAl-Bedak, O. \u0026amp; Moubasher, A. \u003cem\u003eAspergillus gaarensis\u003c/em\u003e, a new addition to section Circumdati from soil of Lake El-Gaar in Wadi-El-Natron, Egypt. Studies in Fungi \u003cb\u003e5\u003c/b\u003e, 59\u0026ndash;65 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVaidya, G., Lohman, D. J. \u0026amp; Meier, R. SequenceMatrix: concatenation software for the fast assembly of multi-gene datasets with character set and codon information. Cladistics \u003cb\u003e27\u003c/b\u003e, 171\u0026ndash;180 (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKatoh, K. \u0026amp; Standley, D. M. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular biology and evolution \u003cb\u003e30\u003c/b\u003e, 772\u0026ndash;780 (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCriscuolo, A. \u0026amp; Gribaldo, S. BMGE (Block Mapping and Gathering with Entropy): a new software for selection of phylogenetic informative regions from multiple sequence alignments. BMC evolutionary biology \u003cb\u003e10\u003c/b\u003e, 210 (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuindon, S. \u003cem\u003eet al.\u003c/em\u003e New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Systematic biology \u003cb\u003e59\u003c/b\u003e, 307\u0026ndash;321 (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFelsenstein, J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution \u003cb\u003e39\u003c/b\u003e, 783\u0026ndash;791 (1985).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLefort, V., Longueville, J.-E. \u0026amp; Gascuel, O. SMS: smart model selection in PhyML. Molecular biology and evolution \u003cb\u003e34\u003c/b\u003e, 2422\u0026ndash;2424 (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar, S., Stecher, G., Li, M., Knyaz, C. \u0026amp; Tamura, K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular biology and evolution \u003cb\u003e35\u003c/b\u003e, 1547\u0026ndash;1549 (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiller, G. L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical chemistry \u003cb\u003e31\u003c/b\u003e, 426\u0026ndash;428 (1959).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJayani, R. S., Saxena, S. \u0026amp; Gupta, R. Microbial pectinolytic enzymes: a review. Process Biochemistry \u003cb\u003e40\u003c/b\u003e, 2931\u0026ndash;2944 (2005).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1.\u0026nbsp;\u003c/strong\u003eEffect of metal ions and EDTA (5 mmol mL\u003cem\u003e\u003csup\u003e-\u003c/sup\u003e\u003c/em\u003e\u003csup\u003e1\u003c/sup\u003e) on pectinase activity produced by \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u0026nbsp;\u003c/em\u003e(mean \u003cem\u003e\u0026plusmn;\u0026nbsp;\u003c/em\u003eSD, n = 3).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe results are expressed as the activity in the tested inhibitory conditions compared to the pectinase activity in the control without inhibitors (in bold).\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"0\" cellpadding=\"0\" cellspacing=\"0\" width=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eMetal ions and inhibitors\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e\u003cem\u003eC. parasphaerospermum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e\u003cem\u003eC. chlamydosporum\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e\u003cem\u003eC. compactisporum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"34.883720930232556%\"\u003e\n \u003cp\u003eSpecific activity\u003c/p\u003e\n \u003cp\u003eU g\u003csup\u003e-1\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.55813953488372%\"\u003e\n \u003cp\u003eSpecific activity\u003c/p\u003e\n \u003cp\u003eU g\u003csup\u003e-1\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"32.55813953488372%\"\u003e\n \u003cp\u003eSpecific activity\u003c/p\u003e\n \u003cp\u003eU g\u003csup\u003e-1\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5758\u0026plusmn;159\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3716\u0026plusmn;3.8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3482\u0026plusmn;50\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eNa\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e4027\u0026plusmn;22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e3600\u0026plusmn;40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e2025\u0026plusmn;18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eK\u003csup\u003e+\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e3272\u0026plusmn;24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e3086\u0026plusmn;26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e4287\u0026plusmn;85\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eFe\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e3068\u0026plusmn;71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e759\u0026plusmn;55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e1865\u0026plusmn;15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eCu\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e3406\u0026plusmn;49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e2851\u0026plusmn;38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e1336\u0026plusmn;14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eCa\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e3875\u0026plusmn;52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e2734\u0026plusmn;15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e3379\u0026plusmn;67\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eMg\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e2231\u0026plusmn;45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e1682\u0026plusmn;15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e3215\u0026plusmn;33\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eZn\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e4185\u0026plusmn;30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e913\u0026plusmn;15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e2182\u0026plusmn;26\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eNi\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e2106\u0026plusmn;19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e2954\u0026plusmn;31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e3432\u0026plusmn;30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eCo\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e3473\u0026plusmn;106\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e1553\u0026plusmn;31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e2390\u0026plusmn;36\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eMn\u003csup\u003e+2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e2955\u0026plusmn;26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e1553\u0026plusmn;29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e5992\u0026plusmn;51\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"18.095238095238095%\"\u003e\n \u003cp\u003eEDTA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"28.571428571428573%\"\u003e\n \u003cp\u003e2383\u0026plusmn;15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e2165\u0026plusmn;100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"26.666666666666668%\"\u003e\n \u003cp\u003e1253\u0026plusmn;22\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2.\u003c/strong\u003e List of specimens and GenBank accession numbers of sequences used in this study. Bold accession numbers were generated from this study.\u0026nbsp;\u003c/p\u003e\n \u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eCladosporium\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003especies\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003e\u003cstrong\u003eStrain\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003e\u003cstrong\u003eLocality\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eITS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eACT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eLSU\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. parasphaerospermum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eAUMC 10865\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eEgypt\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMN826828\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eOL514008\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMW205008\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. chlamydosporigenum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eAUMC 11340\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eEgypt\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMN826919\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eOL514009\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMW205067\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. compactisporum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eAUMC 11366\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eEgypt\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMN826822\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eOL514010\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMW205086\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. aerium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eDTO:323-B4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eChina\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF472897\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF473747\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO:323-G7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eChina\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF472899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473749\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO:323-G6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eChina\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF472898\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473748\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. allicinum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 121624\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eBelgium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH863126\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679502\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH874678\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO 111-A5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eDenmark\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP701924\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP702047\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO 249-G3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP701975\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP702097\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. antarcticum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 690.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eAntarctica\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_121332\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679484\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. cladosporioides\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 112388\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_119839\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eHM148490\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 15167\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eSlovenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148052\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148539\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eGZYQ-08-01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eChina\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMK852271\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMK852272\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. colombiae\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 274.80B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eColombia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH861262\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eFJ936166\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. cycadicola\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 137970\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eAustralia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_156279\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eKJ869227\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNG_058881\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. delicatulum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eDTO 145-C4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eGermany\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eKP701940\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eKP702062\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 14372\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eDenmark\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148089\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148578\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 14363\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eDenmark\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148088\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148577\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. domesticum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCPC 22307\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_156348\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF473805\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO 308-B1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF472966\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473816\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC:22413\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF472961\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473811\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. exasperatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 125986\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eAustralia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH863865\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eHM148579\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH875326\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. floccosum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eDTO 323-H6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eChina\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF472979\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF473829\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 22968\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF472978\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473828\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 22399\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF472977\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473827\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. halotolerans\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 119416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_119605\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF101397\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO 257-F4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP701989\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP702111\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. herbaroides\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 121626\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eIsrael\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_119655\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679509\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. herbarum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 289.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eSwitzerland\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH856530\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679558\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH868058\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 12183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eEF679368\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eEF679521\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. iridis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 107.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH854682\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679522\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH866199\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCBS 138.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eNR_111271\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eEF679523\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. kenpeggii\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCPC 19248\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eAustralia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eKY646222\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eKY646225\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. macrocarpum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCPC 12759\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679380\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679534\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 14305\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eDenmark\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473141\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473990\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. ossifragi\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 842.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eNorway\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH862342\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679535\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. parahalotolerans\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eDTO 307-H4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eMexico\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF473161\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF474009\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO 324-B7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eChina\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF473169\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMF474017\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. perangustum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 167.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eHM148124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eHM148613\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCBS 126365\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMH863940\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148612\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eMH875401\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. sphaerospermum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 193.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eSlovenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_111222\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEU570269\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eDTO 255-H7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP701988\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eKP702110\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. spinulosum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 119907\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eSlovenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_119660\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679542\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. subcinereum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eUTHSC DI-13-257\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_148193\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eLN834617\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. subinflatum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 121630\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eSlovenia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH863129\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679543\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH874681\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. tenuissimum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 125995\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH864840\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eHM148687\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH876286\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. tuberosum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eUTHSC DI-13-217\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eSpain\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eLN834417\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eLN834601\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH878168\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. uredinicola\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCPC 5390\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eAY251071\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eHM148712\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEU019264\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. variabile\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 121635\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH863131\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eEF679557\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH874683\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. verrucocladosporioides\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 126363\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eSouth Korea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMH863939\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eHM148717\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. versiforme\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 140491\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eNetherlands\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_152297\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eKT600613\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. vicinum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCPC 22316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF473311\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF474161\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.38888888888889%\"\u003e\n \u003cp\u003eCPC 15457\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.444444444444443%\"\u003e\n \u003cp\u003eNew Zealand\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148060\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003eHM148547\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.055555555555557%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eC. wyomingense\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCPC 22310\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF473315\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eMF474165\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cem\u003eCercospora beticola\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"19.587628865979383%\"\u003e\n \u003cp\u003eCBS 116456\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.43298969072165%\"\u003e\n \u003cp\u003eUSA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNR_121315\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eAY840458\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.402061855670103%\"\u003e\n \u003cp\u003eNG_068999\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eAbbreviations used:\u003c/strong\u003e AUMC = Assiut University Mycological Centre, Assiut University, Assiut, Egypt, CBS = Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, CPC = Culture Collection of Pedro Crous, housed at CBS, Utrecht, The Netherlands, DTO = Culture Collection of Jos Houbraken, UTHSC = The University of Tennessee Health Science Center, Monroe Avenue, Memphis, Tennessee, USA.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\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":"Biodiversity, Fungi, Pectinase, Phylogeny, Taxonomy","lastPublishedDoi":"10.21203/rs.3.rs-1291889/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1291889/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBased on phenotypic characters and sequencing of the internal transcribed spacers (ITS), 28S Large Subunit (LSU) and partial actin (ACT), three novel species of \u003cem\u003eCladosporium\u003c/em\u003e were described in this study as \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e. Activities of pectinases produced by the three strains were 5700, 3720, and 3480 U mg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e enzymes, at the optimum pH (6.0, 7.0, and 8.0) and temperature (35, 30, and 35 \u0026ordm;C), respectively. The ions potassium and manganese had a strong activating impact on the activity of \u003cem\u003eC. compactisporum\u003c/em\u003e pectinase (124.4 and 172%, respectively). When tested under optimal conditions, EDTA and the other metal ions inhibited the activity of the pectinases in varied degrees. The strongest inhibitory effects of \u003cem\u003eC. parasphaerospermum\u003c/em\u003e, \u003cem\u003eC. chlamydosporum\u003c/em\u003e, and \u003cem\u003eC. compactisporum\u003c/em\u003e pectinases were with Ni\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e, Zn\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e, and EDTA, respectively. \u003cem\u003eCladosporium\u003c/em\u003e species have a long history of biotechnological uses, and here we add another: pectinase makers. We offer three unique suppliers of low-temperature active fungal pectinases for biotechnological applications such as the management of Pectinacious wastes in the juice industry.\u003c/p\u003e","manuscriptTitle":"Production and Partial Purification of Pectinases Produced by Three Novel Cladosporium Species Isolated from Egypt","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-02-18 17:12:30","doi":"10.21203/rs.3.rs-1291889/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2022-04-11T06:29:17+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2022-04-10T04:47:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"d1670389-72b5-40cf-8cbd-ced2b728f471","date":"2022-03-24T08:38:04+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2022-03-16T11:15:46+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"17972541-2fa5-42e1-bf2e-624a61650bdd","date":"2022-03-16T10:55:21+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2022-03-16T10:47:01+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2022-03-16T10:44:02+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2022-02-16T19:22:23+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2022-02-16T17:31:03+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2022-01-24T13:10:38+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":"ebd50826-2ec3-4eb5-b04b-1cd1a43859ed","owner":[],"postedDate":"February 18th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2022-09-05T09:29:15+00:00","versionOfRecord":[],"versionCreatedAt":"2022-02-18 17:12:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1291889","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1291889","identity":"rs-1291889","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}