Transferability and Polymorphism of Cymbidium Hybrid EST-SSR Markers in Chinese Cymbidium Cultivars
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摘要:目的 探索杂交兰EST-SSR标记在国兰中的通用性，为国兰寻找新的分子标记，并利用这些新标记分析国兰的种间遗传多样性，以期为国兰的品种鉴定和种质资源的合理利用奠定基础。方法 以建兰、墨兰、春兰等国兰44个品种为试验材料，对240对杂交兰EST-SSR标记引物的通用性进行研究。在此基础上，通过筛选的引物对国兰的遗传多样性进行分析。结果 杂交兰240对EST-SSR引物在国兰中的通用性比例分别为春兰（67.92%）、建兰（66.25%）、墨兰（71.25%）。利用其中的17对引物对44份国兰品种资源进行分子系统发育研究，有10对引物表现出多态性，10对引物共扩增出52个条带，多态性条带数为3～9个，多态性信息含量分布范围为0.504～0.850。聚类分析结果表明，44个国兰品种可分为4大聚类群，分类结果与传统的植物学分类相吻合。结论 本研究发掘的杂交兰EST-SSR引物可以有效用于国兰的分子遗传多样性研究。Abstract:Objective Cymbidium hybrid EST-SSR markers were studied to determine their transferability for applications on the cultivar identification and germplasm utilization of Chinese cymbidium cultivars.Method Genetic diversity of 240 pairs EST-SSR primers in 44 cultivars of Cymbidium goeringii, C. ensifolium, C. sinense and others were analyzed, and their transferability among Chinese cymbidium cultivars examined.Result The rates of transferability were found to be 67.92% for C. goeringii, 66.25% for C. ensifolium, and 71.25% for C. sinense. Among the markers, 17 EST-SSR primer pairs were selected to study the molecular phylogeny of the 44 cultivars. Out of them, 10 primer pairs showed polymorphisms with expanded 52 bands. The bands numbered between 3 to 9 with a range of polymorphism information content between 0.504 and 0.850. These cultivars were clustered into 4 groups, same as concluded by the conventional botanical classification.Conclusion The Cymbidium hybrid EST-SSR markers selected from this study effectively showed the molecular genetic diversity of the various Chinese cymbidium cultivars.
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Keywords:
- Cymbidium hybrid /
- EST-SSR marker /
- Chinese cymbidium /
- transferability /
- polymorphism
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表 1 国兰供试材料信息
Table 1 Information on Chinese cymbidium
cultivars 编号
No.品种名
Variety name种类
Species1 锦旗 Jinqi 建兰 C. ensifolium 2 四季玉妃 Sijiyufei 建兰 C. ensifolium 3 四季虎斑 Sijihuban 建兰 C. ensifolium 4 宝岛仙女 Baodaoxiannv 建兰 C. ensifolium 5 市长红 Shizhanghong 建兰 C. ensifolium 6 素君荷 Sujunhe 建兰 C. ensifolium 7 新品荷 Xinpinhe 建兰 C. ensifolium 8 红荷冠 Hongheguan 建兰 C. ensifolium 9 花叶建兰 Huayejianlan 建兰 C. ensifolium 10 春剑大富贵 Chunjiandafugui 春剑 C. longibractium 11 红荷梅 Honghemei 春兰 C. goeringii 12 红荷 Honghe 春兰 C. goeringii 13 碧龙玉素 Bilongyusu 莲瓣兰 C. lianpan 14 碧玉奇素 Biyuqisu 莲瓣兰 C. lianpan 15 晃辉 Huanghui 建兰 C. ensifolium 16 大唐盛世 Datangshengshi 春兰 C. goeringii×莲瓣兰 C. lianpan 17 瑞梅 Ruimei 春兰 C. goeringii 18 高山春色 Gaoshanchunse 建兰 C. ensifolium 19 国色天香 Guosetianxiang 建兰 C. ensifolium 20 铁骨素梅 Tiegusumei 建兰 C. ensifolium 21 铁金刚 Tiejingang 建兰 C. ensifolium 22 马耳 Maer 建兰 C. ensifolium 23 红梅 Hongmei 建兰 C. ensifolium 24 素心爪 Suxinzhua 建兰 C. ensifolium 25 地荷 Dihe 建兰 C. ensifolium 26 金丝马尾爪 Jinsimaweizhua 建兰 C. ensifolium 27 金针 Jinzhen 建兰 C. ensifolium 28 桃娇 Taojiao 建兰 C. ensifolium 29 小桃红 Xiaotaohong 建兰 C. ensifolium 30 萨摩锦 Samojin 建兰 C. ensifolium 31 青山玉泉 Qingshanyuquan 建兰 c. ensifolium 32 凤 Feng 建兰 c. ensifolium 33 丹霞建 Danxiajian 建兰 C. ensifolium 34 大凤素 Dafengsu 建兰 C. ensifolium 35 达摩爪 Damozhua 墨兰 C. sinense 36 龙梅 Longmei 墨兰 C. sinense 37 四季达摩 Sijidamo 墨兰 C. sinense 38 红宝石 Hongbaoshi 墨兰 C. sinense 39 玉兰蔻 Yulankou 墨兰 C. sinense 40 贵夫人 Guifuren 墨兰 C. sinense 41 黄荷梅 Huanghemei 春兰 C. goeringii 42 金边报岁兰 Jinbianbaosuilan 墨兰 C. sinense 43 银托 Yintuo 墨兰 C. sinense 44 企黑 Qihei 墨兰 C. sinense 
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表 2 杂交兰EST-SSR标记多态性分析
Table 2 Information on Cymbidium hybrid EST-SSR primer pairs
引物名
Primer引物序列
Primer sequence长度
Length/bp重复单元
Repeat motifs总条带
Total band多态性条带
Polymorphism band多态性比率
Polymorphism percentage/%多态性信息含量
Polymorphism information contentCYM81 5′-CTTCCTTCTCTGCTGCCATT-3′
5′-AAACAGAATCCGGCTCACAC-3′181 （CT）8 9 9 100.00 0.850 CYM107 5′-CGGTGGGATAAGGCGTATAA-3′
5′-TTGATTCCGGCAATTAAAGC-3′201 （GA）8 6 6 100.00 0.765 CYM108 5′-AAAGATGACACTGTGCGTCG-3′
5′-CCTCGCCCACTCTAACTTGA-3′189 （GA）8 7 6 85.71 0.807 CYM128 5′-ATGCAAGGGCGTACAAAAAC-3′
5′-GAATCTCCGATCCCGTAACA-3′239 （GT）8 5 5 100.00 0.692 CYM153 5′-CGAGCGAGATATGTGGATGA-3′
5′-GGCATCCTTTGTACAATTTTGA-3′243 （TA）8 5 5 100.00 0.744 CYM165 5′-CCATCGCAATGTCTTTCCTT-3′
5′-AACAATGGGGTCACTCCCTA-3′183 （TA）6 3 3 100.00 0.606 CYM174 5′-CAAGATCAGCTGGCATTGAA-3′
5′-TTGGACAACCTGTCTCTATCCA-3′270 （TC）9 4 3 75.00 0.504 CYM270 5′-ATACCCACTGCCATAGCTGC-3′
5′-GGATTACGTCATCGGAGGAA-3′223 （ACC）5 4 3 75.00 0.652 CYM278 5′-AGGCACATAGGAGAGCCTGA-3′
5′-CTGAGCAGGAACTTGAAGCC-3′269 （AGA）6 4 3 75.00 0.570 CYM287 5′-CATCAACGCGGTGTATGAAC-3′
5′-CCGAGATTTGAGTGTCGGAT-3′198 （AGC）8 5 5 100.00 0.669
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