| 高羊茅EST-SSR标记开发及遗传多样性分析 |
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| 引用本文： | 李家丽,高 雪,王秀华,石香雪,于鸿翔,赵 岩.高羊茅EST-SSR标记开发及遗传多样性分析[J].园艺学报,2016,43(12):2401-2411. |
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| 作者姓名： | 李家丽 高 雪 王秀华 石香雪 于鸿翔 赵 岩 |
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| 作者单位： | （1土肥资源高效利用国家工程实验室，山东农业大学资源与环境学院，山东泰安 271018；2作物生物学国家重点实验室，山东农业大学农学院，山东泰安 271018） |
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| 摘 要： | 采用从其他作物转移与利用自身EST序列设计两种方法为高羊茅(Festuca arundinacea L.)开发EST-SSR分子标记,并利用所开发标记对高羊茅品种(系)进行遗传多样性分析。利用来自小麦、大麦、玉米、高粱和水稻的732对EST-SSR在高羊茅上进行通用性分析,得到406个有效的扩增引物对(55.46%)。利用高羊茅EST数据库(Gen Bank/db EST)中63 853条EST序列进行微卫星序列的查找,共发现包含微卫星的EST序列420条,占整个EST总数的0.658%。其中三核苷酸基序出现频率最高(43.33%),次之为二核苷酸基序(33.57%)。二核苷酸基序以CT/GA出现频率最高(16.90%),三核苷酸基序以CAG/GTC出现的频率最高(10.63%)。进一步运用Primer 5.0软件设计了108个引物对,并交上海桑尼公司合成。PCR检测表明,92个引物对(85.19%)在高羊茅中均可以扩增出稳定清晰的带型。从498对(其中5种作物的406对,高羊茅的92对)有效扩增的EST-SSR引物中随机选取81对引物,对12个高羊茅品种(系)进行扩增。79个引物对显示出多态性(97.53%)。共检测到133个等位变异,平均每对引物有1.68个等位变异;多态性信息含量(PIC)值最高为0.66,最低为0.14,平均为0.48。聚类分析结果表明,12份材料在相似系数为0.61处可分为5大类:第Ⅰ类为‘爆发力’和‘法思’;第Ⅱ类为‘强劲’、‘家园’和‘翠碧A’;第Ⅲ类为‘可其思3号’和‘凌志’;第Ⅳ类为‘雅典娜2号’、‘美洲虎3号’、‘火凤凰’和‘TF160’;‘球道’单独为第Ⅴ类。
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| 关 键 词： | 高羊茅 EST-SSR 通用性 标记开发 遗传多样性 |
Development of EST-SSR Markers and Analysis of Genetic Diversity in Tall Fescue |
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| LI Jia-li,GAO Xue,WANG Xiu-hua,SHI Xiang-xue,YU Hong-xiang,ZHAO Yan,.Development of EST-SSR Markers and Analysis of Genetic Diversity in Tall Fescue[J].Acta Horticulturae Sinica,2016,43(12):2401-2411. |
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| Authors: | LI Jia-li GAO Xue WANG Xiu-hua SHI Xiang-xue YU Hong-xiang ZHAO Yan |
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| Institution: | （1National Engineering Laboratory for Efficient Utilization of Soil and Fertilizer Resources，College of Resources and Environment，Shandong Agricultural University，Tai’an，Shandong 271018，China；2State Key Laboratory of Crop Biology，College of Agronomy，Shandong Agricultural University，Tai’an，Shandong 271018，China） |
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| Abstract: | In this study，tall fescue EST-SSR markers were developed using two ways of transferring from other crops and designing from EST sequences of tall fescue. Genetic diversity of tall fescue was analysed by these markers. Transferability of 732 EST-SSRs primer pairs from wheat，barley，maize， sorghum and rice was analysed in tall fescue. Of which，406（55.46%）primer pairs showed effective amplification. Besides，in the 63 853 tall fescue EST sequences from GenBank/dbEST，420 EST sequences（0.658%）containing SSRs were found. Among these SSRs of tall fescue，trinucleotide motifs appeared to be the most abundant SSRs（43.33%），followed by dinucleotide（33.57%）. For the motifs of dinucleotide and trinucleotide，CT/GA（16.90%）and CAG/GTC（10.63%）were the most abundant motif respectively. One hundred and eight primer pairs were designed by Primer 5.0 software and synthesized by Shanghai Sunny Company. PCR analysis showed that，92 primer pairs had stable and distinctive bands in tall fescue. From EST-SSRs of 5 crops（406）and tall fescue（92），81 EST-SSR primers were selected randomly to detect the polymorphism in the 12 tall fescue cultivars（strain），and 79 primer pairs（97.53%）showed polymorphism with totally 133 alleles，1.68 alleles per primer in average. The PIC（Polymorphism information contents）for each polymorphic primer varied from 0.14 to 0.66，with an average of 0.48. Clustering analysis indicated that the 12 tall fescue cultivars were clustered into 5 groups based on 0.61 of GS. GroupⅠincludes Explosiveness and Fawn，group Ⅱ includes Powerful，Plantation and Triple A，group Ⅲ includes Kochis 3 and Lexus，group Ⅳ includes Athena 2，Jaguar 3，Exerion and TF160，group Ⅴ includes Fireway. |
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| Keywords: | tall fescue EST-SSR transferability marker development genetic diversity |
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