基于转录组测序信息的水生植物绒球百合SSR标记的开发

Yangchen Yuan, Qing‐Feng Wang, Jin‐Ming Chen
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引用次数: 3

摘要

利用MISA程序,从水生植物peltata Nymphoides的79536条est中共筛选出12319条est - ssrs。二核苷酸型和三核苷酸型是主要的基序类型,分别占57.31%和30.87%。二基序和三基序的丰度分别为AG/CT(29.76%)和AAG/CTT(8.66%)。随机选择130对引物,对其在两个种群中的应用和多态性进行了评价。结果表明,共扩增出78对SSR标记,获得了清晰可辨识的条带,其中37对标记存在多态性,多态性率为47.44%。利用多态性SSR标记,共检测到114个等位基因。每个位点的等位基因数为2 ~ 6个,平均3.08个。每个位点的观察杂合度(Ho)和期望值(He)分别在0.229 ~ 1.000和0.351 ~ 0.756之间变化。多态性信息含量(PIC)范围为0.286 ~ 0.698,平均值为0.495。结果表明,利用水生植物peltata下一代测序产生的EST数据集开发SSR分子标记是有价值和有效的。这些新生成的SSR标记将为研究大鼠茅的遗传多样性和群体遗传结构提供新的工具。
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Development of SSR Markers in Aquatic Plant Nymphoides peltata( Menyanthaceae ) Based on Information from Transcriptome Sequencing
Using the program MISA,a total of 12319EST-SSRs were sought out from 79536ESTs of aquatic plant Nymphoides peltata.Dinucleotide and trinucleotide SSRs were the two main motif types,accounting for 57.31% and 30.87%,respectively.The most abundant di-and tri-motifs were AG/CT(29.76%)and AAG/CTT(8.66%),respectively.We randomly selected 130primer pairs to evaluate their application and polymorphism across two populations of N.peltata.Seventyeight SSR markers were amplified successfully,yielding clear and discernible bands,among which37were found to be polymorphic and the polymorphic rate was 47.44%.Using the polymorphic SSR markers,a total of 114alleles were detected.The number of alleles per locus ranged from 2to 6(mean 3.08).Observed heterozygosity(Ho)and expected value(He)per locus varied from 0.229to 1.000and from 0.351to 0.756,respectively.Polymorphism information content(PIC)ranged from 0.286to 0.698,with an average of 0.495.The results indicated that the development of SSR molecular markers from the EST dataset generated by next generation sequencing in the aquatic plant N.peltata is valuable and effective.These newly generated SSR makers will provide new tools for studying genetic diversity and population genetic structure of N. peltata.
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