Ramalakshmi Oviya Iyappan, Piramanayagam Shanmughavel
Bioinformation. 2010 Nov 27;5(6):240-3. doi: 10.6026/97320630005240.
With the ever increasing number of Expressed Sequence Tags (ESTs) from various sequencing projects, ESTs have become valuable and first-hand source of in-silico mining of simple sequence repeats (SSR) markers. We examined a total of 3419 EST sequences from three bamboo species, namely, Phyllostachys edulis, Bambusa oldhamii and Dendrocalamus sinicus for the presence of di- to hexa- microsatellites. The frequency of SSR containing ESTs varied from 5.36% in B. oldhamii to 13.05% in P. edulis. No SSRs were found in D. sinicus. Tri-nucleotide repeats (49.34%) were most frequent in P. edulis, while not much comparable difference in repeats was found in B. oldhamii. Flanking primer pairs were also designed in-silico for the sequences containing SSRs and their position on the genome hypothesized using similarity searching. SSRs located in open reading frame (ORF) were given functional annotation using Gene Ontology. Polymorphic SSRs were also detected using new pipeline- polySSR. Polymorphism level was very low (2.43%) and the position of the polymorphic SSRs was determined. The development of SSRs and the study of polymorphism will help in the further study of intra- and inter- gene flow, genetic structure, variability, linkage mapping and evolutionary relationships in bamboo.
随着来自各种测序项目的表达序列标签(EST)数量不断增加,EST已成为通过电子挖掘获取简单序列重复(SSR)标记的宝贵且第一手的资源。我们检查了来自三种竹子的总共3419条EST序列,这三种竹子分别是毛竹、绿竹和巨龙竹,以查找二至六核苷酸微卫星的存在情况。含有SSR的EST的频率在绿竹中为5.36%,在毛竹中为13.05%。在巨龙竹中未发现SSR。三核苷酸重复(49.34%)在毛竹中最为常见,而在绿竹中重复情况的差异不大。还通过电子方式为含有SSR的序列设计了侧翼引物对,并使用相似性搜索推测它们在基因组上的位置。使用基因本体论对位于开放阅读框(ORF)中的SSR进行了功能注释。还使用新的流程——polySSR检测了多态性SSR。多态性水平非常低(2.43%),并确定了多态性SSR的位置。SSR的开发以及多态性研究将有助于进一步研究竹子的基因内和基因间流动、遗传结构、变异性、连锁图谱绘制以及进化关系。