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剑麻斑马纹病菌EST—SSR标记开发与评价

材料写作网    时间: 2020-08-01 04:22:31     阅读:次     手机站

摘 要 剑麻斑马纹病是剑麻生产中具有破坏性的一种真菌病害。本研究从GenBank(http://www.ncbi.nlm.nih.gov/nucest)中随机下载10 525条寄生疫霉EST序列,经过去冗余处理后得到5 519条无冗余EST。利用MISA(MIcroSAtellite identification tool)软件进行SSR发掘,从中共搜索到199个1~6碱基SSR,其中三核苷酸重复基元类型最多,共鉴定到55个,占SSR总数的27.6%;其次是二核苷酸重复基元类型和单核苷酸重复基元类型,分别鉴定到51和47个,各占所鉴定SSR总数的25.6%和23.6%。进一步分析表明,AG/CT二核苷酸重复基元类型为优势重复类型,占二核苷酸SSR总数的76.5%。而AAG/CTT和AGC/CTG 2个基元类型在三核苷酸中出现频率最多,分别为15和11次,分别占三核苷酸SSR总数的27.3%及20.0%。从鉴定的199个SSR中,选取含有SSR的合适区段设计了22对引物,并通过18个剑麻斑马纹病菌菌株的基因组DNA进行了评价,结果只有其中9对引物能从剑麻斑马纹病菌基因组DNA中有效扩增,其扩增移效率为40.9%。由此表明,利用此方法来开发剑麻斑马纹病菌的分子标记具有可行性,只是存在一个转移效率问题。

关键词 剑麻斑马纹病菌;表达序列标签(EST);SSR;分子标记

中图分类号 Q78;S563.8 文献标识码 A

Abstract Zebra spot disease caused by Phytophora nicotianae var. parasitica(or Phytophora parasitica var. nicotianae)is the most serious and devastating fungi disease in sisal. In the present study, a total of 5 519 non-redundant P. nicotianae were generated from the 10 525 publicly available EST sequences by CAP3 software, which used to mine potential microsatellites by microsatellite identification tool(MISA). A total of 199 SRR was found in P. nicotianae expressed-squence tags data. Of the 199 SSR, the most abundant SSR was the tri-nucleotide type, with the SSR numbers being 55, accounting for 27.6%, followed by the di-nucleotide and mono-nucleotide type, with the SSR numbers being 51 and 47, accounting for 25.6% and 23.6%. Among the di -nucleotide repeats, AG/CT was the most motifs and accounted for 76.5%....



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