中国野生大豆中大豆花叶病毒的鉴定及演化分析

发布时间：2018-02-21 09:16

  本文关键词： 野生大豆 大豆花叶病毒 全基因组 演化　出处：《南京大学》2017年硕士论文　论文类型：学位论文

【摘要】：大豆花叶病毒(Soybean mosaic virus,SMV)是一种世界性的大豆病害,严重影响大豆的产量和品质。目前,对该病毒的调查主要在栽培大豆中展开,而对中国野生大豆所携带的SMV种类和遗传多样性的研究尚不充分。本研究对2013年采集自中国东北和南方地区共60份野生大豆种子携带病毒的情况进行了调查。结果表明,其中12份野生大豆材料具有典型花叶、皱缩等症状。通过设计两对马铃薯Y病毒属(Potyvirus)兼并引物(PVYG-F1/R1和PVYG-F2/R2)进行RT-PCR检测及测序分析,这12个分离物均为SMV,并可以归类为9种不同的序列类型;对其中7种SMV类型,成功进行了全基因组测序。将这7条SMV全基因组新序列与NCBI数据库中的83条SMV基因组序列共同排序并构建系统演化树后,结果发现,分离于南方野生大豆材料中的4条SMV全基因组序列位于演化支H中,与已经报道的SMV分离物SC7、BYX006和LJZ002亲缘关系相近。这4个新发现的SMV分离物均属于重组型SMV,其5'端含有一段大约900 bp的菜豆普通花叶病毒(Bean common mosaic virus,BCMV)的基因序列。而分离于北方野生大豆材料中的3条SMV全基因组序列分别位于演化支Ⅲ和Ⅳ中。由此可知,中国野生大豆也是SMV的重要宿主,在SMV的传播过程中起着重要作用。本研究进而选取了南、北方SMV分离物SC7-N与HLJBADS001,对所采集野生大豆材料中代表性个体的繁殖后代进行了初步侵染实验。结果显示,33份样品抗HLJBADS001与SC7-N两个SMV分离物,26份样品只抗HLJBADS001。因此,野生大豆中蕴藏着丰富的抗SMV的资源。
[Abstract]:Soybean mosaic virus (Soy bean mosaic virus) is a worldwide soybean disease, which seriously affects the yield and quality of soybean. However, the SMV species and genetic diversity of wild soybean in China were not fully studied. In 2013, 60 wild soybean seed carriers from Northeast and South China were investigated. Twelve wild soybean materials had typical mosaic, shrinkage and other symptoms. By designing two pairs of potato Y virus degenerate primers, PVYG-F1 / R1 and PVYG-F2 / R2), RT-PCR detection and sequencing were performed. The 12 isolates are SMVs and can be classified into 9 different sequence types. The whole genome was sequenced successfully. The seven new SMV genome sequences were sequenced with 83 SMV genome sequences in the NCBI database and the phylogenetic tree was constructed. Four SMV complete genome sequences isolated from southern wild soybean were located in the evolutionary branch H. The four newly discovered SMV isolates all belong to recombinant SMVs, and their 5'ends contain a gene sequence of about 900bp common bean mosaic virus, common mosaic virusBCMV, which is similar to the reported SMV isolates SC7BYX006 and LJZ002. The four newly discovered SMV isolates all belong to the recombinant SMVs, and the 5 'end of the isolates contain the gene sequence of the common bean mosaic virus common mosaic virusBCMV (BCMV). Three SMV complete genome sequences isolated from northern wild soybean were located in evolutionary branch 鈪，

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