摘要
为探究水稻嗜水小核菌中发现的病毒与真菌的相互作用以及明确其是否可用于病原真菌的生物防治,采用修改的单引物拉增法(M-SPAT)对嗜水小核菌[HZ11]菌丝体和病毒粒子(VPL)中分离的10条双链RNA(dsRNA)片段的RNA5和RNA8进行克隆,并对这些克隆产物进行测序和分析。结果表明,dsRNA5的长度为2038 bp,编码RNA依赖RNA聚合酶(RdRp);dsRNA8的长度为1809 bp,编码外壳蛋白(CP);系统进化分析表明,dsRNA5蛋白和dsRNA8蛋白分别与双分病毒科α病毒属的RdRps和CP高度相近,这两条ds RNA的5'-非翻译区(UTR)包含保守序列5'-GAA(G)T-3'、5'-TTTTCTA(/CG)CCTC(/A)GATAT(A)AAATA(CG)GAA-3'和5'-AAACG(A)AA(G)TC(A)TTACCTCT-3';3'-UTR包含保守序列5'-AAAAAAAA-3',5'-AAAAAG(A)AAAAAAAAAAAAC(A)4A-3'和5'-AAA-3'。表明ds RNA5和dsRNA8属于一种新的α病毒的基因组,推定的新病毒被暂命名为嗜水小核菌病毒3(ShV3);嗜水小核菌分离物dsRNA5和dsRNA8分别对应于ShV3的dsRNA1和dsRNA2。
[Objective]In order to explore mycoviruses in Sclerotium hydrophilum(S.hydrophilum),conserve them as biological resources,provide a foundation for studying the interaction between fungi and mycoviruses,and apply mycoviruses to biocontrol,we conducted a study to detect new viruses in S.hydrophilum[HZ11]mycelia.[Method]The modified single-primer amplification technique(M-SPAT)was used to clone RNA5 and RNA8 from 10 double-stranded RNA(dsRNA)segments in S.hydrophilum[HZ11]mycelia and virus-like particles(VPLs)isolated from mycelia.The cloned products were sequenced and analyzed.[Result]dsRNA5 was 2038 bp in length and coded for a predicted RNA-dependent RNA polymerase(RdRp).dsRNA8 was 1809 bp in length and coded for a predicted coat protein(CP).Phylogenetic analysis indicated that the dsRNA5 protein and dsRNA8 protein were highly close to RdRps and CP of viruses in the genus Alphapartitivirus,family Partitiviridae,respectively.The 5′-untranslated regions(UTRs)of the two dsRNAs contained the conserved sequences 5′-GAA(G)T-3′,5′-TTTCTA(/CG)CCTC(/A)GATAT(A)AAATA(CG)GAA-3′,and 5′-AACG(A)AA(G)TC(A)TTACCTCT-3′.The 3′-UTRs of the two dsRNAs contained the conserved sequences 5′-AAAAAAA-3′,5′-AAAAAAG(A)AAAAAAAAC(A)AAAA-3′,and 5′-AAAAAAAAA-3′.[Conclusion]We suggested that dsRNA5 and dsRNA8 belonged to the genome of a new Alphapartitivirus.The putative new virus was designated to be S.hydrophilum virus 3(ShV3).dsRNA5 and dsRNA8,named from the S.hydrophilum isolation,correspond to dsRNA1 and dsRNA2 of ShV3,respectively.
作者
陈宇辰
王春兰
谭欣宇
吴娟
郭珍珍
竺锡武
CHEN Yu-chen;WANG Chun-lan;TANG Xin-yu;WU Juan;GUO Zhen-zhen;ZHU Xi-wu(Institute of Agriculture and Biotechnology,Hunan University of Humanities,Science and Technology,Loudi 417000,PCR;Institute of Bioengineering,Zhejiang Sci-Tech University,Hangzhou 310018,PCR)
基金
Supported by Open Fund for Innovation Platform of Education Department in Hunan Province(18K100)。
