梨砧木主要病毒RT-PCR检测
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摘要
砧木作为果树病毒传播的主要载体之一,其病毒检测研究亟待开展。本研究以梨nad5基因作为内标系统,利用RT-PCR技术对青岛农业大学砧木资源圃保存的52份梨砧木资源进行了病毒检测。研究发现梨砧木中均可以检测到1种以上病毒,但不同病毒对梨砧木的侵染率存在差异,ASGV(苹果茎沟病毒)、ASPV(苹果茎痘病毒)、ASSVd(苹果锈果类病毒)、ACLSV(苹果褪绿叶斑病毒)四种病毒的携带率分别为100%、15.4%、3.8%、0%。对检测到的ASGV和ASSVd病毒片段序列分析证明了该病毒检测方法的准确性。本研究为梨砧木病毒检测和产业化应用奠定了基础。
It was urgent to develop the detection technique of virus in pear rootstock,that might serve as an important vehicle for virus transmission.In this study,the nad5 gene was used as an internal standard gene,virus of 52 pear rootstock germplasm resources or varieties,which were preserved in resources nursery of pear rootstock of Qingdao agricultural university,were detected by using RT-PCR technique.The result indicated that pear rootstock was infected at least one virus,and differences of virus infection rate was found in different pear rootstock.Rate of virus carrier of ASGV,ASPV,ASSVd and ACLSV was100%,15.4%,3.8%and 0%respectively.And the results also were proved by sequence analysis of ASGV and ASSVd.This study laid a foundation for detection virus and industrialization applications of pear rootstock in the future.
It was urgent to develop the detection technique of virus in pear rootstock,that might serve as an important vehicle for virus transmission.In this study,the nad5 gene was used as an internal standard gene,virus of 52 pear rootstock germplasm resources or varieties,which were preserved in resources nursery of pear rootstock of Qingdao agricultural university,were detected by using RT-PCR technique.The result indicated that pear rootstock was infected at least one virus,and differences of virus infection rate was found in different pear rootstock.Rate of virus carrier of ASGV,ASPV,ASSVd and ACLSV was100%,15.4%,3.8%and 0%respectively.And the results also were proved by sequence analysis of ASGV and ASSVd.This study laid a foundation for detection virus and industrialization applications of pear rootstock in the future.
引文
[1]中华人民共和国国家统计局.中国统计年鉴-2017(M).北京:中国统计出版社,2017
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[2]Hancock J,Lobos G.Pears(M).In:Hancock J.F.(eds)Temperate Fruit Crop Breeding.Springer,Dordrecht,2008,299-337
[3]王国平,洪霓,张尊平,等.我国北方梨产区主栽品种病毒种类的鉴定研究[J].中国果树,1994(2):1-4
[4]牛建新,马兵钢,何梅,等.库尔勒香梨主要病毒多重RT-PCR检测技术研究[J].植物病理学报,2006,36(01):14-23
[5]邓晓云,王国平.梨病毒病研究新进展[J].果树学报,2002,19(5):321-325
[6]王国平,洪霓.果树病毒检测与脱除技术的研究进展[J].武汉:华中农业大学学报,2004,23(6):685-691
[7]王利平.两种线状病毒分子变异的研究及热处理对梨基因表达的影响[D].武汉:华中农业大学,2009
[8]王钰婷.苹果和梨树的主要类病毒检测及序列分析[D].石河子:石河子大学,2013
[9]沙守峰,张绍铃,李俊才.梨矮化砧木的选育及其应用研究进展[J].北方园艺,2009(8):140-143
[10]蔡忠民,李俊才,王家珍,等.梨矮化砧木研究进展[J].北方果树,2013(5):1-4
[11]梁成林,赵玲玲,宋来庆,等.几种苹果实生砧木种子传毒潜力检测[J].果树学报,2014,31(6):1164-1169
[12]唐科志.柑桔碎叶病毒RT-PCR检测技术研究及应用[D].重庆:西南大学,2006
[13]甘琴华,王英超,魏晓棠,等.法国进境葡萄砧木中啤酒花矮化类病毒的检测与序列分析[J].植物保护学报,2014,41(3):346-351
[14]刘聪利,李明,赵改荣,等.樱桃病毒病脱毒检测技术及抗病毒基因工程研究进展[J].果树学报,2014(s1):7-13
[15]张绍铃.梨学[M].第一版.北京:中国农业出版社,2013,599-619