长春地区马铃薯纺锤块茎类病毒检测研究
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摘要
马铃薯纺锤块茎类病毒(PSTVd)是世界范围内影响马铃薯生产的主要病害之一。利用精准的检测技术对其进行检测可有效的控制病害的传播。RT-PCR技术具有快速、灵敏、简便、特异性强等优点,该技术可检测出植物组织中含量极低的病毒RNA,RT-PCR技术已经越来越广泛地应用在RNA病原鉴定上。通过RT-PCR技术在检测马铃薯纺锤块茎类病毒上的应用,可使马铃薯种薯在脱毒过程中杜绝病害的传播,提高马铃薯种薯质量,以达到提高马铃薯产量的目的。
     本文针对长春地区马铃薯主栽品种春薯4号,克新1号,克新2号,春薯3号,尤金,富金,延薯4号,东农303,春薯5号,大西洋和费乌瑞它11个品种,18份材料,根据马铃薯纺锤块茎类病毒基因序列,设计合成三对引物,以阳性对照、阴性对照和未知品种的植株叶片的cDNA为模板,进行RT-PCR扩增,检测马铃薯纺锤块茎类病毒,为RT-PCR检测方法在本区域的广泛应用提供技术指导。
     结果表明18份试验材料中Y217-2和09-Y217-1带有马铃薯纺锤块茎类病毒,其余品种:4-6,5-2,Y10-217,春5-7,F3,F-5,F-7,F15,(P),4-6苗,5苗,Y10-217苗,(P)苗,F-7苗,F15苗,F-7盘均不含有马铃薯纺锤块茎类病毒。且扩增出来的片段长度与预期目的片段大小相同。并证明植株感染病毒与培养时期和培养条件不存在相关性。
Potato spindle tuber viroid (PSTVd) is a main disease of potato in the world. Detection techniqueis an effective method to control diseases. RT-PCR has the advantage of rapid, sensitive, simple,specificity, etc. It can detect low content RNA in pant tissue and it has widely used in pathogenidentification. Using RT-PCR detect PSTVd can produce virus-free potato, increase potato quality andyield.
     In this paper,we collected Changchun area18samples include11varieties, including "Chunshu4","Kexin1","Kexin2","Kexin3","Youjin","Fujin","Yanshu4","Dongnong303","Chunshu5","Atlantic" and "Favorite". Based on the homologous nucleotide sequences of PSTVd3pairs of primerswere designed. Using the method of RT-PCR detect PSTVd, and provide technology instruction forChangchun area.
     The results showed that "Y217-2" and "09-Y217-1" were PSTVd positive, others were negative.Amplified fragment were highly homologous with PSTVd. There was no evidence show that they haverelationship of PSTVd infect with culture date and culture condition
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