甘蔗杂交种子传播甘蔗花叶病毒、高粱花叶病及黄叶病的检测(英文)

Detection of Sugarcane mosaic virus,Sorghum mosaic virus and Yellow leaf virus transmission through sugarcane hybrid seeds

【目的】探究甘蔗杂交种子对甘蔗花叶病毒、高粱花叶病及黄叶病的传播性,为甘蔗抗病育种提供参考。【方法】采集父本或母本感染花叶病或黄叶病的甘蔗杂交种子样品10份,分别以SCMV、SrMV和SCYLV特异性引物进行一步法RT-PCR扩增、克隆和测序分析。对田间实生苗进行生物学观察,并采用RT-PCR检测其叶片SCMV、SrMV和SCYLV携带性。【结果】经特异性引物检测,10份杂交种子中仅有样品H4显示SrMV阳性,扩增的特异片段大小为828bp,其核苷酸序列与GenBank中已报道的SrMV基因序列同源性达99%。聚类分析结果表明,样品H4的SrMV和GenBank中SrMV外壳蛋白序列的自举水平达100%,证实H4样本感染的病毒为SrMV。田间连续45d观察均未发现出现病毒症的甘蔗幼苗;经RT-PCR检测,均未在10个样品实生苗叶片中发现此3种病毒。【结论】甘蔗种子可携带高粱花叶病(SrMV),但未能通过种子传播至实生苗。

[Objective ]The present study was conducted to determine the transmission of sugarcane pathogenic virus viz., Sugarcane mosaic virus （SCMV）, Sorghum mosaic virus （SrMV） and Sugarcane yellow leaf virus （SCYLV） through sugarcane hybrid seeds, in order to provide references for sugarcane disease-resistant varieties breeding. [Method]Ten samples of sugarcane hybrid seeds were collected from parent plants which infected by SrMV, SCMV or/and SCYLV in Hainan Breeding Base, China. The seeds were detected by one-step RT-PCR with species-specific primers of SCMV, SrMV and SCYLV, respectively. Amplified fragments were cloned with pGEM^-T Easy and sequenced. The seedlings de- rived from hybrid seeds were planted for further symptomatic observation and virus detection of SCMV, SrMV and SCYLV. [Result]The results showed that only one sample H4 showed SrMV-positive in 10 samples, and a 828 bp of specific fragment was obtained and submitted to GenBank （accession number KM999539）. Sequence aligament results showed that the nucleotide sequence of fragment in sample H4 had a 99% homology with reported SrMV gene from Gen- Bank. It found that the strain H4 was clustered with all coat protein sequences of SrMV from GenBank at 100% boot- strap level, which confirmed that sample H4 was infected by SrMV. For symptomatic observation of 45 days after seeds germination, no any viral disease symptoms could be found in leaves of seedling derived from hybrid seeds. RT-PCR detection showed that, all of 10 samples were virus-negative. [Conclusion]The results suggested that SrMV could be transmitted from sugarcane plant to seed, but failed to spread from seeds to seedlings.