水稻黑条矮缩病毒P7-1基因对水稻的遗传转化及转基因植株的抗病性分析
收稿日期: 2018-01-05
修回日期: 2018-05-08
网络出版日期: 2018-07-10
基金资助
国家重点研发计划资助项目(2016YFD0300706);国家自然科学基金资助项目(31471768)
Genetic Transformation of Rice black streaked dwarf virus P7-1 Gene in Rice and Its Virus Resistance Analysis
Received date: 2018-01-05
Revised date: 2018-05-08
Online published: 2018-07-10
【目的】通过在水稻中过量表达水稻黑条矮缩病毒(Rice black streaked dwarf virus, RBSDV) P7-1基因,分析RBSDV P7-1是否是导致水稻不育的重要致病因子,并明确过量表达P7-1转基因植株对病毒的抗性。【方法】采用RT-PCR方法从感病植物中扩增获得RBSDV P7-1基因,构建pCAMBIA-1300-P7-1载体,通过农杆菌转化法获得转基因水稻,观察水稻是否能够正常结实,并采用人工接种病毒方法分析转基因植株的抗性。【结果】过量表达RBSDV P7-1的转基因水稻生长、结实正常。在接种7 d和14 d时转基因植株中病毒积累量显著低于野生型对照,但在28 d时部分转基因株系中病毒积累量高于对照,接种30 d时转基因植株的发病率与对照无明显差异。【结论】在水稻中过量表达RBSDV P7-1不影响水稻的结实;转基因水稻在RBSDV侵染早期可抑制病毒的积累,但在后期对病毒的抗性与对照没有明显差异。
袁平平, 孙枫, 倪海平, 朱佳慧, 周益军, 蒋选利, 徐秋芳 . 水稻黑条矮缩病毒P7-1基因对水稻的遗传转化及转基因植株的抗病性分析[J]. 中国水稻科学, 2018 , 32(4) : 342 -348 . DOI: 10.16819/j.1001-7216.2018.8014
【Objective】The purpose of this study is to determine whether Rice black streaked dwarf virus (RBSDV) P7-1 is an important pathogenic factor related to infertility in rice and to analyze the resistance of the P7-1 transgenic rice to RBSDV by overexpression of P7-1 in rice. 【Method】 RBSDV P7-1 gene was amplified from virus-infected plants by RT-PCR. The recombination plasmid pCAMBIA-1300-P7-1 was constructed and transformed to rice by Agrobacterium transformation. The fertility of transgenic plants was evaluated and the resistance of the transgenic rice plants to RBSDV was identified by artificial inoculation of virus. 【Result】The transgenic plants showed normal growth and fertility. The virus accumulation in the transgenic plants was significantly lower than the wild type at 7 days and 14 days after inoculation, but it was higher in some transgenic lines than the control at 28 days after inoculation. There were no significant differences in disease incidence between the transgenic plants and the wild type at 30 days after inoculation. 【Conclusion】 Overexpressing RBSDV P7-1 in rice does not affect the fertility. Transgenic rice had lower accumulation of virus in the early stage after RBSDV infection, but did not exhibit resistance to the virus at the later stage.
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