摘要
目的探明马铃薯Y病毒脉坏死株系(PVYN)CP基因3′端序列短片段的不同结构转基因诱导转基因植物产生RNA介导抗性的有效性。方法以PVYN的CP基因cDNA3′端202bp片段构建非翻译的正向重复和反向重复结构的植物表达载体转化烟草。结果攻毒试验表明前者没有一例转基因植株表现为抗病,而转化反向重复结构的转基因植株82.8%表现近似免疫的高度抗病型,Southern印迹杂交证明目的基因已整合到烟草基因组,Northern印迹杂交结果显示反向重复结构转基因植物的抗病性与RNA的表达量呈现负相关。结论抗病性是RNA介导的病毒抗性。3′端短片段诱导产生的转基因抗病株比例比5′端短片段诱导产生的高。
[ Objective]To determine and compare the induction efficiency of RNA silencing triggered by different construction transgene of 3′ -terminal segment derived from the Potato virus Y （veinal necrosis strain, PVY^N ） coat protein gene, [Method] Transgenic tobacco plants expressing untranslated direct repeats （DR） and inverted repeats （IR） of 3′ -terminal 202bp of the PVYN coat protein gene were obtained via Agrobacterium tumefaciens-mediated transformation system, [Result] Resistance assay of the transgenic plants indicated that none of the DR-transgenic plants are resistant to PVY, but 82.8% of IR-transgenic plants showed high resistance. Northern blot analysis revealed an inverse correlation between transgene transcript accumulation and virus resistance. [Conclusion] These results showed that the resistance was RNA-mediated. The proportion of the resistant transgenic plants induced by 3′ -terminal segment transgene ofPVYN is higher than that induced by 5′ -terminal segment.
出处
《中国农业科学》
CAS
CSCD
北大核心
2006年第6期1153-1158,共6页
Scientia Agricultura Sinica
基金
国家自然科学基金资助项目(30471139
30100117)
山东省优秀中青年科学家科研奖励基金资助项目(2004BS02006
2005BS02007)
