摘要
以马铃薯X病毒(potatovirusX,PVX)的RNA为模板,应用反转录聚合酶链式反应(RT PCR)方法分别扩增出长度为681bp的非翻译马铃薯X病毒25kD运动蛋白基因(PVX p25)和长度为714bp的非翻译马铃薯X病毒外壳蛋白基因(PVX CP)。并分别构建植物表达载体pROKⅡp25和pROKⅡCP。利用农杆菌介导方法转化烟草NC89。经卡那霉素筛选、PCR检测,共获得转非翻译PVX p25的转基因植株78株,转非翻译PVX CP的转基因植株83株。抗病性试验表明,转非翻译PVX p25的78株中有31株对PVX的侵染表现高度抗病,抗性比例为39.7%;转非翻译PVX CP的83株中有11株对PVX的侵染表现高度抗病,抗性比例为13.3%。分子生物学检测和抗病性分析表明,抗病性均为RNA介导的病毒抗性。研究结果初步证明,转非翻译PVX p25可以更有效地获得抗PVX转基因植株。
According to the published nucleotide sequence of potato virus X (PVX), the non-translational movement protein gene (681 bp in length) and the non-translational PVX coat protein (CP) gene (714 bp in length) of PVX were synthesized by reverse transcription-polymerase chain reaction (RT-PCR). The synthesized cDNAs were then introduced into the plant expression vector pROKⅡ. The recombinant binary vectors of pROKⅡ-p25 and pROKⅡ-CP were introduced into (tobacco) (NC89) via Agrobacterium tumefaciens-mediated transformation method. The transformed tissues were selected in the presence of Kanamycin, and the regenerated plants were screened by PCR. Seventy-eight and eighty-three plants transformed with PVX-p25 and PVX-CP, respectively, were obtained. Resistance test indicated that 31 and 11 of the plants transformed with PVX-p25 and PVX-CP, respectively, were highly resistant to PVX infection. The proportions of disease resistance were 39.7% and 13.3%, respectively. Molecular evaluation and disease resistance assay demonstrated that the resistance mediated by both cDNAs was RNA-mediated virus resistance. The results also imply that PVX-p25 is more effective in evoking RNA-mediated resistance than PVX-CP in transformed plants.
出处
《作物学报》
CAS
CSCD
北大核心
2005年第7期827-832,共6页
Acta Agronomica Sinica
基金
国家自然科学基金(30270875)。
