摘要
用根癌农杆菌Agrobacterium tumesfaciens介导法，将烟草β-1，3-葡聚糖酶Glucanase基因、苜蓿防御素alfAFP基因及二者双价基因导入番茄Micro—Tom中，以期获得抗灰霉病的转基因株系，并比较验证双价基因是否具有协同作用。绿色荧光蛋白基因（GFP）活性检测、PCR和Southem杂交检测表明，外源基因分别以1—4个拷贝整合到番茄基因组中。对T1代植株进行接种鉴定表明，转基因植株对灰霉病侵染的抵抗能力较对照明显增强，各株系的抗病能力与外源基因的表达相关，且葡聚糖酶基因和防御素基因对抗灰霉菌表现一定的协同作用。T1株系田间抗病鉴定结果表明，转基因株发病程度不同，病情指数在19—28之间，抗病性均高于对照。所得到的稳定表达的株系可作为抗病育种材料进一步研究。
The tobacco β-1, 3-Glucanase gene, the alfalfa defensin gene alfAFP and their bivalent gene GLU-AFP were introduced into tomato line Micro-Tom via Agrobacterium-mediated method. The aim is to gain transgenic lines resistant to Botrytis cinerea, and test the synergistical effects of bivalent gene. Transgenic lines carrying each of the transgenes were confirmed for integration into the tomato genome using GFP, PCR and Southern blot hybridization. One to four copies of the transgene were integrated into the tomato genome. Performance test of resistance analyses to B. cinerea with T1 generation transgenic tomato lines showed the transgenic lines exhibited higher resistance to the pathogens infected than that of the non-transgenic plants, and resistance levels were related to expression levels of the transgene, showing -effect. The transgenic tomato harboring GLU-AFP cassette showed the highest disease resistance. It suggested that the alfAFP and Glucanase gene have some synergistical effects on the resistance to B. cinerea. Performance test of resistance of T2 families with GLU-AFP gene showed that M3, Ms and ME0 had a resistance index of 19 to 22, which showed a high resistance level. Some independent lines with high disease resistance, low variability and stable expression of transgenes could be selected for the further studies and molecular breeding.
出处
《植物保护学报》
CAS
CSCD
北大核心
2006年第4期357-362,共6页
Journal of Plant Protection
基金
国家"863"计划资助项目(2001AA247010)
关键词
转基因番茄
葡聚糖酶基因
苜蓿防御素基因
灰霉病
Transgenic tomato
tobacco glucanase gene
alfalfa defensin gene
Botrytis cinerea
