摘要： 通过改进外植体处理方法、添加乙酰丁香酮提高外植体芽分化频率、筛选芽分化培养基中激素浓度和确定转化后再生体系的筛选物质及其使用浓度等方法建立和优化了马铃薯脱毒微型种薯的根癌农杆菌遗传转化体系。 利用该体系将水稻类甜蛋白基因和水稻几丁质酶基因导入津引薯8号脱毒微型种薯中,并获得转基因株系。通过 PCR和southern杂交分子检测,证明ZGY-298,ZGY-283和ZGY-301 3个株系均整合了这2个基因。采用晚疫病主要生理混合小种温室活体接种和离体接种进行抗病性分析表明,在晚疫病发病高峰期(接种后5-6 d)。转基因株系对晚疫病抗性明显高于对照品种,并可使病情蔓延推迟2-3 d。
关键词:
马铃薯,
农杆菌,
转基因,
晚疫病,
抗病性
Abstract: To obtain transgenic potato varieties with high resistance to late blight disease from two ways of degenerating the cell walls and cell membranes by transferring both gene of rice tomautin-like protein and gene of rice chitinase 11 to virus-free potato minituber. An improved transformation system for agrobacterium mediated transformation of virus-free potato minituber was established using the tube slices as explants and optimized through the way of developing treatment method of explants, adding acetosyringone to the medium to promote the level of shooting, selecting the concentration of hormones in the shooting medium and determining the selective substances and their concentrations. Based on this system, rice thaumatin-like protein gene (TLP) and rice chitinase gene (CHI11)were transferred into virus-free potato minituber named Jinyinshu No.8 and transgenic plants which were detected by PCR and southern blot were obtained. By the greenhouse vivo inoculation and vitro inoculation using mix of outbreak and main physiological races of phytophthora infestans resistances were measured and disease developing was analyzed. It was proved that TLP and CHI11 were combined with three strains of ZGY-298,ZGY-283 and ZGY-301.Resistant experiments showed that transgenic plants significantly expressed higher resistance than control plaints and delaying the development of disease symptoms especial1y during the high peak period of disease (5-6 d after inoculation).For transgenic plants could delay the development of disease symptoms 5-6 d, It is hopeful to control the disease especially in the high peak period of disease.
Key words:
Potato,
Agrobacterium,
Transgenic,
Late-blight,
Disease resistance
中图分类号:
金红, 罗智敏, 陈峥, 李戍彤, 王永, 魏众济, 程奕. 马铃薯抗晚疫病基因工程育种研究[J]. 华北农学报, 2005, 20(1): 46-51. doi: 10.3321/j.issn:1000-7091.2005.01.011.
JIN Hong, LUO Zhi-min, CHEN Zheng, LI Shu-tong, WANG Yong, WEI Zhong-ji, CHENG Yi. Studies on Breeding Varieties of Potato Resistant to Potato Late-blight by Genetic Engineering Techniques[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2005, 20(1): 46-51. doi: 10.3321/j.issn:1000-7091.2005.01.011.