应用浸种法导入CV85DNA转化烤烟遗传性状变异D_2代研究  

Studies On the Genetic Mutation of D2 Generation Tobacco Transformed with CV85 DNA by Seed-Immersion Method

作　　者：杨丽娟[1] 张建光[1] 史芝文[1] 

机构地区：[1]东北农业大学生命科学学院

出　　处：《农业与技术》2002年第6期69-74,79,共7页Agriculture and Technology

摘　　要：提取抗赤星病烤烟CV85的总DNA,采用浸种法导入感赤星病烤烟NC89。D1代筛选出的优良变异株收种并种植后得到D2代植株,对其活动体接种赤星病菌,检测其抗病性,并于接曲前后测定叶片过氧化物酶活性:田间生长性状调查,对其成熟烟叶总糖、蛋白质及总氮含量进行分析,并进行蛋白质聚丙烯酰胺凝胶电泳检测。结果表明:浸种法直接导入外源DNA,在变异株D2代仍可有效转移株高、株型、叶型、叶面积及抗病性等性状;赤星病菌人工接种检抗病性和过氧化物酶活性测定,筛选出高抗赤星病株系Jz9802;经蛋白质SDS-PAGE谱带分析表明,变异株D2代Jz9802出现了受体没有的而供体特有的4条谱带,RrA10.07、A20.1 5、B50.59、C40.87:变异株Jz9802的总糖、蛋白质和总氮与受体基本一致,保持了受体原有的优良品质。The total DNA of resistant red star disease from tobacco CV85 was transformed into Tobacco NC89 which sensitive to red star disease through seed-immersion method. D2 generation was obtained from the cultivar which screened from the target characteristics D1 generation mutant. The living was inoculated with red star pathogen and inspected its resistance, the enzyme activity of leaves was tested around inoculated: Surveyed the quality of field growth, analyzed its total sugar, protein and nitrogen content of mature tobacco leaves and tested by SDS-PAGE. The result indicated that: the phenotypes such as height, type, leaves, leaf area and disease-resistance etc were still effectively transferred into D2 generation through seed-immersion DNA transformation: The high resistance of red star plant Jz9802 was screened by tested resistance and enzyme activity from plant of artificial inoculated red star pathogen: and the D2 generation mutant plant Jz9802 exhibited four specific bands, named RfAl 0.07, A2 0.15, B5 0.59, C, 0.87, that only belong to the donar plants, by SDS-PAGE analysis; The total sugar、protein and nitrogen content of mature tobacco leaves were similar between the mutant plant Jz9802 and the receptor plants, it is to say that the mutants still reserved the good characters of the receptors.

关 键 词：浸种法 CV85DNA 烤烟 遗传性状变异 D2代 赤星病 外源DNA导入 

分 类 号：S572[农业科学—烟草工业]