针对严重威胁烤烟大田生产的赤星病病害,培育抗病品种。以烤烟品种NC89为受体,具有选择标记性状的CV87为供体,采用浸种法将供体总DNA导入受体,获得具有目的性状的Do代变异株收种并种植后得到D1代,对其D1代进行实验检测结果表明浸种法直接导入外源DNA在D1代,仍可有效转移株高,株型,叶色,叶面积,生育期及抗病性等性状,并获得较高转化率D1代为15％。导入后D1代成熟烟叶总糖与蛋白质含量变异株与对照基本一致,说明变异株基本保持了受体原有的优良品质。蛋白质聚丙烯酰胺凝胶电泳检测,表明导入D1代变异株出现了受体没有的而供体所特有的两条谱带,Rr0.19和Rr0.63。具有供体特有的两条谱带的D1代变异株有A14B6.C5C10D3等5个优良株系。经过D1代一系列检测和筛选,选出5株早熟,优质,抗病变异株有A14.B6.C5.C10.D3。为今后进行D2代筛选,通过分子验证,为烟草育种创造新种质,开创新途径。 In view of the serious threat to the production of flue-cured tobacco leaf spot disease, cultivate disease-resistant varieties. Using CV87 with selectable marker traits as the donor, the total DNA of the donor was introduced into the recipient by seed soaking, and the Do-like mutants with the trait of interest were seeded and planted to obtain D1 generation. The results of D1 test showed that direct introduction of exogenous DNA by soaking seeds can still effectively transfer plant height, plant type, leaf color, leaf area, growth period and disease resistance in D1 generation and achieve higher transformation rate D1 On behalf of 15%. The total sugar and protein content of D1 mature tobacco leaves after induction were basically consistent with those of the control, indicating that the mutants basically maintained the original excellent quality of the recipient. Protein polyacrylamide gel electrophoresis showed that there were two donor-specific donor-specific bands, Rr0.19 and Rr0.63, introduced into the D1 generation. There are 5 excellent lines such as A14B6.C5C10D3 in the D1 generation with donor-specific two bands. After a series of tests and screening of D1 generation, five early-maturing, high-quality and disease-resistant mutants were selected as A14.B6.C5.C10.D3. D2 generation screening for the future, through molecular verification, create new germplasm for tobacco breeding, open up new avenues.