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利用生物技术方法对棉花进行遗传改良主要限于有效的遗传转化系统。以新疆主栽优良陆地棉品种‘新陆早33号’为材料,利用下胚轴作为外植体对影响农杆菌介导的棉花遗传转化及体细胞胚胎发生的因素进行研究,成功建立了除草剂Basta筛选的棉花遗传转化技术体系。同时将植物抗病相关基因多聚半乳糖醛酸酶抑制蛋白基因AtPGIP1导入棉花,经过对再生转化植株的PCR鉴定,初步证明外源基因已经整合到棉花基因组。研究发现:Basta是棉花遗传转化中很有效的筛选剂,低浓度Basta(2.5mg/L)就能够获得很好的筛选效果;较低的共培养温度(20℃)及合适的农杆菌浓度(OD600=0.5)有助于提高转化效率。该研究结果表明,‘新陆早33号’具备作为棉花优良遗传转化受体的基本特征,研究中获得的15株AtPGIP1转基因植株经PCR分子检测均为阳性植株。该研究为新疆棉区棉花分子生物学研究及转基因育种研究奠定了重要基础。
The use of biotechnological methods for genetic improvement of cotton is largely limited to efficient genetic transformation systems. Taking Xinluzao 33, a new upland cotton cultivar in Xinjiang, as the material, we studied the factors influencing Agrobacterium-mediated cotton transformation and somatic embryogenesis by using hypocotyls as explants and successfully established weeds Basta Screening Cotton Genetic Transformation Technology System. At the same time, the plant disease-related gene polygalacturonase inhibitor protein gene AtPGIP1 was introduced into cotton. After PCR identification of the regenerated transformed plants, it was initially proved that the exogenous gene has been integrated into the cotton genome. The results showed that Basta was a very effective screening reagent for genetic transformation of cotton, and Basta (2.5 mg / L) could obtain a good screening effect. The lower co-culture temperature (20 ℃) and the suitable Agrobacterium concentration OD600 = 0.5) helps improve conversion efficiency. The results showed that ’Xinluzao 33’ had the basic characteristics as an excellent genetic transformation receptor for cotton. The 15 AtPGIP1 transgenic plants obtained in this study were positive by PCR. This study laid an important foundation for the research on cotton molecular biology and transgenic breeding in Xinjiang cotton area.