提高棉花花粉管通道技术转化率的研究

Study on Promoting the Rate of Pollen-tube Pathway Transformation in Cotton

利用花粉管通道技术将含有IPT基因的双元表达载体质粒 pBG121 导入不同的陆地棉品种(中棉所10号、中棉所24和中棉所36)中,采取正交试验设计,探讨基因型、导入时间、导入DNA的浓度和化学诱变剂的浓度对棉花转化率的影响,筛选出了适宜提高棉花花粉管通道技术转化率的方法。应用结果表明,改进方法的转化率比传统的方法提高两倍。

The plant bivalent expression vector pBG121 carried IPT gene was transformed into different cotton cultivars (CCRI 10, CCRI 24, CCRI 36) using pollen-tube pathway method. Through orthogonal experiment, we discussed the effects of genotype, introduction time (after pollination), DNA concentration and EMS concentration on transformation rate in cotton. The orders of importance on the transformation rate was: EMS concentration > acceptor genotype > DNA concentration > introducing time. Correlated with transformation rate, the acceptor genotype and the EMS concentration had significant differences between different treatments; the DNA concentration and the introducing time didn't make significant differences among different treatments. So an optimal condition, in which the transformation rate was promoted, was obtained: the genotype was CCRI24, the DNA concentration was 0.01 g·L^(-1), the EMS concentration was 0.01 g·L^(-1), the introducing time was 36 hours after pollination. In order to verify the function of the senescence-inhibition chimeric gene and the optimal transformation condition that was screened, we selected CARI10 as the acceptor genotype, introduced 0.01 g·L^(-1) DNA and 0.01 g·L^(-1) EMS at 36 hours after pollination. Another treatment (CK) was the same as above except EMS. The CK had buffer of the same volume as EMS in optimal condition. The applied result showed that compared with the CK, the transformation rate of optimal condition enhances two times.