Developing Herbicide-Resistant Pol CMS Restore Lines Of Brassica Napus By Marker-Assisted Selection

As pest and disease, weed is a great threaten to the development of agriculture, and affects the yield and quality of crops severely. Moreover, the adaptability to environment and the ability of reproduction of weeds are much better than crops as well as their varieties. As a result, the traditional way of weeding can't satisfy the need of agriculture production. Chemistry medicament has become the most efficient and economic way of weeding. So, developing herbicide-resistant crops is the exigent need of chemistry weeding. We can transform the herbicide-resistant gene which exists in the crops that has gained the herbicide-resistant character into the target varieties by routine hybridization and back-cross. The restore lines or sterile lines can be transmitted with herbicide-resistant gene, so that their hybrids can get the character of herbicide-resistance.Our research mainly contains: transforming the glyphosate-resistant gene which exists in the RH into three Pol CMS restore lines by one time hybridization, three times back cross, two times of selfing and two times background selection, and enabling these restore lines having pure glyphosate-resistant gene. Main results are as follows:1.The validation of glyphosate-resistant gene's inheritance mode. By investigating the number of seedling before and after the treatment of glyphosate, we found that the segregation proportion of alive seedling and dead seedling was 1:1 in BC₂F₁ generation, and 3:1 in BC₁F₂ generation. Results showed that glyhosate-resistance gene was controlled by one dominant gene.2.The PCR detection of glyphosate-resistant gene. We used the published primers which have been used to detect the glyphosate-resistant transgenic rapeseed to amplify our samples, and found nothing in the parent restore lines. But in the alive seedlings which were tolerant to glyphosate, two gene fragments, cp4-epsps and gox, were amplified.3.Background selection of BC₂F₁ generation and BC₃F₂ generation. We used 64 random AFLP primers to amplify the BC₂F₁ generation and BC₃F₂ generation respectively, and after clustering analysis we found that the genetic background was mostly consistent with parent restore lines.4.The test of seed quality. Main seed quality traits of three restore lines and their BC₃F₂ generation were analyzed and the results showed that, besides the oil content was slightly lower than their parents respectively, no obvious difference was detected in other traits.