The Genetic Transformation Of Cotton Via Agrobacterium-mediated And Screening Of Homozygous Lines

It is great significant that Improving inorganic phosphorus use efficiency in cotton. This study used the upland cotton Jin mian 14, Shan 724, Zhong 6331 and Zhong mian suo 18 as target materials and transformed the expression vector PhyA gene into the embryogenic callus and hypocotyls of these varieties via Agrobactetium-mediated. The objective of this study is to increase new content and new germplasm for cotton. The aim of the thesis is to find out some homozygous lines by PCR methods. The activity analysis of roots excreted phytase was also be detected. The results were as follows:1. Effect of kanamycin concentration, the growth status of embryogenic callus and the genotypes of the target material on the transformation efficicency had been assayed. Examines the research to indicate that the card that mildew element's screening best density should be 75 mg/L. We used embryogenic callus coloured in yellow as target materials. It helps to improve the transformation efficiency. We selected cotton Jin mian 14 and Shan 724 are two excellent receptor transgenic materials.2. Embryogenic callus of Jin mian 14 and Shan 724 were used as the receptor of Agrobacterim-mediated transformation. PCR analysis using primer sets of the interest gene PhyA indicated that plants which came from different cell lines produced the wanted 1 347 bp fragments. No wanted fragments were observed in the control plants; this study used the upland cotton Zhong mian suo 18 as target materials and transformed the expression vector PhyA gene into the hypocotyls of this variety via Agrobactetium-mediated, and obtained a large amount of kanamycin-resistant callus of cotton.3. All the 3 230 T3 regeneration plants have already been detected by PCR. We have found out one homozygous line by PCR. And also, the result revealed the activity of phytase secreted from roots T3 indivudal plants were increased 1.18-5.13 times. In this study, the foreign gene separated in T3 generation plants. The proporation of the positive plants was lower than 63: 1. It did not accord with Mendel's law.