The Genetic Transformation Of Cotton Via Particle Bombardment And Genetic Analysis Of Generation

This study used the upland cotton GW2031 and KB18 as target materials, andtransformed the expression vector phyA gene into the shoot meristems of these twovarieties via particle bombardment. The transgenic plants of these two varieties and theT₁, T₂ generation were obtained after kanamycin selection culture, PCR, PCR-Southernand Southern blot hybridization analysis, and genetic analysis and activity analysis ofroots excreted phytase. The objective of this study is to increase new content and newgermplasm for cotton molecular breeding and provide the effective method for solvingthe problem of using the phosphorus in soil. The results were as follows:1. The seedling medium was obtained by regulating the contents of media. GA₃ andNAA were found to have notable effect in apex culture, GA₃ had better inhibition tothe basal callus of apex, and NAA had some effect to the rootage of apex.2. Transformed plantlets could be obtained in about 2 months after particlebombardment. A total of 39 Kan resistant plants were obtained, one was GW2031and 38 were KB18. PCR-Southern analysis showed that 32 resistant plantscontained PhyA gene, 1 was GW2031 and 31 was KB18. The transformation ratioof GW2031 was 1.59% and that of KB18 was 5.23%.3. Southern blot hybridization were studied in some individual plants of transgenicplant T₁ and T₂ generation of KB18, the result showed that the hybridization signalintensity of different periods DNA had much difference and DNA of cotton youngleafs had better effect to carriy out Southern blot hybridization.4. Six T₁ indivudal plants and twelve T₂ indivudal plants with Southern blothybridization signal were choosen arbitrarilyand and analyzed the activity ofphytase secreted from roots after culturing for 20 days. The result revealed theactivity of phytase secreted from roots T₁ indivudal plants and T₂ indivudal plantswere increased 1.01～4.4 times.