Study On Exogenous DNA Transformed Colored Cotton Via Ar～+ Ion Beam

Genetic transformation is an important research domain in genetic engineering. Heavy–ion beam mediated gene transferring is a new technique that was developed recently. It has made enormous success in theory and application, But the research of this technology on colored cotton has not been reported. In this thesis, this new methodology has been used to study the transgenic technique focusing on relevant biological effects of botanic seeds after being exposed to radioactive sources. Random amplified polymorphic DNA (RAPD) was used to evaluate DNA variation of these varieties and analyzed each hereditary variation. Seeds of KL1(green) were implanted by Ar⁺, energy was 25 keV, doses were 0,6×1015Ar+/cm2,8×1015Ar+/cm2,10×10¹⁵Ar⁺/cm² ,20×10¹⁵Ar⁺/cm²,40×10¹⁵Ar⁺/cm²,60×10¹⁵Ar⁺/cm². The result showed that: 1) The physiological indexes (MDA,SS,PPO,GTP,POD,TA) were studied under Fuserium oxysporum stress condition and analyzed the relationship among the radiation doses and all the data collected. Through correlation analysis about physiological indexes, the result demonstrated: all physiological indexes except that MDA disposal content all to be higher than CK, and had the remarkable negative correlation. The activity of GTP and PPO are higher in leaves than in stems. Proved that leaf tissues were more sensitive than stems. Indirectly explained extraneous DNA entered into acceptor and the acceptor gene group caused its biochemistry index to change. 2) The main economic traits in M1 were detected, the results indicated that by using 10×10¹⁵Ar⁺/cm² the fiber length can increase 22.5%,fiber length's improvement can be increased 17.1% by using 8×10¹⁵Ar⁺/cm². but micronaire has actually presented the obvious negative effect. 3) The genomic DNA of white cotton was delivered into colored cotton with the seeds transformation system established above. The germinated seeds were sowed in pathogenic soil and plants with resistance to wilt diseases were survived. DNA extracted from the survivals were used for RAPD analysis .The result of the RAPD analysis showed that two primers, S53, S176 can be used as molecular marker in assistance of selecting the transgenic plants after the preliminary study. This finding, for the first time, identifies DNA fragment in white cotton had integrated into the recipients'genome. And proved further the ion beam lies between the feasibility of the transgenic method.