The Stady On Agrobacterium-mediated Transformation Of Sugarbeet(beta Vulgaris) With Vacuolar H~+-PPase (AVP1)Gene

Sugar beet is one of the important sugar crops in our country, and it will play an important role in developing the sugar industry to meet rising demand. But over the past 10 years, some unfavorable to plant survival factors has emerged by natural disasters and human factors, such as drought, salinization, flood , low temperature and barren ,etc. In expanded beet production scale, soil drought, salinization, barren will become the main environmental factor of restricting output, Enhance anti-reversibility is sustained development important basis of production promotion in the current and future. Most experiments show that,AVP1 gene over-expression could improve drought tolerance, salt tolerance and barren tolerance of transgenic plants, it is providing the method and thinking for production of drought tolerance, salt tolerance, barren tolerance, high quality, high yield, high sugar beet new strains.In this study, AVP1 gene was successfully transformed into the explants of cotyledon node. by using A grobacterium-mediated transformation methord, and established the stability and the high efficient genetic transformation system of suger beet. Meanwhile, the transgenic plants with drought stress in handle and physiological function identification.The major results studied as follows:1.The methods of sugar beet explants are acquired:The kind of shell with sand paper burnish,use NaClO, sulfuric acid and HgCl₂ soaking 10min in each one, this during with sterile water flush multiple, pollution rate is only 3.3%, germination rate for 98.9%, To save time, save seed resources ,and laid a tissue culture foundation for sugar beet genetic engineering.2.An efficient genetic transformation system was established:The results show that, cotyledon node 24h for pre-culture, bacterium concentration OD₆₀₀=0.4-0.5, bacterium infection 6-8min,36-48h of co-culture, Cef concentration 500mg/L, Kan concentration 100mg/L has obtain the lightest transformation rate .3.This study realized the genetic transformation on AVP1gene of sugar beet, as the experimental result, 72 resistant seedlings were obtained, PCR assaying of plant DNA confirmed 37 transgenic plants, conversion rate for 11.31%.4.For detection of transgenic beet physiological resistance function, and transgenic plants drought-resistance analysis,the results show that,The transgenic plants'relative water content of leaves and roots,the content of MDA and the content of proline has been changed, the AVP1gene was successfully transformed into the explants that the drought resistance was improved of the plant.