| Presently, the main method of the biological prevention and cure of plant disease was chemical prevention and cure, but thus may lead to some new problem, such as environment pollution, pathogenic bacterium producing drug-resistant. In order to avoid these problems effectively we can adopt biological prevention and cure method. Numb mildew Trichoderma spp was one of the most microorganisms applying for the biological to prevent and cure of plant disease in the world. So it was very important to improve ability of tolerance of alfalfa to abiotic bacteria. Cultivated alfalfa (Medicago sativa L.), a perennial legume crop, was widely planted all over the world and was studied frequently. With the development of plant gene engineering, an important approach for alfalfa bacteria-resistant breeding was founded in the prensent work, CHI gene was transferred to alfalfa by Agrobacterium-mediated transformation to improve bacteria-resistant of alfalfa Peru plant.The main achievements of our studies are as following:1,Regeneration ability tissiue culture establishment:MS+2,4-D(2.0mg/L)+6-BA(0.5mg/L)+candy0.3%+agaragar0.08%;MS+KT(0.5mg/L)+NAA(0.01mg/L)+ candy 0.3%+ agaragar0.08% ; 1/2MS+NAA(0.01mg/L)+ candy 0.15%+ agaragar 0.08% are in order inducing callus ,sprout and root of alfalfa. Induced frenquency of hypocotyls of Callus was higher than that of cotyledon. And Peru was much higher than and Long-dong purple medic in different breeds. To repeat six same experiments, we can find that callus inducing frenquency of hypocotyls was higher than that of cotyledon, but differentiatal rate was relatively low on A3 culture medium. So culture medium having different concentration ratio have different effect on regeneration system establishment. In four differentiatal culture medium, differentiatal frenquency of culture medium No.B1 can reach 96%, that of B4 was lowest.Hypocotyls:B4 |
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