| Soybean(Glycine max L.Merr.) is widely planted in China as one of the most important resources of oil and protein. In soybean production,drought,high salt, low temperature and other abiotic stresses are the reasons which lead to lower quality and yield reduction. Especially in the coastal areas, salinization restricted the development of local agricultural production. Although the agricultural technology has made great development, drought and salinization of soil is still one of the most serious factors restricting of crop production. With the rapid development of molecular biology and genetic engineering, transforming the resistance gene into soybean is one of the effective approaches to enhance the resistance of varieties. Soybean genetic transformation is very difficult because of the low plant regeneration rate. Therefore,this is necessary to establish a simple and efficient soybean regeneration and transformation system.In this research, study on soybean genotype, phytohormone, induction time and other factors on the soybean hypocotyls adventitious shoots induction, in order to further optimize the soybean hypocotyls regeneration system; To determine the concentration of the selective reagent Ka and NaCl of three genotype. Discussing the effect of soybean genotypes, selective reagent on HaBADH transformation of soybean hypocotyls, to further optimization of Agrobacterium-mediated genetic transformation system, with a view to using transgenic technology improved varieties of soybean resistance to lay experimental basis. Betaine aldehyde dehydrogenase (BADH) is a key enzyme for the synthesis of glycine betaine, Betaine is not only plays a role of non-toxic osmoprotectants, and it can protect the cellular proteins and enzymes activity, thereby improving plant drought, salt tolerance.The main results of this research as follows:1. There were large differences of induction rate of adventitious shoots between soybean genotypes. Induction rate of adventitious shoots was between15.63%-87.50%in eight soybean genotypes that were used in this research. The results in this research had shown that induction rate of adventitious shoots was high of Hefeng35and Huaidou9, which were84.72%and87.50%respectively, which higher than the commonly used Heinong44.2. When germination medium supplemented with1.0mg/L6-BA, induction medium with different concentrations of IBA and6-BA had different effects on the adventitious shoot of soybean hypocotyl. when the concentration of IBA was0.1mg/L in induction medium had a better induction effect, and when the concentration of IBA was0.1mg/L, inducting9d or11d would be better.3. The selection genes of HaBADH were NTPⅡ, and the selective reagent were Ka or NaCl.When induction medium supplemented with0.1mg/L IBA, the suitable concentration of Ka and NaCl for Heinong44, Hefeng35, Huaidou9was100mg/L,150mg/L,100mg/L and100mmol/L,100mmol/L,75mmol/L respectively.4. Studies had shown that Heinong44and Heifeng35may more suitable as explant than Huaidou9in soybean hypocotyl transformation, because the induction rate of Heinong44and Hefeng35were significantly higher than Huaidou9.5. Ka as the selective regeant, the PCR positive rate of Heinong44and Hefeng35were36.26%and29.75%. NaCl as the selective regeant, the resisitance seedling rate of Heinong44and Hefeng35were47.37%and61.76%.NaCl therefore,may more suitable as a selective regeant in this study. But the concentration of NaCl screening in this test may be too high, it inhibited the growth of adventitious shoot, then we should properly reduce the concentration of NaCl in screening process. |
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