Cloning And Functional Analysis On Halotolerance3Gene Gmhal3A And Gmhal3B From Soybean (Glycine Max)

Crops will inevitably subject to a different environmental stress, such as drought and high salinity impact on plants growth and productivity. Plants have evolved complex mechanisms to control the expression of various genes to adapt to such environmental stresses. But this is not enough. We need to continue to explore the use of resilience genetic resources, cultivating adverse resistant and high yielding variety to cope with stress. n this study, we cloned GmHAL3a and GmHAL3b gene from soybean. Cloned Two RNAi fragment each of the two genes to construct four RNAi vectors, and successfully transferred to Agrobacterium tumefaciens strain EHA105; Analyzed expression pattern of the two genes with real-time quantitative PCR tissue expression analysis and abiotic stress analysis; studied on functional analysis of transgenic GmHAL3a and GmHAL3b gene Arabidopsis which were transformed via Agrobacterium mediation after treated with various stress. Our results are mainly as follows:1. The protein sequences of these two genes contain similar conserved sites with other species. The phylogenetic relationship appeared that GmHAL3a and GmHAL3b genes were more similar to Arabidopsis HAL3gene than others.2. Tissue expression analysis found, the expression of GmHAL3a and GmHAL3b genes were highest at the root from seedling to30days after flowering and lowest in leaves during the flowering stage. The expression level of the two genes in the stem were progressively upward with the growth of plants. Before55days after flowering, they expression higher in pods than seeds; Since then, the opposite, when65days after flowering, the expression of the two genes reached the highest value in seeds.3. Abiotic stress analysis of the GmHAL3a and GmHAL3b genes showed that they were induced by salt, sorbitol and LiCl, and most intense by LiCl. 4, After transformed GmHAL3a and GmHAL3b genes into Arabidopsis via Agrobacterium mediation, transgenic Arabidopsis.After Arabidopsis were treated with various stress. We found that AtHAL3a,AtHAL3b and AtHIPl genes expression levels were higher than that of the control Arabidopsis.These results indicate that overexpression of GmHAL3a and GmHAL3b genes can activates AtHAL3a, AtHAL3b and AtHIPl genes in Arabidopsis.5. After stress induced on transgenic pure lines by salt, sorbitol and LiCl, we found that the germination rate of seeds and root elongation of transgenic plants were increased. Also proline content in transgenic Arabidopsis was significantly higher than wild Arabidopsis, but active oxygen content in was lower. The results suggested that GmHAL3a and GmHAL3b genes enhanced the tolerance to salt and osmotic stress in Arabidopsis.