Transfers The Soybean WRKY Gene Tobacco Descendant's Appraisal And The Pphysiological Characterstic Research

WRKY family is a special class of transcription factors of plants discovered in recent years. They were deeply involved in biotic and abiotic stress response. The WRKY genes were induced in biotic stress, defense response, drought, the cold, trauma and metal of environmental stress and play important role in plants metabolism, fruits ripening, skin and root-like body growth and ageing et al.In this research T1 seeds of tobacco transformed by HY1 and HY5 genes from soybean respectively were used as test materials. Antibiotics selection, PCR and RT-PCR were carried out to validate their genetic stability. Physical index of T2 generation under salt stress was tested to provide reference for their function. Our study would benefit for breeding of stress-resistant plants using soybean WRKY genes and investigating the mechanism of plant-resistance in the future. The main results are as follows:T1 seeds of tobacco transformed by HY1 and HY5 genes from soybean respectively were inoculated into solid non-hormone MS culture (appending 3% sugar, 0.7% agar, and pH5.8) after the disinfected treatment and young seedlings were obtained with seedling rate of 56.6% and 56.4%. After a week culturing the young seedlings were transferred to MS medium plus 50mg/L Kanamycin. Total DNA of T1 and T2 generation were extracted by CTAB method. After PCR detection 600bp and 530bp fragments were obtained according to anticipation using wild plant as negative control.Total RNA of T2 generation was extracted by Guanidinium rhodanide method from the leaves. Unique primers of HY1 and HY5 genes were applied in RT-PCR. The result showed that 600bp and 530bp fragments were obtained according to anticipation using wild plant as negative control.POD, SOD activity and soluble protein contents of T2 generation under different concentration of NaCl were detected. POD and SOD activity increased in advance and then fall down. POD activity achieved maximum in 100mmol/L of NaCl. SOD activity and soluble protein content of T2 generation of HY1 transformed tabacco achieved maximum in 50mmol/L of NaCl. SOD activity of T2 generation of HY5 transformed tabacco achieved maximum in 150mmol/L of NaCl and soluble protein content achieved maximum in 100mmol/L. The above three indicators of transgenic plants were somewhat higher than that of the wild plants indicating that the capacity of the salt-resistance of transgenic tobacco improved.