Functional Analysis Of Common Bean 14-3-3 Gene PvGF14n/d Under Abiotic Stress

Common bean is delicious and nutritious as people's popular vegetable crop.However,in the growing area of Phaseolus vulgaris,it was repeatedly attacked by abiotic stresses such as drought,high salinity and low temperature.This seriously affects the growth and yield of common bean.It is very important for us to develop a new variety of common bean with strong resistance.One of the strategies is to tap the stress related genes in common bean and improve resistance to common bean by molecular breeding.In early study,9 common bean 14-3-3 protein genes were excavated and found to be in varying degrees induced by abiotic stresses.In order to study whether they can cause stress tolerance in transgenic plants,this study,the functions of two isoforms Pv GF14 n and Pv GF14 d in drought and salt stress were analyzed.The results were as follows:1.The homology and evolutionary relationship between PvGF14n/d and otherplant 14-3-3 proteins were analyzed by bioinformatic methods.It indicated that Pv GF14 n has a high homology with tomato 14-3-3 protein TFT4,and Pv GF14 d has the highest homology with soybean 14-3-3 protein gene SGF14 d.We speculate that they may have similar functions;2.We cloned the full length of PvGF14n/d gene from common bean and cloned it into expression vector p Early Gate101 using Gateway technology.The target gene was expressed in Arabidopsis by bud soaking method,and PvGF14n/d transgenic Arabidopsis plants were obtained by DNA-PCR and RT-PCR identification.3.Arabidopsis thaliana transgenic plants,which have been obtained from PvGF14n/d,were tested for functional verification under drought and salt conditions.The treatment of mannitol simulated drought stress and soil water stress showed that overexpression of Pv GF14 n and Pv GF14 d in of Arabidopsis plants resulted in a decrease tolerance to drought stress;Under salt stress,Pv GF14 n and Pv GF14 d transgenic lines during seed germination to green seedling stage showed lower tolerance than that of the wild type;However,in the 2-3 week old seedling stage,there was no significant difference in salt tolerance between Pv GF14 n transgenic lines and wild type seedlings,and the tolerance of Pv GF14 d transgenic lines to salt was lower than that of wild type;The results showed that the two isoforms of 14-3-3 protein in common bean were both conserved in function and specific in the drought and salt signaling pathway.To sum up,the stress resistance of PvGF14n/d under abiotic stress provides the basis for the selection of resistance genes and common bean breeding.