The Molecular Mechanism Of Soybean GmPLC10 In The Regulation Of Plant Salt And Alkali Stress

Phospholipid signal,as a significant component of regulating metabolism,growth,cell activities and signal transduction in plant cells,and has been playing an irreplaceable role in response to abiotic stress.As an important hydrolase in vivo,Phospholipase C（PLC）is a kind of specific enzyme that hydrolyzes phosphatidylinositol4,5-bisphosphate,PIP₂ into two second messengers inositol-1,4,5-trisphosphate,IP₃ and 1,2-diacylglycerol,DAG,which are the third messengers.They participate in various salt alkali signal transduction pathways in eukaryotic cell synthesis.PLC activity-specific inhibitor U73122 can inhibit the accumulation of IP₃during the growth of seeds and seedlings under salt and alkali stress,thus greatly reducing the damage of salt,salt and alkali,cold and drought to plant cells.Because IP₃ is the specific product of PI-PLC,the discovery and research of these specificity show that PI-PLC is widely involved in the regulation of various salt and alkali signal stress responses of eukaryotic cells.In this experiment,based on the previous laboratory basis and the T3 generation of GmPLC10 transgenic Arabidopsis plants,the germination rate,main root length and corresponding physiological indexes of transgenic and wild-type Arabidopsis plants were determined after salt alkali stress treatment.Based on the statistical analysis of salt tolerance data of transgenic Arabidopsis plants with GmPLC10 gene,the following conclusions were obtained:1.By using 150 mM NaCl,145 mM NaCl+5 mM NaHCO₃ was used to simulate salt alkali stress.The germination rate of wild-type Arabidopsis and transgenic Arabidopsis with GmPLC10 gene in the control group were measured,and the main root length of wild-type Arabidopsis and transgenic Arabidopsis was measured.It was found that the germination rate and main root length of transgenic Arabidopsis GmPLC10 were higher than those of wild-type Arabidopsis.2.In this study,the process of DAG phosphorylation to PA was specifically inhibited by the hydrolysate DAG inhibitor R59022 of PLC,which was added to the solution containing 150mM NaCl,145 mM NaCl+5 mM NaHCO₃ The germination and root length of wild-type Arabidopsis and GmPLC10 transgenic Arabidopsis were compared.It was found that the germination rate and root length of transgenic Arabidopsis tended to be the same under salt stress compared with wild-type Arabidopsis.However,due to the failure of normal rooting under alkali stress,the transformation of leaf albinism could be observed Gene plants are better than wild type.3.Three The physiological indexes of transgenic Arabidopsis GmPLC10 and wild-type Arabidopsis were determined 20 days after the plants were cultured in the soil for 14 days and were irrigated with salt and alkali gradually.We found that compared with wild-type Arabidopsis,transgenic Arabidopsis GmPLC10 significantly increased the dry weight and water content of the plants,while the fresh weight slightly increased,while the fresh weight slightly increased In the determination of physical indexes,salt alkali stress reduced the content of malondialdehyde,increased the content of proline,slightly decreased the content of Na⁺,increased the content of K⁺,and decreased the content of Na⁺/K⁺,in conclusion,GmPLC10 can improve the salt alkali tolerance of plants.4.In the early stage of our experiment,we found that the PLC protein is located in the cytoplasm,has a transmembrane structure,and has no signal peptide at the N-terminus.Therefore,we chose the membrane system yeast two-hybrid system,constructing a soybean c DNA library and transforming yeast strains to interact with proteins screening.