Screening And Functional Verification Of Genes Related To Osmotic Stress Signaling Pathway In Arabidopsis Thaliana

Water has a significant impact on plant growth and crop productivity which as an crucial environmental condition.Plant can perceive different environmental signals,and then respond to drought stress at the molecular,cellular,physiological and biochemical levels.Ca2+,a second messenger at the heart of a complex signaling pathway network,is a very important regulator of plant cell physiology and response to external stimuli.But because of the functional redundancy of sensory proteins or that a receptor may be necessary for plant growth,functional deletion mutants will not survive,only a few stresses receptorare currently had been found.The laboratory has mature technology and complete system.Based on the identified OSCA1,we use forward genetic methods to study,hope that we can have a higher probability of identifying the key ingredient in plant osmotic stress signaling pathways.The calcium ion probe aequorin(AQ)expressed in Arabidopsis thaliana was used to detect the changes of cytosolic calcium concentration induced by hyperosmosis.And two map clonal groups of calcium-responsive mutants were established(contain 1200 individual plants).The second-generation sequencing was used to obtain the linked chromosomes of the mutants,and the primers designed by SNP were used to finely map.Finally we obtained two candidate genes(OSCA2 and BST1).The candidate genes were partially verified by genetics.The complementary material of OSCA2 and BST1 can complement the calcium ion fluorescence-deficient phenotype of its mutant and the physiological phenotype sensitive to hypertonicity.At the same time,it was also found that the OSCA2 and BST1 was expressed in the roots and leaves of Arabidopsis thaliana seedlings.And OSCA2 might be a membrane protei which was involved in the perception and response of Arabidopsis to drought signals.In summary,OSCA2 and BST1 may be important factors in plant response to drought stress signaling pathways,and its function is important for us to explain the stress-induced calcium signaling.