Identification And Stress Response Analysis Of Tomato Calcium Channel OSCA Gene Family

Tomato(Solanum lycopersicum)is a crop that is grown in large quantities around the world because it is sweet and nutritious,it is loved by people all over the world.Tomato is also a model crop in scientific research.At present,the whole genome data of tomato has been published,which lays a foundation for the study of the molecular mechanism of tomato.In nature,abiotic stress and biotic stress like drought,high salt,cold and gray mold cause a large reduction in tomato yield.In order to avoid more economic losses,it is the focus of people's efforts to improve the resistance of plants in the work of tomato molecular breeding.When plants are exposed to stress,it will cause the increase of calcium concentration in the plant and trigger the calcium signal.The calcium signal participates in many life activities of the plant,which is an important regulation mechanism after the plant exposed to stress.Calcium channels are an integral part of the calcium signaling pathway.Hypertonic calcium channel OSCA1 first discovered in Arabidopsis in 2014,was found to play an important role in osmotic stress,but the OSCA family has not been discovered and studied in tomato.This study aimed at the bioinformatics analysis of tomato OSCA gene family,as well as gene expression analysis and functional verification.The results are as follows:(1)Using bioinformatics,we found 12 Sl OSCA genes in tomato,which are located on 8 chromosomes of tomato.Subcellular localization predictive analysis revealed that this family of proteins are located on the Golgi and plasma membrane.Through phylogenetic analysis of OSCA gene family members of tomato,soybean,rice and arabidopsis,it can be divided into five subtribes,and tomatoes are located in four subtribes.Structural analysis indicated that members of the OSCA family contained a conserved DUF221 domain.(2)In the tissue-specific analysis of tomato OSCA gene family,it was found that different genes expressed different amounts in different tissues.On the whole,the expression levels of Sl OSCAs in leaves and fruits were generally higher,the overall expression levels of Sl OSCA4 and Sl OSCA12 in tomato were higher than other genes.(3)In the analysis of biotic expression patterns:After the B.cinerea stress and the stress of B.cinerea and C.rosea stress,the expression of OSCA gene family showed different degrees of up-regulation and down-regulation,Sl OSCA10 and Sl OSCA6 responded strongly to the biotic stress,and the expression level reached 30-40 times of the control,and the Sl OSCA10 response stress was the strongest.(4)In the study of abiotic stress expression patterns:This study investigated the expression of OSCA gene family in tomato under drought stress,ABA stress,high salt stress and cold stress.The gene family members showed different degrees of up-regulation and down-regulation after stress.In drought stress,Sl OSCA9,Sl OSCA8 and Sl OSCA7 responded strongly to this stress,and Sl OSCA9 was the strongest.The response in high salt stress is the same as in drought stress.In ABA stress,Sl OSCA9,Sl OSCA8 and Sl OSCA11 responded strongly to stress,and Sl OSCA9 was the strongest.In cold stress,Sl OSCA3 and Sl OSCA11 were responded stronger,but the response of the gene family to cold stress was less than other stresses.We filter out Sl OSCA9,Sl OSCA8,Sl OSCA11,and Sl OSCA7,which responded strongly to abiotic stress.Among them,Sl OSCA9 is the strongest one.(5)Functionalization of Sl OSCA9 and Sl OSCA10 by VIGS silencing vector construction and infection of tumefaciens.The silenced plants and the control plants were subjected to stress treatment,and measured the enzyme activity of APX?POD?PRO?SOD?MDA and CAT in the plants after the stress.It was found that under the conditions of drought,high salt,ABA and cold stress,the physiological and bioc hemical indexes of Sl OSCA9 gene silencing plant were significantly changed,and the overall stress resistance decreased,especially the drought tolerance was the most obvious.The Sl OSCA10 gene silencing plant showed a significant decrease in the ability to resist gray mold.