Protein Interaction Of Karelinia Caspia KcCBLs-KcCIPKs Family And Expression Pattern Analysis Under High Temperature Stress

High temperature is one of the main abiotic stresses suffered by desert plants,and it is also one of the important factors leading to desertification.Karelinia capsica,as a representative of desert plants,has strong stress resistance such as high temperature resistance,drought resistance and saline alkali resistance.It is a pioneer plant for wind prevention and sand fixation in the transition zone between Taklimakan Desert and farmland.In the process of long-term adaptation to desert environment,it has formed a strong broad-spectrum stress resistance,which has high research value of plant stress resistance.At present,there are few reports on the stress resistance genes of Karelinia capsica.Therefore,exploring the stress resistance genes of Karelinia capsica is of great significance to cultivate crops suitable for drought,extreme temperature and saline alkali environment and improve the land use efficiency of Tarim Basin.CBL-CIPK pathway with Ca²⁺as the main signal response means in plants plays an important role in response to abiotic stress.In order to explore the function of CBL-CIPK pathway in Karelinia capsica,In this study,KcCBLs and KcCIPKs gene family related genes involved in high temperature stress response were cloned and identified based on the room temperature high temperature differential transcriptome data of Karelinia capsica and the c DNA sequences of related genes of Arabidopsis and other species.The expression of KcCBLs and KcCIPKs under high temperature stress was analyzed by RT-PCR,and the protein interaction was analyzed by yeast two hybrid experiment.The main results of this study are as follows:(1)Analysis of KcCBLs and KcCIPKs gene family in Karelinia capsicaAccording to the transcriptome data of Karelinia capsica,combined with the c DNA sequences of related genes of Arabidopsis thaliana and other species,3 KcCBLs and 7 KcCIPKs involved in high temperature response were identified in the transcriptome of Karelinia capsica.Gene structure analysis was carried out in species such as Karelinia caspica,Arabidopsis thaliana,Helianthus annuus,Lactuca sativa,Gossypium herbaceum,Glycine max,Solanum lycopersicum,Medicago truncatula,Oryza sativa and Zea mays,The analysis results show that the identified KcCBLs and KcCIPKs are relatively conservative in sequence structure and evolution,which is worthy of further study.(2)Semi quantitative expression pattern analysis of KcCBLs and KcCIPKs in Karelinia capsicaThe expression profiles of KcCBLs and KcCIPKs under high temperature stress were detected by semi quantitative RT-PCR.The results showed that the expression of KcCBLs and KcCIPKs in root,stem and leaf organs of Karelinia capsica were significantly increased under high temperature stress.(3)KcCBLs-KcCIPKs protein interaction validation.Through the study of KcCBLs-KcCIPKs protein interaction,it is preliminarily verified that KcCBL1 protein can interact with 7 KcCIPKs proteins in varying degrees,KcCBL2 protein interacts with 3 KcCIPKs proteins:KcCIPK5,KcCIPK6 and KcCIPK7,and KcCBL4 protein can interact with 6 KcCIPKs proteins except KcCIPK9.The experiment of protein interaction verifies that CBL and CIPK interact in a subset.KcCBL1,KcCBL2 and KcCBL4 can interact with multiple KcCIPKs.To sum up,this study found that the functional domain structures of KcCBLs and KcCIPKs were highly conservative with those of other plants;RT-PCR experiments showed that the screened KcCBLs and KcCIPKs genes related to high temperature tolerance could respond to high temperature stress,and the expression increased significantly;The interaction between KcCBLs and KcCIPKs was preliminarily verified by yeast two hybrid experiment.The related functional mechanisms of KcCBLs and KcCIPKs involved in high temperature stress need to be further studied.This study provides a theoretical basis for the molecular response mechanism of Karelinia capsica under high temperature stress,and provides a high-quality genetic reference for the artificial breeding of local crops with high temperature tolerance.