Response of NADPH Oxidase Gene Family to Abiotic Stress in Pigeonpea (Cajanus cajan)
FAN Yu-Xin1, Li Na1, SONG Zhi-Hua1, LIU Teng-Yue1, DONG Bi-Ying1, CAO Hong-Yan1, DUAN Yu-Sheng1, MENG Dong1,2,*, FU Yu-Jie1,2,*, YANG Qing1,2,*
1 College of Forestry,Beijing Forestry University, Beijing 100083, China; 2 Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing Forestry University, Beijing 100083, China
Abstract Reactive oxygen species (ROS) play an important role in intercellular signal transduction between plant cells, while respiratory burst oxidase homologue (RBOH, also known as NADPH oxidase) is a key enzyme that catalyzes the production of reactive oxygen species and widely participates in mediating intra plant signal transmission.Pigeonpea (Cajanus cajan) is a woody leguminous plant which can be used as medicine and food. In this study, the members of CcRboh gene family were screened from the whole genome of pigeonpea by bioinformatics software, and their basic functions including chromosome location, phylogeny, conserved domains and promoter elements were analyzed.The expression of CcRboh in different tissues was screened by transcriptome data, and the response mode of CcRboh gene under abiotic stress was determined by qPCR. Nine Rboh family members were identified named CcRbohA~CcRbohI according to their position on chromosome. They encoded 818~940 amino acid residues with isoelectric point (pI) ranging from 8.76 to 9.36.According to the sequence similarity, they can be clustered into 5 subfamilies, all of which contained 4 domains such as NADPH oxidase structure. Subcellular localization prediction showed that CcRboh proteins were mainly located on the plasma membrane. The cis-element prediction results showed that the promoter region of CcRboh gene contained a variety of hormone and stress response elements. The widespread jasmonate acid response element and light response element indicated that CcRboh gene could respond to the induction of jasmonate and light. The expression pattern analysis results of Rboh in pigeonpea roots and leaves showed that CcRboh gene had a high expression level in roots, indicated that it played an important role in roots. The transcriptome data of jasmonic acid treatment showed that the expression of CcRbohB and CcRbohG were significantly up-regulated (P<0.05), and increased more than 5 folds after 3 h of treatment, indicating that they were indeed more susceptible to jasmonate acid induction. In addition,CcRboh had a good response to high temperature stress, and its expression was significantly up-regulated after 3 h of treatment (P<0.05), The expression levels of CcRbohC, CcRbohF, CcRbohG, CcRbohH and CcRbohI after 3 h treatment were 6.4, 22.5, 21.0, 70.4 and 15.5 times higher than those of the control, respectively.The expression of CcRbohI increased significantly after 12 h treatment (P<0.05), and the highest expression reached 37.4 times; However, for drought and salt stress, CcRboh was induced after 6 h; Moreover, CcRbohG and CcRbohH had the most significant responses to drought, salt and high temperature stress (P<0.05), which indicated that reactive oxygen species may play a crucial role in the response to high temperature stress, especially CcRbohG and CcRbohH genes. In this study, the genes of NAPDH oxidase family were analyzed and screened by bioinformatics,and the key genes CcRbohG and CcRbohH in response to high temperature and drought stress were obtained, which provides candidate genes and research basis for improving the stress resistance of C. cajan.
FAN Yu-Xin,Li Na,SONG Zhi-Hua, et al. Response of NADPH Oxidase Gene Family to Abiotic Stress in Pigeonpea (Cajanus cajan)[J]. 农业生物技术学报, 2022, 30(2): 284-295.
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