Cloning and Salt-tolerance Analysis of Stress Associated Protein Gene GhSAP8 in Gossypium hirsutum
Wang Yi-Xue1,2, Dong Yan-Hui1, Zhang Huan-Huan1, Hao Yao-Shan1, Sun Yi1, Wu Shen-Jie1,2,*
1 Biotechnology Research Center, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China; 2 Shanxi Key Laboratory of Cotton Germplasm Resources Utilization and Molecular Design Breeding, Yuncheng 044000, China
Abstract Stress associated proteins (SAPs) are zinc-finger proteins containing A20 and (or) AN1 domain which are involved in abiotic stress responses in plants. In this study, a stress associated protein gene, GhSAP8 (GenBank No. XM_016813997.1), was isolated from upland cotton (Gossypium hirsutum) via homology cloning. The full-length sequence of GhSAP8 gene was 519 bp, encoding a protein of 172 amino acids with a relative molecular weight of 18.22 kD and an isoelectric point of 7.51. GhSAP8 contained 1 conserved A20 domain in the N-terminus and 1 AN1 domain in the C-terminus which was typical SAP protein domain combinations. The expression of GhSAP8 gene was identified in different tissues, and qRT-PCR detection showed that the highest expression occurred in seedling leaf and it was induced by treatment of salt which suggested that GhSAP8 gene might be involved in responses to salt stress. The GhSAP8 gene was constructed into the plant expression vector pCAMBIA1300 and transformed into Arabidopsis thaliana. Under the treatment with 150 mmol/L NaCl, the survival rate (53%~70%) of Arabidopsis with overexpressed GhSAP8 gene was significantly higher than that of the wild type (27%), indicating the overexpression of GhSAP8 gene could enhance the tolerance to salt stress. In addition, the expression of salt-stress responsive genes, such as AtP5CS1 (Δ1-pyrroline-5-carboxylate synthetase 1), AtRD29B (responsive to desiccation 29 B), AtSOS1 (salt overly sensitive 1) and AtNHX1 (sodium/hydrogen exchanger 1), were significantly higher in transgenic Arabidopsis than that in wild type under salt stress. However, only a few of transgenic Arabidopsis lines showed increased expression of AtP5CS1, AtRD29B and AtSOS1 in wild type under non-salt stress. The above results indicated that GhSAP8 gene might improve tolerance to salt stress in transgenic Arabidopsis by regulating the expression of salt-stress responsive genes. The present study could provide superior gene resource for cultivation of transgenic salt-tolerant cotton.
Wang Yi-Xue,Dong Yan-Hui,Zhang Huan-Huan, et al. Cloning and Salt-tolerance Analysis of Stress Associated Protein Gene GhSAP8 in Gossypium hirsutum[J]. 农业生物技术学报, 2020, 28(7): 1230-1239.
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