Analysis of the salt-stress responsive element of the promoter of peanut small GTP binding protein gene <i>AhRabG3f</i>.

Guoning DU; Jie Xiang; Shunyu Lin; Xiangyuan Kong; Xiuling WU; Xuedong Guan; Hong Zhu; Jingshan Wang; Lixian Qiao; Jiongming Sui; Chunmei Zhao

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Analysis of the salt-stress responsive element of the promoter of peanut small GTP binding protein gene AhRabG3f.

Author: Guoning DU ¹ ; Jie XIANG ¹ ; Shunyu LIN ¹ ; Xiangyuan KONG ¹ ; Xiuling WU ² ; Xuedong GUAN ² ; Hong ZHU ¹ ; Jingshan WANG ¹ ; Lixian QIAO ¹ ; Jiongming SUI ¹ ; Chunmei ZHAO ³
Author Information

1. Dry-land Farming Technology Laboratory of Shandong Province, Peanut Industry Collaborative Innovation Center of Shandong Province, College of Agronomy, Qingdao Agricultural University, Qingdao 266109, Shandong, China.
2. Zhucheng Agricultural and Rural Bureau, Zhucheng 262200, Shandong, China.
3. College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, Shandong, China.
Publication Type:Journal Article
Keywords: peanut; promoter; salt stress-responsive element; small GTP binding protein
MeSH: Arachis/genetics*; Fabaceae/genetics*; GTP-Binding Proteins/metabolism*; Gene Expression Regulation, Plant; Glucuronidase/metabolism*; Plant Proteins/metabolism*; Plants, Genetically Modified/genetics*; Salt Stress; Stress, Physiological/genetics*; Tobacco/genetics*
From: Chinese Journal of Biotechnology 2022;38(8):2989-2998
CountryChina
Language:Chinese
Abstract: To study the molecular mechanism of salt stress response of peanut small GTP binding protein gene AhRabG3f, a 1 914 bp promoter fragment upstream of the start codon of AhRabG3f gene (3f-P) from peanut was cloned. Subsequently, five truncated fragments (3f-P1-3f-P5) with lengths of 1 729, 1 379, 666, 510 and 179 bp were obtained through deletion at the 5' end, respectively. Plant expression vectors where these six promoter fragments were fused with the gus gene were constructed and transformed into tobacco by Agrobacterium-mediated method, respectively. GUS expression in transgenic tobacco and activity analysis were conducted. The gus gene expression can be detected in the transgenic tobacco harboring each promoter segment, among which the driving activity of the full-length promoter 3f-P was the weakest, while the driving activity of the promoter segment 3f-P3 was the strongest. Upon exposure of the transgenic tobacco to salt stress, the GUS activity driven by 3f-P, 3f-P1, 3f-P2 and 3f-P3 was 3.3, 1.2, 1.9 and 1.2 times compared to that of the transgenic plants without salt treatment. This suggests that the AhRabG3f promoter was salt-inducible and there might be positive regulatory elements between 3f-P and 3f-P3 in response to salt stress. The results of GUS activity driven by promoter fragments after salt treatment showed that elements included MYB and GT1 between 1 930 bp and 1 745 bp. Moreover, a TC-rich repeat between 682 bp and 526 bp might be positive cis-elements responsible for salt stress, and an MYC element between 1 395 bp and 682 bp might be a negative cis-element responsible for salt stress. This study may facilitate using the induced promoter to regulate the salt resistance of peanut.