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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (6): 1573-1583.doi: 10.3724/SP.J.1006.2023.21028

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Biological functional analysis of common buckwheat (Fagopyrum esculentum) FeSGT1 gene in enhancing drought stress resistance

ZHU Xu-Dong1(), YANG Lan-Feng1, CHEN Yuan-Yuan1, HOU Ze-Hao1,2, LUO Yi-Rou1, XIONG Ze-Hao1, FANG Zheng-Wu1,*()   

  1. 1College of Agriculture, Yangtze University/Hubei Center for Collaborative Innovation of Grain Industry, Jingzhou 434025, Hubei, China
    2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2022-04-18 Accepted:2022-09-05 Online:2023-06-12 Published:2022-09-15
  • Contact: *E-mail: fangzhengwu88@163.com
  • Supported by:
    National Natural Science Foundation of China(31671755)

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Abstract:

SGT1 (Suppressor of the G2 Allele of skP1) is an inhibitor of skp1-4, which plays an important role in the abiotic stress response of plants. Based on the early transcriptomics and proteinomics analyses of common buckwheat under drought stress, a FeSGT1 gene was screened and cloned, which contained a 1086 bp open reading frame encoding 361 amino acids and 3 domains including TPR, CS, and SGS. Homologous protein comparison showed that FeSGT1 was closely related to CqSGT1 (XP_021726759.1), BvSGT1 (XP_010671588.1), and SoSGT1 (XP_021839743.1). Besides, FeSGT1 gene encoded membrane localization protein. The relative expression levels revealed that FeSGT1 tended to be up-regulated within 24 hours of drought stress. The expression of FeSGT1 gene peaked at 12 hours and began to decline after 24 hours under salt, low temperature (4℃), and ABA treatments. Overexpression of FeSGT1 gene in transgenic Arabidopsis not only conferred drought and salt tolerance, but also significantly increased root length, fresh weight, and survival rate compared with the wild type (WT) plant, accompanied by the elevated activities of catalase (CAT), the lowered malonaldehyde (MDA) and H2O2 contents, thus allowing plants to better adapt to adverse environments. Our results provided information in the exploring of the molecular regulation mechanism responding to drought tolerance in common buckwheat.

Key words: Common buckwheat (Fagopyrum esculentum), FeSGT1, transgenic Arabidopsis, drought resistance

Table 1

Summary of primer sequence"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 用途
Purpose
FeSGT1-F ATGGCGTCCGATCTCGAAAAG ORF扩增
FeSGT1-R TTAGTATTCCCATTTCTTCACCTC Amplification of ORF
Actin-QF ACCTTGCTGGACGTGACCTTAC 荧光定量内参基因
Actin-QR CCATCAGGAAGCTCATAGTTC Reference gene of qRT-PCR
FeSGT1-QF ATCACGCTGAACTCTTCGCA 荧光定量PCR
FeSGT1-QR TGCGTTCATCACACTCACGA Quantitative real time-PCR
pH-FeSGT1-F CTCGAGATGGCCGGCCCTGATATTTTG 亚细胞定位
pH-FeSGT1-R CCTAGGATCCTCTTGGCTCTCATC Subcellular Localization
pBI-FeSGT1-F CTCGAGATGGCCGGCCCTGATATTTTG 转基因拟南芥
pBI-FeSGT1-R CCCGGGCTAATCCTCTTGGCTCTCATC Transgenic Arabidopsis
p121-SGT1-F TTTGGAGAGGACCTCGACCT 转基因拟南芥鉴定
p121-SGT1-R TTAGTATTCCCATTTCTTCACCTC Identification of transgenic Arabidopsis

Fig. 1

Identification and structural analysis of FeSGT1 in common buckwheat A: multiple alignment of FeSGT1 with other plant SGT1 proteins; B: Phylogenetic tree analysis of FeSGT1 protein; C: 3D structure analysis of FeSGT1 protein."

Fig. 2

Subcellular localization of FeSGT1 Subcellular localization of FeSGT1 protein in tobacco leaves. The green indicates GFP signals, and the red indicates chloroplast autofluorescence."

Fig. 3

Expression analysis of FeSGT1 in common buckwheat * and ** indicate significantly different at the 0.05 and 0.01 probability levels, respectively. The error bars indicate ± SD (n = 3)."

Fig. 4

Phenotype analysis of FeSGT1 transgenic Arabidopsis under drought treatment A: seed germination assays of WT and OE lines under drought stress. B: the germination rates of WT and OE lines under drought stress; the germination rate was calculated for the next 5 days. C: root length assays of WT and OE lines under drought stress. D: The total root lengths and fresh weights of WT and OE lines under drought stress; * indicates significantly different at the 0.05 probability level."

Fig. 5

Ectopic expression of FeSGT1 enhanced drought tolerance in transgenic Arabidopsis A: phenotypes of 3-week-old WT and OE lines under drought stress. B: the survival rates of WT and OE lines under drought condition was monitored 7 days after rewatering. C-E: the physiological indicators of WT and OE lines under normal and drought treatments, including MDA (C), H2O2 (D) contents, and CAT (E) activities. * and ** indicate significantly different at the 0.05 and 0.01 probability levels, respectively."

Fig. 6

Phenotype analysis of FeSGT1 transgenic plants under salt treatment A: seed germination assays of WT and OE lines under salt stress. B: the germination rates of WT and OE lines under salt stress; the germination rate was calculated for the next 5 days. C: root length assays of WT and OE lines under salt stress. D: the total root lengths and fresh weights of WT and OE lines under salt stress; * indicates significantly different at the 0.05 probability level."

Fig. 7

Ectopic expression of FeSGT1 enhanced salt tolerance in transgenic Arabidopsis A: phenotypes of 3-week-old WT and OE lines under salt stress. B: the survival rates of WT and OE lines under salt stress. C-E: the physiological indicators of WT and OE lines under normal and salt treatments, including MDA (C), H2O2 (D) contents, and CAT (E) activities; * indicates significantly different at the 0.05 probability level."

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