Overexpression of SlWRKY41 Improves the Tolerance of Tomato Seedlings to Drought

doi:10.13560/j.cnki.biotech.bull.1985.2024-0688

Abstract

Abstract:

Objective To explore the role of SlWRKY41 transcription factor in response to drought stress in tomato, and to provide theoretical value and important basis for the enrichment of tomato germplasm resources and the selection and breeding of drought-resistant varieties of tomato. Method The experiment was conducted with wild-type tomato 'Ailsa Craig' as the material, and its cDNA was used as the template to clone the SlWRKY41 gene to construct the OE-SlWRKY41 transgenic tomato lines, and the wild-type and OE-SlWRKY41 transgenic tomato lines were subjected to drought stress treatments, and the phenotypic observations and related physiological and biochemical indexes were determined at the day 4 and 8 of the treatment. Result Compared with wild-type plants, after 8 d of drought stress, OE-SlWRKY41 transgenic plants had curled leaves with less wilting and significantly lower levels of membrane lipid peroxidation. Stomatal conductance and transpiration rates were significantly lower, and the net photosynthetic rate was significantly higher. The root morphology indexes showed an overall upward trend, and the leaf relative water content, antioxidant enzyme activities and osmotic regulating substances (soluble sugars and proline content) increased significantly, and relative conductivity decreased significantly. The stress response genes (SlSOD, SlPOD and SlP5CS1) in the plants overexpressing SlWRKY41 were significantly up-regulated. Conclusion SlWRKY41 respond to drought, and the overexpression of SlWRKY41 can improve the tolerance of tomato seedlings to drought stress.

Key words: tomato, transcription factors, SlWRKY41, drought stress, photosynthesis, antioxidant system, osmoregulation

LIU Jie, WANG Fei, TAO Ting, ZHANG Yu-jing, CHEN Hao-ting, ZHANG Rui-xing, SHI Yu, ZHANG Yi. Overexpression of SlWRKY41 Improves the Tolerance of Tomato Seedlings to Drought[J]. Biotechnology Bulletin, 2025, 41(2): 107-118.

Figures/Tables 10

Table 1 Primer sequences for RT-qPCR

引物名称Primer name	引物序列Primer sequence（5′-3′）
SlActin-F	ACCACTGAGCACAATGTTACCG
SlActin-R	GTCCTCTTCCAGCCATCCA
SlWRKY41-F	GGCTTGAAGGTCCATGTGAAGATG
SlWRKY41-R	ACCCCTGTGTGTTCCTATGAGTAC
SlSOD-F	AATTCATCATTGTGGCAGCA
SlSOD-R	GCCCTTAAGGACAGCAACAG
SlPOD-F	GGTCCAACATGGCAAGTTCT
SlPOD-R	ACATCTTGCCCTTCCAAATG
SlP5CS1-F	TGATGGAAGATTAGCACTTGGAA
SlP5CS1-R	CAACACCTACAGCACCAGAA

Fig. 1 Genetic transformation of SlWRKY41-transgenic tomato plants and PCR and RT-qPCR detection of transgenic tomato plantsA: Preparation of exosomes. B: Differentiation of healing tissues. C: Generation of resistant buds. D: Rooting culture. E: SlWRKY41 transgenic tomato genomic DNA PCR assay for the T1 generation. M: DNA maker DL2000; 1: negative control; 2: wild-type plants; 3-8: SlWRKY41 transgenic lines; 9: plasmid control. F: RT-qPCR detection of WRKY41 expression in T2 generation transgenic tomato plants, and the different lowercase letters indicate significant levels of difference among treatments(P<0.05), the same below

Fig. 2 Phenotypic observations of wild-type and SlWRKY41-overexpressed plants under drought stressOE-WRKY41 from left to right is OE-WRKY41-1，and OE-1WRKY41-2

Table 2 Effects of drought stress on the biomasses of tomato seedlings overexpressing SlWRKY41

处理

Treatment

株高

Plant height/cm

茎粗

Stem diameter/mm

地上鲜重Shoot fresh weight/g

地下鲜重 Root fresh weight/g

地上干重Shoot dry weight/g

地下干重 Root dry weight/g

总干重 Total dry weight/g

总鲜重 Total fresh weight/g

Table 3 Effects of drought stress on the root system indexes of tomato seedlings overexpressing SlWRKY41

处理

Treatment

根长

Root length/cm

根系表面积

Root surface area/cm²

根平均直径

Mean root diameter/mm

根系体积

Root volume/ cm³

根尖数

Root tips

根分叉数

Root forks

交叉数

Root crosses

Fig. 3 Effects of drought stress on photosynthetic indexes in the leaves of tomato seedlings overexpressing SlWRKY41

Fig. 4 Effects of drought stress on the relative electrical conductivity and relative water content in the leaves of tomato seedlings overexpressing SlWRKY41

Fig. 5 Effects of drought stress on malondialdehyde， reactive oxygen species content and antioxidant enzyme activities in the leaves of tomato seedlings overexpressing SlWRKY41

Fig. 6 Effects of drought stress on soluble sugar content and proline in the leaves of tomato seedlings overexpressing SlWRKY41

Fig. 7 Relative expression variations of stress-related genes in the tomato seedlings overexpressing SlWRKY41 under drought stress

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