Function of the SmMYB13 gene in drought stress response in eggplant (Solanum melongena L.)

doi:10.3969/j.issn.1004-1524.20241009

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (8): 1666-1679.DOI: 10.3969/j.issn.1004-1524.20241009

• Horticultural Science • Previous Articles Next Articles

Function of the SmMYB13 gene in drought stress response in eggplant (Solanum melongena L.)

LI Yujing¹^,²(), HUANG Qianru², ZHANG Aidong², WU Xuexia², ZHU Dongxing³, XIAO Kai²^,^*()

1. College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China
2. Protected Horticultural Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
3. Agriculture and Rural Service Center, Zhangyan Town, Jinshan District, Shanghai, Shanghai 201514, China

Received:2024-11-21 Online:2025-08-25 Published:2025-09-03
Contact: XIAO Kai

Abstract

Abstract:

To explore the function of the SmMYB13 gene in eggplant (Solanum melongena L.) under drought stress, the transcription factor SmMYB13 was cloned from eggplant, and bioinformatics analysis and expression detection under drought stress were conducted. The SmMYB13 gene was heterologously overexpressed in Arabidopsis thaliana using the inflorescence infiltration method, and the drought resistance and the expression of abscisic acid (ABA)-related genes in transgenic plants were analyzed. The results showed that the full length of the SmMYB13 gene was 777 bp, encoding 258 amino acids. It was a hydrophilic unstable protein without signal peptide and transmembrane structure, and was mainly located in the nucleus and mitochondria. Phylogenetic tree analysis revealed that the SmMYB13 protein was closely related to MYB13 proteins from Solanum tuberosum L. (potato) and Solanum lycopersicum L.(tomato). qRT-PCR analysis showed that the SmMYB13 gene was expressed in the root, stem, leaf, pericarp, flower and flesh of eggplant, with the highest expression level in the pericarp. Under drought stress, compared with wild-type A. thaliana, SmMYB13-overexpression A. thaliana exhibited reduced cell membrane permeability in leaf, was less stained by toluidine blue (TB), the water loss rate significantly decreased, while the activities of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), were significantly enhanced, malondialdehyde (MDA) content decreased, and drought resistance improved. Under ABA treatment, compared with wild-type A. thaliana, SmMYB13-overexpression A. thaliana showed upregulated expression of ABA-negative regulator genes and downregulated expression of ABA-positive regulator genes, resulting in reduced sensitivity to ABA. The above results indicated that the SmMYB13 gene in eggplant was a drought stress response factor and played a positive regulatory role in drought stress.

Key words: Solanum melongena L., transcription factor, SmMYB13 gene, drought stress, Arabidopsis thaliana

CLC Number:

S641.1

LI Yujing, HUANG Qianru, ZHANG Aidong, WU Xuexia, ZHU Dongxing, XIAO Kai. Function of the SmMYB13 gene in drought stress response in eggplant (Solanum melongena L.)[J]. Acta Agriculturae Zhejiangensis, 2025, 37(8): 1666-1679.

Figures/Tables 13

Table 1 Primers used in this study

引物名称Primer name	引物序列Primer sequences(5'-3')	用途Usage
SmMYB13-F SmMYB13-R	GCTGTGAGAAGAAGGGATTGAA GCCCATAGTACCTCGGACCAAA	基因克隆 Gene cloning
pEarleyGate 103-MYB13-F pEarleyGate 103-MYB13-R	CTATCTGTCACTTCATCGAAAGGAC CTTGGTTGTTGCTGAATCTCAATAC	载体构建 Vector construction
EF1α-F EF1α-R	CCACACTTCTCATATTGCTGTCA ACCAGCATCACCATTCTTCAAAA	实时荧光定量PCR qRT-PCR
qSmMYB13-F qSmMYB13-R	GCTGTGAGAAGAAGGGATTGAA GCCCATAGTACCTCGGACCAAA	实时荧光定量PCR qRT-PCR

Table 2 Primers for ABA signal regulatory factor genes

基因名称 Gene name	正向引物序列 Forward primer sequences(5'-3')	反向引物序列 Reverse primer sequences(5'-3')
ACTIN2	TTACCCGATGGGCAAGTCG	CTCATACGGTCAGCGATACC
ABI1	AGAGTGTGCCTTTGTATGGTTTTA	CATCCTCTCTCTACAATAGTTCGCT
ABI2	GATGGAAGATTCTGTCTCAACGAT	TGTTTCTCCTTCACTATCTCCTCCG
ABI5	CAATAAGAGAGGGATAGCGAACG	AGCGTCCATTGCTGTCTCCTCCA
ABF4	AACAACTTAGGAGGTGGTGGTC	CTTCAGGAGTTCATCCATGTTC
SnRK2.2	ATATGCCATCGGGATCTGAAT	TGGTTGGGAATGAAGAACAG
SnRK2.3	GTTGGATGGAAGTCCTGCTC	TGCCATCATATTCCTGACGA
PYR1	GAACAAACTTCGAGAGCGCGCG	CGGGAGTTACGAGCCATAGCTTC
PYL2	ATGAAGAGCAGAAAACCCTC	TTCAAGAACTCATTGACCGA
GTG1	GAGAGTCTTATGATTGTTTGGCG	TATAAGAAATTTTGAACTGATC
GTG2	TCAAGATGACCAAAAGGAGCAA	ACACACAGTAAATGGAACACGCA
RD29A	GGCGTAACAGGTAAACCTAG	AGTCCGATGTAAACGTCGTCC
RD22	GGTTCGGAAGAAGCGGAGG	AAACAGCCCTGACGTGATAT
COR47	GGAGTACAAGAACAACGTTCCCG	ATGTCGTCGCTGGTGATTCCTCT
NCED3	CAGCTTGTAGCTTTTGGGCTGTA	TAACAGAAACCAGCTGAGCTCGA
COR15A	GGCCACAAAGAAAGCTTCA	GCTTGTTTGCGGCTTCTTTTC
KIN1	AACAAGAATGCCTTCCAAGC	CGCATCCGATACACTCTTTCC

Fig.1 The full-length PCR product of the SmMYB13 gene

Fig.2 Prediction of hydrophilicity (A) and tertiary structure (B and C) of SmMYB13 protein

Fig.3 Phylogenetic tree of MYB13 in different species

Table 3 The similarity of SmMYB13 protein sequence between eggplant and other species

物种 Species	蛋白质序列号 Protein serial number	相似度 Similarity/%
马铃薯Solanum tuberosum L.	XP_—006352742.	82
番茄Solanum lycopersicum L.	XP_—015078812.2	77
辣椒Capsicum annuum	XP_—016576820.1	72
烟草Nicotiana tabacum	XP_—016496658.1	74
樟树Cinnamomum camphora L.	XP_—009770485.1	73
巴西橡胶树Hevea brasiliensis	XP_—021640273.1	49
莴苣Lactuca sativa	XP_—023769776.1	49
牵牛花Ipomoea nil	XP_—019189153.1	51
铁皮石斛	XP_—020684786.1	51
Dendrobium officinale Kimura et Migo.

Fig.4 Verification of the transcriptional activation activity of the SmMYB13 gene (A) and its relative expression levels in different organs of Solanum melongena L. (B) The bars marked without the same lowercase letter indicated significant differences (p<0.05).

Fig.5 Relative expression levels of the SmMYB13 gene in eggplant seedling leaves under different abiotic stresses “****” indicates significant differences compared with the wild type at the p<0.001 level,“ns” indicates no significant difference compared with the wild type (p>0.05). Different lowercase letters for the same treatment at different sampling times indicate significant differences (p<0.05). The same as below.

Fig.6 Identification of SmMYB13 overexpression Arabidopsis thaliana lines by Basta resistance screening (A), PCR (B), and qRT-PCR (C) “+” indicates the positive control (plasmid); “-” indicates the negative control (wild-type plant); Numbers 1-10 represent ten transgenic positive lines. Col-0 indicates the wild-type plant, and OE represents the SmMYB13 overexpression line.

Fig.7 Water loss rate and Trypan blue (TB) staining results of Arabidopsis thaliana leaves, and phenotypes of Arabidopsis thaliana plants under drought stress A, Water loss rate of Arabidopsis thaliana leaves during 0-6 h under drought treatment, “**” and “****” indicate significant differences compared with the wild type at p <0.01 and p<0.001 levels, respectively; B, Trypan blue (TB) staining results of Arabidopsis thaliana leaves after drought treatment; C, Phenotype of Arabidopsis thaliana plants after 10 days of drought treatment. Col-0 indicates the wild-type plant, and OE represents the SmMYB13 overexpression lines. The same as below.

Fig.8 Activities of POD (A), CAT (B), SOD (C) and MDA content (D) in Arabidopsis thaliana leaves after drought stress POD, Peroxidase; CAT, Catalase; SOD, Superoxide dismutase; MDA, Malondialdehyde.

Fig.9 Phenotype (A-C) and primary root length (D) of Arabidopsis thaliana roots after 7-day treatment with different culture media The bars marked without the same lowercase letter in the same culture medium indicate significant differences (p<0.05).

Fig.10 The relative expression levels of ABA-responsive genes and regulatory genes in Arabidopsis thaliana under ABA treatment A-F, Relative expression levels of ABA-responsive genes within 0-12 h of ABA treatment; G, Relative expression levels of regulatory genes after 3 h of ABA treatment. “*” and “**” indicate significant differences compared with wild type at p<0.05 and p<0.01 levels, respectively.

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Function of the SmMYB13 gene in drought stress response in eggplant (Solanum melongena L.)

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