Functional Study of Transcription Factor VcMYB17 in Regulating Drought Tolerance in Blueberry

doi:10.13560/j.cnki.biotech.bull.1985.2024-1031

Abstract

Abstract:

【Objectve】This study aims to clarify the functional mechanism of the transcription factor VcMYB17 in blueberry drought stress response, providing potential genetic resources for blueberry cultivar improvement. Method Bioinformatics methods were employed to characterize the basic properties of VcMYB17. RT-qPCR was used to analyze the tissue specificity of VcMYB17, the response of VcMYB17 to different hormones and drought stress, and the effect of overexpression of VcMYB17 on drought resistance genes in Arabidopsis thaliana. Subcellular localization of VcMYB17 was examined in transiently transformed tobacco, and its transcriptional activating activity was verified in yeast. The VcMYB17-OE transgenic blueberry callus and A. thaliana were obtained through Agrobacterium infection. Drought stress phenotypes were evaluated, including measurements of survival rate, fresh weight, water loss rate, relative conductivity, malondialdehyde (MDA) content, and peroxidase (POD), and superoxide dismutase (SOD) activities. Result VcMYB17 is an R2R3 type MYB transcription factor that is highly expressed in the leaves of blueberry. Exogenous treatment with ABA, GA, MeJA, SA and PEG causes the expressions of VcMYB17 to be downregulated. VcMYB17 is localized in the nucleus and has transcriptional activating activity. Under drought stress, the overexpression of VcMYB17 inhibits plant growth, makes plants accumulate more MDA content and decrease POD and SOD activities. Furthermore, the expressions of drought response genes such as AtERD1, AtERD15, AtP5CS1, AtRD29A, AtRD22 and AtDREB1A in VcMYB17-OE plants are significantly lower than that in wild type plants. Conclusion VcMYB17 is an R2R3 MYB transcription factor localized in the nucleus. The expression of VcMYB17 is inhibited by ABA, GA, MeJA, SA and PEG. Overexpression of VcMYB17 decreases the expression of drought response genes, resulting in reduced drought resistance in A. thaliana and blueberry callus.

Key words: VcMYB17, gene function, blueberry, resistance to drought, drought stress, transgenic plant

TIAN Qin, LIU Kui, WU Xiang-wei, JI Yuan-yuan, CAO Yi-bo, ZHANG Ling-yun. Functional Study of Transcription Factor VcMYB17 in Regulating Drought Tolerance in Blueberry[J]. Biotechnology Bulletin, 2025, 41(4): 198-210.

Figures/Tables 8

Table 1 Sequences of primers for RT-qPCR

基因名称 Gene name	上游引物 Forward primer sequence （5′‒3′）	下游引物 Reverse primer sequence （5′‒3′）
VcMYB17	ATTCTGAGGAACTTCTAATGCCTT	CCAATCACCGCCATACCCAA
VcUBC28	CCATCCACTTCCCTCCAGATTATCCAT	ACAGATTGAGAGCAGCACCTTGGA
Actin	GGTAACATTGTGCTCAGTGGTGG	AACGACCTTAATCTTCATGCTGC
AtERD1	CTTTCTCTATCAGCACGAAACG	CGGTGCGATATATTGACAATCC
AtERD15	TTCGACTTGGTACCCTGATTAC	GGAAGAAGATCAGCTACATCGA
AtP5CS1	AGCTTGATGACGTTATCGATCT	AGATTCCATCAGCATGACCTAG
AtRD22	GACTTTCGATTTTACCGACGAG	CGCTACCGGTTTTACCTTTATG
AtRD29A	TTCTGTAAGGACGACGTTTACA	CGTACTCGTTACATCCTCTGTT
AtDREB1A	CGTTTCAGGATGAGATGTGTGA	CTCATCGTGCATATAAAACGCA

Table 2 Sequences of primers for vector construction

引物名称 Primer name	序列 Sequence （5′‒3′）
VcMYB17-T-F	ATGGGGAGAAGCCCATGTTGC
VcMYB17-T-R	TTAATTATCTGCTAGCCAATCACCGCC
VcMYB17-pCAMBIA1205-F	CGCGGATCCATGGGGAGAAGCCCA
VcMYB17-pCAMBIA1205-R	CGGGGTACCGGCGGTGATTGGCTA
VcMYB17-pGBKT7-F	TGCATATGGCCATGGATGGGGAGAAGCCCA
VcMYB17-pGBKT7-R	CCGCTGCAGGTCGACATTATCTGCTAGCCA
VcMYB17-pCAMBIA1300-F	AGCTCGGTACCCGGGATGGGGAGAAGCCCA
VcMYB17-pCAMBIA1300-R	CATGTCGACTCTAGAATTATCTGCTAGCCA

Fig. 1 Identification and phylogenetic analysis of VcMYB17A: VcMYB17 transcriptome data analysis. B: VcMYB17 domain analysis. C: Multiple sequence alignment of VcMYB17. D: Phylogenetic tree analysis of VcMYB17. Different letters indicate the use of Turkey test methods significantly different (P<0.05), the same below

Fig. 2 Expression pattern analysis of VcMYB17A: Tissue expression specificity analysis of VcMYB17. B-E: The relative expression of VcMYB17 after ABA, GA, MeJA and SA treatment. F: Response of VcMYB17 to 10% PEG

Fig. 3 Subcellular localization and transcriptional activity analysis of VcMYB17A: Subcellular localization analysis of VcMYB17, scale bar=25 μm. B: Transcriptional activity analysis of VcMYB17

Fig. 4 Drought phenotypic analysis of VcMYB17-OE A. thaliana and determination of related indexesA: Identification of transgenic Arabidopsis thaliana at DNA level. B: Identification of transgenic A. thaliana by RT-qPCR. C: Drought phenotype analysis of A.thaliana with different genotypes. D‒I: Results of physiological indexes of A.thaliana with different genotypes under normal and drought conditions

Fig. 5 Phenotypic analysis of callus and drought in VcMYB17-OE blueberry and determination of related indexesA: Identification of VcMYB17-OE blueberry callus. B: Phenotypic analysis of transgenic callus drought, bar=2 cm. C‒F: Results of physiological indexes under normal and 5% PEG treatment

Fig. 6 Expressions of drought-tolerant genes in A. thaliana with different genotypes under normal and drought treatment

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