The cotton MYB33 gene is a hub gene regulating the trade-off between plant growth and defense in Verticillium dahliae infection

Hu Guang; Ge Xiaoyang; Wang Zhian; Wang Ye; Wang Peng; Shi Linfang; Wang Bingting; Zhang Anhong; Li Fuguang; Wu Jiahe

doi:10.1016/j.jare.2023.08.017

The cotton MYB33 gene is a hub gene regulating the trade-off between plant growth and defense in Verticillium dahliae infection

J Adv Res. 2023 Aug 28:S2090-1232(23)00233-3. doi: 10.1016/j.jare.2023.08.017. Online ahead of print.

Authors

Hu Guang¹, Ge Xiaoyang², Wang Zhian³, Wang Ye², Wang Peng², Shi Linfang⁴, Wang Bingting⁴, Zhang Anhong³, Li Fuguang⁵, Wu Jiahe⁶

Affiliations

¹ National Key Laboratory of Cotton Bio‑breeding and Integrated Utilization, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, China; State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
² National Key Laboratory of Cotton Bio‑breeding and Integrated Utilization, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, China.
³ Institute of Cotton Research, Shanxi Agricultural University, Yuncheng 044000, China.
⁴ State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
⁵ National Key Laboratory of Cotton Bio‑breeding and Integrated Utilization, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, China. Electronic address: aylifug@caas.cn.
⁶ State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. Electronic address: wujiahe@im.ac.cn.

PMID: 37648022
DOI: 10.1016/j.jare.2023.08.017

Abstract

Introduction: Sessile plants engage in trade-offs between growth and defense capacity in response to fluctuating environmental cues. MYB is an important transcription factor that plays many important roles in controlling plant growth and defense. However, the mechanism behind how it keeps a balance between these two physiological processes is still largely unknown.

Objectives: Our work focuses on the dissection of the molecular mechanism by which GhMYB33 regulates plant growth and defense.

Methods: The CRISPR/Cas9 technique was used to generate mutants for deciphering GhMYB33 functions. Yeast two-hybrid, luciferase complementary imaging, and co-immunoprecipitation assays were used to prove that proteins interact with each other. We used the electrophoretic mobility shift assay, yeast one-hybrid, and luciferase activity assays to analyze GhMYB33 acting as a promoter. A β-glucuronidase fusion reporter and 5' RNA ligase mediated amplification of cDNA ends analysis showed that ghr-miR319c directedly cleaved the GhMYB33 mRNA.

Results: Overexpressing miR319c-resistant GhMYB33 (rGhMYB33) promoted plant growth, accompanied by a significant decline in resistance against Verticillium dahliae. Conversely, its knockout mutant, ghmyb33, demonstrated growth restriction and concomitant augmentation of V. dahliae resistance. GhMYB33 was found to couple with the DELLA protein GhGAI1 and bind to the specific cis-elements of GhSPL9 and GhDFR1 promoters, thereby modulating internode elongation and plant resistance in V. dahliae infection. The ghr-miR319c was discovered to target and suppress GhMYB33 expression. The overexpression of ghr-miR319c led to enhanced plant resistance and a simultaneous reduction in plant height.

Conclusion: Our findings demonstrate that GhMYB33 encodes a hub protein and controls the expression of GhSPL9 and GhDFR1, implicating a pivotal role for the miR319c-MYB33 module to regulate the trade-offs between plant growth and defense.

Keywords: GhMYB33; Gossypium hirsutum; Trade-off; Verticillium dahliae; ghr-miR319c.