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INTRODUCTION: Verticillium wilt (VW), caused by Verticillium dahliae, severely reduces cotton yield and fiber quality. Previous transcriptomic analysis in V. dahliae-inoculated Arabidopsis thaliana identified the pathogen-induced DIR1-like gene AT3G53980.2. In cotton, we discovered a homologous gene, GhDIR1 (Gh_A09G180700.1), encoding a lipid transfer protein. This study investigates its role in cotton resistance to V. dahliae.
RATIONALE: We characterized GhDIR1’s molecular features, expression patterns under pathogen stress, and functional impact using bioinformatics, subcellular localization, qRT-PCR, and virus-induced gene silencing (VIGS) analyses. Transcriptomic analysis of wild-type and GhDIR1-silenced plants were conducted to unravel downstream regulatory networks, focusing on metabolic pathways linked to plant immunity.
RESULTS: The results showed that GhDIR1 contains a 351 bp ORF encoding 116 amino acids. Subcellular localization confirmed its presence on the cell membrane. qRT-PCR showed rapid induction of GhDIR1 by V. dahliae. Silencing GhDIR1 increased cotton susceptibility to the pathogen. Transcriptomic data revealed that differentially expressed genes in silenced plants were enriched in flavonoid biosynthesis, sesquiterpene/triterpene biosynthesis, and α-linolenic acid metabolism. Key genes (GhCHS, GhDFR, GhCAD, GhSEQ, GhLOX, and GhAOC) in these pathways were downregulated, suggesting impaired synthesis of protective metabolites.
CONCLUSION: It is speculated that GhDIR1 positively regulates cotton resistance to VW by modulating flavonoid and terpenoid biosynthesis and jasmonic acid-related signaling. Its silencing disrupts critical defense pathways, highlighting its role in coordinating immune responses. These findings propose GhDIR1 as a potential target for enhancing disease resistance in cotton.
The induced expression pattern of GhDIR1 and related genes after inoculation with Verticillium dahliae.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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