Functional Analysis of Autophagy Genes via Agrobacterium-Mediated Transformation in the Vascular Wilt Fungus Verticillium dahliae

• Published in: Journal of genetics and genomics Vol. 40; no. 8; pp. 421 - 431
• Main Authors: Zhou, Lei, Zhao, Jun, Guo, Wangzhen, Zhang, Tianzhen
• Format: Journal Article
• Language: English
• Published: China Elsevier Ltd 20.08.2013
• Subjects: Agrobacterium; Agrobacterium tumefaciens - genetics; Arabidopsis; ATMT; Autophagy; conidia; conidiation; Fungal Proteins - genetics; Fungal Proteins - metabolism; fungus physiology; Gene Deletion; genes; Gossypium - microbiology; hyphae; mutants; organelles; Pathogenicity; plant development; Plant Diseases - microbiology; plant pathogenic fungi; proteins; roots; Spores, Fungal - genetics; Spores, Fungal - growth & development; Spores, Fungal - metabolism; Targeted gene replacement; Transformation, Genetic; vascular wilt; Verticillium - cytology; Verticillium - genetics; Verticillium - growth & development; Verticillium - metabolism; Verticillium dahliae; 农杆菌介导转化; 基因功能分析; 枯萎病; 植物病原真菌; 病菌感染; 缺失突变体; 自噬作用; 黄萎病菌; Pathogenicity; Targeted gene replacement; Verticillium dahliae; ATMT; Autophagy
• ISSN: 1673-8527
• DOI: 10.1016/j.jgg.2013.04.006

Autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in tbliar infection by many plant pathogenic fungi. However, the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood. Here, we report the establishment of an Agro- bacterium tumefaciens-mediated transformation （ATMT） system and its application to investigate two autophagy genes, VdATG8 and VdATG12, by means of targeted gene replacement and complementation. Transformation of a cotton-infecting Verticillium dahliae strain Vd8 with a novel binary vector pCOM led to the production of 384 geneticin-resistant translbnnants per 1 × 10^4 conidia. V. dahliae mutants lacking either VdATG8 or VdATGI2 exhibited reduced conidiation and impaired aerial hyphae production. Disease development on Arabidopsis plants was slightly delayed when inoculated with VdATG8 or VdATG12 gene deletion mutants, compared with the wild- type and gene complemented strains. Surprisingly, in vitro inoculation with unimpaired roots revealed that the abilities of root invasion were not affected in gene deletion mutants. These results indicate that autophagy is necessary for aerial hyphae development and plant colonization but not for root infection in E dahliae.