Identification of Genetic and Chemical Factors Affecting Type III Secretion System Expression in Pseudomonas syringae pv. actinidiae Biovar 3 Using a Luciferase Reporter Construct

Author:

Zhi Taihui1,Liu Quanhong1,Xie Ting1,Ding Yue1,Hu Renjian1,Sun Yu1,Fan Rong2,Long Youhua2,Zhao Zhibo1ORCID

Affiliation:

1. Department of Plant Pathology, College of Agriculture, Guizhou University, Guiyang 550025, People’s Republic of China

2. Kiwifruit Engineering and Technology Research Center, College of Agriculture, Guizhou University, Guiyang 550025, People’s Republic of China

Abstract

The type III secretion system (T3SS) is a key factor in the pathogenesis of Pseudomonas syringae pv. actinidiae biovar 3 (Psa3), the causal agent of a global kiwifruit bacterial canker pandemic. To monitor the T3SS expression levels in Psa3, we constructed a luciferase reporter plasmid-expressing HrpAPsa3-NLuc fusion protein. The expression of HrpA-NLuc was induced in hrp-inducing conditions whereas the level of luciferase activity correlated with the expression of hrp/hrc genes in Psa3 confirmed the reliability of the reporter construct. Based on the readout of the NLuc reporter construct, three small molecule compounds 4-methoxy-cinnamic acid, sulforaphane, and ferulic acid were determined as T3SS inhibitors in Psa3, whereas sodium acetate was determined to be a T3SS inducer. Moreover, the aqueous extract of fruit inhibited the accumulation of HrpA-NLuc in Psa3 in medium and in planta. Additionally, the T3SS inhibitors suppress Psa3 virulence, whereas the T3SS inducer promotes Psa3 virulence on kiwifruit. Thus, our findings may provide clues to why the fruit is not infected by Psa3, and the Psa3 T3SS inhibitors have potential as alternatives to current nonspecific antimicrobials for disease management.

Funder

National Natural Science Foundation of China

Guizhou Provincial Science and Technology Foundation

Publisher

Scientific Societies

Subject

Plant Science,Agronomy and Crop Science

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