Arabidopsis Argininosuccinate Lyase and Argininosuccinate Synthase are important for resistance against Pseudomonas syringae

Shahbaz Anwar, Muhammad Amjad Ali, Amjad Abbas, Krzysztof Wieczorek


Background: Arginine is one of the important amino acids and is involved in a variety of plant responses against biotic and abiotic stresses. Two important genes Argininosuccinate Synthase (AS) and Argininosuccinate Lyase (AL) are associated with the production of arginine from ornithine. Here we report molecular characterization of Arabidopsis thaliana artificial micro RNA (amiRNA) mutants for the genes AL and AS in response to infection with Pseudomonas syringae DC3000.

Methods: Quantitative real time PCR (qRT-PCR) and Promoter::GUS plant lines were used for expression analysis. Artificial micro RNAs (amiRNA) mediated gene silencing was used to generate mutant plant lines. Flood inoculation technique was used for infection test essays with Pseudomonas syringae.

Results: Expression analysis of A. thaliana plants harboring promoter AS::GUS construct showed strong promoter activity upon P. syringae infection. Quantitative real time PCR (qRT-PCR) analysis showed that AL and AS expression was strongly induced upon infection with P. syringae. Infection essays for P. syringae showed enhanced susceptibility to virulent (Pto) as well as avirulent (∆avrPto/∆avrPtoB) strains of P. syringae. However, mutant plants infiltered with infiltration medium containing 1 mM L-arginine regain their resistance in comparison with wild type (Col-0) plants.

Conclusion: Our findings suggest that genes related to arginine metabolism play a key role in plant defenses during P. syringae infection on A. thaliana. This study revealed that proper functioning of arginine related genes is required to deploy defense response against P. syringae. Decrease in the expression of these genes improves conditions for the growth of pathogen.

Keywords: Arabidopsis thaliana; arginine; Argininosuccinate Lyase; Argininosuccinate Synthase; Gene silencing; Pseudomonas syringae

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