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

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

Abstract


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


Full Text:

PDF

References


Thordal-Christensen, H. Fresh insights into processes of nonhost resistance. Current Opinion in Plant Biology, (2003); 6(4):351-357.

Chisholm ST, Coaker G, Day B, Staskawicz BJ. Host-microbe interactions: shaping the evolution of the plant immune response. Cell, (2006); 124(4):803-814.

Zipfel C, Robatzek S, Navarro L, Oakeley EJ, Jones JD, et al. Bacterial disease resistance in Arabidopsis through flagellin perception. Nature, (2004); 428(6984):764-767.

Arasimowicz M, Floryszak-Wieczorek J. Nitric oxide as a bioactive signalling molecule in plant stress responses. Plant Science, (2007); 172(5):876-887.

Ali MA, Abbas A, Kreil DP, Bohlmann H. Overexpression of the transcription factor RAP2.6 leads to enhanced callose deposition in syncytia and enhanced resistance against the beet cyst nematode Heterodera schachtii in Arabidopsis roots. BMC Plant Biology, (2013); 13:

Ali MA, Anjam MS, Nawaz MA, Lam HM, Chung G. Signal Transduction in Plant-Nematode Interactions. International Journal of Molecular Sciences, (2018); 19(6):1648.

Rai VK. Role of Amino Acids in Plant Responses to Stresses. Biologia Plantarum, (2002); 45(4): 481-487.

Hofmann J, El Ashry AEN, Anwar S, Erban A, Kopka J, et al. Metabolic profiling reveals local and systemic responses of host plants to nematode parasitism. Plant Journal, (2010); 62(6):1058-1071.

Anwar S, Wieczorek K, Inselsbacher E. Analysis of Arabidopsis amino acid metabolism in response to Heterodera schachtii infection. Pakistan Journal of Nematology, (2018); 36(2): 131-150.

Delledonne M, Xia Y, Dixon RA, Lamb C. Nitric oxide functions as a signal in plant disease resistance. Nature, (1998); 394(6693):585-588.

Imran QM, Hussain A, Mun BG, Lee SU, Asaf S, et al. Transcriptome wide identification and characterization of NO-responsive WRKY transcription factors in Arabidopsis thaliana L. Environmental and Experimental Botany, (2018); 148:128-143.

Modolo LV, Augusto O, Almeida IMG, Pinto-Maglio CAF, Oliveira HC, et al. Decreased arginine and nitrite levels in nitrate reductase-deficient Arabidopsis thaliana plants impair nitric oxide synthesis and the hypersensitive response to Pseudomonas syringae. Plant Science, (2006); 171(1):34-40.

Micallef BJ, Shelp BJ. Arginine metabolism in developing soybean cotyledons .2. Biosynthesis. Plant Physiology, (1989); 90(2):631-634.

Slocum RD. Genes, enzymes and regulation of arginine biosynthesis in plants. Plant Physiology and Biochemistry, (2005); 43(8):729-745.

Anwar S, Inselsbacher E, Grundler FMW, Hofmann J. Arginine metabolism of Arabidopsis thaliana is modulated by Heterodera schachtii infection. Nematology, (2015); 17(9): 1027-1043.

Ishiga Y, Ishiga T, Uppalapati SR, Mysore KS. Arabidopsis seedling flood-inoculation technique: a rapid and reliable assay for studying plant-bacterial interactions. Plant Methods, (2011); 7:32.

Curtis MD, Grossniklaus UA. Gateway Cloning Vector Set for High-Throughput Functional Analysis of Genes in Planta. Plant Physiology, (2003); 133(2):462-469.

Schwab R, Ossowski S, Riester M, Warthmann N, Weigel D. Highly specific gene silencing by artificial microRNAs in Arabidopsis. Plant Cell, (2006); 18:1121-1133.

Szakasits D, Siddique S, Bohlmann H. An improved pPZP vector for Agrobacterium-mediated plant transformation. Plant Molecular Biology Reporter, (2007); 25(3-4):115-120.

Clough SJ, Bent AF. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant Journal, (1998); 16(6):735-743.

Brownfield D L, Todd C D, Deyholos MK. Analysis of Arabidopsis arginase gene transcription patterns indicates specific biological functions for recently diverged paralogs. Plant Molecular Biology, (2008);

(4):429-440.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-DDCt Method. Methods, (2001); 25(4):402-408.

Das P, Lahiri A, Lahiri A, Chakravortty D. Modulation of the Arginase Pathway in the Context of Microbial Pathogenesis: A Metabolic Enzyme Moonlighting as an Immune Modulator. Plos Pathogens, (2010); 6(6):e1000899.

Bouchereau A, Aziz A, Larher F, Martin-Tanguy J. Polyamines and environmental challenges: recent development. Plant Science, (1999); 140:103-125.

Walters DR. Polyamines and plant disease. Phytochemistry, (2003); 64(1):97-107.

Anwar S, Wieczorek K. Serendipita indica modulates expression of arginine metabolism genes during colonization of Arabidopsis thaliana roots. Pure and Applied Biology, (2018); http://dx.doi.org/10.19045/bspab.2018.700203

Haines RJ, Pendleton LC, Eichler DC. Argininosuccinate synthase: at the center of arginine metabolism. International Journal of Biochemistry and Molecular Biology, (2011); 2(1): 8-23.

Erez A, Nagamani SC, Shchelochkov OA, Premkumar MH, Campeau PM, et al. Requirement of argininosuccinate lyase for systemic nitric oxide production. Nature Medicine, (2011); 17(12): 1619-1626.

Chen H. Regulation of Plant Arginase by Wounding, Jasmonate, and the Phytotoxin Coronatine. Journal of Biological Chemistry, (2004); 279(44):45998-46007.

Jubault M, Hamon C, Gravot A, Lariagon C, Delourme R, et al. Differential regulation of root arginine catabolism and polyamine metabolism in clubroot-susceptible and partially resistant Arabidopsis genotypes. Plant Physiology, (2008); 146(4):2008-2019.

Flores T, Todd CD, Tovar-Mendez A, Dhanoa PK, Correa-Aragunde N, et al. Arginase-negative mutants of Arabidopsis exhibit increased nitric oxide signaling in root development. Plant Physiology, (2008); 147(4):1936-1946.




DOI: http://dx.doi.org/10.62940/als.v7i1.803

Refbacks

  • There are currently no refbacks.