scholarly journals Bacterial Leaf Infiltration Assay for Fine Characterization of Plant Defense Responses using the Arabidopsis thaliana-Pseudomonas syringae Pathosystem

10.3791/53364 ◽  
2015 ◽  
Author(s):  
Xiaoyu Liu ◽  
Yali Sun ◽  
Camilla J Kørner ◽  
Xinran Du ◽  
Marie E. Vollmer ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Defense Responses ◽  
Plant Defense Responses

scholarly journals An efficient direct screening system for microorganisms that activate plant immune responses based on plant–microbe interactions using cultured plant cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mari Kurokawa ◽  
Masataka Nakano ◽  
Nobutaka Kitahata ◽  
Kazuyuki Kuchitsu ◽  
Toshiki Furuya
Keyword(s):  
Immune Responses ◽  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Large Scale ◽  
Plant Cells ◽  
Defense Responses ◽  
Root Tip ◽  
General Strategy ◽  
Plant Defense Responses ◽  
Microbe Interactions

AbstractMicroorganisms that activate plant immune responses have attracted considerable attention as potential biocontrol agents in agriculture because they could reduce agrochemical use. However, conventional methods to screen for such microorganisms using whole plants and pathogens are generally laborious and time consuming. Here, we describe a general strategy using cultured plant cells to identify microorganisms that activate plant defense responses based on plant–microbe interactions. Microbial cells were incubated with tobacco BY-2 cells, followed by treatment with cryptogein, a proteinaceous elicitor of tobacco immune responses secreted by an oomycete. Cryptogein-induced production of reactive oxygen species (ROS) in BY-2 cells served as a marker to evaluate the potential of microorganisms to activate plant defense responses. Twenty-nine bacterial strains isolated from the interior of Brassica rapa var. perviridis plants were screened, and 8 strains that enhanced cryptogein-induced ROS production in BY-2 cells were selected. Following application of these strains to the root tip of Arabidopsis seedlings, two strains, Delftia sp. BR1R-2 and Arthrobacter sp. BR2S-6, were found to induce whole-plant resistance to bacterial pathogens (Pseudomonas syringae pv. tomato DC3000 and Pectobacterium carotovora subsp. carotovora NBRC 14082). Pathogen-induced expression of plant defense-related genes (PR-1, PR-5, and PDF1.2) was enhanced by the pretreatment with strain BR1R-2. This cell–cell interaction-based platform is readily applicable to large-scale screening for microorganisms that enhance plant defense responses under various environmental conditions.

scholarly journals Jasmonic Acid at the Crossroads of Plant Immunity and Pseudomonas syringae Virulence

2020 ◽  
Vol 21 (20) ◽  
pp. 7482
Author(s):  
Aarti Gupta ◽  
Mamta Bhardwaj ◽  
Lam-Son Phan Tran
Keyword(s):  
Jasmonic Acid ◽  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Regulatory Networks ◽  
Genetic Manipulation ◽  
Plant Immunity ◽  
Stomatal Closure ◽  
Defense Responses ◽  
Plant Defense Responses ◽  
Callose Deposition

Sensing of pathogen infection by plants elicits early signals that are transduced to affect defense mechanisms, such as effective blockage of pathogen entry by regulation of stomatal closure, cuticle, or callose deposition, change in water potential, and resource acquisition among many others. Pathogens, on the other hand, interfere with plant physiology and protein functioning to counteract plant defense responses. In plants, hormonal homeostasis and signaling are tightly regulated; thus, the phytohormones are qualified as a major group of signaling molecules controlling the most widely tinkered regulatory networks of defense and counter-defense strategies. Notably, the phytohormone jasmonic acid mediates plant defense responses to a wide array of pathogens. In this review, we present the synopsis on the jasmonic acid metabolism and signaling, and the regulatory roles of this hormone in plant defense against the hemibiotrophic bacterial pathogen Pseudomonas syringae. We also elaborate on how this pathogen releases virulence factors and effectors to gain control over plant jasmonic acid signaling to effectively cause disease. The findings discussed in this review may lead to ideas for the development of crop cultivars with enhanced disease resistance by genetic manipulation.

Characterization of three Arabidopsis thaliana immunophilin genes involved in the plant defense response against Pseudomonas syringae

Gene ◽  
2014 ◽  
Vol 538 (1) ◽  
pp. 12-22 ◽  
Author(s):  
Gennady V. Pogorelko ◽  
Maria Mokryakova ◽  
Oksana V. Fursova ◽  
Inna Abdeeva ◽  
Eleonora S. Piruzian ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Defense Response ◽  
Plant Defense Response

scholarly journals Suppression of plant defense responses by extracellular metabolites from Pseudomonas syringae pv. tabaci in Nicotiana benthamiana

2013 ◽  
Vol 13 (1) ◽  
pp. 65 ◽  
Author(s):  
Seonghee Lee ◽  
Dong Sik Yang ◽  
Srinivasa Rao Uppalapati ◽  
Lloyd W Sumner ◽  
Kirankumar S Mysore
Keyword(s):  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Nicotiana Benthamiana ◽  
Defense Responses ◽  
Plant Defense Responses ◽  
Extracellular Metabolites

Corrigendum to “Characterization of three Arabidopsis thaliana immunophilin genes involved in the plant defense response against Pseudomonas syringae” [Gene 538 (2014) 12–22]

Gene ◽  
2014 ◽  
Vol 539 (2) ◽  
pp. 283
Author(s):  
Gennady V. Pogorelko ◽  
Maria Mokryakova ◽  
Oksana V. Fursova ◽  
Inna Abdeeva ◽  
Eleonora S. Piruzian ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Defense Response ◽  
Plant Defense Response

scholarly journals Effectors from Wheat Rust Fungi Suppress Multiple Plant Defense Responses

2017 ◽  
Vol 107 (1) ◽  
pp. 75-83 ◽  
Author(s):  
Sowmya R. Ramachandran ◽  
Chuntao Yin ◽  
Joanna Kud ◽  
Kiwamu Tanaka ◽  
Aaron K. Mahoney ◽  
...  
Keyword(s):  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Hypersensitive Response ◽  
Defense Responses ◽  
Effector Proteins ◽  
R Gene ◽  
Plant Defense Responses ◽  
Functional Studies ◽  
Rust Diseases ◽  
Suppression Assay

Fungi that cause cereal rust diseases (genus Puccinia) are important pathogens of wheat globally. Upon infection, the fungus secretes a number of effector proteins. Although a large repository of putative effectors has been predicted using bioinformatic pipelines, the lack of available high-throughput effector screening systems has limited functional studies on these proteins. In this study, we mined the available transcriptomes of Puccinia graminis and P. striiformis to look for potential effectors that suppress host hypersensitive response (HR). Twenty small (<300 amino acids), secreted proteins, with no predicted functions were selected for the HR suppression assay using Nicotiana benthamiana, in which each of the proteins were transiently expressed and evaluated for their ability to suppress HR caused by four cytotoxic effector‐R gene combinations (Cp/Rx, ATR13/RPP13, Rpt2/RPS‐2, and GPA/RBP‐1) and one mutated R gene—Pto(Y207D). Nine out of twenty proteins, designated Shr1 to Shr9 (suppressors of hypersensitive response), were found to suppress HR in N. benthamiana. These effectors varied in the effector-R gene defenses they suppressed, indicating these pathogens can interfere with a variety of host defense pathways. In addition to HR suppression, effector Shr7 also suppressed PAMP-triggered immune response triggered by flg22. Finally, delivery of Shr7 through Pseudomonas fluorescens EtHAn suppressed nonspecific HR induced by Pseudomonas syringae DC3000 in wheat, confirming its activity in a homologous system. Overall, this study provides the first evidence for the presence of effectors in Puccinia species suppressing multiple plant defense responses.

scholarly journals Pseudomonas syringae pv. tomato DC3000 Effector HopG1 is a multi-faceted protein that Triggers Necrotic Cell Death that is attenuated by the Nonhost Resistance 2B (AtNHR2B) Protein

2021 ◽  
Author(s):  
Catalina Rodriguez-Puerto ◽  
Rupak Chakraborty ◽  
Raksha Singh ◽  
Perla Rocha-Loyola ◽  
Clemencia M. Rojas
Keyword(s):  
Cell Death ◽  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Plant Immunity ◽  
Defense Responses ◽  
Necrotic Cell Death ◽  
Plant Defense Responses ◽  
Necrotic Cell ◽  
Tomato Dc3000 ◽  
Type Iii

The plant pathogenic bacterium Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) has become a paradigm in plant-bacteria interactions due to its ability to cause disease in the model plant Arabidopsis thaliana. Pst DC3000 uses the type III secretion system to deliver type III secreted effectors (T3SEs) directly into the plant cytoplasm. Pst DC3000 T3SEs contribute to pathogenicity by suppressing plant defense responses and targeting plant’s physiological processes. Although the complete repertoire of effectors encoded in the Pst DC3000 genome have been identified, the specific function for most of them remains to be elucidated. The mitochondrial-localized T3E HopG1, suppresses plant defense responses and promotes the development of disease symptoms. Here, we show that HopG1 triggers necrotic cell death that enables the growth of non-adapted pathogens. We further showed that HopG1 interacts with the plant immunity-related protein AtNHR2B and that AtNHR2B attenuates HopG1- virulence functions.

scholarly journals Down Regulation of Virulence Factors of Pseudomonas aeruginosa by Salicylic Acid Attenuates Its Virulence on Arabidopsis thaliana and Caenorhabditis elegans

2005 ◽  
Vol 73 (9) ◽  
pp. 5319-5328 ◽  
Author(s):  
B. Prithiviraj ◽  
H. P. Bais ◽  
T. Weir ◽  
B. Suresh ◽  
E. H. Najarro ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Arabidopsis Thaliana ◽  
Salicylic Acid ◽  
Plant Defense ◽  
Virulence Factors ◽  
Target Genes ◽  
Defense Responses ◽  
Infection Model ◽  
Plant Defense Responses ◽  
Wild Type

ABSTRACT Salicylic acid (SA) is a phenolic metabolite produced by plants and is known to play an important role in several physiological processes, such as the induction of plant defense responses against pathogen attack. Here, using the Arabidopsis thaliana-Pseudomonas aeruginosa pathosystem, we provide evidence that SA acts directly on the pathogen, down regulating fitness and virulence factor production of the bacteria. Pseudomonas aeruginosa PA14 showed reduced attachment and biofilm formation on the roots of the Arabidopsis mutants lox2 and cpr5-2, which produce elevated amounts of SA, as well as on wild-type Arabidopsis plants primed with exogenous SA, a treatment known to enhance endogenous SA concentration. Salicylic acid at a concentration that did not inhibit PA14 growth was sufficient to significantly affect the ability of the bacteria to attach and form biofilm communities on abiotic surfaces. Furthermore, SA down regulated three known virulence factors of PA14: pyocyanin, protease, and elastase. Interestingly, P. aeruginosa produced more pyocyanin when infiltrated into leaves of the Arabidopsis transgenic line NahG, which accumulates less SA than wild-type plants. This finding suggests that endogenous SA plays a role in down regulating the synthesis and secretion of pyocyanin in vivo. To further test if SA directly affects the virulence of P. aeruginosa, we used the Caenorhabiditis elegans-P. aeruginosa infection model. The addition of SA to P. aeruginosa lawns significantly diminished the bacterium's ability to kill the worms, without affecting the accumulation of bacteria inside the nematodes' guts, suggesting that SA negatively affects factors that influence the virulence of P. aeruginosa. We employed microarray technology to identify SA target genes. These analyses showed that SA treatment affected expression of 331 genes. It selectively repressed transcription of exoproteins and other virulence factors, while it had no effect on expression of housekeeping genes. Our results indicate that in addition to its role as a signal molecule in plant defense responses, SA works as an anti-infective compound by affecting the physiology of P. aeruginosa and ultimately attenuating its virulence.

scholarly journals Isolation of Arabidopsis Mutants With Enhanced Disease Susceptibility by Direct Screening

Genetics ◽  
1996 ◽  
Vol 143 (2) ◽  
pp. 973-982 ◽  
Author(s):  
Jane Glazebrook ◽  
Elizabeth E Rogers ◽  
Frederick M Ausubel
Keyword(s):  
Plant Defense ◽  
Pseudomonas Syringae ◽  
Disease Susceptibility ◽  
Bacterial Pathogen ◽  
Defense Responses ◽  
Screening Strategy ◽  
Genetic Dissection ◽  
Plant Defense Responses ◽  
Defense Genes ◽  
Increased Susceptibility

Abstract To discover which components of plant defense responses make significant contributions to limiting pathogen attack, we screened a mutagenized population of Arabidopsis thalzana for individuals that exhibit increased susceptibility to the moderately virulent bacterial pathogen Pseudomonas syringae pv. maculicola ES4326 (Psm ES4326). The 12 enhanced disease susceptibility (eds) mutants isolated included alleles of two genes involved in phytoalexin biosynthesis (pad2, which had been identified previously, and pad4, which had not been identified previously), two alleles of the previously identified npr1 gene, which affects expression of other defense genes, and alleles of seven previously unidentified genes of unknown function. The npr-1 mutations caused greatly reduced expression of the PRI gene in response to PsmES4326 infection, but had little effect on expression of two other defense genes, BGL2 and PR5, suggesting that PR1 expression may be important for limiting growth of PsmES4326. While direct screens for mutants with quantitative pathogen-susceptibility phenotypes have not been reported previously, our finding that mutants isolated in this way include those affected in known defense responses supports the notion that this type of screening strategy allows genetic dissection of the roles of various plant defense responses in disease resistance.

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