HSP70s Enhance a <i>Phytophthora infestans</i> Effector-Induced Cell Death via an MAPK Cascade in <i>Nicotiana benthamiana</i>

Lee, Joo Hyun,Lee, So Eui,Oh, Soohyun,Seo, Eunyoung,Choi, Doil APS Press 2018 Molecular plant-microbe interactions Vol.31 No.3

Potato virus Y HCPro localization at distinct, dynamically related and environment-influenced structures in the cell cytoplasm.

del Toro, Francisco,Fern?ndez, F?tima Tena,Tilsner, Jens,Wright, Kathryn M,Tenllado, Francisco,Chung, Bong Nam,Praveen, Shelly,Canto, Tomas APS Press 2014 Molecular plant-microbe interactions Vol.27 No.12

<P>Potyvirus HCPro is a multifunctional protein that, among other functions, interferes with antiviral defenses in plants and mediates viral transmission by aphid vectors. We have visualized in vivo the subcellular distribution and dynamics of HCPro from Potato virus Y and its homodimers, using green, yellow, and red fluorescent protein tags or their split parts, while assessing their biological activities. Confocal microscopy revealed a pattern of even distribution of fluorescence throughout the cytoplasm, common to all these modified HCPros, when transiently expressed in Nicotiana benthamiana epidermal cells in virus-free systems. However, in some cells, distinct additional patterns, specific to some constructs and influenced by environmental conditions, were observed: i) a small number of large, amorphous cytoplasm inclusions that contained α-tubulin; ii) a pattern of numerous small, similarly sized, dot-like inclusions distributing regularly throughout the cytoplasm and associated or anchored to the cortical endoplasmic reticulum and the microtubule (MT) cytoskeleton; and iii) a pattern that smoothly coated the MT. Furthermore, mixed and intermediate forms from the last two patterns were observed, suggesting dynamic transports between them. HCPro did not colocalize with actin filaments or the Golgi apparatus. Despite its association with MT, this network integrity was required neither for HCPro suppression of silencing in agropatch assays nor for its mediation of virus transmission by aphids.</P>

A systems approach for identifying resistance factors to Rice stripe virus.

Kim, Kangmin,Choi, Daeseok,Kim, Sang-Min,Kwak, Do-Yeon,Choi, Jaemyung,Lee, Seungchul,Lee, Bong-Choon,Hwang, Daehee,Hwang, Ildoo APS Press 2012 Molecular plant-microbe interactions Vol.25 No.4

<P>Rice stripe virus (RSV) causes disease that can severely affect the productivity of rice (Oryza sativa). Several RSV-resistant cultivars have been developed. However, host factors conferring RSV resistance in these cultivars are still elusive. Here, we present a systems approach for identifying potential rice resistance factors. We developed two near-isogenic lines (NIL), RSV-resistant NIL22 and RSV-susceptible NIL37, and performed gene expression profiling of the two lines in RSV-infected and RSV-uninfected conditions. We identified 237 differentially expressed genes (DEG) between NIL22 and NIL37. By integrating with known quantitative trait loci (QTL), we selected 11 DEG located within the RSV resistance QTL as RSV resistance factor candidates. Furthermore, we identified 417 DEG between RSV-infected and RSV-uninfected conditions. Using an interaction network-based method, we selected 20 DEG highly interacting with the two sets of DEG as RSV resistance factor candidates. Among the 31 candidates, we selected the final set of 21 potential RSV resistance factors whose differential expression was confirmed in the independent samples using real-time reverse-transcription polymerase chain reaction. Finally, we reconstructed a network model delineating potential association of the 21 selected factors with resistance-related processes. In summary, our approach, based on gene expression profiling, revealed potential host resistance factors and a network model describing their relationships with resistance-related processes, which can be further validated in detailed experiments.</P>

Use of a Secretion Trap Screen in Pepper Following Phytophthora capsici Infection Reveals Novel Functions of Secreted Plant Proteins in Modulating Cell Death.

Yeom, Seon-In,Baek, Hyang-Ku,Oh, Sang-Keun,Kang, Won-Hee,Lee, Sang Jik,Lee, Je Min,Seo, Eunyoung,Rose, Jocelyn K C,Kim, Byung-Dong,Choi, Doil APS Press 2011 Molecular plant-microbe interactions Vol.24 No.6

<P>In plants, the primary defense against pathogens is mostly inducible and associated with cell wall modification and defense-related gene expression, including many secreted proteins. To study the role of secreted proteins, a yeast-based signal-sequence trap screening was conducted with the RNA from Phytophthora capsici-inoculated root of Capsicum annuum 'Criollo de Morelos 334' (CM334). In total, 101 Capsicum annuum secretome (CaS) clones were isolated and identified, of which 92 were predicted to have a secretory signal sequence at their N-terminus. To identify differences in expressed CaS genes between resistant and susceptible cultivars of pepper, reverse Northern blots and real-time reverse-transcription polymerase chain reaction were performed with RNA samples isolated at different time points following P. capsici inoculation. In an attempt to assign biological functions to CaS genes, we performed in planta knock-down assays using the Tobacco rattle virus-based gene-silencing method. Silencing of eight CaS genes in pepper resulted in suppression of the cell death induced by the non-host bacterial pathogen (Pseudomonas syringae pv. tomato T1). Three CaS genes induced phenotypic abnormalities in silenced plants and one, CaS259 (PR4-l), caused both cell death suppression and perturbed phenotypes. These results provide evidence that the CaS genes may play important roles in pathogen defense as well as developmental processes.</P>

The Hypersensitive Induced Reaction and Leucine-Rich Repeat Proteins Regulate Plant Cell Death Associated with Disease and Plant Immunity

Choi, H.W.,Kim, Y.J.,Hwang, B.K. APS PRESS 2011 Molecular plant-microbe interactions Vol.24 No.1

Whole-genome expression profiling of Bradyrhizobium japonicum in response to hydrogen peroxide.

Jeon, Jeong-Min,Lee, Hae-In,Donati, Andrew J,So, Jae-Seong,Emerich, David W,Chang, Woo-Suk APS Press 2011 Molecular plant-microbe interactions Vol.24 No.12

<P>Bradyrhizobium japonicum, a nitrogen-fixing bacterium in soil, establishes a symbiotic relationship with the leguminous soybean plant. Despite a mutualistic association between the two partners, the host plant produces an oxidative burst to protect itself from the invasion of rhizobial cells. We investigated the effects of H(2)O(2)-mediated oxidative stress on B. japonicum gene expression in both prolonged exposure (PE) and fulminant shock (FS) conditions. In total, 439 and 650 genes were differentially expressed for the PE and FS conditions, respectively, at a twofold cut-off with q < 0.05. A number of genes within the transport and binding proteins category were upregulated during PE and a majority of those genes are involved in ABC transporter systems. Many genes encoding ? factors, global stress response proteins, the FixK(2) transcription factor, and its regulatory targets were found to be upregulated in the FS condition. Surprisingly, catalase and peroxidase genes which are typically expressed in other bacteria under oxidative stress were not differentially expressed in either condition. The isocitrate lyase gene (aceA) was induced by fulminant H(2)O(2) shock, as was evident at both the transcriptional and translational levels. Interestingly, there was no significant effect of H(2)O(2) on exopolysaccharide production at the given experimental conditions.</P>

FSR1 is essential for virulence and female fertility in Fusarium verticillioides and F. graminearum.

Shim, Won-Bo,Sagaram, Uma Shankar,Choi, Yoon-E,So, Jinny,Wilkinson, Heather H,Lee, Yin-Won APS Press 2006 Molecular plant-microbe interactions Vol.19 No.7

<P>Fusarium verticillioides (teleomorph Gibberella moniliformis) and F. graminearum (teleomorph G. zeae) are well known to cause devastating diseases on cereal crops. Despite their importance, our understanding of the molecular mechanisms involved in these host-pathogen interactions is limited. The FSR1 locus in F. verticillioides was identified by screening REMI mutants for loss of virulence in maize stalk rot inoculation studies. FSR1 encodes an 823-codon open reading frame interrupted by two introns. The Fsr1 protein shares 60% sequence identity with the Sordaria macrospora Pro11, a multimodular protein with four putative protein-protein binding domains (caveolin-binding domain, coiled-coil structure, calmodulin-binding motif, and seven-WD40 repeats), which plays a regulatory role in cell differentiation and ascocarp development. Our data demonstrate that FSR1 is essential for female fertility and virulence in F. verticillioides. Significantly, targeted disruption of the FSR1 ortholog in F. graminearum (FgFSR1) reduced virulence on barley and deterred perithecia formation. Cross-complementation experiments demonstrated that the gene function is conserved in the two Fusarium species. FSR1 is expressed constitutively, and we hypothesize that Fsr1 regulates virulence by acting as a scaffold for a signal transduction pathway. A survey of available genome databases indicates Fsr1 homologs are present in a number of filamentous fungi and animal systems but not in budding yeast or plants. A maximum likelihood analysis of this gene family reveals well-supported monophyletic clades associated with fungi and animals.</P>

2R,3R-butanediol, a bacterial volatile produced by Pseudomonas chlororaphis O6, is involved in induction of systemic tolerance to drought in Arabidopsis thaliana.

Cho, Song Mi,Kang, Beom Ryong,Han, Song Hee,Anderson, Anne J,Park, Ju-Young,Lee, Yong-Hwan,Cho, Baik Ho,Yang, Kwang-Yeol,Ryu, Choong-Min,Kim, Young Cheol APS Press 2008 Molecular plant-microbe interactions Vol.21 No.8

<P>Root colonization of plants with certain rhizobacteria, such as Pseudomonas chlororaphis O6, induces tolerance to biotic and abiotic stresses. Tolerance to drought was correlated with reduced water loss in P. chlororaphis O6-colonized plants and with stomatal closure, indicated by size of stomatal aperture and percentage of closed stomata. Stomatal closure and drought resistance were mediated by production of 2R,3R-butanediol, a volatile metabolite of P. chlororaphis O6. Root colonization with bacteria deficient in 2R,3R-butanediol production showed no induction of drought tolerance. Studies with Arabidopsis mutant lines indicated that induced drought tolerance required the salicylic acid (SA)-, ethylene-, and jasmonic acid-signaling pathways. Both induced drought tolerance and stomatal closure were dependent on Aba-1 and OST-1 kinase. Increases in free SA after drought stress of P. chlororaphis O6-colonized plants and after 2R,3R-butanediol treatment suggested a primary role for SA signaling in induced drought tolerance. We conclude that the bacterial volatile 2R,3R-butanediol was a major determinant in inducing resistance to drought in Arabidopsis through an SA-dependent mechanism.</P>

Xanthomonas campestris pv. vesicatoria Effector AvrBsT Induces Cell Death in Pepper, but Suppresses Defense Responses in Tomato.

Kim, Nak Hyun,Choi, Hyong Woo,Hwang, Byung Kook APS Press 2010 Molecular plant-microbe interactions Vol.23 No.8

<P>A type III effector protein, AvrBsT, is secreted into plant cells from Xanthomonas campestris pv. vesicatoria Bv5-4a, which causes bacterial spot disease on pepper (Capsicum annuum) and tomato (Solanum lycopersicum). To define the function and recognition of AvrBsT in the two host plants, avrBsT was introduced into the virulent pepper strain X. campestris pv. vesicatoria Ds1. Expression of AvrBsT in Ds1 rendered the strain avirulent to pepper plants. Infection of pepper leaves with Ds1 (avrBsT) expressing AvrBsT but not with near-isogenic control strains triggered a hypersensitive response (HR) accompanied by strong H(2)O(2) generation, callose deposition, and defense-marker gene expressions. Mutation of avrBsT, however, compromised HR induction by X. campestris pv. vesicatoria Bv5-4a, suggesting its avirulence function in pepper plants. In contrast, AvrBsT acted as a virulence factor in tomato plants. Growth of strains Ds1 (avrBsT) and Bv5-4a DeltaavrBsT was significantly enhanced and reduced, respectively, in tomato leaves. X. campestris pv. vesicatoria-expressed AvrBsT also significantly compromised callose deposition and defense-marker gene expression in tomato plants. Together, these results suggest that the X. campestris pv. vesicatoria type III effector AvrBsT is differentially recognized by pepper and tomato plants.</P>

The pepper calmodulin gene CaCaM1 is involved in reactive oxygen species and nitric oxide generation required for cell death and the defense response.

Choi, Hyong Woo,Lee, Dong Hyuk,Hwang, Byung Kook APS Press 2009 Molecular plant-microbe interactions Vol.22 No.11

<P>Calcium signaling has emerged as an important signal transduction pathway of higher plants in response to biotic and abiotic stresses. Ca2+-bound calmodulin (CaM) plays a critical role in decoding and transducing stress signals by activating specific targets. Here, we isolated and functionally characterized the pathogen-responsive CaM gene, Capsicum annuum calmodulin 1 (CaCaM1), from pepper (C. annuum) plants. The cellular function of CaCaM1 was verified by Agrobacterium spp.-mediated transient expression in pepper and transgenic overexpression in Arabidopsis thaliana. Agrobacterium spp.-mediated transient expression of CaCaM1 activated reactive oxygen species (ROS), nitric oxide (NO) generation, and hypersensitive response (HR)-like cell death in pepper leaves, ultimately leading to local acquired resistance to Xanthomonas campestris pv. vesicatoria. CaCaM1-overexpression (OX) Arabidopsis exhibited enhanced resistance to Pseudomonas syringae and Hyaloperonospora parasitica, which was accompanied by enhanced ROS and NO generation and HR-like cell death. Treatment with the calcium-channel blocker suppressed the oxidative and NO bursts and HR-like cell death that were triggered by CaCaM1 expression in pepper and Arabidopsis, suggesting that calcium influx is required for the activation of CaCaM1-mediated defense responses in plants. Upon treatment with the CaM antagonist, virulent P. syringae pv. tomato-induced NO generation was also compromised in CaCaM1-OX leaves. Together, these results suggest that the CaCaM1 gene functions in ROS and NO generation are essential for cell death and defense responses in plants.</P>