Homologous Expression and T3SS-Dependent Secretion of TAP-Tagged Xo2276 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract and Its Direct In Vitro Recognition of Putative Target DNA Sequence

Kim, Seunghwan; Nguyen, Thi-Dieu-Hanh; Joo-Hee, Lee; Hong, Myoung-Ki; Pham, Tan‐Viet; Ahn, Yeh‐Jin; Lee, Byoung-Moo; Han, Ye; Kim, Dong-Eun; Kim, Jeong-Gu; Kang, Lin‐Woo

doi:10.4014/jmb.1207.07039

Cited by 10 publications

(12 citation statements)

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“…Mutant Xoo strains are usually used to study the pathogenic role of target genes by the comparison with wild-type Xoo strains. In this study, we used an in vitro assay system that activates Xoo pathogenicity by treating Xoo cells with rice leaf extracts (RLX) [ 28 , 29 ]. The in vitro assay system using RLX could induce the expression and secretion of putative effectors of XoAvrBs2 and Xo2276 (AvrBs3-type TAL effector-like protein) and the T3SS-dependent secretions of XoAvrBs2 and Xo2276 were confirmed at 4 h after RLX treatment.…”

Section: Introductionmentioning

confidence: 99%

Time-resolved pathogenic gene expression analysis of the plant pathogen Xanthomonas oryzae pv. oryzae

et al. 2016

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BackgroundPlant-pathogen interactions at early stages of infection are important to the fate of interaction. Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight, which is a devastating disease in rice. Although in vivo and in vitro systems have been developed to study rice-Xoo interactions, both systems have limitations. The resistance mechanisms in rice can be better studied by the in vivo approach, whereas the in vitro systems are suitable for pathogenicity studies on Xoo. The current in vitro system uses minimal medium to activate the pathogenic signal (expression of pathogenicity-related genes) of Xoo, but lacks rice-derived factors needed for Xoo activation. This fact emphasizes the need of developing a new in vitro system that allow for an easy control of both pathogenic activation and for the experiment itself.ResultsWe employed an in vitro system that can activate pathogenicity-related genes in Xoo using rice leaf extract (RLX) and combined the in vitro assay with RNA-Seq to analyze the time-resolved genome-wide gene expression of Xoo. RNA-Seq was performed with samples from seven different time points within 1 h post-RLX treatment and the expression of up- or downregulated genes in RNA-Seq was validated by qRT-PCR. Global analysis of gene expression and regulation revealed the most dramatic changes in functional categories of genes related to inorganic ion transport and metabolism, and cell motility. Expression of many pathogenicity-related genes was induced within 15 min upon contact with RLX. hrpG and hrpX expression reached the maximum level within 10 and 15 min, respectively. Chemotaxis and flagella biosynthesis-related genes and cyclic-di-GMP controlling genes were downregulated for 10 min and were then upregulated. Genes related to inorganic ion uptake were upregulated within 5 min. We introduced a non-linear regression fit to generate continuous time-resolved gene expression levels and tested the essentiality of the transcriptionally upregulated genes by a pathogenicity assay of lesion length using single-gene knock-out Xoo strains.ConclusionsThe in vitro system combined with RNA-Seq generated a genome-wide time-resolved pathogenic gene expression profile within 1 h of initial rice-Xoo interactions, demonstrating the expression order and interaction dependency of pathogenic genes. This combined system can be used as a novel tool to study the initial interactions between rice and Xoo during bacterial blight progression.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2657-7) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Time-resolved pathogenic gene expression analysis of the plant pathogen Xanthomonas oryzae pv. oryzae

et al. 2016

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“…vesicatoria, the role of AvrBs3 is that of a transcription activator-like (TAL) effector that activates the expression of plant immunity genes (Kay, Hahn, Marois, Wieduwild, & Bonas, 2009). The secretion of effectors XoAvrBs2 ( Xoo0168 )—a Xoo ortholog of AvrBs2—and XoAvrBs3 ( Xoo2276)— a Xoo ortholog of AvrBs3—through T3SS was confirmed upon interaction with RLX (S. H. Kim et al, 2011; S. Kim et al, 2013). In addition to the transcriptome and proteome, the expression and secretion of effectors were assessed using dot blots of Xoo cells transformed with the TAP-tagged effector genes (Figure 6).…”

Section: Resultsmentioning

confidence: 96%

“…We had previously developed an in vitro pathogenicity assay to recapitulate Xoo-rice interactions at the site of infection by treating Xoo cells with the rice homogenate (RLX), and assessed the time-resolved changes in the transcriptome (S. Kim et al, 2016; S. H. Kim et al, 2011; S. Kim et al, 2013). The in vitro pathogenicity assay provides high signal to noise data with Xoo cells synchronized with respect to the timing of pathogenicity activation.…”

Section: Introductionmentioning

confidence: 99%

Combined analysis of the time-resolved transcriptome and proteome of plant pathogen Xanthomonas oryzae pv. oryzae

Kim

Jang

Kim

et al. 2020

Preprint

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Xanthomonas oryzae pv. oryzae (Xoo) is a plant pathogen responsible for causing bacterial blight in rice. It is responsible for blight outbreaks in most rice-producing countries and can reduce the rice yield by 50% due to the lack of an effective pesticide. The immediate alterations in Xoo upon initial contact with rice at the infection site are essential for pathogenesis. Here, we studied time-resolved gene expression in pathogenicity-activated Xoo cells at the trans criptome and proteome levels. The early response genes of Xoo include genes related to cell motility, inorganic ion transport, and effectors, which aid the Xoo cells to invade damaged rice leaf tissues, obtain scarce cofactors, and evade rice immune responses, respectively. Alteration of gene expression is initiated as early as few minutes after the initial interaction and changes with time. Although there was a correlation between the overall expression patterns of mRNAs and proteins for many genes, some genes also exhibited differences with respect to the expression timing and level of mRNAs and proteins, suggesting an important role of translational regulation in the early stages of pathogenesis. Gene expression analysis using time-resolved transcriptome and proteome data provided valuable information regarding Xoo pathogenesis during the initial stages of Xoo-rice interaction and revealed translational regulation of gene expression in bacteria.

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“…This phenomenon was also found in Honey bees and Ae. albopictus (Guo et al, 2013; Kim et al, 2013). The low expression levels suggested that the novel miRNAs were expressed in specific developmental stages and organs.…”

Section: Resultsmentioning

confidence: 99%

Identification of differentially expressed microRNAs in Culex pipiens and their potential roles in pyrethroid resistance

Hong

Guo

Wang

et al. 2014

Insect Biochemistry and Molecular Biology

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Pyrethroids are the major class of insecticides used for mosquito control. Excessive and improper use of insecticides, however, has resulted in pyrethroid resistance, which has become a major obstacle for mosquito control. The development of pyrethroid resistance is a complex process involving many genes, and information on post-transcription regulation of pyrethroid resistance is lacking. In this study, we extracted RNA from mosquitoes in various life stages (fourth-instar larvae, pupae, male and female adult mosquitoes) from deltamethrin-sensitive (DS) and resistant (DR) strains. Using Illumina sequencing, we obtained 13760296 and 12355472 reads for DS-strains and DR-strains, respectively. We identified 100 conserved miRNAs and 42 novel miRNAs derived from 21 miRNA precursors in Culex pipiens. After normalization, we identified 28 differentially expressed miRNAs between the two strains. Additionally, we found that cpp-miR-71 was significant down regulated in female adults from the DR-strain. Based on microinjection and CDC Bottle Bioassay data, we found that cpp-miR-71 may play a contributing role in deltamethrin resistance. The present study provides the firstly large-scale characterization of miRNAs in Culex pipiens and provides evidence of post-transcription regulation. The differentially expressed miRNAs between the two strains are expected to contribute to the development of pyrethroid resistance.

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Homologous Expression and T3SS-Dependent Secretion of TAP-Tagged Xo2276 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract and Its Direct In Vitro Recognition of Putative Target DNA Sequence

Cited by 10 publications

References 18 publications

Time-resolved pathogenic gene expression analysis of the plant pathogen Xanthomonas oryzae pv. oryzae

Time-resolved pathogenic gene expression analysis of the plant pathogen Xanthomonas oryzae pv. oryzae

Combined analysis of the time-resolved transcriptome and proteome of plant pathogen Xanthomonas oryzae pv. oryzae

Identification of differentially expressed microRNAs in Culex pipiens and their potential roles in pyrethroid resistance

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