Native and Foreign Proteins Secreted by the Cupriavidus metallidurans Type II System and an Alternative Mechanism

( Houjuan Xu ),( Timothy P. Denny ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.4

The type II secretion system (T2SS), which transports selected periplasmic proteins across the outer membrane, has rarely been studied in nonpathogens or in organisms classified as Betaproteobacteria. Therefore, we studied Cupriavidus metallidurans (Cme), a facultative chemilithoautotroph. Gel analysis of extracellular proteins revealed no remarkable differences between the wild type and the T2SS mutants. However, enzyme assays revealed that native extracellular alkaline phosphatase is a T2SS substrate, because activity was 10-fold greater for the wild type than a T2SS mutant. In Cme engineered to produce three Ralstonia solanacearum (Rso) exoenzymes, at least 95% of their total activities were extracellular, but unexpectedly high percentages of these exoenzymes remained extracellular in T2SS mutants cultured in rich broth. These conditions appear to permit an alternative secretion process, because neither cell lysis nor periplasmic leakage was observed when Cme produced a Pectobacterium carotovorum exoenzyme, and wild-type Cme cultured in minimal medium secreted 98% of Rso polygalacturonase, but 92% of this exoenzyme remained intracellular in T2SS mutants. We concluded that Cme has a functional T2SS despite lacking any abundant native T2SS substrates. The efficient secretion of three foreign exoenzymes by Cme is remarkable, but so too is the indication of an alternative secretion process in rich culture conditions. When not transiting the T2SS, we suggest that Rso exoenzymes are probably selectively packaged into outer membrane vesicles. Phylogenetic analysis of T2SS proteins supports the existence of at least three T2SS subfamilies, and we propose that Cme, as a representative of the Betaproteobacteria, could become a new useful model system for studying T2SS substrate specificity.

AbSte7, a MAPKK Gene of Alternaria brassicicola, Is Involved in Conidiation, Salt/Oxidative Stress, and Pathogenicity

Xu, Houjuan,Zhang, Qianqian,Cui, Wenjuan,Zhang, Xiaofei,Liu, Weiyang,Zhang, Li,Islam, Md. Nurul,Baek, Kwang-Hyun,Wang, Yujun The Korean Society for Microbiology and Biotechnol 2016 Journal of microbiology and biotechnology Vol.26 No.7

Alternaria brassicicola (Schwein.) invades Brassicaceae and causes black spot disease, significantly lowering productivity. Mitogen-activated protein kinases (MAPKs) and their upstream kinases, including MAPK kinases (MAPKKs) and MAPKK kinases (MAPKKK), comprise one of the most important signaling pathways determining the pathogenicity of diverse plant pathogens. The AbSte7 gene in the genome of A. brassicicola was predicted to be a homolog of yeast Ste7, a MAPKK; therefore, the function was characterized by generating null mutant strains with a gene replacement method. AbSte7 replacement mutants (RMs) had a slower growth rate and altered colony morphology compared with the wild-type strain. Disruption of the AbSte7 gene resulted in defects in conidiation and melanin accumulation. AbSte7 was also involved in the resistance pathways in salt and oxidative stress, working to negatively regulate salt tolerance and positively regulate oxidative stress. Pathogenicity assays revealed that AbSte7 RMs could not infect intact cabbage leaves, but only formed very small lesions in wounded leaves, whereas typical lesions appeared on both intact and wounded leaves inoculated with the wild-type strain. As the first studied MAPKK in A. brassicicola, these data strongly suggest that the AbSte7 gene is an essential element for the growth, development, and pathogenicity of A. brassicicola.

AbSte7, a MAPKK Gene of Alternaria brassicicola, Is Involved in Conidiation, Salt/Oxidative Stress, and Pathogenicity

( Houjuan Xu ),( Qianqian Zhang ),( Wenjuan Cui ),( Xiaofei Zhang ),( Weiyang Liu ),( Li Zhang ),( Md. Nurul Islam ),( Kwang-hyun Baek ),( Yujun Wang ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.6

Alternaria brassicicola (Schwein.) invades Brassicaceae and causes black spot disease, significantly lowering productivity. Mitogen-activated protein kinases (MAPKs) and their upstream kinases, including MAPK kinases (MAPKKs) and MAPKK kinases (MAPKKK), comprise one of the most important signaling pathways determining the pathogenicity of diverse plant pathogens. The AbSte7 gene in the genome of A. brassicicola was predicted to be a homolog of yeast Ste7, a MAPKK; therefore, the function was characterized by generating null mutant strains with a gene replacement method. AbSte7 replacement mutants (RMs) had a slower growth rate and altered colony morphology compared with the wild-type strain. Disruption of the AbSte7 gene resulted in defects in conidiation and melanin accumulation. AbSte7 was also involved in the resistance pathways in salt and oxidative stress, working to negatively regulate salt tolerance and positively regulate oxidative stress. Pathogenicity assays revealed that AbSte7 RMs could not infect intact cabbage leaves, but only formed very small lesions in wounded leaves, whereas typical lesions appeared on both intact and wounded leaves inoculated with the wild-type strain. As the first studied MAPKK in A. brassicicola, these data strongly suggest that the AbSte7 gene is an essential element for the growth, development, and pathogenicity of A. brassicicola.

The Mitogen-Activated Protein Kinase Signal Transduction Pathways in Alternaria Species

Houjuan Xu,Xiaoxue Xu,Yujun Wang,비벡바지파이,Lisha Huang,Yongfang Chen,백광현 한국식물병리학회 2012 Plant Pathology Journal Vol.28 No.3

Mitogen-activated protein kinase (MAPK) cascades are conserved signaling modules in the eukaryotic cells. They are involved in many major cell processes in fungi such as stress responses, vegetative growth, pathogenicity,secondary metabolism and cell wall integrity. In this review, we summarized the advances of research on the MAPK signaling pathways in Alternaria species. As major phytopathogenic fungi, Alternaria species reduce crop production. In contrast to the five MAPK pathways known in yeast, only three MAPK pathways as Fus3/Kss1-type, Hog1-type, and Slt2-type have been characterized in Alternaria. The Fus3/Kss1-type MAPK pathway participates in regulation of vegetative growth,conidiation, production of some cell-wall-degrading enzymes and pathogenicity. The Hog1-type pathway is involved in osmotic and oxidative stress, fungicides susceptibility and pathogenicity. The Slt2-type MAP kinases play an important role on maintaining cell wall integrity, pathogenicity and conidiation. Although recent advances on the MAPK pathways in Alternaria spp. reveal many important features on the pathogenicity,there are many unsolved problems regarding to the unknown MAP kinase cascade components and network among other major signal transduction. Considering the economic loss induced by Alternaria spp., more researches on the MAPK pathways will need to control the Alternaria diseases.

The Mitogen-Activated Protein Kinase Signal Transduction Pathways in Alternaria Species

Xu, Houjuan,Xu, Xiaoxue,Wang, Yu-Jun,Bajpai, Vivek K.,Huang, Lisha,Chen, Yongfang,Baek, Kwang-Hyun The Korean Society of Plant Pathology 2012 Plant Pathology Journal Vol.28 No.3

Mitogen-activated protein kinase (MAPK) cascades are conserved signaling modules in the eukaryotic cells. They are involved in many major cell processes in fungi such as stress responses, vegetative growth, pathogenicity, secondary metabolism and cell wall integrity. In this review, we summarized the advances of research on the MAPK signaling pathways in Alternaria species. As major phytopathogenic fungi, Alternaria species reduce crop production. In contrast to the five MAPK pathways known in yeast, only three MAPK pathways as Fus3/Kss1-type, Hog1-type, and Slt2-type have been characterized in Alternaria. The Fus3/Kss1-type MAPK pathway participates in regulation of vegetative growth, conidiation, production of some cell-wall-degrading enzymes and pathogenicity. The Hog1-type pathway is involved in osmotic and oxidative stress, fungicides susceptibility and pathogenicity. The Slt2-type MAP kinases play an important role on maintaining cell wall integrity, pathogenicity and conidiation. Although recent advances on the MAPK pathways in Alternaria spp. reveal many important features on the pathogenicity, there are many unsolved problems regarding to the unknown MAP kinase cascade components and network among other major signal transduction. Considering the economic loss induced by Alternaria spp., more researches on the MAPK pathways will need to control the Alternaria diseases.

Potential Roles of Essential Oils on Controlling Plant Pathogenic Bacteria Xanthomonas Species: A Review

Bajpai, Vivek K.,Kang, So-Ra,Xu, Houjuan,Lee, Soon-Gu,Baek, Kwang-Hyun,Kang, Sun-Chul The Korean Society of Plant Pathology 2011 Plant Pathology Journal Vol.27 No.3

Diseases caused by plant pathogenic bacteria constitute an emerging threat to global food security. Xanthomonas is a large genus of Gram-negative bacteria that cause disease in several host plants leading to considerable losses in productivity and quality of harvests. Despite the ranges of controlling techniques available, the microbiological safety of economically important crops and crop plants including fruits and vegetables continues to be a major concern to the agriculture industry. On the other hand, many of the currently available antimicrobial agents for agriculture are highly toxic, non-biodegradable and cause extended environmental pollution. Besides, the use of antibiotics has provoked an increased resistance among the bacterial pathogens and their pathovars. Thus, novel efficient and safe remedies for controlling plant bacterial diseases are necessary. There has been an increasing interest worldwide on therapeutic values of natural products such as essential oils, hence the purpose of this review is to provide an overview of the published data on the antibacterial efficacy of essential oils that could be considered suitable for application in agriculture as biocontrol measures against plant pathogenic bacteria of Xanthomonas species. The current knowledge on the use of essential oils to control Xanthomonas bacteria in vitro and in vivo models has been discussed. A brief description on the legal aspects on the use of essential oils against bacterial pathogens has also been presented. Through this review, a mode of antibacterial action of essential oils along with their chemical nature and the area for future research have been thoroughly discussed.

Potential Roles of Essential Oils on Controlling Plant Pathogenic Bacteria Xanthomonas Species: A Review

Vivek K. Bajpai,Sora Kang,Houjuan Xu,이순구,백광현,강선철 한국식물병리학회 2011 Plant Pathology Journal Vol.27 No.3

Diseases caused by plant pathogenic bacteria constitute an emerging threat to global food security. Xanthomonas is a large genus of Gram-negative bacteria that cause disease in several host plants leading to considerable losses in productivity and quality of harvests. Despite the ranges of controlling techniques available, the microbiological safety of economically important crops and crop plants including fruits and vegetables continues to be a major concern to the agriculture industry. On the other hand, many of the currently available antimicrobial agents for agriculture are highly toxic, non-biodegradable and cause extended environmental pollution. Besides, the use of antibiotics has provoked an increased resistance among the bacterial pathogens and their pathovars. Thus, novel efficient and safe remedies for controlling plant bacterial diseases are necessary. There has been an increasing interest worldwide on therapeutic values of natural products such as essential oils,hence the purpose of this review is to provide an overview of the published data on the antibacterial efficacy of essential oils that could be considered suitable for application in agriculture as biocontrol measures against plant pathogenic bacteria of Xanthomonas species. The current knowledge on the use of essential oils to control Xanthomonas bacteria in vitro and in vivo models has been discussed. A brief description on the legal aspects on the use of essential oils against bacterial pathogens has also been presented. Through this review, a mode of antibacterial action of essential oils along with their chemical nature and the area for future research have been thoroughly discussed.

The MAP Kinase Kinase Gene AbSte7 Regulates Multiple Aspects of Alternaria brassicicola Pathogenesis

Lu, Kai,Zhang, Min,Yang, Ran,Zhang, Min,Guo, Qinjun,Baek, Kwang-Hyun,Xu, Houjuan The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.2

Mitogen-activated protein kinase (MAPK) cascades in fungi are ubiquitously conserved signaling pathways that regulate stress responses, vegetative growth, pathogenicity, and many other developmental processes. Previously, we reported that the AbSte7 gene, which encodes a mitogen-activated protein kinase kinase (MAPKK) in Alternaria brassicicola, plays a central role in pathogenicity against host cabbage plants. In this research, we further characterized the role of AbSte7 in the pathogenicity of this fungus using ${\Delta}AbSte7$ mutants. Disruption of the AbSte7 gene of A. brassicicola reduced accumulation of metabolites toxic to the host plant in liquid culture media. The ${\Delta}AbSte7$ mutants could not efficiently detoxify cruciferous phytoalexin brassinin, possibly due to reduced expression of the brassinin hydrolase gene involved in detoxifying brassinin. Disruption of the AbSte7 gene also severely impaired fungal detoxification of reactive oxygen species. AbSte7 gene disruption reduced the enzymatic activity of cell walldegrading enzymes, including cellulase, ${\beta}$-glucosidase, pectin methylesterase, polymethyl-galacturonase, and polygalacturonic acid transeliminase, during host plant infection. Altogether, the data strongly suggest the MAPKK gene AbSte7 plays a pivotal role in A. brassicicola during host infection by regulating multiple steps, and thus increasing pathogenicity and inhibiting host defenses.