Large-Scale Gene Disruption in Magnaporthe oryzae Identifies MC69, a Secreted Protein Required for Infection by Monocot and Dicot Fungal Pathogens
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{"title"=>"Large-scale gene disruption in Magnaporthe oryzae identifies MC69, a secreted protein required for infection by monocot and dicot fungal pathogens", "type"=>"journal", "authors"=>[{"first_name"=>"Hiromasa", "last_name"=>"Saitoh", "scopus_author_id"=>"7202529028"}, {"first_name"=>"Shizuko", "last_name"=>"Fujisawa", "scopus_author_id"=>"10039563900"}, {"first_name"=>"Chikako", "last_name"=>"Mitsuoka", "scopus_author_id"=>"6701476013"}, {"first_name"=>"Akiko", "last_name"=>"Ito", "scopus_author_id"=>"7403722269"}, {"first_name"=>"Akiko", "last_name"=>"Hirabuchi", "scopus_author_id"=>"36189917200"}, {"first_name"=>"Kyoko", "last_name"=>"Ikeda", "scopus_author_id"=>"55228161500"}, {"first_name"=>"Hiroki", "last_name"=>"Irieda", "scopus_author_id"=>"14121470000"}, {"first_name"=>"Kae", "last_name"=>"Yoshino", "scopus_author_id"=>"55203316500"}, {"first_name"=>"Kentaro", "last_name"=>"Yoshida", "scopus_author_id"=>"55725246300"}, {"first_name"=>"Hideo", "last_name"=>"Matsumura", "scopus_author_id"=>"7202025347"}, {"first_name"=>"Yukio", "last_name"=>"Tosa", "scopus_author_id"=>"7003430030"}, {"first_name"=>"Joe", "last_name"=>"Win", "scopus_author_id"=>"7801494299"}, {"first_name"=>"Sophien", "last_name"=>"Kamoun", "scopus_author_id"=>"16157156400"}, {"first_name"=>"Yoshitaka", "last_name"=>"Takano", "scopus_author_id"=>"7401622081"}, {"first_name"=>"Ryohei", "last_name"=>"Terauchi", "scopus_author_id"=>"7004222954"}], "year"=>2012, "source"=>"PLoS Pathogens", "identifiers"=>{"issn"=>"15537366", "scopus"=>"2-s2.0-84863709162", "pui"=>"365220895", "doi"=>"10.1371/journal.ppat.1002711", "isbn"=>"1553-7374", "sgr"=>"84863709162", "pmid"=>"22589729"}, "id"=>"3acbf0e5-6020-32a3-9adb-e3684f7dda90", "abstract"=>"To search for virulence effector genes of the rice blast fungus, Magnaporthe oryzae, we carried out a large-scale targeted disruption of genes for 78 putative secreted proteins that are expressed during the early stages of infection of M. oryzae. Disruption of the majority of genes did not affect growth, conidiation, or pathogenicity of M. oryzae. One exception was the gene MC69. The mc69 mutant showed a severe reduction in blast symptoms on rice and barley, indicating the importance of MC69 for pathogenicity of M. oryzae. The mc69 mutant did not exhibit changes in saprophytic growth and conidiation. Microscopic analysis of infection behavior in the mc69 mutant revealed that MC69 is dispensable for appressorium formation. However, mc69 mutant failed to develop invasive hyphae after appressorium formation in rice leaf sheath, indicating a critical role of MC69 in interaction with host plants. MC69 encodes a hypothetical 54 amino acids protein with a signal peptide. Live-cell imaging suggested that fluorescently labeled MC69 was not translocated into rice cytoplasm. Site-directed mutagenesis of two conserved cysteine residues (Cys36 and Cys46) in the mature MC69 impaired function of MC69 without affecting its secretion, suggesting the importance of the disulfide bond in MC69 pathogenicity function. Furthermore, deletion of the MC69 orthologous gene reduced pathogenicity of the cucumber anthracnose fungus Colletotrichum orbiculare on both cucumber and Nicotiana benthamiana leaves. We conclude that MC69 is a secreted pathogenicity protein commonly required for infection of two different plant pathogenic fungi, M. oryzae and C. orbiculare pathogenic on monocot and dicot plants, respectively.", "link"=>"http://www.mendeley.com/research/largescale-gene-disruption-magnaporthe-oryzae-identifies-mc69-secreted-protein-required-infection-mo", "reader_count"=>79, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>7, "Student > Doctoral Student"=>7, "Researcher"=>23, "Student > Ph. D. Student"=>24, "Student > Postgraduate"=>3, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>5, "Lecturer"=>2, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>7, "Student > Doctoral Student"=>7, "Researcher"=>23, "Student > Ph. D. Student"=>24, "Student > Postgraduate"=>3, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>5, "Lecturer"=>2, "Professor"=>1}, "reader_count_by_subject_area"=>{"Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>72, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>72}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "United States"=>3, "Denmark"=>1, "United Kingdom"=>1, "Uganda"=>1, "France"=>1, "Germany"=>1, "Ethiopia"=>1}, "group_count"=>1}

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  • {"files"=>["https://ndownloader.figshare.com/files/640563"], "description"=>"<p>(A) <i>MC69</i> is required for pathogenicity of <i>M. oryzae</i> strain Ina72. Conidial suspension of the wild-type strain Ina72 (Ina72 WT) and the <i>mc69</i> mutants (<i>mc69-9</i>, <i>-12</i>, <i>-87</i>) were inoculated on barley (cv. Nigrate) and rice (cv. Shin No. 2) leaves, and incubated for 4 and 7 days, respectively. (B) Germination, appressorium formation and appressorial penetration of Ina72 WT and the <i>mc69</i> mutants. The ratio of germination was calculated as the mean percentage of conidia germinated after 24 h on rice (cv. Shin No. 2) leaf sheath cells. The mean percentage of appressorium formation on rice leaf sheath cells among the germinated conidia is presented. Three replicates of ∼50 conidia were counted for each observation. The mean percentage of appressorial penetration by the <i>mc69</i> mutants 32 h after inoculation is presented. Standard errors are indicated by the vertical bars. (C) Appressorial penetration assays on rice leaf sheath cells. Conidia from Ina72 WT and <i>mc69-87</i> germinated and formed melanized appressoria. All appressoria formed by Ina72 WT penetrated and produced infectious hyphae but no appressoria formed by <i>mc69-87</i> produced infectious hyphae in most of area. Photographs were taken 32 h after inoculation. Scale bar = 40 µm. (D) Invasive growth rating of rice leaf sheath cells 32 h after inoculating with Ina72 WT and <i>mc69-87</i>. The levels for invasive growth rating are given above. For details of the invasive growth levels and rating see <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002711#s4\" target=\"_blank\">Materials and Methods</a>. Scale bar = 20 µm. (E), (F) Complementation of <i>mc69</i> mutant with the wild type allele of <i>MC69</i>. (E) Appressorial penetration by Ina72 WT, <i>mc69-87</i> (<i>mc69</i>) and the <i>MC69</i> re-introduced strain (<i>mc69</i>+<i>MC69</i>). Mean percentage of appressorial penetration is recorded 32 h after inoculation in rice leaf sheath cells. Four replicates of ∼50 appressoria were counted for each observation. (F) Blast symptoms caused by Ina72 WT, <i>mc69</i> and <i>mc69</i>+<i>MC69</i> on barley (cv. Nigrate) and on rice (cv. Shin No 2) 3 and 4 days after inoculation, respectively.</p>", "links"=>[], "tags"=>["appressorial", "penetration", "pathogenicity"], "article_id"=>311038, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MC69_is_involved_in_appressorial_penetration_and_pathogenicity_of_M_oryzae_/311038", "title"=>"<i>MC69</i> is involved in appressorial penetration and pathogenicity of <i>M. oryzae</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:17:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/641221"], "description"=>"<p>Conidial suspension of wild-type strain TH68-141 (TH68-141 WT), the three independent <i>mc69</i> mutants (<i>mc69-49</i>, <i>mc69-55</i> and <i>mc69-66</i>), wild-type strain Hoku1 (Hoku1 WT) and the two independent <i>mc69</i> mutants (<i>mc69-75</i> and <i>mc69-82</i>) were inoculated on barley (cv. Nigrate) leaves and incubated for 5 days.</p>", "links"=>[], "tags"=>["pathogenicity", "japanese", "isolates", "th68-141"], "article_id"=>311692, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g006", "stats"=>{"downloads"=>2, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MC69_is_required_for_pathogenicity_of_other_two_different_Japanese_field_isolates_TH68_141_and_Hoku1_/311692", "title"=>"<i>MC69</i> is required for pathogenicity of other two different Japanese field isolates TH68-141 and Hoku1.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:28:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/640863"], "description"=>"<p>Merged DIC and mCherry images of rice leaf sheath cells infected by <i>M. oryzae</i> Sasa2 strain harboring (A) <i>PWL2p::PWL2::mCherry::NLS</i>, (B) <i>PWL2p::MC69::mCherry::NLS</i>, and (C) <i>PWL2p::MC69::mCherry</i> 24, 27 and 32 h after inoculation as observed by confocal laser scanning microscopy. Arrows indicate BICs and triangles indicate rice nuclei. Pinhole setting is 240 µm for all panels. Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["translocated"], "article_id"=>311337, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g003", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MC69_mCherry_is_not_translocated_into_the_rice_cytoplasm_/311337", "title"=>"MC69::mCherry is not translocated into the rice cytoplasm.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:22:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/641432"], "description"=>"<p>(A) Sequence alignment of MC69 between <i>M. oryzae</i> (Mo) and <i>C. orbiculare</i> (Co). Amino acid sequences were aligned using Clustal W program <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002711#ppat.1002711-Tompson1\" target=\"_blank\">[52]</a>. Identical amino acids are indicated as white letters on a black background. Similar residues are shown by gray background. Gaps introduced for alignment are indicated by hyphens. (B) Pathogenicity test of the <i>Comc69</i> mutants on cucumber. Conidial suspensions were inoculated on detached cotyledons of cucumber (<i>Cucumis sativa</i>). On the left half of the cotyledons, the wild-type strain 104-T was inoculated as positive control. On the right half, the <i>Comc69</i> strains (DMC1 and DMC2) were inoculated. Inoculated cotyledons were incubated for 7 days. (C) Pathogenicity test of the <i>Comc69</i> mutants on <i>N. benthamiana</i>. On the left half of the detached leaves of <i>N. benthamiana</i>, the strain 104-T was inoculated as positive control. On the right half, the <i>Comc69</i> strains (DMC1 and DMC2) were inoculated. Inoculated leaves were incubated for 7 days. (D) mCherry-based reporter assay for expression of the <i>CoMC69</i> gene. Conidia from the <i>C. orbiculare</i> strain carrying the <i>CoMC69</i> promoter-<i>mCherry</i> fusion gene (<i>CoMC69p::mCherry</i>) was inoculated onto the lower surfaces of cucumber cotyledons, and the inoculated plant was incubated for 4 days. a, appressorium; ih, intracellular hypha. Scale bars = 10 µm.</p>", "links"=>[], "tags"=>["fungal", "pathogenicity"], "article_id"=>311911, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g008", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CoMC69_is_involved_in_fungal_pathogenicity_of_C_orbiculare_/311911", "title"=>"<i>CoMC69</i> is involved in fungal pathogenicity of <i>C. orbiculare</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:31:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/330618", "https://ndownloader.figshare.com/files/330649", "https://ndownloader.figshare.com/files/330694", "https://ndownloader.figshare.com/files/330750", "https://ndownloader.figshare.com/files/330798", "https://ndownloader.figshare.com/files/330851", "https://ndownloader.figshare.com/files/330909", "https://ndownloader.figshare.com/files/330943", "https://ndownloader.figshare.com/files/330981", "https://ndownloader.figshare.com/files/331003", "https://ndownloader.figshare.com/files/331043", "https://ndownloader.figshare.com/files/331079"], "description"=>"<div><p>To search for virulence effector genes of the rice blast fungus, <em>Magnaporthe oryzae</em>, we carried out a large-scale targeted disruption of genes for 78 putative secreted proteins that are expressed during the early stages of infection of <em>M. oryzae</em>. Disruption of the majority of genes did not affect growth, conidiation, or pathogenicity of <em>M. oryzae</em>. One exception was the gene <em>MC69</em>. The <em>mc69</em> mutant showed a severe reduction in blast symptoms on rice and barley, indicating the importance of MC69 for pathogenicity of <em>M. oryzae</em>. The <em>mc69</em> mutant did not exhibit changes in saprophytic growth and conidiation. Microscopic analysis of infection behavior in the <em>mc69</em> mutant revealed that MC69 is dispensable for appressorium formation. However, <em>mc69</em> mutant failed to develop invasive hyphae after appressorium formation in rice leaf sheath, indicating a critical role of MC69 in interaction with host plants. <em>MC69</em> encodes a hypothetical 54 amino acids protein with a signal peptide. Live-cell imaging suggested that fluorescently labeled MC69 was not translocated into rice cytoplasm. Site-directed mutagenesis of two conserved cysteine residues (Cys36 and Cys46) in the mature MC69 impaired function of MC69 without affecting its secretion, suggesting the importance of the disulfide bond in MC69 pathogenicity function. Furthermore, deletion of the <em>MC69</em> orthologous gene reduced pathogenicity of the cucumber anthracnose fungus <em>Colletotrichum orbiculare</em> on both cucumber and <em>Nicotiana benthamiana</em> leaves. We conclude that MC69 is a secreted pathogenicity protein commonly required for infection of two different plant pathogenic fungi, <em>M. oryzae</em> and <em>C. orbiculare</em> pathogenic on monocot and dicot plants, respectively.</p> </div>", "links"=>[], "tags"=>["large-scale", "disruption", "identifies", "secreted", "monocot", "dicot", "fungal", "pathogens"], "article_id"=>125284, "categories"=>["Cell Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1002711.s001", "https://dx.doi.org/10.1371/journal.ppat.1002711.s002", "https://dx.doi.org/10.1371/journal.ppat.1002711.s003", "https://dx.doi.org/10.1371/journal.ppat.1002711.s004", "https://dx.doi.org/10.1371/journal.ppat.1002711.s005", "https://dx.doi.org/10.1371/journal.ppat.1002711.s006", "https://dx.doi.org/10.1371/journal.ppat.1002711.s007", "https://dx.doi.org/10.1371/journal.ppat.1002711.s008", "https://dx.doi.org/10.1371/journal.ppat.1002711.s009", "https://dx.doi.org/10.1371/journal.ppat.1002711.s010", "https://dx.doi.org/10.1371/journal.ppat.1002711.s011", "https://dx.doi.org/10.1371/journal.ppat.1002711.s012"], "stats"=>{"downloads"=>4, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Large_Scale_Gene_Disruption_in_Magnaporthe_oryzae_Identifies_MC69_a_Secreted_Protein_Required_for_Infection_by_Monocot_and_Dicot_Fungal_Pathogens/125284", "title"=>"Large-Scale Gene Disruption in <em>Magnaporthe oryzae</em> Identifies MC69, a Secreted Protein Required for Infection by Monocot and Dicot Fungal Pathogens", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-05-10 01:28:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/641134"], "description"=>"<p>(A) Schematic diagram of MC69(36A)::mCherry fusion protein expression construct. (B) Merged DIC and mCherry images of rice leaf sheath cells infected by the MC69(C36A)::mCherry-expressing transformants 24 h and 48 h after inoculation. Scale bar = 20 µm. (C) Western blot probed with an anti-DsRed antibody. Samples were loaded as follows: lane 1, culture filtrate from mCherry-expressing strain; lane 2, culture filtrate from MC69::mCherry-expressing strain; lane 3, culture filtrate from MC69(C36A)::mCherry-expressing strain.</p>", "links"=>[], "tags"=>["fusion"], "article_id"=>311612, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g005", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Secretion_of_MC69_C36A_mCherry_and_MC69_mCherry_fusion_proteins_/311612", "title"=>"Secretion of MC69(C36A)::mCherry and MC69::mCherry fusion proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:26:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/641551"], "description"=>"a<p>Expressed gene in the cAMP-treated <i>M. oryzae</i> on dialysis membrane at 6 hpi in the <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002711#ppat.1002711.s012\" target=\"_blank\">Table S1</a>.</p>b<p>Expressed gene in the M. oryzae-infected rice leaf sheath at 40 hpi in the Supplemental data set 1 <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002711#ppat.1002711-Yoshida1\" target=\"_blank\">[20]</a>.</p><p>(+) or (−) indicate the expressed gene or not, respectively in the cAMP-treated or invasive hyphae of <i>M. oryzae</i>.</p>", "links"=>[], "tags"=>["disruption", "78", "putative", "secreted"], "article_id"=>312023, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.t001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Gene_disruption_analysis_of_78_putative_secreted_protein_genes_/312023", "title"=>"Gene disruption analysis of 78 putative secreted protein genes.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-05-10 00:33:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/640705"], "description"=>"<p>(A) and (D) Schematic diagrams of mCherry and MC69::mCherry fusion protein expression constructs. (B) and (E) Conidia from WT+<i>mCherry</i> and <i>mc69</i>+<i>MC69::mCherry</i> were harvested, and appressorium development was observed for 12 h on glass coverslips. Merged DIC and mCherry (red) images were taken. Scale bars = 10 µm. (C) and (F) Merged DIC and mCherry images of the rice leaf sheath cells infected with mCherry- and MC69::mCherry-expressing transformants 24 h and 48 h after inoculation. Scale bars = 20 µm. (G) Blast symptoms caused by Ina72 WT, <i>mc69</i> and <i>mc69</i>+<i>MC69::Cherrry</i> on rice (cv. Shin No. 2) 7 days after inoculation.</p>", "links"=>[], "tags"=>["suggests", "promoter", "constitutively", "fusion"], "article_id"=>311180, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Microscopic_analysis_suggests_that_MC69_promoter_is_constitutively_active_and_MC69_mCherry_fusion_protein_is_secreted_/311180", "title"=>"Microscopic analysis suggests that <i>MC69</i> promoter is constitutively active and MC69::mCherry fusion protein is secreted.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:19:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/641342"], "description"=>"<p>(A) <i>MC69</i> is required for pathogenicity of <i>M. oryzae</i> strain 70-15. Conidial suspension of the wild-type strain 70-15 (70-15 WT) and the <i>mc69</i> mutants (<i>mc69-119</i>, <i>-31</i>) were inoculated on barley (cv. Nigrate) and rice (cv. Shin No. 2) leaves, and incubated for 7 days. (B) Germination, appressorium formation and appressorial penetration of 70-15 WT and the <i>mc69</i> mutants. The ratio of germination was calculated as the mean percentage of conidia germinated after 32 h on rice (cv. Shin No. 2) leaf sheath cells. The mean percentage of appressorium formation on rice leaf sheath cells among the germinated conidia is presented. Three replicates of ∼50 conidia were counted for each observation. The mean percentage of appressorial penetration by the <i>mc69</i> mutants is presented 32 h after inoculation. Standard errors are indicated by the vertical bars.</p>", "links"=>[], "tags"=>["appressorial", "penetration", "pathogenicity"], "article_id"=>311812, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g007", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MC69_is_necessary_for_appressorial_penetration_and_pathogenicity_of_a_laboratory_strain_70_15_/311812", "title"=>"<i>MC69</i> is necessary for appressorial penetration and pathogenicity of a laboratory strain 70-15.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:30:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/641020"], "description"=>"<p>(A) DISULFIND (<a href=\"http://disulfind.dsi.unifi.it/\" target=\"_blank\">http://disulfind.dsi.unifi.it/</a>) output of the mature form of MC69 protein. (B) Amino acid sequences of disulfide mutants of MC69. The predicted signal peptide is indicated in blue. (C) Appressorial penetration by <i>M. oryzae</i> wild-type strain Ina72 (WT) and transformants with various versions of MC69. Mean percentage of invasion in rice (cv. Shin No. 2) leaf sheath cells 30 h after inoculation is presented. Four replicates of ∼50 appressoria were counted for each observation. (D) Blast symptoms caused by WT, the <i>mc69</i> mutant (<i>mc69</i>), the <i>MC69</i>-, <i>MC69(C36A)</i>-, <i>MC69(C46A)</i>- and the <i>MC69(C36A,C46A)</i>-re-introduced strains [<i>mc69</i>+<i>MC69</i>, <i>mc69</i>+<i>MC69(C36A)</i>, <i>mc69</i>+<i>MC69(C46A)</i> and <i>mc69</i>+<i>MC69(C36A,C46A)</i>] on barley (cv. Nigrate) and on rice (cv. Shin No. 2) 4 and 7 days after inoculation, respectively.</p>", "links"=>[], "tags"=>["cysteine", "residues", "mc69", "virulence"], "article_id"=>311489, "categories"=>["Plant Biology"], "users"=>["Hiromasa Saitoh", "Shizuko Fujisawa", "Chikako Mitsuoka", "Akiko Ito", "Akiko Hirabuchi", "Kyoko Ikeda", "Hiroki Irieda", "Kae Yoshino", "Kentaro Yoshida", "Hideo Matsumura", "Yukio Tosa", "Joe Win", "Sophien Kamoun", "Yoshitaka Takano", "Ryohei Terauchi"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002711.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Two_cysteine_residues_are_important_for_MC69_virulence_activity_/311489", "title"=>"Two cysteine residues are important for MC69 virulence activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:24:49"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"20", "full-text"=>"23", "pdf"=>"27", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"9"}
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  • {"unique-ip"=>"18", "full-text"=>"16", "pdf"=>"6", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"3"}
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Relative Metric

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