Systematic Analysis of Zn2Cys6 Transcription Factors Required for Development and Pathogenicity by High-Throughput Gene Knockout in the Rice Blast Fungus
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{"title"=>"Systematic analysis of Zn2Cys6 transcription factors required for development and pathogenicity by high-throughput gene knockout in the rice blast fungus", "type"=>"journal", "authors"=>[{"first_name"=>"Jianping", "last_name"=>"Lu"}, {"first_name"=>"Huijuan", "last_name"=>"Cao"}, {"first_name"=>"Lilin", "last_name"=>"Zhang"}, {"first_name"=>"Pengyun", "last_name"=>"Huang"}, {"first_name"=>"Fucheng", "last_name"=>"Lin"}], "year"=>2014, "source"=>"PLoS pathogens", "identifiers"=>{"doi"=>"10.1371/journal.ppat.1004432", "issn"=>"15537374", "isbn"=>"1553-7374 (Electronic)\\r1553-7366 (Linking)", "pmid"=>"25299517"}, "id"=>"fcee359b-7c57-380a-b3b4-b6c98a7d26e2", "abstract"=>"Because of great challenges and workload in deleting genes on a large scale, the functions of most genes in pathogenic fungi are still unclear. In this study, we developed a high-throughput gene knockout system using a novel yeast-Escherichia-Agrobacterium shuttle vector, pKO1B, in the rice blast fungus Magnaporthe oryzae. Using this method, we deleted 104 fungal-specific Zn(2)Cys(6) transcription factor (TF) genes in M. oryzae. We then analyzed the phenotypes of these mutants with regard to growth, asexual and infection-related development, pathogenesis, and 9 abiotic stresses. The resulting data provide new insights into how this rice pathogen of global significance regulates important traits in the infection cycle through Zn(2)Cys(6)TF genes. A large variation in biological functions of Zn(2)Cys(6)TF genes was observed under the conditions tested. Sixty-one of 104 Zn(2)Cys(6) TF genes were found to be required for fungal development. In-depth analysis of TF genes revealed that TF genes involved in pathogenicity frequently tend to function in multiple development stages, and disclosed many highly conserved but unidentified functional TF genes of importance in the fungal kingdom. We further found that the virulence-required TF genes GPF1 and CNF2 have similar regulation mechanisms in the gene expression involved in pathogenicity. These experimental validations clearly demonstrated the value of a high-throughput gene knockout system in understanding the biological functions of genes on a genome scale in fungi, and provided a solid foundation for elucidating the gene expression network that regulates the development and pathogenicity of M. oryzae.", "link"=>"http://www.mendeley.com/research/systematic-analysis-zn2cys6-transcription-factors-required-development-pathogenicity-highthroughput", "reader_count"=>39, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Researcher"=>5, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>6, "Student > Bachelor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Researcher"=>5, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>6, "Student > Bachelor"=>3}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>33}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>33}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Germany"=>1, "India"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1711756"], "description"=>"<p>Green or red arrows are the directions of gene expression regulation in Δ<i>gpf1</i> or Δ<i>cnf2</i>.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199373, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g008", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_differentially_expressed_genes_between_gpf1_and_cnf2_/1199373", "title"=>"Comparison of differentially expressed genes between Δ<i>gpf1</i> and Δ<i>cnf2</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711752"], "description"=>"<p>(<b>A</b>) Growth inhibition rate. 5-mm mycelial blocks of <i>M. oryzae</i> strains were inoculated on CM medium and medium with stress conditions for 8 days, and the diameter of colonies was then measured to calculate the growth inhibition rate. The experimental strains were the wild-type strain, mutants (Δ<i>Monit4</i>, Δ<i>gpf1</i>, Δ<i>gcc1</i>, Δ<i>tas1</i> and Δ<i>gta1</i>), and complemented strains (<i>Monit4-c</i>, <i>gpf1-c</i>, <i>gcc1-c</i>, <i>tas1-c</i> and <i>gta1-c</i>). Same capital letters in same stress item indicate non-significant difference estimated by Duncan's test (<i>P</i>≤0.05). (<b>B</b>) Mycelial growth of Δ<i>Monit4</i> on MM medium. The wild-type strain, Δ<i>Monit4</i>, and complemented strain <i>Monit4-c</i> were grown on MM medium at 25°C for 8 days. Bar = 5 mm.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199369, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g007", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Growth_of_the_M_oryzae_strains_under_stress_conditions_/1199369", "title"=>"Growth of the <i>M. oryzae</i> strains under stress conditions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711690"], "description"=>"<p>(<b>A</b>) Number of TFs showing multiple mutant phenotypes. (<b>B</b>) Number of mutants showing mutant phenotypes in each developmental stage. (<b>C</b>) Venn diagram showing the number of mutant phenotypes. The phenotypes included vegetative growth (conidial germination, colony growth, pigmentation and mycelial appearance), conidiation (asexual reproduction), appressorium formation and pathogenicity to rice and barley.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199322, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g002", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_Zn_2_Cys_6_transcription_factor_mutant_phenotypes_in_fungal_development_stages_/1199322", "title"=>"Analysis of Zn<sub>2</sub>Cys<sub>6</sub> transcription factor mutant phenotypes in fungal development stages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711702"], "description"=>"<p>(<b>A</b>) Colony growth of the wild-type strain 70-15, mutants (Δ<i>gcc1</i>, Δ<i>gpf1</i> and Δ<i>gta1</i>), and complemented strains (<i>gcc1</i>-c, <i>gta1</i>-c, <i>gta1</i>-c) on CM medium for 8 days. Bar = 1 cm. (<b>B</b>) Conidiophores of the wild-type, mutants (Δ<i>cnf1</i>, Δ<i>cnf2</i>, Δ<i>pcf1</i>, Δ<i>cca1</i>, Δ<i>conx1</i>, Δ<i>gcc1</i> and Δ<i>Mocod1</i>), and complemented strains (<i>cnf1-c</i>, <i>cnf2-c</i>, <i>pcf1-c</i>, <i>cca1-c</i>, <i>conx1-c</i>, <i>gcc1-c</i> and <i>Mocod1-c</i>). Bar = 20 µm.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199332, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g003", "stats"=>{"downloads"=>2, "page_views"=>25, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Colony_growth_and_conidiophores_of_M_oryzae_strains_/1199332", "title"=>"Colony growth and conidiophores of <i>M. oryzae</i> strains.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711758"], "description"=>"<p>Notes: The strains (5-mm mycelial blocks) were grown on CM medium for 8 days, and the diameter of colonies was then measured (growth) or conidia were collected and counted (conidiation). 40-µl (1×10<sup>5</sup> conidia/ml) conidial suspensions were incubated on plastic slides for 4 hpi (conidial germination assay) and 24 hpi (appressorium formation assay).</p><p>*Same capital letters indicate non-significant difference estimated by Duncan's test in each developmental item (<i>P</i>≤0.05).</p><p>The experimental strains were the wild-type strain, the mutants (Δ<i>cca1</i>, Δ<i>conx1</i>, Δ<i>cnf1</i>, Δ<i>cnf2</i>, Δ<i>fzc16</i>, Δ<i>gcc1</i>, Δ<i>gpf1</i>, Δ<i>gta1</i>, Δ<i>Mocod1</i>, Δ<i>Monit4</i> and Δ<i>pcf1</i>) and complemented strains (<i>cca1-c</i>, <i>conx1-c</i>, <i>cnf1-c</i>, <i>cnf2-c</i>, <i>fzc16-c</i>, <i>gcc1</i>-c, <i>gpf1-c</i>, <i>gta1-c</i>, <i>Mocod1-c</i>, <i>Monit4-c</i> and <i>pcf1-c</i>).</p><p>Developmental characteristics of <i>M. oryzae</i> strains.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199375, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.t002", "stats"=>{"downloads"=>2, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Developmental_characteristics_of_M_oryzae_strains_/1199375", "title"=>"Developmental characteristics of <i>M. oryzae</i> strains.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711759"], "description"=>"<p>Note:</p>a<p>) The expression of genes increased or decreased in Δ<i>gpf1</i> and Δ<i>cnf2</i>.</p><p>The number means the fold change, and the symbol “−” means no significant change (FDR<0.05) afterwards compared with the wild-type strain.</p><p><i>GPF1</i>- or <i>CNF2</i>-dependent genes encoding known virulence factors.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199376, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.t003", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GPF1_or_CNF2_dependent_genes_encoding_known_virulence_factors_/1199376", "title"=>"<i>GPF1</i>- or <i>CNF2</i>-dependent genes encoding known virulence factors.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711757"], "description"=>"<p>Notes: The phenotypes of the mutants in colony growth, conidiation, conidial germination, appressorium formation, virulence and response to stress were compared with the wild-type strain 70-15. The genes are named as <i>FZC1</i>∼<i>FZC87</i> (<u>F</u>ungal-specific <u>Z</u>n<sub>2</sub><u>C</u>ys<sub>6</sub> transcription factor 1 to 87) if not designated in the succeeding text. The detailed phenotypic information of mutants was shown in Table S4 in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004432#ppat.1004432.s005\" target=\"_blank\">Text S1</a>.</p><p>Phenotypic summary of 104 Zn<sub>2</sub>Cys<sub>6</sub> TF gene-deleted mutants.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199374, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.t001", "stats"=>{"downloads"=>3, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phenotypic_summary_of_104_Zn_2_Cys_6_TF_gene_deleted_mutants_/1199374", "title"=>"Phenotypic summary of 104 Zn<sub>2</sub>Cys<sub>6</sub> TF gene-deleted mutants.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711689"], "description"=>"<p>(<b>A</b>) Features of a new binary yeast-<i>Escherichia</i>-<i>Agrobacterium</i> shuttle vector, pKO1B. (<b>B</b>) Building of gene-deletion cassettes in pKO1B by yeast recombinational cloning. The 5′ and 3′ flanking fragments of the targeted genes were separately amplified from genomic DNA with primers 5f/5r and 3f/3r. Primers 5r and 3f have 5′ tails homologous to the <i>SUR</i> cassette, whereas those for 5f and 3r are homologous to the vector. The two flanks were cotransformed into yeast along with the <i>SUR</i> cassette and gapped pKO1B. Homologous recombination created the circular knockout vector, and the final knockout vector was subsequently transformed into <i>A. tumefaciens</i>. (<b>C</b>) Deletion of the targeted gene. The gene-deletion cassette was transformed into the fungal cells via ATMT. Homologous recombination created three types of transformants: null mutants, ectopic insertion transformants, and null and ectopic insertion mutants. The <i>GFP</i> gene was discarded in the null mutants. Primers p1/p2 or p3/p4 were used to identify the unique recombinational DNA fragment, indicating a knockout event. (<b>D</b>) The transformants are screened for GFP fluorescence under a microscope. Putative null mutants do have not GFP fluorescence, but ectopic transformants do. (<b>E</b>) The transformants are screened by double PCR for the targeted gene using the <i>β-tubulin</i> gene as a positive control. The wild-type strain or ectopic transformants produced a characteristic band, indicating the targeted gene, while the null mutants did not. (<b>F</b>) The transformants are screened by PCR for a unique recombinational DNA fragment marked as a knockout event. The null mutants have a 1.2–2.0 kb band on an electrophoretic gel, while the wild-type strain and the ectopic transformants do not.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199321, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g001", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Overview_of_the_high_throughput_gene_knockout_system_in_fungus_/1199321", "title"=>"Overview of the high-throughput gene knockout system in fungus.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711769", "https://ndownloader.figshare.com/files/1711770", "https://ndownloader.figshare.com/files/1711771", "https://ndownloader.figshare.com/files/1711772", "https://ndownloader.figshare.com/files/1711773"], "description"=>"<div><p>Because of great challenges and workload in deleting genes on a large scale, the functions of most genes in pathogenic fungi are still unclear. In this study, we developed a high-throughput gene knockout system using a novel yeast-<i>Escherichia</i>-<i>Agrobacterium</i> shuttle vector, pKO1B, in the rice blast fungus <i>Magnaporthe oryzae</i>. Using this method, we deleted 104 fungal-specific Zn<sub>2</sub>Cys<sub>6</sub> transcription factor (TF) genes in <i>M. oryzae</i>. We then analyzed the phenotypes of these mutants with regard to growth, asexual and infection-related development, pathogenesis, and 9 abiotic stresses. The resulting data provide new insights into how this rice pathogen of global significance regulates important traits in the infection cycle through Zn<sub>2</sub>Cys<sub>6</sub>TF genes. A large variation in biological functions of Zn<sub>2</sub>Cys<sub>6</sub>TF genes was observed under the conditions tested. Sixty-one of 104 Zn<sub>2</sub>Cys<sub>6</sub> TF genes were found to be required for fungal development. In-depth analysis of TF genes revealed that TF genes involved in pathogenicity frequently tend to function in multiple development stages, and disclosed many highly conserved but unidentified functional TF genes of importance in the fungal kingdom. We further found that the virulence-required TF genes <i>GPF1</i> and <i>CNF2</i> have similar regulation mechanisms in the gene expression involved in pathogenicity. These experimental validations clearly demonstrated the value of a high-throughput gene knockout system in understanding the biological functions of genes on a genome scale in fungi, and provided a solid foundation for elucidating the gene expression network that regulates the development and pathogenicity of <i>M. oryzae</i>.</p></div>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199386, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1004432.s001", "https://dx.doi.org/10.1371/journal.ppat.1004432.s002", "https://dx.doi.org/10.1371/journal.ppat.1004432.s003", "https://dx.doi.org/10.1371/journal.ppat.1004432.s004", "https://dx.doi.org/10.1371/journal.ppat.1004432.s005"], "stats"=>{"downloads"=>20, "page_views"=>40, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Systematic_Analysis_of_Zn_2_Cys_6_Transcription_Factors_Required_for_Development_and_Pathogenicity_by_High_Throughput_Gene_Knockout_in_the_Rice_Blast_Fungus/1199386", "title"=>"Systematic Analysis of Zn<sub>2</sub>Cys<sub>6</sub> Transcription Factors Required for Development and Pathogenicity by High-Throughput Gene Knockout in the Rice Blast Fungus", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711722"], "description"=>"<p>The conidia (0 h) of the wild-type strain, mutant Δ<i>cca1</i>, and its complemented strain <i>cca1-c</i> were grown in H<sub>2</sub>O on plastic cover slides for 4 h (conidial germination) and 24 h (appressorium formation). Bar = 10 µm.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199352, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g005", "stats"=>{"downloads"=>1, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Conidia_conidial_germination_and_appressorium_formation_of_the_M_oryzae_strain_cca1_/1199352", "title"=>"Conidia, conidial germination and appressorium formation of the <i>M. oryzae</i> strain Δ<i>cca1</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711744"], "description"=>"<p>(<b>A</b>) Pathogenicity assay of 5 mutants and their complementation strains on rice. These 5 mutants are Δ<i>gpf1</i>, Δ<i>gta1</i>, Δ<i>cnf2</i>, Δ<i>cnf1</i>, Δ<i>cca1</i> and their complementation strains <i>gpf1-c</i>, <i>gta1-c</i>, <i>cnf2-c</i>, <i>cnf1-c</i>, and <i>cca1-c</i>. The rice seedlings were sprayed with a conidial suspension of <i>M. oryzae</i> strains and cultured for 7 days. (<b>B</b>) Pathogenicity assay of the mutants on rice leaf explants. The mycelial agar plugs of the mutants Δ<i>Mocod1</i> and Δ<i>conx1</i>, their complementation strains <i>Mocod1-c</i> and <i>conx1-c</i>, and the wild-type strain 70-15 were placed on intact rice leaves for 4 days. (<b>C</b>) Penetration assay. 10-or 20-µl (5×10<sup>4</sup> conidia/ml) conidial suspensions were inoculated on onion cuticle or barley leaves (intact or abraded) and incubated for 24 or 48 h. The experimental strains were the wild-type strain 70-15, mutants (Δ<i>gpf1</i>, Δ<i>gta1</i>, Δ<i>cnf2</i>, Δ<i>cnf1</i> and Δ<i>cca1</i>) and complemented strains (<i>gpf1-c</i>, <i>gta1-c</i>, <i>cnf2-c</i>, <i>cnf1-c</i> and <i>cca1-c</i>). Same capital letters in same treatment item indicate non-significant difference estimated by Duncan's test (<i>P</i>≤0.05). (<b>D</b>) Penetration of Δ<i>gpf1</i>. 20 µl (5×10<sup>4</sup> conidia/ml) conidial suspensions of Δ<i>gpf1</i>, complemented strain <i>gpf1-c</i> or wild-type strain were inoculated on barley leaf explants (intact or abraded) and incubated for 24 or 48 h. Arrows indicate the <i>in planta</i> hyphae invaded. Bar = 25 µm.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199364, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g006", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pathogenicity_assay_of_the_M_oryzae_strains_/1199364", "title"=>"Pathogenicity assay of the <i>M. oryzae</i> strains.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/1711716"], "description"=>"<p>(<b>A</b>) The mycelial appearance of the wild-type strain, Δ<i>cnf1</i> and its complemented strain <i>cnf1-c</i>. Stars indicate non-sporulating hyphae. Bar = 1 cm. (<b>B</b>) Spore-bearing aerial hyphae of the wild-type strain, Δ<i>cnf1</i> and its complemented strain <i>cnf1-c</i> on CM medium. Bar = 20 µm. (<b>C</b>) Conidial development on conidiophores of the <i>M. oryzae</i> strains Δ<i>cnf1</i> and Δ<i>cca1</i>. The conidiophore pictures of the wild-type, mutants (Δ<i>cnf1</i> and Δ<i>cca1</i>), and their complemented strains (<i>cnf1-c</i> and <i>cca1-c</i>) were taken on the strains grown on CM medium on microscope slides for 1 day. Bar = 40 µm.</p>", "links"=>[], "tags"=>["104 Zn 2Cys TF genes", "GPF", "gene expression network", "Zn 2Cys genes", "rice blast fungus Magnaporthe oryzae", "9 abiotic stresses", "cnf", "TF genes", "1b", "Rice Blast Fungus", "Zn 2Cys Transcription Factors"], "article_id"=>1199346, "categories"=>["Biological Sciences"], "users"=>["Jianping Lu", "Huijuan Cao", "Lilin Zhang", "Pengyun Huang", "Fucheng Lin"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1004432.g004", "stats"=>{"downloads"=>4, "page_views"=>226, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mycelial_appearance_and_spore_bearing_aerial_hyphae_of_the_M_oryzae_strains_/1199346", "title"=>"Mycelial appearance and spore-bearing aerial hyphae of the <i>M. oryzae</i> strains.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-09 03:35:23"}

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Mycology", "average_usage"=>[348, 543]}, {"subject_area"=>"/Biology and life sciences/Plant science", "average_usage"=>[278, 438]}, {"subject_area"=>"/Medicine and health sciences/Pathology and laboratory medicine", "average_usage"=>[287]}]}
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