Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus
Following the previous isolation of CLPG1, a gene encoding an endopolygalacturonase (endoPG) secreted into the culture filtrate of Colletotrichum lindemuthianum, we have isolated and sequenced an additional endoPG gene, CLPG2. This gene is present as a single copy in the genome of the fungus. At the...
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| Format: | Article |
| Language: | English |
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The American Phytopathological Society
1997-08-01
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| Series: | Molecular Plant-Microbe Interactions |
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| Online Access: | https://apsjournals.apsnet.org/doi/10.1094/MPMI.1997.10.6.769 |
| _version_ | 1828259964973481984 |
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| author | Sylvie Centis Isabelle Guillas Nathalie Séjalon Marie-Thérèse Esquerré-Tugayé Bernard Dumas |
| author_facet | Sylvie Centis Isabelle Guillas Nathalie Séjalon Marie-Thérèse Esquerré-Tugayé Bernard Dumas |
| author_sort | Sylvie Centis |
| collection | DOAJ |
| description | Following the previous isolation of CLPG1, a gene encoding an endopolygalacturonase (endoPG) secreted into the culture filtrate of Colletotrichum lindemuthianum, we have isolated and sequenced an additional endoPG gene, CLPG2. This gene is present as a single copy in the genome of the fungus. At the amino acid level, CLPG2 shows 61% identity to CLPG1 and between 37 to 59% identity to other fungal endoPGs. RNA blot analyses of endoPG gene expression were followed with specific probes during in vitro culture of the fungus. When conidia were used to inoculate a synthetic medium containing pectin as sole carbon source, only CLPG1 was found to be expressed after 3 days of culture. However, transferring the mycelium grown on glucose for 4 days to a pectin-containing medium allowed the detection of CLPG1 and CLPG2 transcripts as early as 12 h after transfer on this substrate. Expression of CLPG2 was transient while that of CLPG1 was more prolonged. Immunocytological localization of endoPG in C. lindemuthianum-infected bean tissues with antibodies against CLPG1 confirmed that the protein is produced in planta and is associated with extensive degradation of the host cell wall. Detection of endoPG transcripts by reverse transcription-polymerase chain reaction revealed that CLPG1, but not CLPG2, is expressed at the beginning of the necrotrophic stage of infection. These results show that the two endoPG genes are differentially expressed and that CLPG1 encodes the major secreted endoPG both during saprophytic growth and during plant infection. |
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| format | Article |
| id | doaj.art-6402fe249c6c4c319bfba3756b7c25f2 |
| institution | Directory Open Access Journal |
| issn | 0894-0282 1943-7706 |
| language | English |
| last_indexed | 2024-04-13T03:16:27Z |
| publishDate | 1997-08-01 |
| publisher | The American Phytopathological Society |
| record_format | Article |
| series | Molecular Plant-Microbe Interactions |
| spelling | doaj.art-6402fe249c6c4c319bfba3756b7c25f22022-12-22T03:04:54ZengThe American Phytopathological SocietyMolecular Plant-Microbe Interactions0894-02821943-77061997-08-0110676977510.1094/MPMI.1997.10.6.769Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the FungusSylvie CentisIsabelle GuillasNathalie SéjalonMarie-Thérèse Esquerré-TugayéBernard DumasFollowing the previous isolation of CLPG1, a gene encoding an endopolygalacturonase (endoPG) secreted into the culture filtrate of Colletotrichum lindemuthianum, we have isolated and sequenced an additional endoPG gene, CLPG2. This gene is present as a single copy in the genome of the fungus. At the amino acid level, CLPG2 shows 61% identity to CLPG1 and between 37 to 59% identity to other fungal endoPGs. RNA blot analyses of endoPG gene expression were followed with specific probes during in vitro culture of the fungus. When conidia were used to inoculate a synthetic medium containing pectin as sole carbon source, only CLPG1 was found to be expressed after 3 days of culture. However, transferring the mycelium grown on glucose for 4 days to a pectin-containing medium allowed the detection of CLPG1 and CLPG2 transcripts as early as 12 h after transfer on this substrate. Expression of CLPG2 was transient while that of CLPG1 was more prolonged. Immunocytological localization of endoPG in C. lindemuthianum-infected bean tissues with antibodies against CLPG1 confirmed that the protein is produced in planta and is associated with extensive degradation of the host cell wall. Detection of endoPG transcripts by reverse transcription-polymerase chain reaction revealed that CLPG1, but not CLPG2, is expressed at the beginning of the necrotrophic stage of infection. These results show that the two endoPG genes are differentially expressed and that CLPG1 encodes the major secreted endoPG both during saprophytic growth and during plant infection.https://apsjournals.apsnet.org/doi/10.1094/MPMI.1997.10.6.769Phaseolus vulgaris |
| spellingShingle | Sylvie Centis Isabelle Guillas Nathalie Séjalon Marie-Thérèse Esquerré-Tugayé Bernard Dumas Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus Molecular Plant-Microbe Interactions Phaseolus vulgaris |
| title | Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus |
| title_full | Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus |
| title_fullStr | Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus |
| title_full_unstemmed | Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus |
| title_short | Endopolygalacturonase Genes from Colletotrichum lindemuthianum: Cloning of CLPG2 and Comparison of Its Expression to That of CLPG1 During Saprophytic and Parasitic Growth of the Fungus |
| title_sort | endopolygalacturonase genes from colletotrichum lindemuthianum cloning of clpg2 and comparison of its expression to that of clpg1 during saprophytic and parasitic growth of the fungus |
| topic | Phaseolus vulgaris |
| url | https://apsjournals.apsnet.org/doi/10.1094/MPMI.1997.10.6.769 |
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