Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 Gene
<i>Pratylenchus loosi</i> is an important root-lesion nematode that causes damage to tea plantations in Iran and all over the world. The present study reports on the characterization and evolution of three ß-1,4-endoglucanase genes: <i>Pl-eng</i>-2, <i>Pl-eng</i>-...
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MDPI AG
2021-03-01
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| Online Access: | https://www.mdpi.com/2223-7747/10/3/568 |
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| author | Negin Mirghasemi Elena Fanelli Salar Jamali Mohammed Mehdi Sohani Francesca De Luca |
| author_facet | Negin Mirghasemi Elena Fanelli Salar Jamali Mohammed Mehdi Sohani Francesca De Luca |
| author_sort | Negin Mirghasemi |
| collection | DOAJ |
| description | <i>Pratylenchus loosi</i> is an important root-lesion nematode that causes damage to tea plantations in Iran and all over the world. The present study reports on the characterization and evolution of three ß-1,4-endoglucanase genes: <i>Pl-eng</i>-2, <i>Pl-eng</i>-3 and <i>Pl-eng</i>-4. The gene structure of <i>Pl-eng</i>-2 was fully determined with the predicted signal peptide and devoid of the linker domain and carbohydrate-binding domain, while <i>Pl-eng</i>-3 and <i>Pl-eng</i>-4 were only partially sequenced. The transcription of <i>Pl-eng</i>-2 was localized in the secretory esophageal glands of all life stages, but it was upregulated in male and female stages. The exon/intron structures of <i>Pl-eng</i>-2, <i>Pl-eng</i>-3 and <i>Pl-eng</i>-4 confirmed that they resulted from gene duplication followed by sequence and gene structure diversification with loss of the linker domain and carbohydrate-binding domain during evolution. A phylogenetic analysis further confirmed that nematode endoglucanases resulted from the horizontal gene transfer of a bacterial gene, as <i>Pl-eng</i>-3 showed sister relationships with the <i>Cel</i>B cellulase of <i>Bacillus subtilis</i>. Silencing <i>Pl-eng</i>-2 by in vitro RNA interference produced a 60% decrease of the transcript level. The reproductive ability of silenced <i>P. loosi</i> showed a 35% reduction of eggs and larval stages compared to untreated nematodes, suggesting that this gene is involved in the early steps of invasion. |
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| spelling | doaj.art-383013f39a9b4f4d97ffbc972c78ef5b2023-11-21T10:51:52ZengMDPI AGPlants2223-77472021-03-0110356810.3390/plants10030568Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 GeneNegin Mirghasemi0Elena Fanelli1Salar Jamali2Mohammed Mehdi Sohani3Francesca De Luca4Plant Protection Department, Faculty of Agricultural Sciences, University of Guilan, Rasht 4199613769, IranIstituto per la Protezione Sostenibile delle Piante (IPSP), Consiglio Nazionale delle Ricerche (CNR), S.S. Bari, Via G. Amendola 122/D, 70126 Bari, ItalyPlant Protection Department, Faculty of Agricultural Sciences, University of Guilan, Rasht 4199613769, IranDepartment of Biotechnology, Faculty of Agricultural Sciences, University of Guilan, Rasht 4199613769, IranIstituto per la Protezione Sostenibile delle Piante (IPSP), Consiglio Nazionale delle Ricerche (CNR), S.S. Bari, Via G. Amendola 122/D, 70126 Bari, Italy<i>Pratylenchus loosi</i> is an important root-lesion nematode that causes damage to tea plantations in Iran and all over the world. The present study reports on the characterization and evolution of three ß-1,4-endoglucanase genes: <i>Pl-eng</i>-2, <i>Pl-eng</i>-3 and <i>Pl-eng</i>-4. The gene structure of <i>Pl-eng</i>-2 was fully determined with the predicted signal peptide and devoid of the linker domain and carbohydrate-binding domain, while <i>Pl-eng</i>-3 and <i>Pl-eng</i>-4 were only partially sequenced. The transcription of <i>Pl-eng</i>-2 was localized in the secretory esophageal glands of all life stages, but it was upregulated in male and female stages. The exon/intron structures of <i>Pl-eng</i>-2, <i>Pl-eng</i>-3 and <i>Pl-eng</i>-4 confirmed that they resulted from gene duplication followed by sequence and gene structure diversification with loss of the linker domain and carbohydrate-binding domain during evolution. A phylogenetic analysis further confirmed that nematode endoglucanases resulted from the horizontal gene transfer of a bacterial gene, as <i>Pl-eng</i>-3 showed sister relationships with the <i>Cel</i>B cellulase of <i>Bacillus subtilis</i>. Silencing <i>Pl-eng</i>-2 by in vitro RNA interference produced a 60% decrease of the transcript level. The reproductive ability of silenced <i>P. loosi</i> showed a 35% reduction of eggs and larval stages compared to untreated nematodes, suggesting that this gene is involved in the early steps of invasion.https://www.mdpi.com/2223-7747/10/3/568cellulaseevolutiongene duplicationintronRNA interferenceroot-lesion nematode |
| spellingShingle | Negin Mirghasemi Elena Fanelli Salar Jamali Mohammed Mehdi Sohani Francesca De Luca Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 Gene Plants cellulase evolution gene duplication intron RNA interference root-lesion nematode |
| title | Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 Gene |
| title_full | Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 Gene |
| title_fullStr | Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 Gene |
| title_full_unstemmed | Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 Gene |
| title_short | Molecular Characterization of Three B-1,4-Endoglucanase Genes in <i>Pratylenchus loosi</i> and Functional Analysis of <i>Pl-eng</i>-2 Gene |
| title_sort | molecular characterization of three b 1 4 endoglucanase genes in i pratylenchus loosi i and functional analysis of i pl eng i 2 gene |
| topic | cellulase evolution gene duplication intron RNA interference root-lesion nematode |
| url | https://www.mdpi.com/2223-7747/10/3/568 |
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