Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum.
Identifieur interne : 000117 ( Main/Exploration ); précédent : 000116; suivant : 000118Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum.
Auteurs : Rustam Aghazadeh [Iran] ; Mohammadreza Zamani [Iran] ; Mostafa Motallebi [Iran] ; Mehdi Moradyar [Iran] ; Zahra Moghadassi Jahromi [Iran]Source :
- World journal of microbiology & biotechnology [ 1573-0972 ] ; 2016.
Descripteurs français
- KwdFr :
- Ascomycota (effets des médicaments et des substances chimiques), Ascomycota (pathogénicité), Brassica napus (génétique), Brassica napus (microbiologie), Chitinase (génétique), Chitinase (métabolisme), Chitinase (pharmacologie), Maladies des plantes (génétique), Maladies des plantes (microbiologie), Oryza (génétique), Oryza (métabolisme), Protéines de fusion recombinantes (pharmacologie), Protéines fongiques (génétique), Protéines fongiques (métabolisme), Protéines fongiques (pharmacologie), Protéines végétales (génétique), Protéines végétales (métabolisme), Protéines végétales (pharmacologie), Résistance à la maladie (MeSH), Trichoderma (enzymologie), Trichoderma (génétique), Végétaux génétiquement modifiés (microbiologie).
- MESH :
- effets des médicaments et des substances chimiques : Ascomycota.
- enzymologie : Trichoderma.
- génétique : Brassica napus, Chitinase, Maladies des plantes, Oryza, Protéines fongiques, Protéines végétales, Trichoderma.
- microbiologie : Brassica napus, Maladies des plantes, Végétaux génétiquement modifiés.
- métabolisme : Chitinase, Oryza, Protéines fongiques, Protéines végétales.
- pathogénicité : Ascomycota.
- pharmacologie : Chitinase, Protéines de fusion recombinantes, Protéines fongiques, Protéines végétales.
- Résistance à la maladie.
English descriptors
- KwdEn :
- Ascomycota (drug effects), Ascomycota (pathogenicity), Brassica napus (genetics), Brassica napus (microbiology), Chitinases (genetics), Chitinases (metabolism), Chitinases (pharmacology), Disease Resistance (MeSH), Fungal Proteins (genetics), Fungal Proteins (metabolism), Fungal Proteins (pharmacology), Oryza (genetics), Oryza (metabolism), Plant Diseases (genetics), Plant Diseases (microbiology), Plant Proteins (genetics), Plant Proteins (metabolism), Plant Proteins (pharmacology), Plants, Genetically Modified (microbiology), Recombinant Fusion Proteins (pharmacology), Trichoderma (enzymology), Trichoderma (genetics).
- MESH :
- chemical , genetics : Chitinases, Fungal Proteins, Plant Proteins.
- drug effects : Ascomycota.
- enzymology : Trichoderma.
- genetics : Brassica napus, Oryza, Plant Diseases, Trichoderma.
- chemical , metabolism : Chitinases, Fungal Proteins, Oryza, Plant Proteins.
- microbiology : Brassica napus, Plant Diseases, Plants, Genetically Modified.
- pathogenicity : Ascomycota.
- chemical , pharmacology : Chitinases, Fungal Proteins, Plant Proteins, Recombinant Fusion Proteins.
- Disease Resistance.
Abstract
Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T2 generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant.
DOI: 10.1007/s11274-016-2104-6
PubMed: 27430511
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<term>Chitinases (metabolism)</term>
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<term>Plant Proteins (pharmacology)</term>
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<front><div type="abstract" xml:lang="en">Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T2 generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant. </div>
</front>
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<Month>03</Month>
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<ArticleTitle>Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum.</ArticleTitle>
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<Abstract><AbstractText>Canola (Brassica napus) plants were co-transformed with two pathogenesis-related protein genes expressing a Trichoderma atroviride chitinase with a chitin-binding domain (chimeric chitinase) and a thaumatin-like protein (tlp) from Oryza sativa conferring resistance to phytopatogenic fungi by Agrobacterium-mediated transformation. The putative transgenic plants were confirmed by PCR. After measuring the specific activity of the chimeric chitinase and glucanase activity for tlp genes, transgenic plants with high specific activity were selected for southern blot analysis to confirm the copy number of the genes. In vitro assays, the antifungal activity of crude extracted protein against Sclerotinia sclerotiorum showed that the inhibition percentage in double transgenic plants was between 55 and 62, whereas the inhibition percentage in single-gene transformants (chimeric chitinase) ranged from 35 to 45 percent. Importantly, in greenhouse conditions, the double transgenic plants showed significant resistance than the single-gene transformant and wild type plants. The results in T2 generation using the intact leaf inoculation method showed that the average lesion diameters were 10, 14.7 and 29 mm for the double transformant, single-gene transformant and non-transgenic plants, respectively. Combined expression of chimeric chitinase and tlp in transgenic plants showed significantly enhanced resistance against S. sclerotiorum than the one that express single-gene transformant plants. These results suggest that the co-expression of chimeric chitinase and tlp can confer enhanced disease resistance in canola plant. </AbstractText>
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<ForeName>Rustam</ForeName>
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<ForeName>Mohammadreza</ForeName>
<Initials>M</Initials>
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<Author ValidYN="Y"><LastName>Motallebi</LastName>
<ForeName>Mostafa</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, Iran.</Affiliation>
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<ForeName>Mehdi</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, Iran.</Affiliation>
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<ForeName>Zahra</ForeName>
<Initials>Z</Initials>
<AffiliationInfo><Affiliation>Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, Iran.</Affiliation>
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<MeshHeading><DescriptorName UI="D029688" MajorTopicYN="N">Brassica napus</DescriptorName>
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