Journal of Crop Protection
Journal of Crop Protection
JCP
Agriculture
http://jcp.modares.ac.ir
1
admin
2251-9041
2251-905X
en
jalali
1392
12
1
gregorian
2014
3
1
3
1
online
1
fulltext
en
Beta glucanase (Bgn13.1) expressed in transgenic Brassica napus confers antifungal activity against Sclerotinia sclerotiorum
<em>Brassica napus </em>is an important oilseed crop and the yield loss due to fungal disease stem rot caused by <em>Sclerotinia sclerotiorum </em>is a serious problem in cultivation of this crop. The pathogenesis-related (PR) protein, glucanase, hydrolyzes a major cell wall component, glucan, of the pathogenic fungi and acts as a plant defense barrier. In this study, a β-1,3-glucanase (<em>bgn</em>13.1) gene was isolated from the biocontrol fungus <em>Trichoderma virens</em>-10 (showing the high β-glucanase activity) and cloned in pUC19 cloning vector. The cloned fragment was confirmed by molecular analysis and showed to contain two short introns, 52 and 57 bp and an open reading frame coding 761 amino acids. The <em>bgn</em>13.1 gene was over-expressed under the CaMV35S promoter in <em>B. napus</em>, R line Hyola 308. Transformation of cotyledonary petioles was achieved by pBIKH1 containing <em>bgn</em>13.1 gene via <em>Agrobacterium</em> <em>tumefaciens </em>LBA4404. The insertion of transgene was verified by the polymerase chain reaction (PCR) and genomic DNA Southern dot blotting in T0 generation. RT-PCR analysis indicated that the transgenic canola plants were able to transcribe the β-1,3 glucanase gene. Also, we used transgenic over-expression approach in order to investigate antifungal activity of expressed Bgn13.1 on <em>S. sclerotiorum</em>. The heterologous expressed Bgn13.1 of line # 7 and line # 10 compared with other lines showed stronger inhibition against hyphal growth of <em>S. sclerotiorum</em> with
bgn13.1,Brassica napus,glucanase enzyme,Sclerotinia sclerotiorum,Transgenic plants,Trichoderma virens
31
42
http://jcp.modares.ac.ir/browse.php?a_code=A-3-1000-6754&slc_lang=en&sid=3
Hamid-Reza
Kheiri
Hamid-Reza
Kheiri
100319475328460049665
100319475328460049665
No
National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Mostafa
Motallebi
Mostafa
Motallebi
100319475328460041075
100319475328460041075
Yes
National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Mohammad Reza
Zamani
Mohammad Reza
Zamani
100319475328460041076
100319475328460041076
No
National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Ali
Deljo
Ali
Deljo
100319475328460049666
100319475328460049666
No
Department. of Agricultural Biotechnology, Faculty of Agriculture, Bu-Ali Sina University, Hamadan, Iran.
Department. of Agricultural Biotechnology, Faculty of Agriculture, Bu-Ali Sina University, Hamadan, Iran.