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Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis

Valsa mali is a causal agent of apple and pear trees canker disease, which is a destructive disease that causes serious economic losses in eastern Asia, especially in China. The lack of an efficient transformation system for Valsa mali retards its investigation, which poses difficulties to control t...

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Autores principales: Wang, Caixia, Guan, Xiangnan, Wang, Hanyan, Li, Guifang, Dong, Xiangli, Wang, Guoping, Li, Baohua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3867955/
https://www.ncbi.nlm.nih.gov/pubmed/24381526
http://dx.doi.org/10.1155/2013/968432
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author Wang, Caixia
Guan, Xiangnan
Wang, Hanyan
Li, Guifang
Dong, Xiangli
Wang, Guoping
Li, Baohua
author_facet Wang, Caixia
Guan, Xiangnan
Wang, Hanyan
Li, Guifang
Dong, Xiangli
Wang, Guoping
Li, Baohua
author_sort Wang, Caixia
collection PubMed
description Valsa mali is a causal agent of apple and pear trees canker disease, which is a destructive disease that causes serious economic losses in eastern Asia, especially in China. The lack of an efficient transformation system for Valsa mali retards its investigation, which poses difficulties to control the disease. In this research, a transformation system for this pathogen was established for the first time using A. tumefaciens-mediated transformation (ATMT), with the optimal transformation conditions as follows: 10(6)/mL conidia suspension, cocultivation temperature 22°C, cocultivation time 72 hours, and 200 μM acetosyringone (AS) in the inductive medium. The average transformation efficiency was 1015.00 ± 37.35 transformants per 10(6) recipient conidia. Thirty transformants were randomly selected for further confirmation and the results showed the presence of T-DNA in all hygromycin B resistant transformants and also revealed random and single gene integration with genetic stability. Compared with wild-type strain, those transformants exhibited various differences in morphology, conidia production, and conidia germination ability. In addition, pathogenicity assays revealed that 14 transformants had mitigated pathogenicity, while one had enhanced infection ability. The results suggest that ATMT of V. mali is a useful tool to gain novel insight into this economically important pathogen at molecular levels.
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spelling pubmed-38679552013-12-31 Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis Wang, Caixia Guan, Xiangnan Wang, Hanyan Li, Guifang Dong, Xiangli Wang, Guoping Li, Baohua ScientificWorldJournal Research Article Valsa mali is a causal agent of apple and pear trees canker disease, which is a destructive disease that causes serious economic losses in eastern Asia, especially in China. The lack of an efficient transformation system for Valsa mali retards its investigation, which poses difficulties to control the disease. In this research, a transformation system for this pathogen was established for the first time using A. tumefaciens-mediated transformation (ATMT), with the optimal transformation conditions as follows: 10(6)/mL conidia suspension, cocultivation temperature 22°C, cocultivation time 72 hours, and 200 μM acetosyringone (AS) in the inductive medium. The average transformation efficiency was 1015.00 ± 37.35 transformants per 10(6) recipient conidia. Thirty transformants were randomly selected for further confirmation and the results showed the presence of T-DNA in all hygromycin B resistant transformants and also revealed random and single gene integration with genetic stability. Compared with wild-type strain, those transformants exhibited various differences in morphology, conidia production, and conidia germination ability. In addition, pathogenicity assays revealed that 14 transformants had mitigated pathogenicity, while one had enhanced infection ability. The results suggest that ATMT of V. mali is a useful tool to gain novel insight into this economically important pathogen at molecular levels. Hindawi Publishing Corporation 2013-12-04 /pmc/articles/PMC3867955/ /pubmed/24381526 http://dx.doi.org/10.1155/2013/968432 Text en Copyright © 2013 Caixia Wang et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Wang, Caixia
Guan, Xiangnan
Wang, Hanyan
Li, Guifang
Dong, Xiangli
Wang, Guoping
Li, Baohua
Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis
title Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis
title_full Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis
title_fullStr Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis
title_full_unstemmed Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis
title_short Agrobacterium tumefaciens-Mediated Transformation of Valsa mali: An Efficient Tool for Random Insertion Mutagenesis
title_sort agrobacterium tumefaciens-mediated transformation of valsa mali: an efficient tool for random insertion mutagenesis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3867955/
https://www.ncbi.nlm.nih.gov/pubmed/24381526
http://dx.doi.org/10.1155/2013/968432
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