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Genetic Transformation and Green Fluorescent Protein Labeling in Ceratocystis paradoxa from Coconut

文献类型： 外文期刊

作者： Niu, Xiaoqing 1 ; Pei, Mengtian 1 ; Liang, Chenyu 1 ; Lv, Yuexiao 1 ; Wu, Xinyi 1 ; Zhang, Ruina 1 ; Lu, Guodong 1 ; Yu, 1 ;

作者机构： 1.Fujian Agr & Forestry Univ, Key Lab Biopesticide & Chem Biol, Minist Educ, Fuzhou 350002, Fujian, Peoples R China

2.Chinese Acad Trop Agr Sci, Coconut Res Inst, Wenchang 571339, Peoples R China

关键词： stem-bleeding disease of coconut; Ceratocystis paradoxa; gfp gene; transformation; protoplast

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）

ISSN： 1422-0067

年卷期： 2019 年 20 卷 10 期

页码：

收录情况： SCI

摘要： Ceratocystis paradoxa, the causal agent of stem-bleeding disease of the coconut palm, causes great losses to the global coconut industry. As the mechanism of pathogenicity of C. paradoxa has not been determined, an exogenous gene marker was introduced into the fungus. In this study, pCT74-sGFP, which contains the green fluorescent protein (GFP) gene, and the hygromycin B resistance gene as a selective marker, was used as an expression vector. Several protoplast release buffers were compared to optimize protoplast preparation. The plasmid pCT74-sGFP was successfully transformed into the genome of C. paradoxa, which was verified using polymerase chain reaction and green fluorescence detection. The transformants did not exhibit any obvious differences from the wild-type isolates in terms of growth and morphological characteristics. Pathogenicity tests showed that the transformation process did not alter the virulence of the X-3314 C. paradoxa strain. This is the first report on the polyethylene glycol-mediated transformation of C. paradoxa carrying a reporter' gene GFP that was stably and efficiently expressed in the transformants. These findings provide a basis for future functional genomics studies of C. paradoxa and offer a novel opportunity to track the infection process of C. paradoxa.

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