Agrobacterium-mediated transformation of the ascomycete mushroom Morchella importuna using polyubiquitin and glyceraldehyde-3-phosphate dehydrogenase promoter-based binary vectors.

Publisher: Springer Country of Publication: Germany NLM ID: 9012472 Publication Model: Electronic Cited Medium: Internet ISSN: 1573-0972 (Electronic) Linking ISSN: 09593993 NLM ISO Abbreviation: World J Microbiol Biotechnol Subsets: MEDLINE

Publication: 2005- : Berlin : Springer
Original Publication: Oxford, OX, UK : Published by Rapid Communications of Oxford Ltd in association with UNESCO and in collaboration with the International Union of Microbiological Societies, c1990-

Transformation, Genetic*; Agaricales/*genetics ; Agrobacterium tumefaciens/*genetics ; Ascomycota/*genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases/*genetics ; Molecular Biology/*methods ; Polyubiquitin/*genetics ; Promoter Regions, Genetic/*genetics; Agaricales/metabolism ; Ascomycota/cytology ; Ascomycota/metabolism ; Cinnamates/pharmacology ; DNA, Bacterial/genetics ; Gene Expression Regulation, Fungal ; Genes, Reporter ; Genetic Vectors/genetics ; Genetic Vectors/metabolism ; Glucuronidase/genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Hygromycin B/analogs & derivatives ; Hygromycin B/pharmacology ; Plasmids ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism

Morchella importuna is a worldwide distributed edible mushroom with high ecological and economic values, but the molecular and genetic research about this mushroom has been hindered due to lack of an efficient transformation method. Here, we report for the first time the successful transformation of M. importuna by using a hypervirulent Agrobacterium tumefaciens strain bearing the constructed binary plasmid p1391-U-GUS. The selectable markers used were the genes for hygromycin resistance under the control of the polyubquitin promoter from M. importuna. The reporter genes were those for enhanced green fluorescent protein (EGFP) and the β-Glucuronidase (GUS) under the control of glyceraldehyde-3-phosphate dehydrogenase promoter and polyubquitin promoter respectively. The presence of the reporter gene EGFP in the transformants was confirmed by the fluorescence and confocal microscope and molecular analysis and that of the reporter gene GUS was verified by enzyme activity and molecular analysis. The analysis results of both reporter genes indicated that Agrobacterium-mediated transformation was successfully performed in M. importuna.

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2662015PY058 Fundamental Research Funds for the Central Universities; 2662014BQ016 Huazhong Agricultural University Scientific & Technological Self-innovation Foundation

Keywords: Agrobacterium-mediated transformation; GPD promoter; Morchella importuna; Polyubiquitin; True morel

0 (Cinnamates)
0 (DNA, Bacterial)
0 (Recombinant Fusion Proteins)
0 (enhanced green fluorescent protein)
120904-94-1 (Polyubiquitin)
147336-22-9 (Green Fluorescent Proteins)
3XQ2233B0B (Hygromycin B)
3YJY415DDI (hygromycin A)
EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
EC 3.2.1.31 (Glucuronidase)

Date Created: 20180916 Date Completed: 20181024 Latest Revision: 20181114

20231215

10.1007/s11274-018-2529-1

30218324