title: Characterization of the β-1,3-glucanase gene in peanut (Arachis hypogaea L.) by cloning and genetic transformation.
abstract: Plant β-1,3-glucanases are commonly involved in disease resistance. This report describes the cloning and genetic transformation of a β-1,3-glucanase gene from peanut. The gene was isolated from both the genomic DNA and cDNA of peanut variety Huayu20 by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR), respectively. The DNA sequence contained 1471 bp including two exons and one intron, and the coding sequence contained 1047 bp that coded for a 348-amino acid protein with a calculated molecular weight of 38.8 kDa. The sequence was registered in NCBI (GenBank accession No. JQ801335) and was designated as Ah-Glu. As determined by BLAST analysis, the Ah-Glu protein has 42-90% homology with proteins from Oryza sativa (BAC83070.1), Zea mays (NP_001149308), Arabidopsis thaliana (NP_200470.1), Medicago sativa (ABD91577.1), and Glycine max (XP_003530515.1). The over-expression vector pCAMBIA1301-Glu containing Ah-Glu was constructed, confirmed by PCR and restriction enzyme digestion, and transformed into peanut variety Huayu22 by Agrobacterium EHA105-mediated transformation. The putative transformed plants (T0) were confirmed by PCR amplification. RT-PCR analysis and β-glucuronidase (GUS) staining showed that the transferred Ah-Glu was expressed as mRNA and protein. In a laboratory test, the transgenic plants were found to be more resistant to the fungal pathogen Cercospora personata than the non-transgenic plants were.
authors.affiliation: Key Lab of Plant Biotechnology in Universities of Shandong, College of Life Science, Qingdao Agriculture University, Qingdao, China.
authors.forename: L X
authors.initials: LX
authors.lastname: Qiao
authors.affiliation: Key Lab of Plant Biotechnology in Universities of Shandong, College of Life Science, Qingdao Agriculture University, Qingdao, China.
authors.forename: X
authors.initials: X
authors.lastname: Ding
authors.affiliation: Key Lab of Plant Biotechnology in Universities of Shandong, College of Life Science, Qingdao Agriculture University, Qingdao, China.
authors.forename: H C
authors.initials: HC
authors.lastname: Wang
authors.affiliation: Key Lab of Plant Biotechnology in Universities of Shandong, College of Life Science, Qingdao Agriculture University, Qingdao, China.
authors.forename: J M
authors.initials: JM
authors.lastname: Sui
authors.affiliation: Key Lab of Plant Biotechnology in Universities of Shandong, College of Life Science, Qingdao Agriculture University, Qingdao, China jswang319@126.com.
authors.forename: J-S
authors.initials: JS
authors.lastname: Wang
mesh_terms: D010367:Arachis / Q000201:enzymology*; D060467:Disease Resistance / Q000235:genetics*; D043326:Glucan 1,3-beta-Glucosidase / Q000235:genetics* / Q000378:metabolism; D005966:Glucuronidase / Q000235:genetics; D008969:Molecular Sequence Data; D012275:Oryza; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000096:biosynthesis; D014170:Transformation, Genetic
chemical_list: D012333:RNA, Messenger; D005966:Glucuronidase; D043326:Glucan 1,3-beta-Glucosidase