Characterization of the β-1,3-glucanase gene in peanut (Arachis hypogaea L.) by cloning and genetic transformation
| Autor: | Ding X, L X Qiao, Jingshan Wang, Wang Hc, Sui Jiongming |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Arachis
Agrobacterium Molecular Sequence Data Biology law.invention Transformation Genetic law Complementary DNA Genetics RNA Messenger Molecular Biology Gene Polymerase chain reaction Disease Resistance Glucuronidase food and beverages Oryza Glucan 1 3-beta-Glucosidase General Medicine Plants Genetically Modified biology.organism_classification Molecular biology Reverse transcription polymerase chain reaction genomic DNA Transformation (genetics) GenBank |
| Zdroj: | Genetics and Molecular Research. 13:1893-1904 |
| ISSN: | 1676-5680 |
| DOI: | 10.4238/2014.march.17.17 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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