Cloning and Characterization of a Plant Defensin VaD1 from Azuki Bean

• Published in: Journal of agricultural and food chemistry Vol. 53; no. 4; pp. 982 - 988
• Main Authors: Chen, Gan-Hong, Hsu, Ming-Pin, Tan, Chi-Hsing, Sung, Hsien-Yi, Kuo, C. George, Fan, Ming-Jen, Chen, Huei-Mei, Chen, Shu, Chen, Ching-San
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 23.02.2005
• Subjects: Amino Acid Sequence; amino acid sequences; Anti-Bacterial Agents; antibacterial properties; antifungal properties; Base Sequence; Biological and medical sciences; Callosobruchus chinensis; Cloning, Molecular; complementary DNA; defensins; Defensins - chemistry; Defensins - genetics; Defensins - pharmacology; DNA, Complementary - genetics; DNA, Plant - chemistry; DNA, Plant - genetics; Fabaceae - chemistry; Fabaceae - genetics; Food industries; Food microbiology; food pathogens; Fundamental and applied biological sciences. Psychology; Fungicides, Industrial; Fusarium oxysporum; Fusarium oxysporum f. sp. pisi; genes; insecticidal properties; Molecular Sequence Data; mung beans; nucleotide sequences; plant pathogenic fungi; plant proteins; Recombinant Proteins; Salmonella typhimurium; seeds; Seeds - chemistry; Staphylococcus epidermidis; Vigna angularis; Vigna radiata; Antimicrobial activity; protein synthesis Inhibition; azuki bean (Vigna angularis); Antimicrobial agent; Characterization; Resistance; Leguminosae; Vigna angularis; Protein synthesis; Dicotyledones; defensin; Angiospermae; Spermatophyta; Inhibition; bruchid resistance
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf0402227

A recombinant mungbean defensin VrD1 was previously shown to exhibit antifungal and bruchid-resistant activity. To study the function and regulation of VrD1, genomic DNAs of plant defensins were isolated from Vigna radiata VC6089A and azuki bean Vigna angularis Kao Hsiung No. 6. The azuki bean defensin genomic DNA VaD1 was sequenced and converted to VaD1 cDNA. VaD1 defensin was purified from Vigna angularis Kao Hsiung No. 6 to apparent homogeneity. The complete amino acid sequence of the purified VaD1 was determined and was found to be exactly the same as the sequence deduced from VaD1 cDNA. VaD1 is a basic protein containing 46 amino acids with four conserved disulfide bonds and shares high sequence homology (78.3%) with VrD1. VaD1 inhibited the growth of Fusarium oxysporum, Fusarium oxysporum f. sp. pisi, Staphylococcus epidermidis, and Salmonella typhimurium. VaD1 also inhibited in vitro protein synthesis and bruchid larval development, but was less active than the recombinant VrD1. Keywords: Antimicrobial activity; azuki bean (Vigna angularis); bruchid resistance; defensin; protein synthesis inhibition