Cloning and characterization of lipoxygenase gene from germinating seedlings of green gram (Vigna radiata L.)

• Published in: Indian journal of plant physiology Vol. 20; no. 4; pp. 345 - 352
• Main Authors: Kotapati, Kasi Viswanath, Palaka, Bhagath Kumar, Anangi, Raveendra, Kedam, Thyagaraju, Ampasala, Dinakara Rao
• Format: Journal Article
• Language: English
• Published: New Delhi Springer India 01.12.2015; Springer Nature B.V
• Subjects: active sites; Amino acids; Bioinformatics; Biomedical and Life Sciences; Cell Biology; Cloning; complementary DNA; gene expression; genes; germination; Glycine max; isoelectric point; Life Sciences; linoleate 13S-lipoxygenase; messenger RNA; methyl jasmonate; molecular dynamics; molecular weight; mung beans; open reading frames; Original Article; Plant Biochemistry; Plant Ecology; Plant Genetics and Genomics; Plant Physiology; Plant Sciences; rapid amplification of cDNA ends; reverse transcriptase polymerase chain reaction; Seedlings; Vigna radiata; Green gram; Plant lipoxygenases; Molecular dynamics simulations; Homology modeling
• ISSN: 0019-5502; 0974-0252
• DOI: 10.1007/s40502-015-0186-z

Plant LOXs play a significant role in plant physiological processes such as growth, senescence, defence and stress-related responses. In the present study a full length lipoxygenase cDNA ( VrLOX ) has been cloned from green gram germinating seedlings using 5′- and 3′-RACE PCR method and nomenclatured VrLOX . The ORF (2601) encodes 867 amino acid residues with an approximate molecular weight of 97.43 kDa and an isoelectric point of 5.93. The VrLOX comprises of a putative PLAT domain and an iron-binding catalytic domain, and the sequence displayed a significant identity to known plant LOXs deposited in Protein Data Bank. Bioinformatics analysis demonstrated that this gene is most probably a LOX putative protein of type 13-LOX, and shared highest identity (91 %) with the Glycine max LOX-10. Enzymatic assay showed LOX activity was more on day 3 at pH 6.5 and higher MDA levels were observed on day 5 after germination. Semi-quantitative RT-PCR analysis indicated highest VrLOX mRNA expression on day 3 after germination, and could be induced by methyl jasmonate (MeJA). The 3-dimensional structure of the VrLOX was elucidated using in silico approach and molecular dynamics simulations.