Hairy roots biochemical characteristics of vegetable pea’s morphotype with modified leaf

Abstract

BACKGROUND: In the literature, data on the successful production of high-protein pea root cultures with mutant alleles tl and af tl and their biochemical characteristics are not presented.

AIM: The aim of this study is the biochemical characteristic of the obtained transformed cultures of mutant pea lines with a modified leaf morphotype was. Rol genes clarification of transformed pea mutant lines by wild strains of Agrobacterium rhizogenes and amino acid analysis of the total protein of the obtained roots cultures were objectives of the research.

MATERIALS AND METHODS: Total DNA has been isolated from pea mutant root cultures. The studies were carried out on the equipment of the thermal cycler Tertsik of the company DNA-Technology (Russia). Amplicons were detected by electrophoresis in a 2% agarose gel. The gel was visualized and photographed under ultraviolet light (λ = 312 nm). The quantitative and qualitative amino acid composition of root cultures was determined by ion exchange chromatography on the amino acid analyzer AAA-339 (Microtechna, Czech Republic).

RESULTS: PCR analysis revealed the absence of agrobacterial contamination in the transformed cultures and their stable growth on liquid and agar hormone-free media for 5 years. PCR assay showed the presence of rol C and D genes in culture with the tltl genotype and four rol genes A, B, C, and D in culture with the afaftltl genotype. A differentiated content of a number of amino acids in the biomass of transformed cultures was found depending on the genotype of the culture and the insertion of rol genes. Seven essential amino acids were identified in both cultures.

CONCLUSIONS: Tryptophan turned out to be the limiting essential amino acid for both cultures. In terms of the sum of essential, ketogenic and sulfur-containing amino acids, the culture with the rol genes A, B, C, and D turned out to be the richest and most balanced.

Keywords

transformation / Pisum sativum / mutants / Agrobacterium rhizogenes / hairy roots / PCR / amino acid composition

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