Identification, Characterization and Gene Expression of Zip Gene Family in Phaseolus Vulgaris

Abstract

Common bean (Phaseolus vulgaris) is an important warm-season legume and plays a significant role in human nutrition, being an economically viable source of protein, dietary fiber, minerals and vitamins. In recent years common bean has become an alternative to dietary supplements as a way to improve human health. Basic nutritional requirement is essential for every human but still meeting Fe and Zn dietary requirements is challenge for many people. ZIP metal transporters are one of the most important gene families for Zn and Fe cellular uptake and translocation in plants. To increase the understanding of uptake, transportation, and storage of Zn in P.vulgaris a detailed knowledge of ZIP gene is required. In this present study, primer for metal transporter gene namely ZIP was designed by IDT Primer designing tool and validated by Primer-BLAST online tool. Basic nutrient requirements were given to P.vulgaris seedlings through Hoagland solution. Total RNA was isolated followed by cDNA synthesis. The ZIP gene was identified through PCR which resulted in amplicon size of ~133bp. The PCR product was sequenced. The resulted sequence was further analyzed through bioinformatics software’s viz. BioEdit, Mega5. Nucleotide composition, multiple sequence alignments were obtained and phylogenetic tree were constructed. This nucleotide sequence was also translated to protein sequence to know their total amino acid composition. The expression level of ZIP gene was measured by Q-PCR using cox gene as an internal control. Gene expression was assessed by using cDNA from all the zinc treated and untreated P.vulgaris cotyledons when subjected to zinc treatment at different concentrations i.e., 100 mM, 150 mM, and 200 mM for 3, 6, and 12 h respectively. The relative expression of ZIP of zinc treated versus control (untreated) of P.vulgaris cotyledons was calculated using BioRad IQ-5 software. ZIP gene expression was highest at 200 mM for 12 hr followed by zinc treatment at 200 mM for 6 hr and lowest at 150 mM for 3hr. From these studies it can be concluded that ZIP gene present in P.vulgaris and characterization of role of such genes will be useful in biofortification and overcoming zinc deficiency.