Molecular and Biochemical Analysis of Hydroponically Grown Cicer arietinum with Different Levels of Zinc

Abstract

Chickpea has high protein content & minerals including Ca, P, Zn, Fe and Mg. Zn is required for activity of approximately 300 enzymes. The ZIP family is well characterized for its role in Zn transport. In the present study, molecular and biochemical analysis of hydroponically grown Cicer arietinum with different levels of zinc is done. Primers specific to zip gene were designed by IDT Tool and then total RNA from cotyledons was extracted followed by cDNA synthesis. The zip gene was identified by PCR with amplicon length ~775bp and send for sequencing. The obtained sequence was submitted to NCBI with Accession number, further in silico analysis was done. The expression analysis was performed by Q-PCR using cDNA from Cicer arietinum cotyledons using Himedia Insta Q96 software which revealed that the level of zip gene expression was highest at 9μM for 18 hrs followed by 6μM for 18 hrs. The effect of Zn uptake on structure and activity of regulatory enzymes was checked. Zn uptake was measured from seedlings and two enzymes ADH and CA were isolated, protein and enzymatic activity were estimated. The treatment (9μM + 18 hrs) resulted in higher crude enzyme content & enzyme activity in both ADH and CA as compared to all other treatments. It was inferred that on increasing zinc concentration, the crude enzyme content and enzyme activity increased up to a certain Zn concentration with time after which Zn start accumulating, which lead to the formation of a bigger oligomer of enzymes and this oligomer formation causes the inactivation of enzyme. On the basis of experimental findings, it is suggested that incorporating 9μM of Zn up to 18 hrs will probably results in higher protein content and higher enzyme activity. From our study, it can be concluded that inducing such minerals and identifying and characterizing such genes will be helpful in biofortification studies