Analysis of the VIT1 Promoter Activity in Developing Arabidopsis thaliana Plants

Abstract

Iron (Fe) deficiency in plants is one of the widespread problems limiting agricultural production. Generating crops more tolerant to Fe deficiency by genetic engineering or breeding is of great interest but challenging due to the knowledge gaps in general plant Fe homeostasis. Although several genes involved in Fe homeostasis have been identified, characterization of their roles is mainly limited to specific organs at specific developmental stages of the plant, where their mutants show the most striking phenotype. Vacuolar Iron Transporter 1 (VIT1) is a well-known gene that has been characterized for its function in the mature seed of Arabidopsis thaliana. VIT1 is an Fe transporter that determines the correct distribution of Fe storage in this organ. The study aimed to explore new physiological functions for VIT1. As a first step, Arabidopsis thaliana plants that contain PromoterVIT1: GUS constructs were used to study the temporal and spatial expression of the gene throughout the plant‘s lifecycle. GUS histochemical staining revealed the VIT1 promoter is active in the mature leaves and mature reproductive organs. VIT1 promoter activity in the stamen increased developmentally and was limited to tapetum and guard cells in the pollen sac. In the female organ, VIT1 promoter activity increased as the pistil developed into a silique. Although all the silique exhibited staining, staining density was higher in the peduncle, replum, and stigma regions. Inside the developing silique, funicles were heavily stained. Furthermore, in silico analyses of VT1 transcriptome and protein levels confirmed flower and the silique are hot spots for VT1 activity. Thus, the results may suggest a possible involvement of VT1 protein in several stages of the reproductive system, specifically in the flowering and in the fruit development.