Cloning, Expression and Function of Iron Regulated Transporter VvIRT1 in Wine Grape（Vitis vinifera L.）

doi:10.13560/j.cnki.biotech.bull.1985.2022-1552

Abstract

Abstract:

The genes VvIRT of iron regulated transporters（IRT）family from wine grape was cloned, its expression pattern and potential biological function were analyzed, which may provide theoretical foundation for Fe absorption and efficient utilization mechanisms of fruit trees. Homology cloning technology was used to clone the VvIRT1 gene from wine grape, real-time quantitative PCR was applied to analyze tissue-specific expression characteristics and their responses to different Fe supplies, and yeast heterologous expression system was to analyze the functional verification. The iron regulated transporter VvIRT1 from diploid wine grape ‘Marselan’ was isolated and determined. Phylogenetic tree analysis showed that VvIRT1 was closely clustered with citrus CsIRT1（Rutaceae）and upland cotton GhIRT1（Malvaceae）that the genetic distance among them was relatively close. VvIRT1 was specifically expressed in the roots of both 5-year-old mature trees and tissue-cultured seedlings. Fe depletion significantly induced the expressions of VvIRT1 in the whole tested seedlings, while high Fe toxicity significantly reduced the expressions of VvIRT1 in all tested seedlings in the roots of tissue-cultured seedlings. In addition, VvIRT1 recovered the normal growth of yeast mutant DEY1453 and had ability of transporting external Fe²⁺.

Key words: grape, Fe uptake and transport, iron regulated transporter, yeast functional complementation

SONG Zhi-zhong, XU Wei-hua, XIAO Hui-lin, TANG Mei-ling, CHEN Jing-hui, GUAN Xue-qiang, LIU Wan-hao. Cloning, Expression and Function of Iron Regulated Transporter VvIRT1 in Wine Grape（Vitis vinifera L.）[J]. Biotechnology Bulletin, 2023, 39(8): 234-240.

Figures/Tables 5

Fig. 1 Amino acid sequence identity analysis of IRT homologs from different plant species Ah: Arachis hypgaea; At: Arabidopsis thaliana; Cs: Citrus sinensis; Gh: Gossypium hirsutum; Mx: Malus xiaojinensis; Os: Oryza sativa; Pb: Prunus betulaefolia;Pm: Prunus mume; Pp: Prunus persica; Pt: Populus trichocarpa; Sl: Solanum lycopersicum; Vv: Vitis vinifera; Zm: Zea mays. The same below

Fig. 2 Phylogenetic tree of IRT homologs from different plant species

Fig. 3 Tissue-specific expression analysis of VvIRT1 in grape

Fig. 4 Responses of VvIRT1 under different Fe supplies t-test analysis between iron treatment and control conditions（**P<0.01）

Fig. 5 Verification on the functional complementation of VvIRT1 in yeast

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