Cloning and identification of IRT genes and their response to amino acid-iron compound fertilizer in grape
In order to reveal the molecular mechanism of iron absorption and transport in grape,and to clarify the expression difference of iron regulatory transporter gene VvIRT in different development stages of grape fruit and its response characteristics to foliar application of amino acid-iron compound fertilizer,Yanniang 1 was used as the mate-rial in this study.The IRT encoding iron regulatory transporter was isolated and cloned,and the distribution and struc-ture of IRT gene and conserved motifs of proteins encoded by IRT gene were analyzed.The expression difference of IRT gene in different development stages of Yanniang 1 fruit and its response characteristics to foliar spraying treatment were analyzed by real-time fluorescence quantitative PCR.The results showed that 10 VvIRT genes(VvIRT1-VvIRT10)were cloned from the grape genome and distributed on seven chromosomes.The encoded proteins contained Fe2+transporters or zinc/iron transporters(PF02535),which were typical iron regulatory proteins.The proteins enco-ded by VvIRT1,VvIRT2,VvIRT4,VvIRT5 and VvIRT8 were alkaline proteins,while the other five proteins were acid-ic proteins.VvIRT5 protein and VvIRT8 protein were unstable proteins,and the other eight proteins were stable pro-teins.The VvIRT7 protein was mainly located in the vacuolar membrane,and the other proteins were mainly located in the plasma membrane.The overall expression level of VvIRT7 was the highest in all tissues at different developmental stages of grape fruit,followed by VvIRT9 and VvIRT6,while VvIRT1,VvIRT3,VvIRT5,VvIRT8 and VvIRT10 were not expressed during the development of grape fruit.Under foliar application of iron fertilizer,the expression levels of VvIRT7 in phloem and VvIRT9 in fruit and phloem increased significantly from 35 d after flowering(young fruit stage)to 70 d after flowering(veraison stage),while the expression levels of VvIRT7 and VvIRT9 in phloem and leaves de-creased significantly from 85 d after flowering(second expansion stage)to 115 d after flowering(mature stage).Therefore,the response characteristics of VvIRT gene to foliar application of iron fertilizer are closely related to fruit development stage and organ type.The proteins encoded by VvIRT7 and VvIRT9 genes are two important iron regulatory transporters during grape fruit development.
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