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Zinc/iron-regulated transporter genes ZmZIPs from zea mays and application thereof

A transporter, zinc and iron technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problem of few research reports

Inactive Publication Date: 2016-07-06
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is known that many transporters are involved in the zinc and iron ion balance network system in plants, among which the ZIP (Zinc-regulatedtransporters, Iron-regulatedtransporter-likeproteins, ZIP) gene family absorbs, transports and stores divalent metal ions such as zinc and iron. It plays an important role. In Arabidopsis, rice, barley, and soybean, some studies on ZIP family genes have been reported, but the specific mechanism of action of ZIP family genes in plants is not yet fully understood, and the ZIP family of maize There are few studies on genes

Method used

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  • Zinc/iron-regulated transporter genes ZmZIPs from zea mays and application thereof
  • Zinc/iron-regulated transporter genes ZmZIPs from zea mays and application thereof
  • Zinc/iron-regulated transporter genes ZmZIPs from zea mays and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 Cloning of ZmZIPs Gene of the Zinc and Iron Regulatory Transporter in Corn

[0078] 1. Treatment of plant materials

[0079] First soak the vermiculite with Hoagland nutrient solution, sow the corn inbred line X178 seeds in the seedling tray, cover it with a layer of dry vermiculite, cultivate in the greenhouse (16h light / 8h dark, 26℃), 12 days seedling When it grows to 2 leaves and one core, it is transferred to the standard Hoagland nutrient solution and grown for 6 days to 3 leaves and one core (the nutrient solution is changed every 3 days). The corn seedlings with three leaves and one core are in the standard nutrient solution and without zinc, iron, copper, manganese, After 0, 6, 12, 24, 48, 96 hours of treatment under high zinc and iron conditions, the shoots and roots of the seedlings were harvested, and then quickly frozen in liquid nitrogen and stored at -80°C for total RNA extraction.

[0080] 2. Extraction of corn total RNA

[0081] Trizol method was used...

Embodiment 2

[0094] Example 2 Expression pattern of ZmZIPs in seedling, embryo and endosperm

[0095] Dilute the reverse-transcribed cDNA in step 3 of Example 1 by 10 times as a template for the PCR reaction. The PCR reaction system is as follows:

[0096] cDNA2.0μL, ExTaq0.1μL, 2×GCI buffer 10.0μL, 10mMdNTPmix0.8μL, upstream primer RTZmZIP1F / RTZmZIP2F / RTZmZIP3F / RTZmZIP7F / RTZmZIP8F(10μM / μL) 1.0μL, downstream primer RZmZRZRZRZRZRZRZRZR / RTZMIP2R / RTZmZIP1 (10μM / μL)1.0μL, ddH 2 O5.1μL, total 20.0μL;

[0097] RTZmZIP1F5'-CCTCTCTGCGTTGGTTGCTCT-3'

[0098] RTZmZIP1R5'-TTGATGGTTGTTTTCTGGTCGT-3'

[0099] RTZmZIP2F5'-CCACAAATGGCACGAGGTCT-3'

[0100] RTZmZIP2R5'-CGAAGACGGAGTGGAAGCAAA-3'

[0101] RTZmZIP3F5'-GCCTCTTGTTGGTGCCCTTA-3'

[0102] RTZmZIP3R5'-TCAACAATGAACGCTGTAGTGCT-3'

[0103] RTZmZIP7F5'-ACTAGGTGGGTGCATTGCTCAG-3'

[0104] RTZmZIP7R5'-TGCCAGCAGATACCGAGTCAA-3'

[0105] RTZmZIP8F5'-CGTGTCATCGCTCAGGTTCTTG-3'

[0106] RTZmZIP8R5'-CCCTCGAACATTTGGTGGAAG-3'

[0107] PCR reaction conditions: 94°C pre-denaturation f...

Embodiment 3

[0113] Example 3 Bioinformatics analysis of ZmZIPs

[0114] ZmZIPs are composed of 367-483 amino acids and contain 6-9 transmembrane domains. There is a histidine-rich variable region between the third and fourth transmembrane regions, which may be related to the binding and transport of metal ions. Phylogenetic tree analysis shows that ZmZIP1 has a close evolutionary relationship with AtIAR1 and OsIAR1. In addition, ZmZIP3 and ZmZIP4 form a gene cluster with OsZIP3 and OsZIP4, ZmZIP2 is adjacent to OsZIP2, and the zinc transporter OsZIP1, AtZIP2 and AtZIP11 are on the same branch, ZmZIP5 and ZmZIP7 are on the same branch, and the relationship between ZmZIP8 and OsZIP6 and OsZIP6 is evolved. More recently, these results show that the five ZmZIPs isolated in the present invention may be zinc-iron transporters.

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Abstract

The invention discloses zinc / iron-regulated transporter genes ZmZIPs from zea mays and an application thereof. Five zinc / iron-regulated transporter genes ZmZIPs are separated from zea mays. cDNA (complementary deoxyribonucleic acid) sequences of the zinc / iron-regulated transporter genes ZmZIPs are respectively selected from nucleotide sequences shown in SEQ ID No.1, SEQ ID No.3, SEQ ID No.5, SEQ ID No.7 or SEQ ID No.9. Subcellular localization shows that zinc / iron-regulated transporters coded by the genes are localized in plasma membranes and endoplasmic reticulum of cells. Yeast complementation tests show that the five separated genes ZmZIPs have the function of transporting zinc / iron. The genes ZmZIPs have important application prospects in regulating and controlling the zinc / iron uptake, transport and storage capacities of plants, promoting development and maturation of embryos and endosperm and increasing the content of zinc / iron in crop seeds.

Description

[0001] This application is a divisional application with the filing date of "2014-05-30", the application number of "201410240908.X", and the invention titled "ZmZIPs gene and its application of the zinc-iron regulated transporter in corn". Technical field [0002] The present invention relates to metal ion transporter genes isolated from plants, in particular to a regulatory transporter ZmZIPs gene isolated from maize that is related to the absorption, transport or storage of zinc and iron. The present invention further relates to a regulatory transporter ZmZIPs gene in regulating plants The application of the ability to absorb, transport or store zinc or iron, promote the development or maturity of embryos and endosperm, and increase the zinc and iron content of grain crop seeds, belongs to the field of separation and application of plant metal ion regulation transporter genes. Background technique [0003] Zinc and iron are trace elements necessary for organisms, and they play a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/8243C12N15/8261C12N15/8267
Inventor 李素贞陈景堂李宏博赵永锋郭晋杰
Owner HEBEI AGRICULTURAL UNIV.