Zea mays zinc iron-regulated transporter ZmZIPs genes and applications thereof

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

Active Publication Date: 2014-12-17
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 them, the ZIP (Zinc-regulated transporters, Iron-regulated transporter-like proteins, ZIP) gene family can absorb divalent metal ions such as zinc and iron, Transportation and storage play 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 has not yet been fully understood. There are few research reports on ZIP family genes

Method used

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  • Zea mays zinc iron-regulated transporter ZmZIPs genes and applications thereof
  • Zea mays zinc iron-regulated transporter ZmZIPs genes and applications thereof
  • Zea mays zinc iron-regulated transporter ZmZIPs genes and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Cloning of ZmZIPs gene of maize zinc and iron regulated transporter

[0077] 1. Processing of plant material

[0078] First soak the vermiculite with Hoagland nutrient solution, sow the seeds of the corn inbred line X178 on the seedling tray, cover the top with a layer of dry vermiculite, and cultivate them in a greenhouse (16h light / 8h dark, 26°C). When it grows to 2 leaves and one heart, it is moved into the standard Hoagland nutrient solution and grows for 6 days to 3 leaves and one heart (the nutrient solution is changed every 3 days). After treatment under high zinc and iron conditions for 0, 6, 12, 24, 48, and 96 hours, the shoots and roots of the seedlings were harvested, respectively, and stored at -80°C for total RNA extraction after quick freezing in liquid nitrogen.

[0079] 2. Extraction of corn total RNA

[0080] The total RNA of maize was extracted by Trizol method.

[0081] 3. Synthesis of cDNA

[0082] (1) To remove DNA, prepare the reactio...

Embodiment 2

[0100] Example 2 Expression patterns of ZmZIPs in seedlings, embryos and endosperms

[0101] The reverse transcribed cDNA in Step 3 of Example 1 was diluted 10 times as a template for the PCR reaction, and the PCR reaction system was as follows:

[0102] cDNA 2.0 μL, ExTaq 0.1 μL, 2×GCI buffer 10.0 μL, 10 mM dNTP mix 0.8 μL, upstream primer RTZmZIP1F / RTZmZIP2F / RTZmZIP3F / RTZmZIP7F / RTZmZIP8F (10 μM / μL) 1.0 μL, downstream primer RTZmZIP1R / RTZmRT7R / RTZmZIP3 / RTZmZIP8R (10 μM / μL) 1.0 μL, ddH 2 O5.1 μL, 20.0 μL in total;

[0103] RTZmZIP1F 5'-CCTCTCTGCGTTGGTTGCTCT-3'

[0104] RTZmZIP1R 5'-TTGATGGTTGTTTTCTGGTCGT-3'

[0105] RTZmZIP2F 5'-CCACAAATGGCACGAGGTCT-3'

[0106] RTZmZIP2R 5'-CGAAGACGGAGTGGAAGCAAA-3'

[0107] RTZmZIP3F 5'-GCCTCTTGTTGGTGCCCTTA-3'

[0108] RTZmZIP3R 5'-TCAACAATGAACGCTGTAGTGCT-3'

[0109] RTZmZIP7F 5'-ACTAGGTGGGTGCATTGCTCAG-3'

[0110] RTZmZIP7R 5'-TGCCAGCAGATACCGAGTCAA-3'

[0111] RTZmZIP8F 5'-CGTGTCATCGCTCAGGTTCTTG-3'

[0112] RTZmZIP8R 5'-CCCTCGAACATTTG...

Embodiment 3

[0119] Example 3 Bioinformatics analysis of ZmZIPs

[0120] ZmZIPs consist of 367-483 amino acids and contain 6-9 transmembrane domains. There is a histidine-rich variable region between the third and fourth transmembrane domains, which may be related to the binding and transport of metal ions. Phylogenetic tree analysis showed that ZmZIP1 was closely related to AtIAR1 and OsIAR1. In addition, ZmZIP3 and ZmZIP4 form a gene cluster with OsZIP3 and OsZIP4, ZmZIP2 is adjacent to OsZIP2, and zinc transporters OsZIP1, AtZIP2 and AtZIP11 are on a branch, ZmZIP5 is on a branch with ZmZIP7, ZmZIP8 is evolutionary with OsZIP8, ZmZIP6 and OsZIP6 More recently, these results indicated that the five ZmZIPs isolated in the present invention may be zinc-iron transporters.

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Abstract

The invention discloses a Zea mays zinc iron-regulated transporter ZmZIPs genes and applications thereof. Five Zea mays zinc iron-regulated transporter ZmZIP4 and ZmZIP6 genes are separated from Zea mays, and the cDNA sequences of the genes are selected from nucleotide sequences represented by SEQ ID No.1, SEQ ID No.3, SEQ ID No.5, SEQ ID No.7 and SEQ ID No.9 respectively. Subcellular localization shows that a zinc iron-regulated transporter encoded by the genes is localized in the plasma membrane and endoplasmic reticulum of a cell. Yeast complementation experiments show that each of the five ZmZIPs has a zinc and iron transportation function. The separated ZmZIPs genes have important application prospects in the regulation of zinc and iron absorption, transportation and storage of plants, promotion of the embryo and endosperm growth and increase of the content of zinc and iron in grains of food crops.

Description

technical field [0001] The present invention relates to metal ion transporter genes isolated from plants, in particular to the regulatory transporter ZmZIPs genes isolated from corn and related to the absorption, transport or storage of zinc and iron. The present invention further relates to the regulation of transporter ZmZIPs genes in regulating plant The ability to absorb, transport or store zinc or iron, promote the development or maturation of embryos and endosperms, and increase the zinc and iron content of grain crop seeds belongs to the field of isolation and application of plant metal ion-regulated transporter genes. Background technique [0002] Zinc and iron are essential trace elements for organisms and play an important role in the growth and development of plants (Wintz H, Fox T, Wu YY, et al. Expression profiles of Arabidopsis thaliana in mineral deficiencies reveal novel transporters involved in metal homeostasis . The Journal of biological chemistry 2003, 27...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00
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.
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