Application method of stress resistance-related gene ZmHDZIV13 in regulation of plant stress resistance

A technology of plant stress resistance and related proteins, applied in the field of genes that enhance plant drought resistance and stress resistance, can solve the problems of unreported physiological functions and few HD-ZipIV gene families in maize

Inactive Publication Date: 2016-10-12
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still few studies on the maize HD-Zip IV gene family, mainly on its expression patterns in different plant tissues and in the develo...

Method used

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  • Application method of stress resistance-related gene ZmHDZIV13 in regulation of plant stress resistance
  • Application method of stress resistance-related gene ZmHDZIV13 in regulation of plant stress resistance
  • Application method of stress resistance-related gene ZmHDZIV13 in regulation of plant stress resistance

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1; The acquisition of SEQ ID NO: 2 protein and its coding gene

[0043] Tobacco (Nicotiana tobaccocum), ecotype T12, provided by Gansu Academy of Agricultural Sciences.

[0044] Strains: Escherichia coli (Eschrichiacoli) DH5a, Agrobacterium tumefaciens used for genetic transformation of tobacco is LBA4404, both strains are preserved by Gansu Provincial Key Laboratory of Crop Science in Arid Habitat.

[0045] Plasmid: T / A cloning vector pUCm-T was purchased from Sangon Bioengineering (Shanghai) Co., Ltd., and the plant expression vector pCAMBIA3300-35S-PROIIMCS-bar was transformed by Gansu Key Laboratory of Crop Science in Arid Habitats. Kanna resistance was used for T / A cloning.

[0046] 1. The cloning steps of the nucleotide sequence SEQ ID NO: 1 are as follows:

[0047] (1) Extraction and purification of corn total RNA

[0048] Extraction of corn RNA (Trizol method):

[0049] Preparations before the experiment: DEPC water configuration: add 1ml of DEPC ...

Embodiment 2

[0074] Example 2; Functional verification of ZmHDZIV13 and its coding gene

[0075] One, the construction of ZmHDZIV13 gene expression vector

[0076] (1) The target fragment is connected to the pUCm-T vector

[0077] Prepare the following solution according to the instructions of pUCm-T vector (Sangon Biotechnology): 10×Ligation Buffer 1.0 μl, 50% PEG 1.0 μl, pUCm-Tvector 1.0 μl, purified PCR product 4.0 μl, T4 DNA Ligase 1.0 μl, dd H 2 O 2 μl; ligation at 16°C for 6 hours.

[0078] (2) Preparation of Escherichia coli competent cells (prepared in a 1.5ml centrifuge tube)

[0079] Transfer the bacterial solution into a pre-cooled 1.5ml centrifuge tube, centrifuge at 3500rpm at 4°C for 10min, and remove the supernatant. Add 300 μl of ice-cold 0.1 mol / L CaCl to the centrifuge tube 2 Resuspend the bacteria in the solution, and keep in ice bath for 30min. Centrifuge at 3500 rpm for 10 min at 4°C. Remove the supernatant and suspend the cells in 60 μl of ice-cold 0.1 mol / L CaC...

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Abstract

The invention relates to a gene for enhancing plant drought resistance, and provides a related protein for enhancing plant stress resistance. The gene has a is named as ZmHDZIV13, and derived from maize (Zea maysL.) inbred line B73, and has an amino acid sequence shown in a sequence table SEQIDNO:2. A coding gene sequence of the protein is shown in SEQIDNO:1. The invention also relates to an application method of the encoding protein of the related protein for enhancing plant stress resistance. The method is as below: transforming an expression vector carrying the ZmHDZIV13 gene into plant cells or tissues through conventional biological methods including Ti plasmid, Ri plasmid, a plant virus vector, direct DNA conversion, micro injection, conductance and Agrobacterium mediation, and culturing the transformed tissue into plants to obtain plants with improved drought resistance. The ZmHDZIV13 gene can be constructed into the existing plant expression vector by the method used in the prior art, and the encoding gene of the related protein for enhancing plant stress resistance can be converted into other plants by conventional methods to enhance the drought resistance of plants.

Description

technical field [0001] The invention relates to a gene for enhancing the drought resistance of plants; the invention also relates to an application method of the gene in improving the resistance of plants to drought stress. Background technique [0002] Homeodomain-leucine zipper (HD-Zip) is a kind of transcription factor unique to plants, which widely exists in a variety of plants. It not only plays an important role in the growth, development and morphogenesis of higher plants, but also regulates plant Response processes to adversities such as biotic and abiotic stresses. The AtHDG11 gene in Arabidopsis thaliana encodes a transcription factor belonging to the HD-ZipIV family. The expression of this gene can promote root elongation and stomatal closure to improve plant drought tolerance. Previous studies have shown that Arabidopsis transgenic AtHDG11 gene Mustard, tobacco and turfgrass all showed strong drought tolerance. At present, 17 maize HD-Zip IV transcription facto...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00
CPCC07K14/415C12N15/8273
Inventor 彭云玲闫慧萍赵小强武博洋方鹏
Owner GANSU AGRI UNIV
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