Application method of stress tolerance related gene ZmHDZIV14 in regulation and control of plant stress resistance
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A technology for plant stress resistance and related proteins, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, and other methods of inserting foreign genetic materials. and other problems, to achieve the effect of improving stress resistance
Inactive Publication Date: 2016-04-27
GANSU AGRI UNIV
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However, there are still few studies on the maize HD-ZipIV gene family, mainly on its expression patterns in different plant tissues and in the development
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Embodiment 1
[0040] Embodiment 1, the acquisition of ZmHDZIV14 protein and its coding gene
[0041] Seeds of wild-type Arabidopsis thaliana (Arabidopsisthaliana, Col) were preserved by Gansu Key Laboratory of Maize in Arid Habitat Crop Science.
[0042] Bacterial strain: Escherichia coli (Eschrichiacoli) DH5a, Agrobacterium tumefaciens (Agrobacterium tumefaciens) used for genetic transformation of Arabidopsis thaliana is LBA4404, both strains are preserved by Gansu Provincial Key Laboratory of Crop Science in Arid Habitat.
[0043] Plasmid: The T / A cloning vector pUCm-T was purchased from Sangon Bioengineering (Shanghai) Co., Ltd., the plant expression vector pCAMBIA3300-35S-PROIIMCS-bar was transformed by the Gansu Key Laboratory of Crop Science in Arid Habitats, and Kanna resistance was used for T / A cloning.
[0044] Embodiment 1; The acquisition of SEQIDNO:2 protein and its coding gene
[0074] Example 2; Functional verification of ZmHDZIV14 and its coding gene
[0075] One, the construction of ZmHDZIV14 gene expression vector
[0076] (1) The target fragment is connected to the pUCm-T vector
[0077] Configure the following solution according to the instructions of pUCm-Tvector (Sangon Biology): 10×LigationBuffer 1.0 μl, 50% PEG 1.0 μl, pUCm-Tvector 1.0 μl, purified PCR product 4.0 μl, T4DNALigase 1.0 μl, ddH 2 O2 μl. Ligation at 16°C for 6 hours.
[0078] (2) Preparation of Escherichia coli competent cells (prepared in 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 CaCl2 solution to the centrifuge tube to resuspend the bacteria, and ice-bath for 30 minutes. Centrifuge at 3500 rpm for 10 min at 4°C. Remove the supernatant, suspend the bacteria in 60 μl of ice-cold 0.1 mol / L CaCl2 solution, add 10 ...
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Abstract
The invention relates to a plant stress resistance enhancement related protein named as ZmHDZIV14. The protein is derived from a maize (Zea mays L.) inbred line B73, the amino acid sequence of the protein is represented as a sequence table SEQ ID NO:2, and an encoding gene sequence of the protein is a nucleotide sequence represented as SEQ ID NO:1. The invention further provides an application method of an encoding gene of the plant stress resistance enhancement related protein. According to the application method of the encoding gene of the plant stress resistance enhancement related protein, an expression vector carrying the ZmHDZIV14 gene is used for converting plant cells or tissue by the aid of conventional biological methods including application of Ti plasmids, Ri plasmids and plant virus vectors, direct DNA transformation, microinjection, electric conduction, agrobacterium tumefaciens mediated transformation and the like, a plant is cultured from the converted plant through tissue culture, and the plant with improved drought resistance and stress resistance is obtained. The expression vector carrying the ZmHDZIV14 gene is used for converting the plant cells or tissue by the aid of the conventional biological methods including application of Ti plasmids, Ri plasmids and plant virus vectors, direct DNA transformation, microinjection, electric conduction, agrobacterium tumefaciens mediated transformation and the like, the plant is cultured from the converted plant through tissue culture, and the plant with the improved stress resistance is obtained.
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-ZipIV transcription factor...
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