Corn drought-resistant transcription factor gene Zmhdz9 and application thereof

A technology of transcription factor and corn, applied in application, genetic engineering, plant gene improvement, etc., can solve the problem that HD-Zip gene transcription factor has not been reported, and achieve the effect of improving drought tolerance

Inactive Publication Date: 2020-12-01
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

HD-Zip genes have been cloned in plants such as Arabidopsis thaliana, soybean, wheat, rice, etc., and HD-Zip genes in corn have also been reported as transcription factors, such as Zmhdz12 etc. (publication number CN106978424A), which have drought resistance activity, but There are still many HD-Zip gene transcription factors that have not been reported

Method used

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  • Corn drought-resistant transcription factor gene Zmhdz9 and application thereof
  • Corn drought-resistant transcription factor gene Zmhdz9 and application thereof
  • Corn drought-resistant transcription factor gene Zmhdz9 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Cloning of Example 1 Zmhdz9 Gene

[0030] The maize inbred line H8186 was selected as the experimental material, and the three-leaf stage maize leaf tissue was extracted and reverse transcribed for future use.

[0031] The biological information of Zmhdz9 was retrieved from the corn database, and the transcript ID was GRMZM2G041462_P01. Design specific amplification primers using the gene sequence as a template, combine the multiple cloning restriction sites of the cloning vector at the same time, select the BglII and BstEII restriction sites to introduce at both ends of the Zmhdz9 sequence, and the specific amplification primer sequences are as follows:

[0032] Zmhdz9-F: 5'-CACTTTATTGTGAAGATAGTGGAAA-3'

[0033] Zmhdz9-R: 5'-CCATCTAATTCAACAAGAATTGGGACAAC-3'

[0034] Using cDNA as a template, Zmhdz9-F and Zmhdz9-R as primers, the high-fidelity enzyme Primer STAR Max Premix (2×) produced by TAKARA Company was used to perform the amplification reaction. The amplificatio...

Embodiment 2

[0040] Example 2 Bioinformatics analysis of Zmhdz9 gene

[0041] After the sequencing result was compared with the sequence in the database by MEGA6.0 software, it was consistent with the nucleotide sequence shown in SEQ ID NO.1, indicating that the Zmhdz9 gene was successfully cloned. After sequence analysis, it was found that the coding region of Zmhdz9 gene was 1408bp in length, encoding a total of 239 amino acids, with a PI of 5.39 and a MW of 26.24kD. Analysis of its protein sequence revealed that the protein encoded by the Zmhdz9 gene contained two prominent conserved domains, the domain and the domain connected to it, and the domain contained multiple repeated leucine residues. It was predicted that the Zmhdz9 gene Belongs to the HD-Zip I family of transcription factors. The members of the maize HD-ZipI family whose functions have been reported and verified were selected, and the gene members with high homology were screened, and the protein sequences of these gene mem...

Embodiment 3

[0042] Example 3 Construction of Zmhdz9 gene overexpression vector

[0043] According to the multiple cloning site of pCAMBIA3301 and the sequence characteristics of the target gene, BglII and BstEII restriction sites were added to both ends of the target gene Zmhdz9, and the positive plasmid obtained by cloning was used as a template for amplification to obtain PCR amplification fragments. The amplification primer sequences are as follows:

[0044] Zmhdz9-F: 5'-actcttgaccatggtagatctCACTTTATTGTGAAGATAGTGGAAA-3'

[0045] Zmhdz9-R: 5'-ggggaaattcgagctggtcaccCCATCTAATTCAACAAGAATTGGGACAAC-3'

[0046] Transform the recombinant plasmid into Escherichia coli competent cells, shake the bacteria for sequencing and save, extract the plasmid, and use BglII and BstEII to double-enzyme digest the plasmid to obtain a small target fragment. The connection system is as follows:

[0047] Drug Name Dosage (μl) Target gene fragment 6 Pcambia3301 large fragment 2 10...

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Abstract

The invention provides a corn drought-resistant transcription factor gene Zmhdz9. The nucleotide sequence of the corn drought-resistant transcription factor gene Zmhdz9 is shown as a sequence table SEQ ID NO. 1. The invention provides the corn drought-resistant transcription factor gene Zmhdz9 and application thereof. The gene is a novel plant drought resistance related gene, the discovery of thegene enriches a genetic resource library using a corn HD-Zip gene as a transcription factor at present, and the gene has important practical significance for further improving the drought resistance of corn, cultivating novel drought-resistant varieties and solving the drought problem.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a maize drought-resistant transcription factor gene Zmhdz9 and its application. Background technique [0002] Corn (Zeamays L.) is the most important and special crop among the three major food crops in my country, and occupies an important position in the national economy of our country. At present, as people pay more attention to the production and development of corn, the demand for it is increasing day by day. However, according to the statistics of relevant departments, my country's total corn production has shown a downward trend in recent years, and the import volume has been increasing year by year. The main reason is that there are few varieties that are really promoted and it is difficult to form large-scale planting, and the poor quality of corn leads to low product benefits. Lack of market competitiveness. Secondly, a variety of abiotic stress factors have also caused gr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46A01H6/20
CPCC07K14/415C12N15/8273
Inventor 关淑艳焦鹏刘思言
Owner JILIN AGRICULTURAL UNIV
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