Myrothamnus flabellifolia gene MfWRKY7 and application thereof

A kind of Mirosakia, gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem that the gene and its molecular mechanism are not clear, and the anti-stress WRKY transcription factor has not yet been found in Miramu.

Inactive Publication Date: 2019-07-16
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the past, researchers have mainly studied the drought resistance mechanism of the plant through morphological, physiological, biochemical and other aspects, but the genes and molecular mechanisms related to stress resistance are still unclear, and the stress-resistant WRKY transcription factor in Miroma has not yet been found. Related reports on its application

Method used

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  • Myrothamnus flabellifolia gene MfWRKY7 and application thereof
  • Myrothamnus flabellifolia gene MfWRKY7 and application thereof
  • Myrothamnus flabellifolia gene MfWRKY7 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Cloning of Miluo MfWRKY7 gene

[0029] 1. Take the leaves of Miluo wood, quick-frozen them in liquid nitrogen, and store them in the refrigerator at -80℃ for extraction of total RNA; the total RNA was extracted using the Plant Total RNA Isolation Kit purchased by Chengdu Lanbo Biological Company; Miluo cDNA For the synthesis of Reverse Transcriptase M-MLV (RNaseH-) from Dalian Bao Biotechnology Company, the first-strand synthesis was performed according to its product instructions.

[0030] The first strand of cDNA synthesized by the above kit was used as the amplification template to design F: 5'-TCCCCCGGGATGGCGGTTGAGCTTATGTT-3' (SEQ ID NO. 3) and R: 5'-GGACTAGTCTAAGAAGATTCGAGGACCA-3' (SEQ ID NO. 4 ) Is a primer. For subsequent enzyme digestion and recombination and ligation, two enzyme sites, SmaI and SpeI, are added when designing primers. PCR is used for cDNA amplification. The amplification system is shown in Table 1, and the amplification conditions are: 95 ...

Embodiment 2

[0034] Example 2 Sequence homology and homology analysis of MfWRKY7

[0035] According to the results of sequence sequencing, the sequence alignment was performed in the NCBI database, and it was found that the cloned gene sequence had the closest homology relationship with the WRKY family transcription factor. The transcription factor was compared with the protein sequence of other WRKY transcription factor family members and found that it The zinc finger structure type is C2H2. According to the structural characteristics of the DNA binding domain, the MfWRKY7 protein belongs to the second type of WRKY transcription factor family. The amino acid sequence of the aligned species with higher homology is further used to construct a phylogenetic tree (such as figure 1 ), it is found that MfWRKY7 has a long relationship with other species, and its domain is relatively conservative.

Embodiment 3

[0036] Example 3: Construction of Plant Expression Vector of Miluo Gene MfWRKY7

[0037] The plasmid was extracted from the above-sequenced bacteria liquid, the cloning vector plasmid containing the MfWRKY7 gene was digested with SmaI+SpeI, and then a DNA fragment of about 987bp was recovered using a DNA recovery kit, and this fragment was digested with the corresponding digested pGSA1403 expression vector After connecting, the obtained vector was named 35S:MfWRKY7-pGSA1403.

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Abstract

The invention discloses myrothamnus flabellifolia gene MfWRKY7 and an application thereof. The nucleotide sequence of the gene is shown as in SEQID NO.1. The amino acid sequence of gene coded proteinis shown as SEQID NO.2. Myrothamnus flabellifolia drought resisting WRKY transcription factors are researched, and the stress resistance of plants is improved, so that the stress resistance of the plants can be improved.

Description

Technical field [0001] The invention belongs to the technical field of plant genetic engineering, and specifically relates to a milo gene MfWRKY7 and its application. Background technique [0002] Drought and salinization are one of the main non-biological disasters facing plants today, and their main mechanism is to make the internal and external osmotic pressure of the plant inconsistent and cause dehydration. The population growth and the rapid development of urban construction have not only led to increasingly fierce competition for civil, industrial, and agricultural water use, but also increased pollution, destruction of the ecosystem, shortage of water resources, and increasing soil salinization. Therefore, discovering stress resistance genes and cultivating new plant varieties with strong stress resistance is important for accelerating the construction of a resource-saving and environment-friendly society, vigorously developing a green economy, enhancing sustainable devel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 黄卓刘玲蒋才忠陈佳朱培蕾邱嘉睿向香盈易鑫王嘉彤李巧蔡仕珍李西马均孙凌霞
Owner SICHUAN AGRI UNIV
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