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Xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L and application thereof

A drought-inducing, transcription factor technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of unreported MYB transcription factor Xantho sorbifolium drought resistance application, it is difficult to screen out drought-related genes, etc., to improve drought resistance The effect of improving resistance, improving stress resistance and increasing biological yield

Active Publication Date: 2021-04-30
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Due to the specificity of species evolution and the limitation of related genes in specific species, it is difficult to screen out more genes related to drought resistance in some species
And there is no report on the application of MYB transcription factor in improving the drought resistance of X. sorbifolium

Method used

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  • Xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L and application thereof
  • Xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L and application thereof
  • Xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Cloning of XsMYB308L gene of X. sorbifolium

[0026] Construction and Amplification of cDNA Library of Xerifola sorbifolium

[0027] All X. sorbifolium seeds were collected from a single X. sorbifolium tree in Xinjiang Uygur Autonomous Region. The seeds were germinated in the dark at 25±3°C for about 2 weeks until they took root, and then the seedlings were transplanted into pots with mixed soil and grown in the greenhouse. Fresh tissues (roots, stems and leaves) were excised from 2-month-old seedlings to characterize the tissue-specific expression of XsMYB308L. For expression pattern analysis, 2-month-old X. sorbifolium seedlings were treated with PEG6000 (15%, w / v), NaCl (200 mM), ABA (0.1 mM), SA (2 mM) and MeJA (0.1 mM) . The leaves were harvested 144h after treatment, and then stored under liquid nitrogen for RNA extraction. The steps of extracting total RNA from X. sorbifolium are as follows:

[0028] S1. Grind the sorbifolium sorbifolium sample in a liquid n...

Embodiment 2

[0046] Construction of pMD 19T-XsMYB cloning vector

[0047] Recovery and purification of PCR product of XsMYB308L gene

[0048] S1. Use the TIANgel Maxi Purification Kit of Beijing Tiangen Biochemical Technology Co., Ltd. to recover and purify the PCR product of the XsMYB308L gene, dilute the recovered and purified target fragment to 50 ng / μL, and refer to the instructions of pMD 19T to construct the connection system described in Table 3 below :

[0049] Table 3 PCR product recovery and purification connection system

[0050]

[0051] After mixing gently with a pipette gun, connect at 16°C for 12h.

[0052] Sequence Analysis of XsMYB308L

[0053] S2. Analyze it using bioinformatics methods Retrieve the homologous sequence of XsMYB308L from the NCBI database. The XsMYB308L gene contains a 951 bp open reading frame and encodes 316 amino acids. According to the online prediction analysis of ExPASy-ProtParam tool, the relative molecular mass of XsMYB308L protein is 35651....

Embodiment 3

[0055] Construction of Recombinant Virus Vector pTRV2-XsMYB308L

[0056] (1) RT-PCR amplifies the target interference fragment

[0057] Specific primers were designed according to the 3' non-coding region of XsMYB308L gene mRNA sequence. To construct the VIGS vector, EcoRI and BamHI restriction site recognition sequences were added to the 5' ends of the forward primer and reverse primer, respectively. Carry out PCR amplification using cDNA as a template, and react according to the reaction system described in List 4:

[0058] Table 4 RT-PCR amplification target interfering fragment reaction system

[0059]

[0060] The reaction program was: 94°C 5min pre-denaturation, 1 cycle; 94°C 30s denaturation, 55-60°C 30s annealing, 72°C 1min extension, 35 cycles; 72°C 10min extension, 1 cycle; 4°C storage. Use 1% agarose gel electrophoresis to detect PCR products, recover and purify the ligated vector, transform the ligated product into Escherichia coli DH5a, screen transformants ...

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Abstract

The invention belongs to the technical field of transcription factors, and discloses a xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L and application thereof. The invention relates to a xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L, which has a nucleotide sequence as shown in SEQ ID NO.1. The invention also provides a protein coded by the xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L, and the protein has an amino acid sequence shown as SEQ ID NO.2. The invention further provides a pair of primers for amplifying the xanthoceras sorbifolia bunge drought induction transcription factor XsMYB308L. The pair of primers has a nucleotide sequence as shown in SEQ ID NO.3. The XsMYB308L gene provided by the invention has important significance in cultivating transgenic crops of drought-resistant varieties and improving the stress resistance of the crops.

Description

technical field [0001] The invention belongs to the technical field of transcription factors, and the invention relates to XsMYB308L, a drought-induced transcription factor of X. sorbifolium and its application. It specifically relates to XsMYB308L, a drought-inducing transcription factor of X. sorbifolium and its application in improving the drought resistance ability of plants. Background technique [0002] Xanthania sorbifolium, also known as Papaya, belongs to the deciduous trees or shrubs of Sapindaceae Xeriferia genus. It is an important economic tree species in China and has been identified as an important energy source. The layout and planning of the dominant production areas of X. sorbifolium in my country has greatly promoted the rapid development of X. sorbifolium industry. Because the production areas of X. sorbifolium are widely distributed in my country, the growth and development of X. And other factors, such as drought, frost damage, salinity and so on. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/84A01H5/00A01H6/82
CPCC07K14/415C12N15/8273
Inventor 孙颖李景滨赵尚阮成江孙浩张思怡张无梦刘佩鑫
Owner DALIAN NATIONALITIES UNIVERSITY
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