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Tamarix hispida PHD3 transcription factor encoding gene and application thereof

A technology of tamarisk bristles and transcription factors, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problem of not being able to use PHD transcription factor coding genes to cultivate excellent salt-tolerant plants, and achieve the effect of obvious salt tolerance

Active Publication Date: 2020-05-08
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem of being unable to utilize the PHD transcription factor coding gene of Tamarix brix to cultivate excellent salt-tolerant plants, the present invention provides the PHD3 transcription factor coding gene of Tamarix brix and its application

Method used

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  • Tamarix hispida PHD3 transcription factor encoding gene and application thereof
  • Tamarix hispida PHD3 transcription factor encoding gene and application thereof
  • Tamarix hispida PHD3 transcription factor encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Cloning of Tamarix bristles ThPHD3 gene

[0038] The seeds of Tamarix bristles were sown in the artificial soil prepared with peat soil and sand at a mass ratio of 2:1, placed at a relative humidity of 65-75% and a light intensity of 400 μmol m -2 ·s -1 , Grow in a greenhouse with an average temperature of 22±2°C. After 2 months of growth, use 0.3mol·L -1 NaHCO 3 The roots were irrigated with the solution for stress treatment, and the leaves and roots were taken at 0h, 12h, 24h and 48h respectively and put into liquid nitrogen for quick freezing for RNA extraction.

[0039] The CTAB method was used to extract the RNA of Tamarix seedlings in each stress treatment period, and the extracted RNA samples were sent to Shenzhen Huada Gene Technology Co., Ltd. for the construction and sequencing of the transcriptome library. The RNA sequencing results obtained in each processing period were compared and spliced, and the results after sequencing comparison and sp...

Embodiment 2

[0043]Example 2: Using bioinformatics software and online network resources to perform sequence analysis on the gene encoding the DHP3 transcription factor of Tamarix brix

[0044] As shown in SEQ ID No: 1, the coding region of the gene coding for the DHP3 transcription factor of Tamarix biribilis cloned in Example 1 is 759 bp long, encoding 252 amino acids shown in SEQ ID NO: 2. Through BLASTx similarity analysis, it was found that there was a typical PHD domain (595-744bp) at the C-terminal of the gene. Combining ExPASy-PROSITE analysis and figure 1 It was shown that the C-terminus of the ThPHD3 protein sequence contains a conserved PHD domain, which is a typical ZF-PHD (Zinc finger PHD-type profile) protein.

[0045] For the obtained ThPHD3 gene, use ProtParam( http: / / au.expasy.org / tools / protparam.html ) software to calculate and deduce protein molecular weight and theoretical isoelectric point. Select soybean PHD family protein and Tamarix bristles ThPHD3 protein fo...

Embodiment 3

[0047] Example 3: Analysis of the expression pattern of ThPHD3 gene under different abiotic stresses and hormone treatments

[0048] ThPHD3 transcription factor plays an important role in the regulation of plant stress resistance. A single Tamarix ThPHD3 gene with complete open reading frame was cloned and its sequence was analyzed. In order to study the function of Tamarix ThPHD3 gene, real-time fluorescence quantitative RT-PCR was further used to analyze its different abiotic stresses: high salt-400mmol NaCl, drought-20% PEG and hormone stress: ABA-100μmol, GA 3 Expression patterns under -50 μmol treatment condition. Using the same amount of control (0h), the cDNA of the roots and leaf tissues of Tamarix bristles at 6h, 12h, 24h, 48h and 72h of each treatment as templates, real-time quantitative RT-PCR amplification was carried out to detect the ThPHD3 gene of Tamarix bristles under different stresses. The expression situation under treatment, and β-actin, α-tubulin and β-...

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Abstract

The invention relates to a tamarix hispida PHD3 transcription factor encoding gene and an application thereof, and belongs to the technical field of plant genetic engineering breeding. In order to solve the problem that a PHD transcription factor encoding gene of tamarix hispida cannot be used to cultivate excellent salt-tolerant plants, the invention provides the tamarix hispida PHD3 transcription factor encoding gene. The cDNA sequence of the tamarix hispida ThPHD3 gene is shown in SEQ ID No:1, and the amino acid sequence encoded by the cDNA sequence is shown in SEQ ID No:2. Overexpression and suppression expression vectors of the tamarix hispida ThPHD3 gene are constructed, transgenic tamarix hispida is obtained by transient infection, and experiments confirm that overexpression strainshave obvious salt tolerance. The homologous and heterologous overexpression results of the ThPHD3 gene indicate that the gene can obviously improve the salt tolerance of transgenic plants, and can beused to improve plant salt tolerance or to breed salt-tolerant transgenic plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering and breeding, and in particular relates to a gene encoding tamarix bristles PHD3 transcription factor and an application thereof. Background technique [0002] During the growth process, plants will be subjected to abiotic stresses such as drought, high salinity, osmosis, high temperature, low temperature and hormone stimulation. Transcription factors are a class of DNA-binding proteins that can specifically interact with the cis-acting elements of eukaryotic gene promoters, and activate or repress transcription through the interaction between them or with other related proteins. Under stress stimulation, some transcription factors are overexpressed, and then the signal is transmitted and amplified, and the expression of corresponding downstream functional genes is regulated, thereby improving the stress resistance of plants. Since transcription factors can regulate the expressi...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N1/21A01H5/00A01H6/00A01H6/20
CPCC07K14/415C12N15/8273C12N15/8218Y02A40/146
Inventor 高彩球王培龙王媛媛雷晓锦刘中原
Owner NORTHEAST FORESTRY UNIVERSITY
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