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Gene for encoding sweet potato ERF (ethylene responsive factor) transcription factor

A transcription factor and gene technology, applied in the field of genetic engineering, can solve the problem of not being able to comprehensively improve plant stress resistance

Inactive Publication Date: 2016-06-15
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is generally believed that the resistance of plants to stresses such as drought and high salinity is controlled by multiple genes. Therefore, although the introduction of a single functional gene by genetic engineering technology can increase a certain single resistance of a plant, it cannot comprehensively improve the plant's overall resistance. Resilience

Method used

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  • Gene for encoding sweet potato ERF (ethylene responsive factor) transcription factor
  • Gene for encoding sweet potato ERF (ethylene responsive factor) transcription factor
  • Gene for encoding sweet potato ERF (ethylene responsive factor) transcription factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The sequence of the ERF transcription factor IbRAP2.4 gene in sweet potato was obtained, as follows:

[0029] Total RNA was extracted from 100 mg of fresh sweet potato leaves using an OMEGARNA kit, and cDNA was synthesized using a reverse transcription kit (Bioteke).

[0030] Concrete reaction system is as follows:

[0031]

[0032] The reverse transcription reaction was carried out on the PCR instrument according to the following conditions: 1. 50° C., 45 min; 2. 70° C., 10 min; and then cooled on ice.

[0033] The extracted total RNA was sent to Beijing Genomic Institute (BGI), and the sweet potato transcriptome database was obtained through high-throughput sequencing, and the sweet potato Unigene database was obtained through Denovo splicing. After comparing Unigene23081 with the NR database, it was found that the gene may be involved in stress response . Use the online software of OFRFINDER (http: / / www.ncbi.nlm.nih.gov / projects / gorf / ) to predict the complete OR...

Embodiment 2

[0040] IbRAP2.4 protein sequence homology analysis, as follows:

[0041]The full-length cDNA was 907bp, the ORF was 684bp, encoding 227 amino acids of SEQIDNO.2, and the translated protein sequence was compared with the NCBI protein data (http: / / blast.ncbi.nlm.nih.gov / Blast.cgi) , obtained the plant species homologous gene similar to IbRAP2.4 protein sequence. On the basis of multiple comparison analysis, a phylogenetic tree of genes of each homologous plant species was established, see figure 1 . Including tobacco (Nicotianatomentosiformis), potato (Solanum tuberosum), tomato (Solanum lycopersicum), rice (Oryza sativa), grape (Vitis vinifera), soybean (Glycinemax) coffee (Coffea arabica), Arabidopsis thaliana (Arabidopsisthaliana). The phylogenetic tree was constructed using MEGA5.1 software, and the genetic relationship of IbRAP2.4 was closer to that of Arabidopsis thaliana and grapevine.

Embodiment 3

[0043] The expression of IbRAP2.4 after drought stress in different tissues of sweet potato (such as figure 2 , image 3 shown), as follows:

[0044] Different tissue samples, leaf (L), stem (S), root (FR), thickly pigmented root (PR) and storage root (SR) of Ningzi 1 were cut from three-month period for analysis of IbRAP2. Constitutive expression of 4. Cut the 25cm seedlings of Ningzi No. 1, and keep 3 fully expanded leaves, put them into 1 / 2 Hogland nutrient solution and culture them at 25°C for 15 days, treat them with 20% PEG8000 for 3h, 6h, 12h, 24h, 48h, take 3 leaves Repeat, and store in a -80°C freezer after quick freezing in liquid nitrogen. RNA was extracted and reversed into cDNA, the method was as described in Example 1. Fluorescence quantitative kit from TOYOBO Company was used. The reaction was carried out on a quantitative PCR instrument (Applied Biosystems steponeplus), and the expression level of the gene was detected according to a relative quantitative...

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Abstract

The invention discloses a gene for encoding sweet potato ERF (ethylene responsive factor) transcription factor, this gene being a nucleotide sequence shown as in SEQ ID No. 1. A protein encoded by the gene is an amino acid sequence shown as in SEQ ID No. 2. Complete cDNA for encoding ERF transcription factor is isolated from sweet potato and connected to a plant expression vector, a plant is converted by means of Agrobacterium infection to provide a transgenic plant, the transgenic plant has greater root development and higher biomass, drought resistance of the transgenic plant is improved, and this gene is also applicable to plant genetic improvement.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and specifically relates to a gene encoding sweet potato ERF transcription factor, a protein encoded by the gene, a recombinant vector containing the gene and an application of the gene. Background technique [0002] Many genes in plants are induced by adversity stress. According to different modes of action, they are mainly divided into two categories: regulatory genes and functional genes. Regulatory genes are mainly transcription factors that play a regulatory role in signal transduction and gene expression; functional genes mainly encode some enzymes that regulate osmotic potential, scavenge free radicals and active oxygen, etc. It is generally believed that the resistance of plants to stresses such as drought and high salinity is controlled by multiple genes. Therefore, although the introduction of a single functional gene by genetic engineering technology can increase a certain single res...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/84C12N1/21A01H5/00
CPCC07K14/415C12N15/8261C12N15/8273
Inventor 边小峰马佩勇贾赵东谢一芝郭小丁
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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