A kind of iberf4 gene encoding sweet potato erf transcription factor and its application

A technology of transcription factor and sweet potato, applied in the field of genetic engineering, can solve problems such as the inability to comprehensively improve plant stress resistance

Inactive Publication Date: 2017-07-25
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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  • A kind of iberf4 gene encoding sweet potato erf transcription factor and its application
  • A kind of iberf4 gene encoding sweet potato erf transcription factor and its application
  • A kind of iberf4 gene encoding sweet potato erf transcription factor and its application

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Experimental program
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Effect test

Embodiment 1

[0027] The sequence of the ERF transcription factor IbERF4 gene in sweet potato was obtained, as follows:

[0028] Total RNA was extracted from 100 mg of fresh sweet potato leaves using the OMEGA RNA kit, and cDNA was synthesized using a reverse transcription kit (Bioteke).

[0029] Concrete reaction system is as follows:

[0030]

[0031] 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.

[0032] 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 De novo splicing. After comparing Unigene23081 with the NR database, it was found that the gene might Participate in adversity response. Use the OFR FINDER (http: / / www.ncbi.nlm.nih.gov / projects / gorf / ) online software to predict the complete...

Embodiment 2

[0039] IbERF4 protein sequence homology analysis, as follows:

[0040]The full-length cDNA was 907bp, the ORF was 684bp, encoding 227 amino acids of SEQ ID NO.2, and the translated protein sequence was compared with the NCBI protein data (http: / / blast.ncbi.nlm.nih.gov / Blast. cgi), the plant species homologous gene similar to the IbERF4 protein sequence was obtained. On the basis of multiple comparison analysis, a phylogenetic tree of genes of each homologous plant species was established, see figure 1 . Including tobacco (Nicotiana tabacum), potato (Solanum tuberosum), tomato (Solanum lycopersicum), pepper (Capsicum annuum), spirulina (Genlisea aurea), coffee (Coffea arabica), rubber tree (Hevea brasiliensis), chickpea (Cicerarietinum ), Medicago truncatula, Eucalyptus grandis, Arabidopsis thaliana. The phylogenetic tree was constructed using MEGA 5.1 software, and the genetic relationship of IbERF4 was the closest to Solanaceae crops.

Embodiment 3

[0042] The expression of IbERF4 in sweet potato under salt, drought stress and different leaves (such as figure 2 , image 3 shown), as follows:

[0043] 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, and treat them with 20% PEG6000 and 150mM NaCl for 3h, 6h, 12h, 24h, 48h respectively , three replicates were taken, and stored in a -80°C refrigerator 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 stepone plus), and the expression level of the gene was detected according to the relative quantitative method. The reaction procedure was carried out according to the operation manual provided by TOYOBO. The sweet potato Tubulin gene was used as an internal...

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Abstract

The invention discloses an IbERF4 gene for coding a sweet potato ERF transcription factor and an application thereof. A nucleotide sequence of the IbERF4 gene is expressed by SEQ (Sequence) ID (Identity) NO. (Number) 1 and the IbERF4 gene is responsible for regulating the plant stress resistance and promoting the plant senescence. An amino acid sequence of protein coded through the IbERF4 gene is expressed by SEQ ID NO. 2. According to the IbERF4 gene for coding the sweet potato ERF transcription factor and the application thereof, the complete cDNA (complementary Deoxyribose Nucleic Acid) for coding the ERF transcription factor is separated from sweet potatoes and connected onto a plant expression vector, plants are transformed by an agrobacterium infection method, transgenic plants are obtained, the stress resistance analysis is performed on transgenic plants, and the ERF transcription factor can respond to adversity signals, the saline-alkaline resistance of the plants can be negatively regulated, and the pro-senescence of the transgenic plants is achieved due to results. The IbERF4 gene can be applied to plant genetic improvement.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and specifically relates to an IbERF4 gene encoding a sweet potato ERF transcription factor, a protein encoded by the gene, a recombinant vector containing the gene, and an anti-stress 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 increa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/84C12N1/21A01H5/00
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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