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Ethylene response factor gene of cotton transcription factor

A technology of response factors and genes, applied in the field of biotechnology applications, can solve different problems and achieve fast results

Active Publication Date: 2013-09-18
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report about the AP2 domain in this regard, but it is speculated that there may be a similar ERF domain in the AP2 domain, that is, there is a DNA recognition method different from the ERF domain and a possibly different DNA sequence ( Ohme-Takagi and Shinshi, 2000)

Method used

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  • Ethylene response factor gene of cotton transcription factor
  • Ethylene response factor gene of cotton transcription factor
  • Ethylene response factor gene of cotton transcription factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Acquisition of ethylene response factor gene (Gh ERF7):

[0038] The differential EST sequence obtained by mRNA differential display PCR technology was compared with Blastn in NCBI, and it was found that this EST had a high degree of correlation with a part of the 3'-end of a published cotton ethylene-responsive element-binding factor (GhERF1) similarity (93%). According to the characteristics of the ERF gene, using Vitex terrestriale 8891cDNA as a template, the forward degenerate primer Y3954 was designed in its very conserved functionally conserved domain, combined with the reverse primer Y2232R of differential EST RT-PCR for touch-down PCR amplification. The PCR reaction program is as follows: pre-denaturation at 94°C for 5 min; 30 cycles of touch-down program, denaturation at 94°C for 30 sec, annealing at 65°C for 30 sec, and the annealing temperature decreased by 1°C for each subsequent cycle until 56°C, extension at 72°C for 45 sec; ℃ extension 7min. A...

Embodiment 2

[0041] Example 2 The copy number, structure and bioinformatics analysis of ethylene response factor family gene (Gh ERF7)

[0042] Southern blot technology ("Molecular Cloning" second edition, Science Press) analyzes that there are 2 copies of the gene of the present invention in upland cotton ( figure 2 ), in order to analyze the genomic origin of the two genes GhERF7, we used two A-group diploid primitive species African cotton (G.herbaceum, A1) and Asian cotton (G.arboretum, A2), two D-group diploid The original species of Thurberi (G. thurberi, D 1 ) and Raymond cotton (G.raimondii, D 5 ), and the DNA of tetraploid upland cotton cultivars Jing 8891, Xiangzamian 2 and Zhongmian 12 (AADD) were used as templates to amplify their genome sequences using GhERF7 gene-specific primers (Y3172F / Y2232R). The results show( Figure 4), the 808bp GhERF7 gene was amplified in Jing 8891, Xiangzamian 2, and two A-group diploids in African cotton (G.herbaceum) and Asian cotton (G.arbore...

Embodiment 3

[0048] Example 3 Analysis of the expression characteristics of the ethylene response factor gene Gh ERF7

[0049] The primers used are full-length primers (Y3172F / Y2232R):

[0050] Y3172F:5'-ATGGCGATGGGATAGGGAGATT-3'

[0051] Y2232R:5'-CTCCCGTGATTTCTATTTGGTA-3'

[0052] Semi-quantitative RT-PCR detection was carried out, and the results showed that the gene was constitutively expressed in various tissues (roots, stems, leaves) and at different stages of fiber development (0 days, 2, 4, 6 days after flowering) ( figure 1 ), but the expression level is different, except in seedling cotyledons, there are also differences in different stages of fiber development, the expression level at the initial stage is low (0 day), and the expression level is high at the initial elongation stage of fiber (2, 4 , 6 days), but no expression was detected after 10 days. This result reflects that the gene of the present invention is closely related to the initial elongation of fibers, and play...

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Abstract

The invention belongs to the field of application of biotechnology and relates to an ethylene response factor gene of a cotton transcription factor. The ethylene response factor gene is Gh ERF7, a sequence of the ethylene response factor gene in upland cotton vitex 8891 and tetraploid upland cotton (G.hirsutum) TM-1 is shown in SEQ ID NO.1, the sequence of the ethylene response factor gene in a tetraploid sea island cotton (G.barbadense) H7124 is shown in SEQ ID NO.2, and a sequence of the ethylene response factor gene in Africa cotton (G.herbaceum) and Asian cotton (G.arboreum) genome is shown in SEQ ID NO.3. An ethylene response factor family gene (Gh ERF7) has no obvious response on adversity but is related to growth of cotton ovule and early fiber and yield of cotton, and the yield of cotton is obviously increased as the ethylene response factor gene exists in a genome. The ethylene response factor gene can regulate expression of some genes related to control on cotton yield factors, so that great contribution is done to increase of the yield of cotton, and the yield of cotton is hopefully greatly increased when the ethylene response factor gene is excessively expressed.

Description

technical field [0001] The invention belongs to the application field of biotechnology and relates to a cotton transcription factor ethylene response factor gene. technical background [0002] As an important economic crop worldwide, cotton produces natural fibers that are important raw materials for the textile industry. Due to the continuous acceleration of the development of the textile industry, more neat cotton fibers are required to meet various needs, coupled with the enhancement of people's health care awareness and the improvement of living standards, the demand for cotton fiber quality improvement is becoming more and more urgent. [0003] In the past 10 years, cotton breeding has changed from focusing on external quality to focusing on internal quality. At present, in addition to conventional breeding methods at home and abroad, genetic engineering is gradually used to genetically improve cotton fiber quality, and the development of this work first relies on the ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 张天真刘春晓陆可钰郭旺珍
Owner NANJING AGRICULTURAL UNIVERSITY
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