Transcription factor ERF related to soybean stress, coding gene thereof and application of coding gene

A technology of transcription factors and coding genes, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of soybean yield reduction, economic loss, biotic and abiotic stress, etc., and achieve the effect of improving stress resistance

Inactive Publication Date: 2012-02-01
JILIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, biotic and abiotic stresses, including diseases, salt damage, low temperature and drought, have extremely adverse effects on the growth and development of soybeans, resulting in reduced soybean yields and serious economic losses.

Method used

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  • Transcription factor ERF related to soybean stress, coding gene thereof and application of coding gene
  • Transcription factor ERF related to soybean stress, coding gene thereof and application of coding gene
  • Transcription factor ERF related to soybean stress, coding gene thereof and application of coding gene

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Embodiment 1

[0024] Embodiment 1: Cloning and sequence analysis of GmERF6 gene

[0025] Soybean RNA extraction and cDNA synthesis: RNAplant plus Reagent (purchased from TIANGEN) was used to extract total RNA from leaves of soybean Jilin 32, and M-MLV reverse transcriptase (RNase H-) (purchased from TaKaRa) was used to perform reverse transcription to synthesize cDNA.

[0026] Design and synthesis of primers: The sequence obtained from immature embryo expression profile sequencing (completed by Huada Genomics) of soybean variety Jilin 32 at three stages (20, 30 and 50 days) was input into NCBI for Blast comparison to obtain a Unknown cDNA sequence with high homology of ERF transcription factor protein sequence. According to the obtained unknown cDNA sequence, its nucleotide sequence is as described in Sequence NO.1, design synthetic primers, F: 5'-CTTCCTACTCCTCCCTTTCAC-3', R: 5'-CGTAGTAGTGTTCCCAGATGC-3'. Using the above cDNA as a template, carry out PCR amplification according to the follo...

Embodiment 2

[0028] Embodiment 2: the expression of GmERF6 gene in prokaryotic

[0029] Synthetic primers were designed according to the GmERF6 gene sequence, and Sac I and Hind III restriction sites were introduced into the upstream and downstream primers respectively, F: 5′-GAGCTCATGGCCCCAAGAGACAGC-3′; R: 5′-TTCGAATCAGGCAACCTCCGGGAG-3′. The pMD18-T-GmERF6 plasmid was used as a template for PCR amplification. After the amplified product was purified by an agarose gel DNA purification and recovery kit (purchased from TIANGEN), it was double-digested with Sac I and Hind III (all kinds of restriction enzymes were purchased from TaKaRa), and after recovery and purification, it was mixed with Similarly, the pET28a (purchased from Novagen) vector digested with Sac I and Hind III was used for ligation. The ligation product was transformed into Escherichia coli (E.coli) DH5α (purchased from Biovector), and after verification, it was transferred into the host strain Rosetta (DE3) (purchased from ...

Embodiment 3

[0033] Example 3: Expression pattern of GmERF6 gene under stress induction

[0034] Stress treatment of soybean Jilin 32: using Hoagland medium (Ca(NO 3) 2 4H 2 O 0.62g / L, KNO 3 0.34g / L, KH 2 PO 4 0.06g / L, NH 4 NO 3 0.053g / L, MgSO 4 ·7H 2 O 0.493g / L, MgCl 2 0.67mg / L, H 3 BO 3 0.38mg / L, ZnSO 4 ·7H 2 O 0.29mg / L, CuSO 4 ·5H 2 O 0.015625mg / L, FeSO 4 ·7H 2 O0.02785g / L, EDTA-Na 2 0.0373g / L, PH 5.7-5.8), 28°C, 16h light / 8h dark conditions to cultivate soybean hydroponic seedlings, take the four-leaf stage hydroponic seedlings and carry out the following treatments:

[0035] High-salt (NaCl) treatment: Place soybean seedlings in Hoagland culture solution containing 200mM NaCl, take samples after treatment for 0h, 1h, 2h, 5h, 10h and 24h, place them in liquid nitrogen quickly, and store them at -80°C for later use;

[0036] Drought treatment: Soybean seedlings were placed in Hoagland culture solution containing 20% ​​PEG8000, samples were taken after treatment for 0...

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Abstract

The invention relates to a transcription factor ERF related to soybean stress, a coding gene thereof, and application of the coding gene, and belongs to the field of plant gene engineering. The amino acid sequence of the transcription factor ERF related to soybean stress is shown as a Sequence No.2, and the nucleotide sequence of the coding gene of the transcription factor is shown as a Sequence No.1. A soybean GmERF6 transcription factor gene related to stress induction is cloned, the expression mode and transcription regulator activity of the gene are researched, the drought resistance of wild arabidopsis is compared with that of a GmERF6 gene-transformed arabidopsis, a basis is provided for effectively applying the GmERF6 gene, and the GmERF6 gene has important significance for improving the stress resistance of plants, particularly breeding drought-resistant soybean varieties.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to a soybean stress-related transcription factor ERF and its coding gene and application. Background technique [0002] Soybean is an important economic crop and is widely planted all over the world. It is not only the main source of human protein and lipid, but also an important livestock feed and industrial raw material, and it is also of great value in medicine. However, biotic and abiotic stresses, including disease, salt damage, low temperature and drought, have extremely adverse effects on the growth and development of soybean, resulting in a decrease in soybean yield and serious economic losses. Therefore, it is particularly important to elucidate the response mechanism of soybean to stress, identify important genes related to stress and use them to improve the stress resistance of soybean. [0003] Plants can adapt to the environment by regulating the expression of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29A01H5/00
Inventor 王庆钰翟莹李景文王英闫帆雷婷婷苏连泰李艳杰张鑫生王洪预
Owner JILIN UNIV
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