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Soybean ethylene responsive factor (ERF) transcription factor, and coding gene and salt tolerance application thereof

A technology of transcription factors and coding genes, applied in application, genetic engineering, plant gene improvement, etc., can solve the problem of fewer ERF transcription factors and achieve the effect of improving stress resistance

Inactive Publication Date: 2012-06-27
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are relatively few studies on ERF transcription factors in soybean, and only three ERF functions have been reported, all of which are involved in the stress response of soybean and play a positive regulatory role in improving plant stress resistance

Method used

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  • Soybean ethylene responsive factor (ERF) transcription factor, and coding gene and salt tolerance application thereof
  • Soybean ethylene responsive factor (ERF) transcription factor, and coding gene and salt tolerance application thereof
  • Soybean ethylene responsive factor (ERF) transcription factor, and coding gene and salt tolerance application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: GmERF7 Gene cloning and sequence analysis

[0021] Soybean RNA extraction and cDNA synthesis: RNAplant plus Reagent (purchased from TIANGEN) was used to extract total RNA from soybean Jilin 32 leaves, and M-MLV reverse transcriptase (RNase H-) (purchased from TaKaRa) was used to reverse transcribe and synthesize cDNA.

[0022] Design and synthesis of primers: The sequences (completed by Huada Genomics) obtained from immature embryo expression profiles of soybean variety Jilin 32 at three stages (20, 30 and 50 days) were 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, whose nucleotide sequence is as described in Sequence No.1, design synthetic primers, F: 5'-GAGACACAGCCATTGTTTGTTTAC-3', R: 5'-GGTTTTCTTGCTGTGGATTC-3'. Using the above cDNA as a template, carry out PCR amplification according to the following reaction system a...

Embodiment 2

[0024] Example 2: GmERF7 Expression patterns of genes induced by stress

[0025] Stress treatment of soybean Jilin 32: using Hoagland culture medium (Ca(NO 3 ) 2 4H 2 O 0.62 g / L, KNO 3 0.34 g / L, KH 2 PO 4 0.06 g / L, NH 4 NO 3 0.053 g / L, MgSO 4 0.24 g / L, MgCl 2 0.67 mg / L, H 3 BO 3 0.38 mg / L, MnSO 4 0.2 mg / L, ZnSO 4 ·7H 2 O 0.29 mg / L, CuSO 4 0.01 mg / L, FeSO 4 ·7H 2 O 0.02785 g / L, EDTA-Na 2 0.0373 g / L, pH 5.7-5.8), 28°C, 16 h light / 8 h dark conditions to cultivate soybean hydroponic seedlings, and the hydroponic seedlings at the four-leaf stage were subjected to the following treatments:

[0026]High-salt (NaCl) treatment: Place soybean seedlings in Hoagland culture solution containing 200 mmol / L 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 Save for later use;

[0027] Drought treatment: Soybean seedlings were placed in Hoagland culture solution containing 20% ​​PEG80...

Embodiment 3

[0037] Example 3: GmERF7 Gene Expression Patterns in Soybean Tissues

[0038] Total RNA was extracted from the roots, stems, leaves, flowers, and 20-day embryos of soybean Jilin 32 and reverse-transcribed into cDNA. The method was the same as in Example 1. Real-time fluorescent quantitative PCR was carried out using the cDNA of the above-mentioned roots, stems, leaves, flowers and 20-day embryos as templates, and the method was the same as in Example 2.

[0039] GmERF7 The relative expression level (mean value) of genes in different soybean tissues:

[0040] organize leaf 20 day embryo root stem flower relative expression 1 0.71475 3.80061 0.667293 0.131047

[0041] The results show GmERF7 The expression level of the gene was the highest in the root of soybean, and the expression level was in the following order: root﹥leaf﹥20-day embryo﹥stem﹥flower.

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Abstract

The invention relates to a soybean ethylene responsive factor (ERF) transcription factor, and a coding gene and salt tolerance application thereof, and belongs to the field of genetic engineering. The amino acid sequence of the soybean ERF transcription factor is Sequence No.2, the amino acid sequence of the coding gene of the soybean ERF transcription factor is Sequence No.1, and the invention provides application of the soybean ERF transcription factor to salt tolerance of a transgenic plant. A GmERF7 transcription factor gene related with the soybean salt tolerance is cloned, and the expression mode of the gene in soybean, the in-vitro combination of the gene with an adversity element, and transcriptional regulation activity of the gene are researched, the salt tolerance of wild type tobacco and tobacco with trans GmERF7 gene is compared, basis is provided for the effective application of the soybean ERF transcription factor, and the invention has significance for improvement on stress resistance of plants, particularly cultivation of soybean varieties with high salt tolerance.

Description

technical field [0001] The invention relates to the field of plant genetic engineering, in particular to the application of a soybean ERF transcription factor and its coding gene in transgenic plants for salt tolerance. 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. It is also of great value in medicine. However, biotic and abiotic stresses have extremely adverse effects on the growth and development of soybean. Among them, soil salinization seriously affects the yield, quality and benefit of soybean, causing serious economic losses. Therefore, it is particularly important to elucidate the response mechanism of soybean to stress, identify important genes related to salt tolerance and use them to improve soybean salt tolerance. [0003] Under the stimulation of stress, plant transcription f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/84A01H5/00
Inventor 王庆钰翟莹李景文王英闫帆雷婷婷李艳杰张庆林张鑫生王洪预杨旭光唐心隆
Owner JILIN UNIV
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