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Soybean blossoming regulation gene GmCIB4, coding protein and application thereof

A technology for regulating genes and soybeans, applied in the field of genetic engineering, can solve problems such as soybeans that have not fundamentally changed, difficult soybean flowering habits, etc.

Inactive Publication Date: 2010-12-22
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the growth habit of soybean has been changed to a certain extent through traditional cultivation and genetic breeding methods, and some varieties with wider photoperiod adaptability have been obtained, they have not fundamentally changed the flowering habit of soybean (Zhao Cun et al., using photoperiod Screening of light-insensitive soybean varieties (lines) by induction method. Soybean Science. 1996, 15(1): 42-47; ): 225-230; Yang Zhipan and Zhou Xin'an, Research Progress in Soybean Photoperiod Genetic Breeding, Chinese Journal of Oil Crops, 1999, 21(1): 61-73; Luan Xiaoyan, Man Weiqun, Du Weiguang, Chen Yi, Liu Xinlei, Dou Guang Research on insensitive germplasm innovation and photoperiod breeding pathway, Soybean Science, 2004, 23(3): 196-199)
The main reason is that there is little understanding of the molecular mechanism of soybean flowering, which makes it difficult to fundamentally solve the problem of soybean flowering habits

Method used

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  • Soybean blossoming regulation gene GmCIB4, coding protein and application thereof
  • Soybean blossoming regulation gene GmCIB4, coding protein and application thereof
  • Soybean blossoming regulation gene GmCIB4, coding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation and Cloning of GmCIB4 Gene

[0024] According to the CDS (code sequence) sequence of the Arabidopsis CIB1 gene, a homology comparison was performed on the http: / / www.phytozome.net website, and a batch of soybean homologous sequences were found, among which the CDS sequence of GmCIB4 was on the soybean genome The position is Glyma16g10620.1, PCR amplification primers were designed, forward primer 5′-ATGGGAATTCAACTAACAGTTATG-3′ and reverse primer R1: 5′-TCAGAGCTCAACTTTCATCTGTG-3′. Using the total soybean cDNA as a template, PCR was performed to obtain the complete sequence of GmCIB4, the nucleotide sequence of which is shown in SEQ ID NO.1.

[0025] The total PCR reaction system is 25 μL, including soybean cDNA (50ng): 1 μL; dNTP (2.5mM): 2.5 μL; primer F1 (10 μM): 1 μL; primer R1 (10 μM): 1 μL; LATaq enzyme (5U / μL): 0.3 μL; 10× buffer: 2.5 μL; ddH 2 O: 16.7 μL, 25 μL in total. The PCR reaction program was: 94°C pre-denaturation for 5 minutes, 30 cy...

Embodiment 2

[0026] Example 2 Analysis and identification of GmCIB4 gene

[0027] The full-length CDS sequence of GmCIB4 is 1647bp, encoding a 548AA protein, which has 36.1% homology with Arabidopsis CIB1 protein. Protein structure analysis shows that its C-terminus contains a basic helix-loop-helix structure, which is a conserved domain of bHLH family transcriptional regulators, which regulates gene transcription by binding to the E-box in DNA; protein N The end also contains an NLS, which is associated with nuclear import.

[0028] According to the sequence query results on NCBI (www.ncbi.nlm.nih.gov), so far, there is no sequence information similar to CIB1 in soybean; and there are no published papers related to its function research. Therefore, it is considered that GmCIB4 is a new gene in soybean.

Embodiment 3

[0029] Example 3 Obtaining of transgenic Arabidopsis thaliana

[0030] According to the sequence information of GmCIB4, PCR amplification primers were designed at both ends of its CDS, forward primer 5′-ATGGGAATTCAACTAACAGTTATG-3′ and reverse primer R1: 5′-TCAGAGCTCAACTTTCATCTGTG-3′. The complete sequence of GmCIB4 was obtained by PCR using the total soybean cDNA as a template.

[0031] The total PCR reaction system is 25 μL, including soybean cDNA (50ng): 1 μL; dNTP (2.5mM): 2.5 μL; primer F1 (10 μM): 1 μL; primer R1 (10 μM): 1 μL; LATaq enzyme (5U / μL): 0.3 μL; 10× buffer: 2.5 μL; ddH 2 O: 16.7 μL, 25 μL in total. The PCR reaction program was: pre-denaturation at 95°C for 5 minutes, 30 cycles at 94°C for 30s, 52.5°C for 30s, 72°C for 2 minutes, and finally 10 minutes at 72°C. The above PCR procedure was repeated three times, and the three PCR products were combined for agarose gel electrophoresis, and then the purified PCR product was recovered by cutting the gel.

[0032...

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Abstract

The invention discloses a soybean blossoming regulation gene GmCIB4 and a coding protein thereof. The soybean blossoming regulation gene GmCIB4 has a nucleotide sequence shown as SEQ ID No.1 and an amino acid sequence shown as SEQ ID No.2. The blossoming of plants (arabidopsis thaliana) can be obviously promoted through overexpressing the soybean blossoming regulation gene GmCIB4 so as to shorten the blossoming time and the growth period. The invention can be applied to solving the flowering asynchronism problem in crossbreeding, the birth control problems of various crops, vegetables, fruits and flowers, the light period sensitivity problem and the introduction problem.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to soybean flowering regulator gene GmCIB4, its encoded protein and its application in plant photoperiod and flowering time regulation. Background technique [0002] Soybean is one of the important crops, and it is an important source of plant protein, edible oil, biodiesel, and secondary metabolites such as isoflavones and lecithin. Because soybean is a short-day plant, flowering is strictly controlled by photoperiod, so excellent varieties in different regions cannot be introduced to each other, and the growth period is also restricted by environmental photoperiod. If the sensitivity of soybean to photoperiod can be reduced, and the limitation of soybean flowering to photoperiod can be broken, the problem of soybean introduction can be solved, so as to realize the mutual exchange of high-quality varieties in various regions, enrich excellent germplasm resources in various region...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/29C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C07K14/415A01H5/00
Inventor 林辰涛李宏宇孟颖颖
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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