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Soybean abloom time adjusting gene GAL2 and application thereof

A technology of flowering time and gene regulation, applied in the field of genetic engineering, can solve problems such as unclear molecular mechanism, and achieve the effects of reducing breeding cost, prolonging growth period and increasing yield

Inactive Publication Date: 2008-03-26
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

[0005] However, the molecular mechanism of soybean flowering time regulation is still unclear, and the technology to increase the wide adaptability of soybean cultivation by regulating flowering time related genes is still blank.

Method used

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  • Soybean abloom time adjusting gene GAL2 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Isolation and cloning of GAL2 gene

[0018] Using the mRNA sequence of the At4g09960 gene (AGL11) in the Arabidopsis AGL family, a homology comparison was performed in the Genebank database, and a batch of soybean EST sequences were found, among which AW705451 contained the complete 5' end of the candidate gene, from which the PCR was designed. Forward primer F1: TTC ATT TCT TTG GAT TAG AAA TTT TTC C, using total soybean mRNA as a template, and oligo(dT) (GAC TCT GAT GCT GACAAT GAC TTT TTT TTT TTT TTT TT) with a linker as a primer for inversion The cDNA was recorded, and the reverse primer R1 was designed in the linker sequence: GAC TCT GAT GCT GAC AATGAC TTT, and the full sequence of GAL2 was obtained by PCR using F1 and R1 as primers.

[0019] The total PCR reaction system is 25 μL, including 50ng cDNA, 50 μmol / L dNTP, 1 μmol / L primer, 1U rTaq enzyme and 1× buffer. The PCR reaction program was: pre-denaturation at 95°C for 5 min; then 32 cycles of 95°C for ...

Embodiment 2

[0020] Example 2 Analysis and identification of GAL2 gene

[0021] The full length of the GAL2 mRNA sequence is 958bp, of which 71-739 are CDS, encoding a protein of 222 amino acids, and the homology with Arabidopsis SOC1 protein is 70%. Analysis of protein structure shows that its N-terminus contains a MADS-box, which is a conserved domain of AGL family transcriptional regulators, and binds to DNA by forming homologous or heterodimers; the middle of the protein also contains a K-box, This domain may be a helix-loop-helix structure and may be functionally involved in multimer formation.

[0022] According to the sequence query results on NCBI (www.ncbi.nlm.nih.gov), so far, there is no sequence information similar to GAL2 in soybean; and there are no published papers related to its function research so far. Therefore, GAL2 is considered to be a new gene in soybean.

Embodiment 3

[0023] Example 3 Obtaining of overexpressed GAL2 gene Arabidopsis

[0024] According to the sequence information of GAL2, a pair of primers GW-F were designed at both ends of the ORF: ATG GGA AGG GGG AAG ATC G; GW-R: TCA CCC AAG ATG AAGAATC. Using the total soybean mRNA as a template, use this pair of primers to perform RT-PCR to obtain the cDNA sequence of the complete coding region of GAL2. The total PCR reaction system is 25μL, including 50ngcDNA, 50μmol / L dNTP, 1μmol / L primer, 1U rTaq enzyme and 1× buffer. The PCR reaction program was: pre-denaturation at 95°C for 5 min; then 32 cycles of 94°C for 30 s, 55°C for 30 s, and 72°C for 1 min; finally, 72°C for 10 min to blunt the ends. The PCR product was cloned into the pGEM-T vector (Promega Company) (10 μL ligation system, including 75ngcDNA, 25ngpGEM-T vector, 5μL 2XBuffer, 1μL TA ligase, reacted overnight at 16°C.), and the overexpression was constructed after sequencing and identification Vectors 35S::GAL2 and 35S::GFP:...

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Abstract

The present invention provides one kind of soybean flowering time regulating gene GAL2 and its separating and cloning process and application. The soybean flowering time regulating gene GAL2 codes protein with the amino acid sequence as shown in SEQ ID No. 2 or the amino acid sequence of SEQ ID No. 2 through the substitution, deletion or addition of one or several amino acids and with the same function. Over expressing the gene can postpone the soybean flowering time to prolong the growth period of soybean extremely, best utilize the natural conditions and increase the yield. In addition, the present invention makes it easy to cultivating new soybean variety.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a gene GAL2 regulating soybean flowering time and its application in plant breeding. Background technique [0002] Soybean is one of the five major crops in my country, and it plays a very extensive role in the national economy: (1) It is an important source of human and animal plant protein; (2) It is one of the main sources of edible oil; (3) Soybean is a renewable Energy—one of the main raw materials for the production of biodiesel; (4) soybeans contain a large number of biologically active substances, such as isoflavones and D-chiro-inositol. However, for many years, my country's soybean supply has been 50% dependent on imports. Imported soybeans have obviously impacted our soybean production. Increasing soybean yield is the key to change this situation and realize soybean self-sufficiency. [0003] Although my country is rich in soybean resources, s...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82
Inventor 林辰涛傅永福徐通达许娇卉李宏宇
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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