Wheat TaMADS6 gene and application thereof

A wheat and gene technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of little understanding of the structure and function analysis of the MADS-box gene family, and achieve the effect of improving and improving germplasm resources.

Inactive Publication Date: 2017-06-30
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

Wheat is an important food crop for humans, but little is known about the structural and functional analysis of the MADS-box gene family in wheat

Method used

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  • Wheat TaMADS6 gene and application thereof
  • Wheat TaMADS6 gene and application thereof
  • Wheat TaMADS6 gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Cloning of embodiment 1 wheat TaMADS6 gene

[0034] 1. Extraction of total RNA from young ears of wheat

[0035] Extraction of total RNA from Chinese spring (Triticum aestivum L.) young ears, using Plant miniKit (QIAGEN).

[0036] 2. Synthesis of the first cDNA fragment of TaMADS6 gene

[0037] Follow the instructions of Reverse Transcriptase M-MLV (Promega).

[0038] 3. Clone the coding region sequence of TaMADS6 from the cDNA of young panicle of Chinese spring (Triticum aestivum L.); forward primer sequence such as SEQ ID NO.4 and SEQ ID NO.5, reverse primer sequence such as SEQ ID NO.6 .

[0039] PCR program: 94°C, 10min; 94°C, 30s; 56°C, 30s; 72°C, 1min; repeated 35 times; 72°C, 10 minutes.

[0040] PCR system (full gold company):

[0041]

[0042]

[0043] The PCR products were gel-cut, recovered and purified according to the protocol provided by Beijing Quanshijin Biotechnology Co., Ltd. -Blunt Zero Cloning Kit cloning method clones connected to -...

Embodiment 2

[0044] Chromosomal location of embodiment 2 wheat TaMADS6 gene

[0045] According to the TaMADS6 gene obtained in Example 1, the homologous chromosome sequences of A, B, and D were compared and analyzed, and it was found that there were polymorphisms in the nucleotide sequences of the three. Three pairs of specific primers were designed, and the DNA of Chinese spring deficiency-tetrasomies was used as a template for PCR reaction, and the position of TaMADS6 gene in the chromosome was located by agarose gel electrophoresis.

[0046] The 3 pairs of specific primers are:

[0047] The detection of wheat TaMADS6-A gene is realized by the following primers: SEQ ID NO.7-8;

[0048] The detection of wheat TaMADS6-B gene is realized by the following primers: SEQ ID NO.9-10;

[0049] The detection of wheat TaMADS6-D gene is realized by the following primers: SEQ ID NO.11-12.

[0050] Using any pair of primers among the above three pairs of primers for detecting wheat TaMADS6 gene to PC...

Embodiment 3

[0052] Embodiment 3 Wheat TaMADS6-D gene transforms Brachypodium

[0053] Callus induction medium (pH=5.8): 100×MS macroelements 100ml, 100×MS trace elements 10ml, 100×Fe-EDTA 10ml, Sucrose 30g, 2,4-D (1mg / ml) 2.5ml, CuSO 4 (1mg / ml) 0.6ml, Phytagel 2g, make up to 990ml with sterile water, 100×M5vitamins (added after sterilization) 10ml.

[0054] MGL medium (pH=7.2): Tryptone 5g, Yeast extract 2.5g, NaCl 5.2g, Mannitol 10g, L-glutamic acid sodium salt 2.32g, KH 2 PO 4 0.5g, MgSO 4 ·7H 2 O 0.2g, Agar 10g, make up to 1L with sterile water, Biotin (1mg / ml, added after sterilization) 2μl, Acetosyringone (30mg / ml, added after sterilization) 1ml.

[0055] Agrobacterium suspension medium (pH=5.5): 100ml of 10×MS macroelements, 10ml of 100×MS trace elements, 10ml of 100×Fe-EDTA, 10g of Sucrose, 10g of Mannitol, make up to 1L with sterile water, Acetosyringone (30mg / ml , add after sterilization) 1.5ml.

[0056] Co-cultivation medium (pH=5.8): 100×MS macroelements 100ml, 100×MS tr...

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Abstract

The invention provides a wheat TaMADS6 gene and an application thereof, and belongs to the field of crop molecular biology. Primers are designed, TaMADS6 encoding region sequences are cloned from China spring young spike cDNA, TaMADS6 is located in a wheat sixth homologous group by using a China spring nullisomic-tetrasome, and the TaMADS6 gene encoding region sequences in sequences of A, B and D homologous chromosomes are shown in SEQ ID NO.1-3 respectively. The encoding region sequence of the D homologous chromosome of the TaMADS6 gene transforms Brachypodium Bd21-3, the gene can make nrachypodium significantly flower in advance, and flowering is promoted by increasing expression of BdFT1, BdFT2 and BdVRN1.

Description

technical field [0001] The invention relates to the field of crop molecular biology and molecular breeding, more specifically, relates to wheat TaMADS6 gene, its location in chromosome, and the application of the gene. Background technique [0002] Wheat is an important food crop, and about 40% of people in the world take wheat as their main food. At the same time, as one of the three major grains, almost all of its output is used for food, and it is the food crop with the world's total output second only to corn. With the completion of the whole genome sequencing of a variety of plants including rice, corn, wheat, etc., human beings have entered the era of functional genes. Sequence comparison and analysis of target genes with known functions in species, and primer design and homologous sequence cloning for research species using highly conserved sequences are one of the common methods for discovering new genes. [0003] MADS-box genes are a class of genes that play an im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11C12N15/10C12Q1/68C12N15/82A01H5/02
CPCC07K14/415C12N15/1096C12N15/11C12N15/827C12Q1/6895C12Q2600/13C12Q2600/158C12Q2531/113
Inventor 耿帅锋孔星辰赵俊敏陈凯李爱丽毛龙
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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