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Application of wheat WLSH1 gene in regulation and control of spike and grain development of plants

A plant and grain technology, applied in the application field of ear and grain development, can solve the problems of little analysis and research on structure and function, unclear biological function and molecular mechanism, etc.

Active Publication Date: 2019-12-13
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

[0004] At present, the analysis and research on the structure and function of MADS-box gene family in wheat are very little, and the biological function and molecular mechanism of this gene family in wheat are still unclear

Method used

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  • Application of wheat WLSH1 gene in regulation and control of spike and grain development of plants
  • Application of wheat WLSH1 gene in regulation and control of spike and grain development of plants
  • Application of wheat WLSH1 gene in regulation and control of spike and grain development of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Cloning of embodiment 1 wheat WLSH1 gene

[0054] Using the cDNA of the young ear of Triticum aestivum L. as a template, the WLSH1 gene (WLSH1-D) located on the homologous chromosome of wheat 4D was cloned, and the specific method was as follows:

[0055] (1) Extraction of total RNA from young ears of wheat: use miniKit kit (QIAGEN) to extract total RNA from young ears of Chinese spring;

[0056] (2) Synthesis of the first cDNA fragment of WLSH1 gene: according to the operating instructions of reverse transcriptase M-MLV (Promega);

[0057] (3) Cloning the coding region sequence of WLSH1; using the young ear cDNA of Chinese spring (Triticum aestivum L.) obtained in step (2) as a template, using specific primers (SEQ ID NO.7-8) for amplifying the WLSH1 gene PCR amplification of the WLSH1 gene.

[0058] SEQ ID NO.7: forward primer: GAGATGGGTCGGGGGAAG;

[0059] SEQ ID NO. 8: Reverse primer: GTCACCGAAGTACAAATACATTAGC.

[0060] The reaction program of PCR amplification: ...

Embodiment 2

[0063] Construction of the overexpression intermediate vector of embodiment 2 WLSH1 gene

[0064] Before constructing the overexpression vector of the WLSH1-D gene, first combine the WLSH1 gene with the pAHC25-TiDPK1-6×MYC vector (plasmid map as shown in figure 1 Shown) connection, construct the overexpression intermediate vector of WLSH1-D gene, specific method is as follows:

[0065] (1) Primer design: use Primer Premier 5.0 software to design the full-length cloning primers of WLSH1 cDNA, in which, the forward primer ch-WLSH1-F1 has a SpeI restriction site, and the reverse primer ch-WLSH1-R1 has EcoKI enzyme cleavage site;

[0066] SEQ ID NO. 9: ch-WLSH1-F1: GGACTAGTATGGGTCGGGGGAAGGTG;

[0067] SEQ ID NO. 10: ch-WLSH1-R1: CCGAGCTCTCAGAAGCCACGTGATCT;

[0068] (2) Amplification of WLSH1 fragments: use TRIzol Reagent to extract total RNA from young ears of Chinese spring wheat, measure the concentration and reverse transcribe it into cDNA with the reverse transcription kit ...

Embodiment 3

[0079] Example 3 Construction of the overexpression vector plasmid of the WLSH1 gene On the basis of the WLSH1 gene overexpression intermediate vector WLSH1-pAHC25-6×MYC constructed in Example 2, a CUB vector (plasmid map as shown in figure 2 Shown) is the starting vector to construct the overexpression vector plasmid of WLSH1 gene, the specific method is as follows:

[0080] (1) Using the overexpression intermediate vector WLSH1-pAHC25-6×MYC of the WLSH1 gene constructed in Example 2 as a template, primers ch-WLSH1-F2 and ch-WLSH1-R2 (ch-WLSH1-F2 with BamHI digestion site, ch-WLSH1-R2 does not carry a restriction site) to amplify the fusion gene fragment with 6×MYC and WLSH1-D gene fragments;

[0081] SEQ ID NO. 13: ch-WLSH1-F2: GCGGATCCGACGGTATCGATTTAAAGC;

[0082] SEQ ID NO. 14: ch-WLSH1-R2: TCAGAAGCCACGTGATCTCTGTTTG;

[0083] (2) Enzyme digestion, ligation, transformation and screening, identification and sequencing of positive clones are the same as in the steps (3)-(8...

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Abstract

The invention relates to the technical field of plant molecular biology, in particular to application of a wheat WLSH1 gene in regulation and control of spike and grain development of plants. It is found that the WLHS1 gene has the function of regulating and controlling wheat spike and grain development, and can regulate and control the number of spikelets per spike of wheat and the number of grains per spike of wheat. Overexpression of the WLHS1 gene in wheat can significantly increase the number of spikelets per spike of wheat and the number of grains per spike of wheat. Thereby, a new thought is provided for increasing the yield of wheat.

Description

technical field [0001] The invention relates to the technical field of plant molecular biology, in particular to the application of wheat WLSH1 gene in regulating the ear and grain development of plants. Background technique [0002] Wheat is an important food crop, and about 40% of people in the world use wheat as their main food. At the same time, wheat, as one of the three major grains, produces almost all of it for food, and is the food crop with the world's total output second only to corn. With the improvement of the consumption level of Chinese residents, higher requirements are put forward for the yield and quality of wheat. The three elements of wheat yield are the number of ears per unit area, the number of grains per ear, and the weight of grains. The coordination of the relationship between the three is of great significance in improving wheat yield. Among them, the number of grains per panicle is mainly controlled by the development of wheat floral organs, and...

Claims

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Application Information

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IPC IPC(8): C12N15/82C07K14/415C12N15/29C12N15/11C12N15/66A01H5/10A01H5/00A01H6/46
CPCC07K14/415C12N15/66C12N15/8261
Inventor 李爱丽贾美玲耿帅锋毛龙
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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