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Gene and method for altering maize flowering period

A technology of flowering and mutating genes, applied in the field of molecular genetics, can solve problems such as incomplete clarity

Active Publication Date: 2022-04-26
HUAZHONG AGRI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are a total of 53 genes with CCT structure in the maize B73 reference genome. The applicant previously used an association group consisting of 368 maize inbred lines to locate 34 CCT genes related to maize flowering (Jin Minliang. Maize pan-transcriptome Construction of ZmCOL3, a CCT gene represses flowering inmaize by interfering with the circadian clock and functional analysis [D]. Hubei: Huazhong Agricultural University, 2018; Jin M, Liu X, Jia W, et al. activating expression of ZmCCT[J].J Integr Plant Biol.,2018,60(6):465-480.), most of the genes have no specific functional identification results, so which genes will actually control the flowering of maize Traits are not yet fully understood

Method used

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  • Gene and method for altering maize flowering period
  • Gene and method for altering maize flowering period
  • Gene and method for altering maize flowering period

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Identification of changes in maize flowering period after knockout of candidate genes by gene editing

[0059] The present invention uses the CRISPR-Cas9 gene editing technology to perform fixed-point editing on 15 genes among the 34 genes related to maize flowering period shown by the association analysis results.

[0060] Embodiments include the construction of gene editing vectors, genetic transformation of maize, and functional verification of editing effects. details as follows:

[0061] 1. Construction of gene editing vector

[0062] The gene editing vector of the present invention is G08943-CPB-ZmUbi-hspCas9, and its vector diagram is as follows figure 1 shown. The basic vector of this vector is CPB-ZmUbi-hspCas9. In the present invention, the dual-target U6-sgRNA is obtained by OverlapPCR and then cloned into the basic vector by homologous recombination. The specific construction process is as follows:

[0063] (1) Cloning of U6 promoter. The U6 ...

Embodiment 2

[0075] Example 2 In-depth analysis of corn flowering traits and identification of mutant genes

[0076] Conduct more in-depth trait identification of gene-edited materials with altered flowering traits and analyze specific editing sites. After the gene editing of GRMZM2G148453, the maize flowering period of the T0 material was earlier than that of the recipient control KN5585. The anthesis of T1 material from this transformant was further characterized and analyzed for specific editing sites.

[0077] There are two target sites designed during gene editing of GRMZM2G148453, as shown in SEQ ID NO.2 and SEQ ID NO.3 respectively. The gRNA sequences expressed by the vectors containing these two targets are shown in SEQ ID NO.4 and SEQ ID NO.5, respectively.

[0078] Extract seedling DNA to detect gene editing. Primers were designed, and the primer sequences are shown in SEQ ID NO.7 and SEQ ID NO.8. Amplify the target editing segment, the amplification system is: DNA: 3 μL, bid...

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Abstract

The present invention belongs to the field of molecular genetics. It specifically relates to a gene and a method for changing the flowering period of maize. The invention provides a gene sequence controlling the flowering period of maize, and discloses a method for changing the flowering period by mutating the gene by means of genetic engineering. The invention also provides a maize mutant gene sequence with altered flowering period, and the mutant gene sequence can be used to improve the traits of maize flowering period.

Description

technical field [0001] The present invention belongs to the field of molecular genetics. It specifically relates to the gene controlling the florescence of corn and its application in changing the traits of the florescence of corn. The invention provides a gene sequence controlling the flowering period of maize, and discloses a method for shortening the flowering period by mutating the gene by means of genetic engineering. The invention also provides a maize mutant gene sequence with shortened florescence, and the mutant gene sequence can be used to improve the florescence traits of maize. Background technique [0002] In higher plants, flowering represents the transition process from vegetative growth to reproductive growth, which plays an important role in the whole growth and development stages of plants. The biological trait of when to flower is affected by the plant's own genetic factors and external environmental factors. Under the influence of this dual effect, a s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/113A01H5/02A01H6/46C12Q1/6895
CPCC07K14/415C12N15/827C12N15/8216C12N15/113C12Q1/6895C12N2310/20C12Q2600/158C12Q2600/13
Inventor 金敏亮刘相国许洁婷刘海军尹悦佳黄磊郭嘉韩宝柱严建兵
Owner HUAZHONG AGRI UNIV