GhA0749 and GhD0744 transcription factors of upland cotton and application thereof in flowering regulation

A technology of upland cotton, positive regulation, applied in angiosperms/flowering plants, applications, chemical instruments and methods, etc., can solve problems such as unreported regulation of upland cotton functions and applications, and achieve the effect of promoting flowering

Active Publication Date: 2021-09-03
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Problems in the prior art: The function and application of the cotton homologous genes GhA0749 and GhD0744 in regulating the flowering of upland cotton have not been reported in the prior art

Method used

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  • GhA0749 and GhD0744 transcription factors of upland cotton and application thereof in flowering regulation
  • GhA0749 and GhD0744 transcription factors of upland cotton and application thereof in flowering regulation
  • GhA0749 and GhD0744 transcription factors of upland cotton and application thereof in flowering regulation

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Experimental program
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Effect test

Embodiment 1

[0030] This embodiment provides the gene sequences of upland cotton GhA0749 (GH_A07G0749) and GhD0744 (GH_D07G0744). The CDS sequence of the GhA0749 gene is shown in SEQ No. 1 in the sequence table; the CDS sequence of the GhD0744 gene is shown in SEQ No. 2 in the sequence table.

Embodiment 2

[0032] This embodiment provides upland cotton GhA0749 and GhD0744 gene cloning and vector construction methods, including:

[0033] 1. Plant material and reagent selection

[0034] The upland cotton jin668 used in the present invention, the early-maturing cotton variety Zhongmiansuo 74 and the wild-type Arabidopsis thaliana are all preserved in this laboratory and planted in the laboratory plant cultivation room and artificial climate incubator. The present invention adopts polysaccharide polyphenol plant total RNA extraction kit, rapid plasmid mini-extraction kit, and general-purpose DNA purification and recovery kit, all of which are purchased from Tiangen Biochemical Technology Co., Ltd. The reverse transcription kit Transcriptor FirstStrand cDNA Synthesis Kit and the fluorescence quantitative kit FastStart Essential DNA Green Master were purchased from Roche. Antibiotics were purchased from Solebol Corporation. Restriction enzymes were purchased from Baoriji Biotechnolog...

Embodiment 3

[0052] This example provides the application of upland cotton GhA0749 and GhD0744 genes in regulating flowering, including:

[0053] 1. Genetic transformation of Arabidopsis, VIGS silencing of upland cotton and qPCR detection

[0054] After the emergence of wild-type Arabidopsis inflorescences, use the flower dipping method to infect Arabidopsis thaliana inflorescences, and receive T 0 generation seeds. T 1 Generation of positive plants, and continue to screen in MS medium containing Kan to obtain T 2 Positive plants. When the dicotyledons of upland cotton (Zhongmian Institute 74) were flattened, VIGS upland cotton plants were obtained by injection. Quantitative fluorescence (qPCR) was used to detect the expression of target genes in transgenic Arabidopsis and VIGS upland cotton, and the relationship between GhA0749 and GhD0744 and other flowering regulatory genes was detected by qPCR. The primers used are listed in Table 2.

[0055] Table 2 Fluorescent quantitative prim...

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Abstract

The invention provides GhA0749 and GhD0744 transcription factors of cotton and application thereof in flowering regulation, GhA0749 and GhD0744 genes in an MADS-box gene related to flowering regulation of upland cotton are cloned, an overexpression vector and a VIGS silence vector are constructed, and genetic transformation of arabidopsis thaliana and VIGS silence in the cotton are carried out to verify the gene function. It is found that the GhA0749 and GhD0744 genes of the upland cotton can promote early blossoming of arabidopsis thaliana for 3-5 days, so that the squaring stage of the VIGS silenced upland cotton is delayed by 5-7 days, and the flowering stage is delayed by 3-5 days. The GhA0749 and GhD0744 transcription factors of cotton and application thereof in flowering regulation can provide effective support for digging important genes related to flowering time regulation and analyzing a cotton flowering regulation way, can be used for improving prematurity of high-quality mid-late-maturing varieties by utilizing the genes, creating precocious and high-quality cotton germplasm resources and realizing collaborative improvement of prematurity and excellent fiber quality, and play an important role in cultivating high-quality cotton varieties with high prematurity.

Description

technical field [0001] The invention belongs to the fields of cotton germplasm resource cultivation and molecular biology, and specifically relates to: cotton GhA0749 and GhD0744 transcription factors and their application in regulating flowering. Background technique [0002] Food is food, clothing is cotton, clothing, food, housing and transportation come first. Cotton is not only an important agricultural economic product in our country, but also a strategic material related to the national economy and people's livelihood. Cotton fibers, cotton seeds, and cotton stalks are important raw materials for the production of my country's textile industry, national defense industry, pharmaceutical industry, and daily life. This shows the importance of cotton in our national economy and people's lives. China is the largest cotton producer and consumer. In the long-term actual production, we found that due to the large differences in climatic conditions in various regions of our c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/84A01H5/02A01H6/20
CPCC07K14/415C12N15/8218C12N15/827C12N15/8205
Inventor 王彩香张爱宿俊吉刘娟娟李美丽史春辉谢晓宇
Owner GANSU AGRI UNIV
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