Application of GhD1119 gene in regulating and controlling flowering of upland cotton

A ghd1119, upland cotton technology, applied in the application, genetic engineering, plant genetic improvement and other directions, to achieve the effect of promoting early flowering

Pending Publication Date: 2022-01-11
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Judging from the existing literature reports, the key genes regulating flowering in cotton still need to be

Method used

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  • Application of GhD1119 gene in regulating and controlling flowering of upland cotton
  • Application of GhD1119 gene in regulating and controlling flowering of upland cotton
  • Application of GhD1119 gene in regulating and controlling flowering of upland cotton

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] This embodiment provides the upland cotton GhD1119 gene cloning and vector construction methods, including:

[0026] 1. Plant material and reagent selection

[0027] The upland cotton middle cotton 113 and jin668 used in the present invention are preserved by the laboratory and planted in the plant cultivation room of the laboratory. 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. Reverse transcription kit Transcriptor FirstStrand cDNASynthesis Kit and fluorescence quantitative kit FastStart Essential DNA Green Master were purchased from Roche. Antibiotics were purchased from Solebol Corporation. Restriction enzymes were purchased from Baoriji Biotechnology Co., Ltd. Taq master mix was purchased from Akerui Company.

[0028] pGM-T cloning kit, TOP10 competent cells we...

Embodiment 2

[0042] This example provides the application of the upland cotton GhD1119 gene in regulating flowering, including:

[0043] 1. Obtaining transgenic lines of upland cotton overexpressing and CRISPR gene editing GhD1119 gene

[0044] The GhD1119 gene overexpression and CRISPR gene editing vectors constructed in Example 1 were used for genetic transformation of cotton hypocotyls: the peeled cotton seeds were sterilized with mercuric chloride, washed with sterile water, and then put into sterile seedling medium , cultured in the dark at 28°C for 1 day, picked off the seed coat, straightened the seedlings, and cultured in the dark at 28°C for another 4-5 days; streaked with the Agrobacterium liquid, picked a single clone after 2 days, shook the bacteria overnight, discarded the supernatant by centrifugation, and added 10 mL MGL and 25μL of AS, shake to suspend the bacterial liquid, shake at 28°C, activate at 200rpm / min for at least 30min; cut the hypocotyls into small stems, infect...

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Abstract

The invention provides application of a GhD1119 gene in regulating and controlling flowering of upland cotton; the GhD1119 gene is constructed into an overexpression vector and a CRISPR gene editing vector for genetic transformation of the cotton; and the function of the gene is verified. It is found that the upland cotton GhD1119 gene can promote an overexpression transgenic cotton line to bud 2-12 days in advance and bloom 8-24 days in advance, so that the squaring stage of a CRISPR cotton edited line is delayed by 3-9 days. The invention can provide effective support for digging important genes related to cotton flowering time regulation and control and analyzing cotton flowering regulation and control ways, and can be used for improving prematurity of high-quality middle and late maturing varieties by utilizing the genes, creating prematurity high-quality cotton germplasm resources and realizing collaborative improvement of prematurity and excellent fiber quality; and thus, the invention plays 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 the application of GhD1119 in regulating the flowering of upland cotton. Background technique [0002] Cotton (Gossypium spp.) is an important economic crop that can provide natural fibers and plays an important role in our national economy. At present, my country's cotton planting is mainly distributed in the northwest inland cotton area and the cotton area of ​​the Yellow River and Yangtze River Basin. The northwestern inland cotton region represented by Xinjiang cools down quickly in autumn, and the first frost appears early, and the temperature rises slowly in spring, and the end of the last frost is late. Under such climatic conditions, planting early-maturing cotton varieties can effectively take advantage of the limited frost-free period in the region for production. At the same time, mechanized harvesting in the Nort...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/84A01H5/02A01H6/60
CPCC07K14/415C12N15/827C12N15/8205C12N15/8218
Inventor 王彩香宿俊吉刘娟娟李美丽张爱史春辉谢晓宇巨吉生
Owner GANSU AGRI UNIV
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