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Application of cotton ghaco gene in promoting plant flowering

A gene and cotton technology, applied in the application field of GhACO gene in promoting plant flowering, can solve the problem of lack of gene resources and achieve the effect of reducing the number of rosette leaves

Inactive Publication Date: 2021-09-21
INST OF COTTON RES CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are still few studies on genes related to cotton precociousness, and there is a lack of effective genetic resources for genetic engineering breeding of cotton precocious varieties

Method used

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  • Application of cotton ghaco gene in promoting plant flowering
  • Application of cotton ghaco gene in promoting plant flowering
  • Application of cotton ghaco gene in promoting plant flowering

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0024] The following examples are used herein to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventors to be employed in the practice of the invention, and thus can be considered preferred modes for its practice. However, those skilled in the art should understand from this specification that many modifications can be made to the specific embodiments disclosed herein, and the same or similar results can still be obtained without departing from the spirit or scope of the present invention.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs, and the disclosures cited herein and their cited materials are all incorporated by reference .

[0026] Those skilled in the art will recognize, or ...

Embodiment 2

[0122] Example 2 Construction of PBI121-GhACO Plant Expression Vector

[0123] 1. Obtaining target gene fragments with specific restriction sites

[0124] For the cDNA sequence of the cloned GhACO gene, primers containing suitable enzyme cutting sites were designed at the start codon ATG and the stop codon respectively. The enzyme cutting sites used are Xba I (T / CTAGA) and Sma I (CCC / GGG).

[0125] The primer sequence of the restriction site of GhACO gene is as follows:

[0126] Upstream primer F (SEQ ID No.5):

[0127] 5'-CTAGTCTAGAATGGCAGAAATAAGCTTAGAACG-3'

[0128] Downstream primer R (SEQ ID No.6):

[0129] 5'-TCCCCCGGGTTAAGGTGGGCTTGCCTGCATCA-3'

[0130] The amplified target fragment with restriction sites was connected to the pGEM-T Easy cloning vector, transformed into DH5α competent cells, and the intermediary recombinants without sequence mutations were screened out by PCR, restriction restriction verification and sequence determination.

[0131] 2 Construction o...

Embodiment 3

[0138] Example 3 Using the Agrobacterium-mediated method to transform the recombinant vector pBI121-GhACO into Arabidopsis thaliana

[0139] 1 Preparation of Competent Agrobacterium LBA4404

[0140] Using CaCl 2 The preparation of competent cells by method is as follows:

[0141] 1) Pick a single colony and inoculate it in 4 mL of LB liquid medium containing antibiotics, and culture overnight at 28°C and 190 rpm;

[0142] 2) Transfer to 80mL LB liquid medium containing antibiotics at a ratio of 1:90, culture at 28°C, 170rpm to OD 600 =0.6;

[0143] 3) After 30 minutes of ice-bathing the bacterial solution, transfer it to a 50 mL centrifuge tube, centrifuge at 5,000 rpm for 10 minutes at 4°C, and discard the supernatant;

[0144] 4) Add 5mL of pre-cooled 70mM CaCl 2 , gently suspend, let stand on ice for 20min, centrifuge at 5,000rpm at 4°C for 5min, and discard the supernatant;

[0145] 5) Add 2 mL of pre-cooled 70 mM CaCl containing 15% glycerol 2 , to resuspend the pe...

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Abstract

The invention discloses cotton CaCO The gene and its application in promoting plant flowering belong to the technical field of plant genetic engineering. the cotton CaCO The gene has the nucleotide sequence shown in SEQ ID No.3 and can encode the amino acid sequence shown in SEQ ID No.4. The transgene obtained by the present invention through transgenic technology CaCO Arabidopsis thaliana plants with the gene significantly reduced the number of rosette leaves when Arabidopsis bolted, and the flowering time was significantly earlier, indicating that CaCO Gene plays key role in promoting flowering in cotton. The invention provides favorable gene resources for short-season cotton cultivation.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to the application of GhACO gene in promoting flowering of plants. Background technique [0002] Cotton is an important economic crop. As an important raw material in textile, medicine, chemical industry and other fields, it occupies a very important position in my country's national economy. Our country has a lot of people and little land, and grain and cotton compete for land. In order to ensure food security, cotton planting area and output are guaranteed at the same time. An effective way to solve this contradiction - early-maturing cotton breeding. The early-maturing cotton varieties can realize the post-seeding of wheat (rape) in the cotton areas of the Yellow River and the Yangtze River Basin; it is conducive to improving the quality and efficiency of the inland cotton areas in Northwest China, and ensuring the safety of national cotton use. Co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12N9/02C12N15/82A01H5/02A01H6/60A01H6/46A01H6/20
CPCC12N9/0071C12N15/827C12Y114/17004
Inventor 魏恒玲薛羽君喻树迅李林王寒涛马亮付小康
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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