A kind of transcription factor cswrky1 isolated from the glandular trichomes of cannabis and its application

A technology of transcription factors and cannabinoid glands, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems of difficult screening and identification of transcription factors

Active Publication Date: 2022-02-01
厦门梓蔓生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, although some transcription factors that regulate the synthesis of cannabinoids have been discovered, there are still many important transcription factors that have not been discovered, and the screening and identification of transcription factors are difficult. Research on the biology of transcription factors involved in the regulation of gene expression process is very important

Method used

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  • A kind of transcription factor cswrky1 isolated from the glandular trichomes of cannabis and its application
  • A kind of transcription factor cswrky1 isolated from the glandular trichomes of cannabis and its application
  • A kind of transcription factor cswrky1 isolated from the glandular trichomes of cannabis and its application

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Experimental program
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Effect test

Embodiment 1

[0038] The 548bp THCAS promoter was cloned with gene-specific primers, and the binary vector pMDC163:ProTHCAS:GUS was constructed using Gateway technology. Next, Agrobacterium was used to transform Arabidopsis thaliana, and 10 independent ProTHCAS:GUS transgenic lines were analyzed. The expression of the GUS reporter gene was analyzed histochemically in different plant organs, and the results were as follows: figure 1 As shown, among them, picture a: cotyledon of 6-day-old seedling; picture c: mature leaf; picture e: stem; picture g: flower; picture b, d, f, h are trichomes in picture a and picture c respectively Close-up views on , e, and g. a, c, e, g=1000 μm; b, d, f, h=500 μm. It can be seen from the figure that the GUS activity is only in the leafy organ ( figure 1 a-h) and stem ( figure 1 e, f) Observed in trichomes. In flowers, GUS activity was detected only in the trichomes of sepals ( figure 1 g, h).

Embodiment 2

[0040] The same THCAS promoter used for the GUS expression assay was cloned in the MCS (KpnI(5') and XhoI(3')) upstream of the AUR1-C gene in the yeast vector pAbAi using yeast one-hybrid technique to screen for targeted binding sequences subfragment. The AUR1-C gene is an antifungal antibiotic resistance gene. The constructed vector structure was ProTHCAS-AbA, digested and cut by BstBI enzyme, and transformed into Y1HGold yeast strain using PEG-mediated transformation method according to the instructions. Using the URA3 gene of pAbAi as a selection marker, it was integrated into the non-functional ura3 site of Y1HGold yeast strain. Transformants were selected on media lacking synthetic glucose and verified in colony PCR. Using Y1HGold-ProTHCAS-ABA as the material, the Y1H cDNA library was constructed using the mRNA of the female flower of Cannabis (Purple Kush). After filtering, the results are as follows figure 2 It can be seen from the figure that CsWRKY1 can interact ...

Embodiment 3

[0042] To determine the subcellular localization of CsWRKY1, the full-length CsWRKY1 coding region was fused to full-length enhanced yellow fluorescent protein (EYFP). The CsWRKY1-EYFP fusion construct as well as EYFP were transiently expressed in protoplasts isolated from Arabidopsis leaves under the control of the 35S promoter. CsWRKY1-EYFP was only observed in the nucleus ( image 3 ), which is consistent with its role as a transcription factor.

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Abstract

The invention provides transcription factor CsWRKY1 and its application, belonging to the technical field of plant genetic engineering. In the present invention, the transcription factor CsWRKY1 is isolated for the first time from the glandular trichomes of the plant Cannabis trichomes, its nucleotide sequence is shown in SEQ ID NO:1, and its encoded amino acid sequence is shown in SEQ ID NO:2. The functional test of the present invention shows that the transcription factor CsWRKY1 can interact with the THCA synthetase (THCAS) gene promoter, thereby exerting an inhibitory effect on the synthesis of cannabinoid compounds, and can effectively realize the regulatory effect on the synthesis of cannabinoid compounds .

Description

technical field [0001] The invention belongs to the technical field of plant genes, and in particular relates to a transcription factor CsWRKY1 isolated from glandular hairs of cannabis and an application thereof. Background technique [0002] Marijuana (Cannabis sativa) is an annual herb of the Cannabis family Cannabis genus, dioecious, and is one of the earliest crops planted by humans. Monoecious. As a traditional economic crop in China, it has important economic value and involves many aspects such as textiles, building materials, food and pharmaceuticals. For example, the flowers, leaves and roots of marijuana can be used to extract medicinal ingredients for pharmaceutical use, and can also be used as soil fertilizers to kill insects and prevent diseases and increase soil organic matter; bast fiber can be made into high-grade clothes, stalks The core fiber can be used as papermaking and building materials; the grain can be used as food and feed, and the extracted oil ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/28C12Q1/6895
CPCC07K14/415C12N15/8243C12N15/8217
Inventor 刘圆圆张熠平吕素娟
Owner 厦门梓蔓生物科技有限公司
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