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Flos chrysanthemi CmTFL1c gene and application thereof

A chrysanthemum and gene technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve problems such as related functions that have not been reported

Active Publication Date: 2018-05-11
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the related functions of chrysanthemum CmTFL1c gene in the regulation of chrysanthemum flowering and branching have not been reported

Method used

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  • Flos chrysanthemi CmTFL1c gene and application thereof
  • Flos chrysanthemi CmTFL1c gene and application thereof
  • Flos chrysanthemi CmTFL1c gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 The acquisition of chrysanthemum CmTFL1c gene

[0035] 1. Test material

[0036] The stem tip tissue of chrysanthemum 'Jin Buwei' is used as material.

[0037] 2. Plasmids, strains and test reagents

[0038] RNA reverse transcription kit (TAKARA) was purchased from Beijing Liuhetong Economic and Trade Co., Ltd.; Trans1-T1DH5α competent cells, DNAmarker, and cloning vector pEASY-Blunt were purchased from Beijing Quanshijin Biotechnology Co., Ltd. DNA Gel Extraction Kit (AXYGEN), LB Broth (Coolaber), Ampicillin (Amp), Kanamycin (Kan), Isopropylthiogalactoside (IPTG), X-Gal, Agar (Agar) , Agarose were purchased from Baierdi Biotechnology Co., Ltd.; ultra-fast plant RNA extraction kit, GelRed nucleic acid dye were purchased from Beijing Huayueyang Biotechnology Co., Ltd.; 2xPCR Mix was purchased from Beijing Qingke Xinye Biotechnology Co., Ltd., Primer synthesis and sequencing services were provided by Beijing Ruibo Xingke Biotechnology Co., Ltd.

[0039] 3. T...

Embodiment 2

[0050] Example 2 Construction of plant expression vector containing chrysanthemum CmTFL1c gene and phenotype analysis of genetic transformation of chrysanthemum

[0051] 1. To construct a safe expression vector (pCAMBIA1301-pmi-CmTFL1c) with mannose (PMI gene) as a screening marker, it is necessary to replace the Hyg site with the target gene to achieve safety without antibiotic markers. The upstream and downstream of the Hyg site are XhoI restriction sites. In order to ensure the correct connection direction of the target gene, the method of PCR fusion was selected to construct the plant expression vector, and the sequence of the pCAMBIA1301-pmi expression vector connection position was introduced into the upstream and downstream of the target gene fragment respectively. 15-20bp, the specific method is:

[0052] (1) Amplification of the target gene

[0053] Using the pEASY-Blunt-CmTFL1c plasmid saved in the previous work of the laboratory as a template, gene-specific primers...

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Abstract

The invention provides a flos chrysanthemi CmTFL1c gene and an application thereof and belongs to the field of plant genetic engineering. The sequence of the flos chrysanthemi CmTFL1c gene is represented as SEQ ID No.1, and an encoded protein sequence of the flos chrysanthemi CmTFL1c gene is represented as SEQ ID No.2. A flos chrysanthemi leaf disc is infected with an agrobacterium-mediated method, the gene is transformed into flos chrysanthemi, a flos chrysanthemi strain with the genetically modified CmTFL1c gene is obtained, and a result shows that the number of lateral braches of the flos chrysanthemi strain genetically modified with the CmTFL1c gene is increased, land surface cover rate is increased, the number of flowers is also increased, and blooming period is postponed for 15-20 days; therefore, the flos chrysanthemi CmTFL1c gene has functions of postponing the blooming period, promoting growth of the lateral braches of the flos chrysanthemi and increasing the number of flowers, and has good application prospect when applied to plant trait improvement.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to the chrysanthemum CmTFL1c gene, its encoded protein and its application in promoting the growth of plant side branches and increasing the amount of flowering. Background technique [0002] Chrysanthemum is one of the top ten famous flowers originating in my country, which comes from long-term artificial cultivation and natural interspecific hybridization. Chrysanthemum has great changes in leaf, flower shape, and petal shape, and has excellent color, fragrance, posture, and rhyme. It is called "Four Gentlemen of Flowers" together with plum, orchid, and bamboo. cultivation. There are many varieties of chrysanthemums. There are more than 3,000 species in my country and more than 7,000 species in the world. However, the natural flowering period of most varieties is relatively concentrated and short, mostly in October-November. The natural flowering period restri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29A01H5/00A01H6/14
CPCC07K14/415C12N15/8261C12N15/8267C12N15/827
Inventor 高亦珂高耀辉吴志苹范敏张启翔程堂仁王佳
Owner BEIJING FORESTRY UNIVERSITY
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