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A kind of Chunlan cgwrky65 gene and its application

A gene and transgenic technology, applied to the Chunlan CgWRKY65 gene and its application fields, can solve the problems of reduced activity, chromosome influence, gene inactivation, etc., and achieve the effect of prolonging the vegetative growth cycle and being widely used.

Active Publication Date: 2022-05-27
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the foreign gene enters the chromosome, the foreign protein encoded by the gene is produced, and the foreign protein or enzyme leads to the enhancement or reduction of the activity of other enzymes or proteins through a series of reactions. In addition, when the foreign gene is embedded in a certain part of the chromosome When a segment is formed, it will definitely affect the entire chromosome, thereby affecting the activity or regulatory mechanism of other genes.
[0005] Even if the same gene is inserted, due to different insertion sites or different chromosome groups, it will inevitably affect the chromosome where it is located, and further lead to an impact on the gene on the chromosome where it is located, and slight changes in the gene may affect its regulatory mechanism , lead to changes such as gene inactivation, activity reduction, activity enhancement, etc., and also cause differences in the role of genes, and the role of genes can only be obtained through verification, not through inference

Method used

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  • A kind of Chunlan cgwrky65 gene and its application
  • A kind of Chunlan cgwrky65 gene and its application
  • A kind of Chunlan cgwrky65 gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The material used in this example is the leaves of Chunlan 'Songmei', which were quickly frozen in liquid nitrogen after harvest and stored in an ultra-low temperature refrigerator (-80°C).

[0024] 1) Extraction of total RNA from Chunlan leaves

[0025] Follow the instructions of the TaKaRa Plant Total RNA Extraction Kit. The specific operations are:

[0026] The ultra-low temperature frozen Chunlan leaves were quickly transferred to a mortar pre-cooled with liquid nitrogen, and the tissue was ground with a pestle, during which liquid nitrogen was added continuously until it was ground into powder; Buffer PE into a 1.5 mL sterilized tube, pipette repeatedly until there is no obvious precipitation in the lysate; centrifuge the lysate at 12,000 rpm at 4°C for 5 min; carefully pipette the supernatant into a new 1.5 mL sterilized tube . Add 1 / 10 volume of Buffer NB to the supernatant, mix with Vortex, centrifuge at 12,000 rpm and 4°C for 5 min; carefully pipette the supe...

Embodiment 2

[0048] Research indicates WRKY65 The gene is expressed in various tissues and organs in Chunlan ( figure 2 a), but the expression of this gene is the highest in spring orchid, indicating that the gene is functionally active in flowers. Through the expression analysis of the leaves treated with ABA stress of Chunlan ( figure 2 b), to prove CgWRKY65 The gene plays an important regulatory role in the ABA stress response of Chunlan leaves.

[0049] The plant material used in this example is Arabidopsis ( Arabidopsis thaliana ) Col (Columbia) wild-type seeds.

[0050] The Escherichia coli strain used in this example was Trans5α; the Agrobacterium strain was GV3101, which were respectively used to transform Arabidopsis thaliana; the plant expression vector used in the experiment was pBI121. The strains used were purchased from Quanjin Bio and Price Bio.

[0051] 1) CgWRKY65 Construction of gene overexpression vector

[0052] obtained in Example 1 CgWRKY65 The full-lengt...

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Abstract

The invention discloses a kind of Chunlan CgWRKY65 coding genes and their applications, CgWRKY65 The nucleotide sequence of the gene is shown in SEQ ID NO.1, and the amino acid sequence of the expressed protein is shown in SEQ ID NO.2. The present invention adopts Chunlan cultivar 'Songmei' CgWRKY65 Gene cloning and identification, gene expression analysis, and verification of its function, discovery CgWRKY65 At the seedling stage of transgenic Arabidopsis, compared with wild-type Arabidopsis WT, CgWRKY65 The transgenic plants are generally short, with slightly yellow leaves and a tendency of premature senility, which shows that the gene may be widely used in the improvement of orchid vegetative reproduction.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to a kind of chunlan CgWRKY65 Genes and their applications. Background technique [0002] Orchidaceae is one of the largest families of flowering plants, containing more than 25,000 species worldwide, accounting for about 10% of all flowering plants. Chunlan ( Cymbidium goeringii ) belongs to the small-flowered terrestrial orchid species in the genus Orchidaceae. Its flower shape is peculiar, the flower color is light and elegant, the flower fragrance is quiet, the leaves are beautiful, and the ornamental value and economic value are extremely high. Chunlan has high requirements on the growth environment, and is easily affected by harsh environments such as high temperature, low temperature, and drought during the growth process. Therefore, it is of great significance to study the molecular mechanism of plant response to abiotic stress and identify genes with s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/12A01H6/20
CPCC07K14/415C12N15/8261
Inventor 胡凤荣刘倩王连平徐子涵黎猛
Owner NANJING FORESTRY UNIV
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