CgWRKY57 gene of cymbidium goeringii, and application of gene

A gene and amino acid technology, applied in Chunlan CgWRKY57 gene and its application field, achieves the effect of wide application prospect

Inactive Publication Date: 2019-04-26
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the stress defense mechanism of orchids by WRKY protein, and only the reports of Dendrobium can be used for reference.

Method used

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  • CgWRKY57 gene of cymbidium goeringii, and application of gene
  • CgWRKY57 gene of cymbidium goeringii, and application of gene
  • CgWRKY57 gene of cymbidium goeringii, and application of gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The material used in this example is the leaves of Chunlan 'Songmei', which are quickly frozen in liquid nitrogen after harvest and stored in an ultra-low temperature refrigerator (-80°C).

[0021] 1) Extraction of total RNA from Chunlan leaves

[0022] According to the instructions of the TaKaRa plant total RNA extraction kit, the specific operation is as follows:

[0023] The ultra-low temperature frozen leaves of Chunlan were quickly transferred to a mortar pre-cooled with liquid nitrogen, and the tissue was ground with a pestle, during which liquid nitrogen was continuously added until it was ground into powder; the powdered sample was added to a 450 μl In a 1.5 mL sterilized tube of Buffer PE, pipette repeatedly until there is no obvious precipitation in the lysate; centrifuge the lysate at 12,000 rpm at 4°C for 5 minutes; pipette the supernatant carefully into a new 1.5 mL sterilized tube middle. Add 1 / 10 volume of Buffer NB to the supernatant, shake and mix wit...

Embodiment 2

[0045] Research indicates WRKY57 The gene was expressed in every tissue and organ of Chunlan ( figure 2 a), but the expression level of the gene is the highest in Chunlan root, indicating that the gene is functionally active in the root. Through the expression analysis of the leaves of Chunlan low temperature stress treatment ( figure 2 b), to prove CgWRKY57 Genes in Chunlan leaves can respond to low temperature stress, and play an important regulatory role in this process.

[0046] The plant material used in this embodiment is Arabidopsis ( Arabidopsis thaliana ) Col (Columbia) wild-type seeds.

[0047] The Escherichia coli strain used in this example was Trans5α; the Agrobacterium strain was GV3101, which were used to transform Arabidopsis thaliana; the plant expression vector used in the experiment was pBI121. The strains used were purchased from Quanshijin Biotechnology Company and Price Biotechnology Company.

[0048] 1) CgWRKY57 Construction of gene overexpres...

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Abstract

The invention discloses a CgWRKY57 encoding gene of cymbidium goeringii, and application of the encoding gene. The nucleotide sequence of the CgWRKY57 gene is shown in SEQ ID NO. 1, the amino acid sequence of the expressed protein of the CgWRKY57 gene is shown in SEQ ID NO. 2. By cloning and identifying the CgWRKY57 gene of 'Song Mei' which is a cultivated variety of the cymbidium goeringii, analyzing the gene expression and verifying the functions of the gene, an arabidopsis thaliana plant with the overexpressed CgWRKY57 gene is found to have phenotypes such as growth retardation and plant dwarf; therefore, the gene has a broad application prospect in the production and breeding of the cymbidium goeringii and other plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to a spring orchid CgWRKY57 Genes and their applications. Background technique [0002] Chunlan ( Cymbidium goeringii ) is the genus Orchidaceae (Orchidaceae) ( Cymbidium ) is a type of ground orchid, with small plants, pseudobulbs, and belt-shaped leaves. There are many varieties and varieties. The flower types include normal flowers and butterfly flowers. The flower colors are mainly green and light yellow-brown, and most of them are fragrant. Great horticultural ornamental value. However, the maintenance of existing varieties is relatively difficult. Chunlan has relatively strict requirements on temperature, light, humidity, nutrition and many other aspects, and the maintenance cost is relatively high. Therefore, the study of its stress-resistant molecular mechanism can lay the foundation for the cultivation of Chunlan stress-resistant varieties, and can a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415
CPCC07K14/415C12N15/8261
Inventor 胡凤荣景袭俊刘倩王连平张鸽香丁彦芬
Owner NANJING FORESTRY UNIV
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