Spring orchid CgWRKY2 gene and application thereof

A gene and amino acid technology, applied in Chunlan CgWRKY2 gene and its application fields, can solve the problems of horticultural ornamental quality decline, plant death, etc.

Pending Publication Date: 2019-04-16
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chunlan has high requirements on the growth environment. During the growth process, it is easily affected by harsh environments such as high tempe

Method used

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  • Spring orchid CgWRKY2 gene and application thereof
  • Spring orchid CgWRKY2 gene and application thereof
  • Spring orchid CgWRKY2 gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 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).

[0022] 1) Extraction of total RNA from Chunlan leaves

[0023] According to the instructions of the TaKaRa plant total RNA extraction kit, the specific operation is as follows:

[0024] 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

[0046] Research indicates WRKY2 The gene was expressed in every tissue and organ of Chunlan ( figure 2 a), but the expression level of this gene is the highest in Chunlan leaves, indicating that this gene is functionally active in leaves. Through the expression analysis of the leaves of Chunlan low temperature stress treatment ( figure 2 b), to prove wxya 2 genes play an important regulatory role in the response of Chunlan leaves to low temperature stress.

[0047] The plant material used in this embodiment is Arabidopsis ( Arabidopsis thaliana ) Col (Columbia) wild-type seeds.

[0048] 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.

[0049] 1) CgWRKY2 Construction of gene overexpression vector

[00...

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PUM

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Abstract

The invention discloses a spring orchid CgWRKY2 encoding gene and application thereof. The nucleotide sequence of the CgWRKY2 gene is shown as SEQ ID NO.1, and the amino acid sequence of an expressedprotein is shown as SEQ ID NO.2. According to cloning and identification of the CgWRKY2 gene of a spring orchid variety 'Song Mei', gene expression analysis and function verification, it's found thatan arabidopsis thaliana plant over expressing the CgWRKY2 gene grows slowly and is dwarfed with the phenotype of premature aging like early blight of leaves, thereof, the gene has a wide range of usesin production and breeding of the orchid and other plants.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to a spring orchid CgWRKY2 Genes and their applications. Background technique [0002] Orchidaceae (Orchidaceae) is one of the largest families of flowering plants, with more than 25,000 species in the world, accounting for about 10% of all flowering plants. Chunlan ( Cymbidium goeringii ) belongs to the small-flowered ground orchid species in the Orchidaceae genus. It has a peculiar flower shape, elegant flower color, quiet flower fragrance, beautiful leaf appearance, and high ornamental and economic value. Chunlan has high requirements on the growth environment. During the growth process, it is easily affected by harsh environments such as high temperature, low temperature, and drought. Therefore, it is of great significance to study the molecular mechanism of plant response to abiotic stress and identify the genes with stress resistance function for Chunlan ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N1/21A01H5/00A01H6/20
CPCC12N15/8241C07K14/415
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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