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Syringa oblata chalcone synthase gene and application thereof

A technology of lilac and chalcone, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of weak basic research, and the research on CHS gene cloning and function has not yet been reported.

Inactive Publication Date: 2017-04-26
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its basic research is very weak, and there is no report on the cloning and function of CHS gene in Lilac chinensis.

Method used

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  • Syringa oblata chalcone synthase gene and application thereof
  • Syringa oblata chalcone synthase gene and application thereof
  • Syringa oblata chalcone synthase gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] This example provides the cloning method of SoCHS gene.

[0043] S11, extraction of total RNA from Lilac chinensis

[0044] The North China lilac materials used in the present invention are all collected from the Forest Germplasm Resource Nursery of Beijing Agricultural College, a resource preservation unit of the National Forest Germplasm Resource Platform (NFGRP). Select 10 North China lilacs that grow well and are free from diseases and insect pests, and collect plant materials from four directions in the southeast, northwest, and east.

[0045] According to the whole gold RNA extraction kit (TransZol TM Total RNA was extracted according to the instructions of Up Plus RNA Kit. Use 1.2% concentration of agarose to detect the integrity of RNA, use a UV spectrophotometer to measure the absorbance of RNA at 230nm, 260nm and 280nm wavelengths, and determine the concentration and purity of RNA

[0046] S12: RACE-PCR

[0047] Take 50ng~5μg of total RNA, and carry out r...

Embodiment 2

[0061] This example provides functional verification of the SoCHS gene.

[0062] 1. Materials and methods:

[0063] 1. Materials

[0064] Escherichia coli Competent Cell Trans1-T1Phage Resistant, TA Cloning Vector Simple were purchased from Beijing Quanshijin Biotechnology Co., Ltd. The overexpression vector pLeela, the YFP fluorescent expression vector pENSG-YFP, and the Agrobacterium tumefaciens strain GV3101+MPK90 were donated by Liu Yongxiu's research group at the Institute of Botany, Chinese Academy of Sciences. Laser confocal fluorescence microscopy was provided by the Beijing Key Laboratory of New Technologies for Agricultural Applications.

[0065] YEB medium (for cultivating Agrobacterium): meat extract (Beef extract) 5g / L peptone (Peptone) 5g / L yeast extract (Yeast extract) 1g / L sucrose (Sucrose) 5g / L, MgSO 4 ·7H 2 O 4g / L, [solid medium plus agar (Agar) 15g / L] adjust the pH to 7.4 with NaOH, and sterilize under high temperature and high pressure.

[0066] Conf...

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Abstract

The invention relates to the technical field of biology, and concretely relates to a syringa oblate chalcone synthase gene. According to the invention, an Unigene sequence fragment of chalcone synthase gene (SoCHS) obtaining by syringa oblate transcriptome sequencing is taken as a template, a SoCHS cDNA full length sequence is obtained; based on bioinformatics analysis on a structure and coding protein, a molecular biology technology is used, an overexpression and YFP positioning expression carrier of the gene is constructed, through agrobacterium mediation, arabidopis thaliana and tobacco are converted, the protein characteristic is analyzed and the function is verified; and a RT-PCR technology is used for analyzing the tissue specific expression mode of the gene. A molecule mechanism formed by SoCHS-participated pattern control is researched and revealed.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a chalcone synthase gene from Lilac chinensis and application thereof. Background technique [0002] Flower color is one of the important ornamental characteristics of garden plants, and flavonoids and their colored derivatives play an extremely important role in the formation of flower color. Chalcone synthase (CHS) is a key enzyme in the metabolic synthesis pathway of flavonoids, which catalyzes the conversion of one molecule of Coumanyl CoA (Coumanyl CoA) and three molecules of malonyl CoA (Malonyl CoA). CoA) generates 4,5,7-trihydroxyflavanone (Narigenin chalcone). In higher plants, flavonoids are secondary metabolites that widely exist and play an important role in the interaction between plants and the environment. They not only play a decisive role in the formation of plant flower colors, but also participate in various physiological processes of plants. Biochemical processe...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/10C12N15/70C12N15/74C12N15/82A01H5/00
CPCC12N9/1037C12N15/8205C12N15/827C12N15/8271C12Y203/01074
Inventor 郑健胡增辉张睿鹂赵亚洲何祥凤冷平生张克中窦德泉关雪莲王羽窦莹鲁仪增
Owner BEIJING UNIV OF AGRI
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