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Screening method and applications of functional marker of carotenoid accumulated gene Br-or in orange-heading Chinese cabbages

A carotene and Chinese cabbage technology, which is applied in the fields of vegetable breeding and molecular genetics, can solve the problems of not obtaining orange leaf ball candidate genes, difficult early selection of orange leaf ball characters, etc., so as to improve the breeding efficiency, shorten the breeding period, good repeatability

Inactive Publication Date: 2013-12-25
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the molecular markers developed by the above researchers are far away from the carotenoid accumulation genes of orange Chinese cabbage, and it is difficult to perform accurate early selection on the traits of orange leaf bulbs, and none of them have obtained candidate genes for orange leaf bulbs.

Method used

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  • Screening method and applications of functional marker of carotenoid accumulated gene Br-or in orange-heading Chinese cabbages
  • Screening method and applications of functional marker of carotenoid accumulated gene Br-or in orange-heading Chinese cabbages
  • Screening method and applications of functional marker of carotenoid accumulated gene Br-or in orange-heading Chinese cabbages

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Early color identification of orange and white Chinese cabbage variety resources using functional markers

[0053] (1) Genomic DNA of orange and white Chinese cabbage seedlings was extracted by CTAB method, and the extraction steps were as follows:

[0054] A. Take 0.2g of fresh young leaves with the main veins removed, stuff them into a 1.5mL centrifuge tube, put them into liquid nitrogen for quick freezing, and quickly grind them to powder with a clean plastic pestle cleaned with 75% alcohol;

[0055] B. Add 700 μL of CTAB extract (CTAB 2%, Tris-HCl (PH8.0) 100 mmol / L, EDTA 20 mmol / L, NaCl 1.4 mol / L) preheated at 65 °C to the centrifuge tube, and then add 8 μL β-mercaptoethanol , mix quickly;

[0056] C. Then put the centrifuge tube into a 65°C water bath, shake it every 5 minutes, and keep the water bath for 30 minutes;

[0057] D. Take out the centrifuge tube, add an equal volume of phenol: chloroform: isoamyl alcohol mixture (phenol: chloroform: isoamyl ...

Embodiment 2

[0075] Example 2: Using functional markers to identify the color of leaf bulbs in an isolated population

[0076] (1) Plant the bulb color separation population Q67, and number the individual plants, and observe the color of the leaf bulbs of individual plants at the end of bulb formation;

[0077] (2) Using the CTAB method to extract the genomic DNA of each individual plant of the isolated population Q67, the extraction steps are as follows:

[0078] A. Take 0.2g of fresh young leaves with the main veins removed, stuff them into a 1.5mL centrifuge tube, put them into liquid nitrogen for quick freezing, and grind them to powder with a clean plastic pestle washed with 75% alcohol by mass fraction;

[0079] B. Add 700 μL of CTAB extract (CTAB 2%, Tris-HCl (PH8.0) 100 mmol / L, EDTA 20 mmol / L, NaCl 1.4 mol / L) preheated at 65 °C to the centrifuge tube, and then add 8 μL β-mercaptoethanol , mix quickly;

[0080] C. Then put the centrifuge tube into a 65°C water bath, shake it every...

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Abstract

The invention discloses screening method and applications of a functional marker of carotenoid accumulated gene Br-or in orange-heading Chinese cabbages. The obtained carotenoid accumulated gene Br-or in orange-heading Chinese cabbages is allele of gene of coded carotenoid isomerase Br-crtiso on cabbage chromosome 1. By comparing the sequences of the gene in orange-heading Chinese cabbages and Chinese cabbages, the fact that the promoter region of the gene in orange-heading Chinese cabbages has the deficiency of 86bp is discovered, therefore, an InDel marker Br-pro is designed according to the structure difference of two promoters, the InDel marker is co-separated from the carotenoid accumulated gene Br-or and allelic gene thereof in orange-heading Chinese cabbage, and is a first nutrition quality type functional marker related to accumulation of carotenoids in the orange-heading Chinese cabbages. Materials with orange gene Br-or can be fast and accurately screened and identified from massive Chinese cabbage germplasm resources by utilizing the functional marker, and the orange gene can be transferred into common Chinese cabbage varieties with targets, and the breeding of high-quality orange-heading Chinese cabbages can be greatly accelerated.

Description

technical field [0001] The invention belongs to the field of vegetable breeding and molecular genetics, and specifically relates to the screening and application of a Br-or functional marker of an orange Chinese cabbage carotenoid accumulation gene. Background technique [0002] Orange leaf Chinese cabbage is very popular because of its unique color, high nutritional value and delicious taste, and the price of orange leaf Chinese cabbage in the market is much higher than that of ordinary white leaf Chinese cabbage. Cabbage has great market potential and economic value. Therefore, the breeding of orange leafy Chinese cabbage has become one of the important contents of modern Chinese cabbage breeding, but the color of orange Chinese cabbage leafy heads cannot be fully expressed until the end of its heading, and it has to be observed in the field one by one, which is time-consuming and laborious. , greatly delaying the breeding process. Therefore, in order to speed up the bre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 张鲁刚张俊祥李会霞
Owner NORTHWEST A & F UNIV
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