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A molecular marker closely related to glucosinolate content in rapeseed and its application

A technology of molecular markers and glucosinolates, which is applied in the field of rapeseed breeding and molecular biology, can solve the problems that the glucosinolate content of seeds can only be identified and selected in the later stage, is easily affected by the environment, and the selection efficiency is low, so as to speed up quality improvement and reduce workload. The effect of reducing and speeding up the pace

Inactive Publication Date: 2016-04-20
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The discovery of this molecular marker solves the problems in conventional breeding methods that the glucosinolate content of seeds can only be identified and selected at a later stage, the cycle is long, it is easily affected by the environment, and the selection efficiency is low, and it improves the genetic improvement of rapeseed quality.

Method used

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  • A molecular marker closely related to glucosinolate content in rapeseed and its application
  • A molecular marker closely related to glucosinolate content in rapeseed and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A molecular marker EX043693-231 closely related to the content of glucosinolates in rapeseed is obtained by the following method:

[0028] In this example, taking 101 rapeseed varieties with different genetic sources and large variations in seed glucosinolate content as an example, a natural population was constructed, and the method for obtaining molecular markers related to seed glucosinolate content was described in detail, as follows:

[0029] (1) Group construction:

[0030] 101 rapeseed varieties with different genetic sources and large differences in seed glucosinolate content (0.8-219.2 μmol / g) were used as research materials (purchased from the market or in the national oil crop germplasm bank), in a greenhouse (temperature 20 ° C; light cycle 16h, grow to 1-2 true leaves under the light intensity of 15000lx).

[0031] (2) Determination of glucosinolate content in seeds:

[0032] Using near-infrared spectroscopy, the glucosinolate content of 101 natural popul...

Embodiment 2

[0062] The application of a functional molecular marker closely related to rapeseed glucosinolate content in the quality breeding of rapeseed comprises the following steps:

[0063] Taking the 101 cultivar resources in Example 1 used for association analysis as research materials, the average seed glucosinolate content of this group was 65.3 μmol / g. Using specific PCR markers for identification, the steps are as follows:

[0064] (1) Using rapeseed total DNA as a template;

[0065] (2) Perform PCR amplification using the functional marker EX043693-231, and the primer sequence is:

[0066] Forward primer: 5′-TTGTAATAGAGTTCATATATATCG-3′

[0067] Reverse primer: 5′-TTCATACATCAAATACCAAAC-3′;

[0068] (3) PCR reaction system: the total volume is 10 μl, and the specific components are as follows:

[0069] DNA template (25ng / μl)

1.0μl

Forward primer (50ng / μl)

0.5μl

Reverse primer (50ng / μl)

0.5μl

10×PCR Buffer

1.0μl

dNTPs (10mmo...

Embodiment 3

[0079] The application of a functional molecular marker closely related to rapeseed glucosinolate content in the quality breeding of rapeseed comprises the following steps:

[0080] (1) Using the high-glucosinolate variety 'Zhongyou 821' (bred by the Oil Crops Research Institute of the Chinese Academy of Agricultural The F 2 The isolated population is used as the research material, and the detection range is further expanded by using the marker EX043693-231, and the detection method is the same as that in Example 2.

[0081] (2) It is predicted that the content of glucosinolate in seeds of individual plants with 242-bp specific band pattern will be significantly increased, while the content of glucosinolate in seeds of individual plants with 231-bp specific fragment will be significantly reduced.

[0082] (3) A total of F 2 362 individual plants of the population, the results showed that F 2 There were 115 individual plants with only 231-bp band type in the population, and ...

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Abstract

The invention discloses a molecular marker closely related to the glucosinolate content of rapeseed and its application. The forward primer sequence of the molecular marker EX043693-231 is 5′-TTGTAATAGAGTTCATATATATCG-3′, and the reverse primer sequence is 5′-TTCATACATCAAATACCAAAC- 3'; use the total DNA of Raisin napus strain as a template, and use the primer sequence to perform PCR amplification. After the amplified product is separated by 2.5% agarose gel electrophoresis, a specific strip with a molecular weight of 242-bp or 231-bp is obtained. band; the rapeseed strains with 231-bp specific band had lower seed glucosinolate content, and could be used for further breeding selection. The molecular marker developed by the invention can be used for the identification and prediction of the glucosinolate content of rapeseeds. The operation is simple and the method is easy to implement, which solves the problems in the conventional breeding methods that selection and identification must be carried out at a later stage, and accelerates the improvement of rapeseed quality.

Description

technical field [0001] The invention belongs to the fields of rapeseed breeding and molecular biology, and more specifically relates to a molecular marker closely related to the glucosinolate content of rapeseed, and also relates to a molecular marker closely related to the glucosinolate content of rapeseed. Application in breeding for glycoside content. Background technique [0002] Rapeseed is the most important winter oil crop in my country, with an annual planting area of ​​about 110 million mu, ranking first in the world in both area and total output. After rapeseed is processed into edible vegetable oil, it can produce 6 million tons of rapeseed cake per year. The cake contains about 40% protein. Its amino acid composition is balanced and reasonable, and its nutritional value and digestibility are higher than soybean protein. It is a high-quality protein source. , can be widely used in the feed industry, and is of great significance for improving the commodity value o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 陆光远张学昆邹锡玲赵永国徐劲松程勇瞿桢付桂萍
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI