SNP molecular markers related to content of linolenic acid in camellia seed kernel oil and application of SNP molecular marker

A technology of molecular markers and linolenic acid, which is applied in the field of molecular biology and genetic breeding, can solve problems such as the late start of assisted breeding research and the obstacles to mining key regulatory genes of Camellia oleifera

Active Publication Date: 2020-07-28
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0004] The research on molecular marker-assisted breeding of camellia oleifera started relatively late, mainly focusing on the identification of varieties, the study of genetic diversity of natural populations or hybrid populations, etc., and the research on molecular marker-assisted breeding related to traits, especially economic traits, was rarely involved.
At the same time, because the common Camellia oleifera widely planted in production is hexaploid, the highly complex genetic background has brought great obstacles to the development of markers related to economic traits of Camellia oleifera and the mining of key regulatory genes.

Method used

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  • SNP molecular markers related to content of linolenic acid in camellia seed kernel oil and application of SNP molecular marker
  • SNP molecular markers related to content of linolenic acid in camellia seed kernel oil and application of SNP molecular marker
  • SNP molecular markers related to content of linolenic acid in camellia seed kernel oil and application of SNP molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0223] Example 1 Construction and Character Determination of Linolenic Acid Content Segregation Population in Camellia Camellia Oil

[0224] In this example, common Camellia oleifera resources are used to collect natural populations of 500 germplasm resources in the nursery, and their origins cover most of the main Camellia oleifera producing areas in my country, including Zhejiang Province, Hunan Province, Jiangxi Province, Guangxi District, Fujian Province, and Guangdong Province. Province etc. After the fruit of 500 individuals is fully mature (5% of the fruit is cracked), the seeds are collected respectively, and the oil is extracted to determine the fatty acid composition and content. The operation steps are as follows:

[0225] (1) Appropriate amount of Camellia oleifera seeds were baked in an oven at 80°C overnight until constant weight, and the hard seed coat was peeled off.

[0226] (2) After the seed kernels are pulverized with a grinder, they are wrapped with mediu...

Embodiment 2

[0229] Example 2 Transcriptome sequencing and annotation analysis of the third generation Camellia oleifera

[0230] 1. Extraction of RNA from three-generation sequencing samples

[0231] The roots, young leaves, mature leaves, petals and immature seeds of camellia oleifera "Changlin No. 4" were collected, and RNAprep Pure polysaccharide polyphenol plant total RNA extraction kit (spin column type, TIANGEN kit Code No. DP441) was used to extract respectively RNA, the specific steps are as follows:

[0232] (1) First add 500 μl of Lysis Solution SL (beta-mercaptoethanol was added in advance) to a 1.5ml centrifuge tube. Take 0.1g of the sample material and add liquid nitrogen to fully grind, quickly add the ground powder into the centrifuge tube, and immediately vortex vigorously to mix.

[0233] (2) Centrifuge at 12000 rpm for 2 minutes.

[0234] (3) Transfer the supernatant to the filter column CS (the filter column CS is placed in the collection tube), centrifuge at 12000rp...

Embodiment 3

[0247] Example 3 Seed Kernel Transcriptome Sequencing and Polymorphic Site Identification During High-speed Synthesis of Oil

[0248] 1. Extraction of total RNA from seeds of 500 Camellia oleifera clones during high-speed oil synthesis

[0249] The total RNA of the immature kernels of each clone was extracted by RNAprep Pure Polysaccharide Polyphenol Plant Total RNA Extraction Kit (spin column type, TIANGEN Kit Code No. DP441) (see Example 2).

[0250] 2. Next-generation transcriptome sequencing

[0251] The total RNA of each sample was tested for purity and concentration, and the ribosomal RNA was removed to maximize the retention of all codingRNA and ncRNA. The obtained RNA is randomly fragmented into short fragments, and then the fragmented RNA is used as a template to synthesize the first strand of cDNA with six base random primers (random hexamers); then buffer, dNTPs (dUTP instead of dTTP), RNase H are added The second strand of cDNA was synthesized with DNA polymerase...

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Abstract

The invention provides SNP molecular markers related to content of linolenic acid in camellia seed kernel oil and application of the SNP molecular markers. The molecular markers comprise 47 SNP markers significantly associated with the content of linolenic acid in camellia oil, and these markers are located in 30 transcription bodies of camellia. The positions of the SNP markers are clear, the detection method is convenient and rapid, is not affected by the environment, and has higher purposes, small workload, higher efficiency and low cost. By detection of these SNP sites, identification andassistant screening can be performed at a seeding stage, and the production cost is greatly saved, and the selection efficiency is improved. The molecular markers can improve the selection efficiencyof camellia breeding and accelerate the breeding process when used for camellia breeding with high linolenic acid content.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and genetic breeding, in particular to a SNP molecular marker related to the content of linolenic acid in Camellia oleifera seed oil and its application. Background technique [0002] The rapid development of the industry in recent years has made Camellia oleifera, as the only woody oil plant, one of the four major oil plants (rape, peanut, soybean and camellia oleifera) in China. Camellia oleifera oil has a high content of unsaturated fatty acids and stable oleic acid as the main component. It is a high-grade health-care edible oil with nutritional value comparable to that of olive oil and adapted to the high-temperature cooking habits of Chinese people. Vigorously developing the camellia oleifera industry can not only effectively alleviate the pressure on cultivated land, increase the supply capacity of edible oil in our country, but also effectively improve the dietary structure of o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 林萍常君王开良
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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