SNP molecular marker related to oleic acid and linoleic acid content in camellia oleifera seed oil and application thereof

A technology of linoleic acid content and molecular markers, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problem that the speed of breeding of improved varieties cannot meet the needs of industrial development, and the breeding of new varieties is slow , long breeding cycle and other issues

Active Publication Date: 2020-08-07
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The conventional breeding cycle of camellia oleifera is very long, and the breeding of new varieties is slow. Therefore, the speed of breeding of improved varieties cannot meet the needs of industrial development, which has become one of the important factors restricting the development of camellia oleifera industry

Method used

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  • SNP molecular marker related to oleic acid and linoleic acid content in camellia oleifera seed oil and application thereof
  • SNP molecular marker related to oleic acid and linoleic acid content in camellia oleifera seed oil and application thereof
  • SNP molecular marker related to oleic acid and linoleic acid content in camellia oleifera seed oil and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Construction and trait determination of oleic acid and linoleic acid content segregated populations in Camellia oleifera seed oil

[0064] In this embodiment, 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 producing areas of Camellia oleifera in China, including Zhejiang Province, Hunan Province, Jiangxi Province, Guangxi District, Fujian Province, and Guangdong Province. Province etc. After the fruits of 500 individuals are fully ripe (5% fruit cracking), the seeds are collected respectively, and the oil is extracted to measure the fatty acid composition and content. The operation steps are as follows:

[0065] (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.

[0066] (2) After the seed kernels are pulverized with a grinder, they are wrapped with ...

Embodiment 2

[0069] Example 2 Transcriptome sequencing and annotation analysis of the third generation Camellia oleifera

[0070] 1. Extraction of RNA from three-generation sequencing samples:

[0071] 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:

[0072] (1) First add 500 μl of lysate SL into a 1.5ml centrifuge tube (check whether β-mercaptoethanol has been added before use). Take 0.1g of sample material and add liquid nitrogen to fully grind, quickly add the ground sample powder into the centrifuge tube, and immediately vortex vigorously to mix.

[0073] (2) Centrifuge at 12000 rpm for 2 minutes.

[0074] (3) Transfer the supernatant to the filter column CS (the filter column CS is placed in the collection tube), centrif...

Embodiment 3

[0086]Example 3 Seed Kernel Transcriptome Sequencing and Polymorphic Site Identification During High-speed Synthesis of Oil

[0087] 1. Total RNA extraction of 500 Camellia oleifera clones during the high-speed oil synthesis period:

[0088] The total RNA of the immature kernels of each clone was extracted by RNAprep Pure Polysaccharide and Polyphenol Plant Total RNA Extraction Kit (spin column type, TIANGEN Kit Code No. DP441) (see Example 2).

[0089] 2. Second-generation transcriptome sequencing:

[0090] For the total RNA of each sample tested for purity and concentration, the ribosomal RNA was removed to maximize the retention of all coding RNA 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 polymeras...

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Abstract

The invention relates to the technical field of molecular markers, in particular to an SNP molecular marker related to oleic acid and linoleic acid content in camellia oleifera seed oil and application thereof. The invention provides three SNP molecular markers PB.34365.1-621, PB.8541.2-812 and PB.35271.1-555 which are related to the content of oleic acid and linoleic acid in camellia oleifera seed oil. The SNP molecular markers respectively contain nucleotide sequences with polymorphisms of C / T, T / C and G / A at the 621st, 812th and 555th sites of the sequences shown as SEQ ID NO.1-3. The threemolecular markers are used for identifying the content phenotypes of oleic acid and linoleic acid of camellia oleifera, seedling stage identification and auxiliary screening can be achieved, the selection efficiency of camellia oleifera breeding is effectively improved, and the breeding process is accelerated.

Description

technical field [0001] The invention relates to the technical field of molecular markers. It specifically relates to SNP molecular markers related to the content of oleic acid and linoleic acid in Camellia oleifera seed oil and its application. Background technique [0002] Camellia oleifera Abel. belongs to the genus Camellia (Camellia L.) of the family Theaceae (Theaceae), and is a woody oil tree species. Camellia oleifera seed oil is rich in nutrients and is a high-quality edible oil. Its unsaturated fatty acid content is more than 90%, mainly oleic acid and linoleic acid. Camellia oleifera seed oil has anti-oxidation, anti-tumor, hypolipidemic and other effects, and has high nutritional and health value. At present, important progress has been made in Camellia oleifera breeding with selection and cross breeding as the main means and fruit yield as the main breeding purpose, but there are still few researches on breeding for the purpose of improving the quality of Camel...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/172
Inventor 林萍姚小华王开良
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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