SNP molecular marker related to palmitoleic acid content in camellia oleifera seed grease and application thereof

A palmitoleic acid, molecular marker technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as source limitation

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

At present, palm oleic acid is mainly derived from seafood such as fish

Method used

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  • SNP molecular marker related to palmitoleic acid content in camellia oleifera seed grease and application thereof
  • SNP molecular marker related to palmitoleic acid content in camellia oleifera seed grease and application thereof
  • SNP molecular marker related to palmitoleic acid content in camellia oleifera seed grease and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Construction and character determination of the isolated population of palmitoleic acid content in Camellia oleifera seed oil

[0060] In this example, the natural population of 500 germplasm resources in the garden is collected using common camellia oleifera resources, and its origin covers most of the main producing areas of camellia oleifera in China, including Zhejiang Province, Hunan Province, Jiangxi Province, Guangxi District, Fujian Province, Guangdong Province Province, etc. After the fruits of 500 individuals are fully mature (5% of the fruits are 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:

[0061] (1) Bake an appropriate amount of Camellia oleifera seeds in an oven at 80°C overnight to constant weight, and peel off the hard seed coat.

[0062] (2) After pulverizing the seed kernels with a pulverizer, wrap them with medium-speed fil...

Embodiment 2

[0065] Example 2 Transcriptome sequencing and annotation analysis of the third generation of Camellia oleifera

[0066] 1. Extraction of RNA from third-generation sequencing samples:

[0067] The roots, young leaves, mature leaves, petals and immature seeds of Camellia oleifera "Changlin No. 4" were collected and extracted using RNAprep Pure polysaccharide and polyphenol plant total RNA extraction kit (spin column type, TIANGEN kit Code No.DP441). RNA, the specific steps are as follows:

[0068] (1) First, add 500 μl of Lysate SL to a 1.5 ml centrifuge tube (check whether β-mercaptoethanol has been added before use). Take 0.1g of the sample material and add it to liquid nitrogen to fully grind it, quickly add the ground sample powder to the centrifuge tube, and immediately vortex vigorously to mix.

[0069] (2) Centrifuge at 12000 rpm for 2 minutes.

[0070] (3) Transfer the supernatant to the filter column CS (the filter column CS is placed in the collection tube), centrif...

Embodiment 3

[0082] Example 3 Seed Kernel Transcriptome Sequencing and Polymorphic Site Identification During the High-speed Synthesis of Oil

[0083] 1. Extraction of total RNA from seeds and kernels of 500 Camellia oleifera clones during the high-speed synthesis of oils and fats:

[0084] Using RNAprep Pure polysaccharide and polyphenol plant total RNA extraction kit (spin column type, TIANGEN kit Code No. DP441), the total RNA of immature seed kernels of each clone was extracted respectively (see Example 2).

[0085] 2. Second-generation transcriptome sequencing:

[0086] Ribosomal RNA was removed from the total RNA of kernel samples tested for purity and concentration to maximize the retention of all coding RNAs and ncRNAs. The obtained RNA is randomly broken into short fragments, and the fragmented RNA is used as a template to synthesize the first strand of cDNA with random hexamers; then add buffer, dNTPs (dUTP instead of dTTP), RNase H The second strand of cDNA was synthesized wit...

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Abstract

The invention relates to the technical field of molecular markers, in particular to an SNP molecular marker related to palmitoleic acid content in camellia oleifera seed grease and application thereof. The invention provides two SNP molecular markers PB.10070.2-202 and PB.7527.1-77 related to palmitoleic acid content in camellia oleifera seed grease. The PB.10070.2-202 contains a nucleotide sequence with polymorphism of G/A at the 202nd site of the sequence shown as SEQ ID NO.1; the PB.7527.1-77 contains a nucleotide sequence with polymorphism of G/A at the 77th site of the sequence shown as SEQ ID NO.2. The two markers are used for identifying the palmitoleic acid content phenotype 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, in particular to a SNP molecular marker related to palmitoleic acid content in Camellia oleifera seed oil and its application. Background technique [0002] Camellia oleifera Abel. belongs to the Camellia family (Theaceae), Camellia L., and is a woody oil tree species. Camellia oil is rich in nutrients and is a high-quality edible oil. Its unsaturated fatty acid content is over 90%, mainly oleic acid and linoleic acid. Camellia seed oil has anti-oxidant, anti-tumor, lowering blood lipid and other effects, and has high nutritional and health care 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 studies on Camellia oleifera breeding for the purpose of improving the quality of Camellia oleifera seed oil. The conventional breeding cycle...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13Y02P60/87
Inventor 王开良林萍姚小华
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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