SNP molecular markers related to stearic acid content in camellia oleifera seed oil and applications of SNP molecular markers

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

Active Publication Date: 2020-09-25
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
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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 var

Method used

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  • SNP molecular markers related to stearic acid content in camellia oleifera seed oil and applications of SNP molecular markers
  • SNP molecular markers related to stearic acid content in camellia oleifera seed oil and applications of SNP molecular markers
  • SNP molecular markers related to stearic acid content in camellia oleifera seed oil and applications of SNP molecular markers

Examples

Experimental program
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Effect test

Embodiment 1

[0061] Example 1 Construction and Character Determination of Stearic Acid Content Segregation Population in Camellia Camellia Oil

[0062] 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 500 individual fruits were fully mature (5% of the fruits were cracked), the seeds were collected respectively, and the oil was extracted to determine the composition and content of fatty acids. The operation steps are as follows:

[0063] (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.

[0064] (2) After the seed kernels are pulverized with a grinder, they are wrapped with m...

Embodiment 2

[0067] Example 2 Transcriptome sequencing and annotation analysis of the third generation Camellia oleifera

[0068] 1. Extraction of RNA from three-generation sequencing samples:

[0069] 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:

[0070] (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.

[0071] (2) Centrifuge at 12000 rpm for 2 minutes.

[0072] (3) Transfer the supernatant to the filter column CS (the filter column CS is placed in the collection tube), centrif...

Embodiment 3

[0085] Example 3 Seed Kernel Transcriptome Sequencing and Polymorphic Site Identification During High-speed Synthesis of Oil

[0086] 1. Total RNA extraction of 500 Camellia oleifera clones during the high-speed oil synthesis period:

[0087] 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).

[0088] 2. Second-generation transcriptome sequencing:

[0089] The total RNA of each sample tested for purity and concentration was removed from the ribosomal RNA 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 polymera...

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Abstract

The invention relates to the technical field of molecular markers, and specifically relates to SNP molecular markers related to stearic acid content in camellia oleifera seed oil and applications of the SNP molecular markers. Two SNP molecular markers PB.35271.1-548 and PB.35271.1-555 related to the stearic acid content in camellia oleifera seed oil are provided; the SNP molecular marker PB.35271.1-548 contains the nucleotide sequence shown as SEQ ID NO.1 and of which polymorphism is C/T at the position 548th site; and the SNP molecular marker PB.35271.1-555 contains the nucleotide sequence shown as the SEQ ID NO.1 and of which polymorphism is G/A at the position 555th site. Through the utilization of the two markers to identify the stearic acid content phenotype of camellia oleifera, theidentification and auxiliary screening of seedling stages can be realized, the selection efficiency of camellia oleifera breeding can be effectively enhanced, and breeding process can be accelerated.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to SNP molecular markers related to stearic acid content in camellia oleifera seed oil and applications thereof. Background technique [0002] Camellia oleifera Abel., belonging to the genus Camellia L. of the family Theaceae, is an important 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 Camellia oleifera seed oil. The conventional ...

Claims

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Application Information

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