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DNA fragment related to palmitic acid content in camellia oleifera seed oil, SNP molecular marker closely linked with DNA fragment and use thereof

A technology of molecular markers and palmitic acid, which is applied in the field of DNA fragments and SNP molecular markers, can solve the problems of unsatisfactory breeding speed of improved varieties, long breeding cycle of camellia oleifera, slow breeding of new varieties, etc.

Active Publication Date: 2021-09-24
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

For a long time, Camellia oleifera breeding with selection and hybrid breeding as the main means has made great progress, but the conventional breeding cycle of Camellia oleifera is long, the selection of new varieties is slow, and the speed of breeding of improved varieties cannot meet the needs of industrial development. One of the important factors restricting the development of camellia oleifera industry

Method used

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  • DNA fragment related to palmitic acid content in camellia oleifera seed oil, SNP molecular marker closely linked with DNA fragment and use thereof
  • DNA fragment related to palmitic acid content in camellia oleifera seed oil, SNP molecular marker closely linked with DNA fragment and use thereof
  • DNA fragment related to palmitic acid content in camellia oleifera seed oil, SNP molecular marker closely linked with DNA fragment and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Construction and Character Determination of Palmitic Acid Content Separation Population in Camellia oleifera Seed Oil

[0065] In this example, Changlin No. 53 and Changlin No. 81 were used as female and male parents respectively, and controlled pollination techniques were used to create hybrid F1 populations with widely separated economic traits. The F1 population was designed with 3 repetitions in randomized blocks and was kept in Dongfanghong Forest Farm, Wucheng District, Zhejiang Province. 180 progeny individuals waited for the fruit to fully mature (5% of the fruit cracked), collected seeds, and determined the content of palmitic acid in the seed oil. The operation steps are as follows:

[0066] (1) Take an appropriate amount of Camellia oleifera seeds and bake them in an oven at 80°C overnight until constant weight, and peel off the hard seed coat.

[0067] (2) After the seed kernels are pulverized with a grinder, they are wrapped with medium-speed fi...

Embodiment 2

[0070] Example 2 Camellia oleifera linkage map construction

[0071] 1. Genomic DNA extraction

[0072] The spring young leaves of 180 individuals of Changlin 53×Changlin 81 family and their parents were collected in March, and the total genomic DNA was extracted by KAC method (TaKaRa kit Code No.9768). Specific steps are as follows:

[0073] (1) First, add 500 μl of Buffer HS II to a 1.5ml centrifuge tube; accurately weigh 100 mg of Camellia oleifera young leaves for liquid nitrogen grinding; quickly add the ground powder into the centrifuge tube and mix well, then add 10 μl of RNase A ( 10mg / ml), shake and mix thoroughly, and incubate in a water bath at 56°C for 10 minutes.

[0074] (2) Add 62.5 μl of Buffer KAC and mix well. Place on ice for 5 minutes and centrifuge at 12,000 rpm for 5 minutes. Take the supernatant, add the same volume of Buffer GB as the supernatant, and mix well.

[0075] (3) Place the Spin Column in the Collection Tube, move the solution to the Spin...

Embodiment 3

[0097] Example 3 Mining of palmitic acid content gene loci and linked SNP loci in camellia oleifera seed oil

[0098] The QTL Icimapping software was used for data analysis, and the QTL mapping of palmitic acid content was carried out by using the complete interval mapping method (ICIM). The scan step was set at 1 cM; the probability of marker entry (PIN) was 0.002 (POUT=2*PIN=0.002) by stepwise regression; the LOD value was 2.5. LOD significance thresholds were determined by running 1000 permutation tests. On Camellia oleifera chromosome 11, the gene locus rzs.11-2 of palmitic acid content was located, with a contribution rate of 11.5%, and the SNP molecular marker closely linked to it was Chr11_39972457 (Table 1, figure 1 ). The molecular marker contains a nucleotide sequence whose polymorphism is C / A at position 240 of the sequence shown in SEQ ID NO.1.

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Abstract

The invention relates to the technical field of camellia oleifera molecular markers and genetic breeding, in particular to a DNA fragment related to palmitic acid content in camellia oleifera seed oil, an SNP molecular marker closely linked to the DNA fragment and use thereof. The provided DNA fragment related to palmitic acid content in camellia oleifera seed oil is located at rzs.11-2 of 167.343cM of a camellia oleifera linkage map LG11. The provided SNP molecular marker closely linked with palmitic acid content in camellia oleifera seed oil comprises an SNP molecular marker Chr11_39972457 containing a nucleotide sequence of C / A polymorphism at the 240th site of the sequence shown as SEQ ID NO.1. The molecular marker can explain phenotypic variance of palmitic acid content in 11.5% of camellia oleifera seed oil. By detecting the SNP molecular marker, identification and auxiliary screening can be carried out in a seedling stage, such that production cost is greatly saved and selection efficiency of selective breeding of an oil character of camellia oleifera is improved.

Description

technical field [0001] The invention relates to the technical field of camellia oleifera molecular markers and genetic breeding, in particular to DNA fragments related to palmitic acid content in camellia oleifera seed oil, closely linked SNP molecular markers and applications thereof. Background technique [0002] Camellia oleifera, as one of the four major oilseeds (rape, peanut, soybean and camellia oleifera), is widely planted in subtropical regions, with a planting area of ​​more than 65 million mu. Camellia oleifera seed oil has an unsaturated fatty acid content of more than 90%, and is rich in nutrients such as squalene and vitamin E. It is a high-quality edible oil and is called "Oriental olive oil". Breeding high-yield (oil) and high-quality Camellia oleifera varieties has always been the basis and guarantee for the healthy development of the Camellia oleifera industry. For a long time, Camellia oleifera breeding with selection and hybrid breeding as the main means...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11A01H1/04
CPCC12Q1/6895C12Q1/6858A01H1/04C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 常君林萍王开良姚小华
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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