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Camellia oleifera self-incompatibility associated gene, SNP (Single Nucleotide Polymorphism) molecular marker and application

A molecular marker, SNP181968 technology, applied in the field of molecular markers, can solve problems such as affecting the agronomic traits of Camellia oleifera

Active Publication Date: 2021-10-15
HUNAN UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention screened CoMAPK9 from the Camellia oleifera pistil transcriptome database as a unique sequence of Camellia oleifera, and it was specifically expressed in the pistil of Camellia oleifera 24 hours after self-pollination and reached the peak value. Therefore, it is speculated that CoMAPK9 participates in the regulation of the self-incompatibility process of Camellia oleifera, thereby affecting the yield of Camellia oleifera The agronomic traits such as fruit set rate, related research has not been reported so far

Method used

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  • Camellia oleifera self-incompatibility associated gene, SNP (Single Nucleotide Polymorphism) molecular marker and application
  • Camellia oleifera self-incompatibility associated gene, SNP (Single Nucleotide Polymorphism) molecular marker and application
  • Camellia oleifera self-incompatibility associated gene, SNP (Single Nucleotide Polymorphism) molecular marker and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1, total DNA extraction and fruit setting rate calculation of Camellia oleifera seed kernel, root or leaf

[0039] The DNA secure new plant genomic DNA extraction kit (TIANGEN Kit Code No. DP320, spin column type) combined with the improved CTAB method in the laboratory was used to extract the total DNA of a single plant. The specific operation steps are as follows:

[0040] (1) Take 600 μL of 2×CTAB solution in a 1.5mL centrifuge tube, mix well and keep the temperature in an air heater at 65°C for later use;

[0041] (2) quickly take out the camellia oleifera seeds, roots or leaves from the liquid nitrogen, and quickly grind them into fine powder in a pre-cooled mortar filled with liquid nitrogen;

[0042] (3) Transfer the ground powder to (1) the prepared centrifuge tube, mix the powder and the solution on a vortex instrument, and put it in a water bath at 65°C for 10 minutes, and shake it every 2 minutes during this period;

[0043] (4) Add 600 μL of chloro...

Embodiment 2

[0049] Example 2. Three-generation sequencing + next-generation sequencing samples and annotation analysis

[0050] 1. Sequencing sample preparation and collection:

[0051] The self-pollination combination (Self -pollination: SP), cross-pollination combination (Cross-pollination: CP) and non-pollination combination (Non-pollination: NP) ('Joajin' and 'Huaxin' emasculation treatment bagging). Cross-pollination was done manually Pollination is complete. Pick the flower buds of 'Huaxin' and 'Huajin' at the dew stage, gently stroke the anthers with tweezers, spread the anthers in a sulfuric acid paper bag, and fully crack the anthers at 25°C to release mature pollen (Pollen: Pn); store the pollen in a 1.5mL centrifuge tube at -80°C; take the pollen out of the refrigerator and equilibrate it at room temperature for 1 hour before pollination. When pollinating, select flower buds in the white stage (after the flower buds are peeled off, the anthers are not powdered and the pistil i...

Embodiment 3

[0059] Example 3: CoMAPK gene acquisition and SNP site analysis

[0060] 1. Analysis of CoMAPK9 gene characteristics related to self-incompatibility

[0061] Transcript annotation analysis, differential transcripts, and differential protein trend analysis and Pearson correlation analysis were performed, and CoMAPK9 was found to be a unique sequence of Camellia oleifera, which strongly responded to self-pollination and out-pollination. The functional characteristics of CoMAPK9 gene in the process of Camellia oleifera self-incompatibility were studied by sequence analysis, expression pattern analysis, prokaryotic expression, subcellular localization and other experiments.

[0062] Using real-time fluorescent quantitative PCR to analyze the expression of CoMAPK9 in different parts of flowers ( figure 2 A), the expression patterns of 2-84h in the styles of different treatments ( figure 2 B).

[0063] CoMAPK9 protein expression was induced by prokaryotic expression experiments...

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Abstract

The invention discloses a camellia oleifera self-incompatibility associated gene, an SNP marker and application. SNP missense mutation exists at the position of SNP071031 in OFR of the genome sequence of CoMAPK9, a basic group is changed from C to A, SNP missense mutation exists at the position of SNP181968, the basic group is changed from A to G, and the SNP mutations of the two sites have significant correlation with the self-incompatibility of camellia oleifera, which is shown in that the fruit setting rate of the SNP 071031 site genotype CC or CA after pollination is significantly higher than the fruit setting rate of the genotype AA; and after pollination, the fruit setting rate of the genotype AA or AG of the SNP181968 site is obviously higher than that of the genotype GG. The gene SNP combined molecular marker can be used for identifying the phenotypic seedling stage of the fruit setting rate of camellia oleifera, so that the variety configuration selection efficiency of a new camellia oleifera planting land and the yield of camellia oleifera per unit area are effectively improved.

Description

Technical field: [0001] The invention belongs to the technical field of molecular markers, and relates to a new gene associated with the self-incompatibility trait of camellia oleifera, its SNP molecular marker and its application. Background technique: [0002] Self-incompatibility widely exists in flowering plants. Selfing refers to the union of male and female gametes from the same individual or mating between individuals with the same genotype or mating between individuals from the same clonal line. Self-incompatibility refers to the phenomenon that hermaphroditic plants that can produce male and female gametes with normal functions and mature at the same time cannot complete fertilization during self-pollination or cross-pollination of the same genotype. This is an important genetic mechanism for plants to prevent inbreeding and degeneration, and it is of great significance in plant reproductive biology and heterosis utilization. Due to the self-incompatibility of pla...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11C12Q1/6895
CPCC07K14/415C12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/172
Inventor 江南谭晓风周俊琴
Owner HUNAN UNIV OF TECH
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