Screening method and application of masson pine high-fat-yield functional SNP (Single Nucleotide Polymorphism) marker
A screening method and masson pine technology, applied in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc., can solve the problems of high R&D costs, consumption, large R&D costs, etc., to reduce R&D costs and simplify The effect of technical process and high development efficiency
Active Publication Date: 2022-03-29
GUANGDONG ACAD OF FORESTRY
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Problems solved by technology
[0003]
Because the SNP development technology needs to sequence each individual in the population, it usually requires a lot of research and development costs
However, the SNP sites developed by enzyme-based simplified genome sequencing are usually located in non-coding regions, which is not conducive to mining functional genes
Whole-genome resequencing is billed according to the genome size and sequencing depth. In the face of species with large genomes (usually more than 20GB) such as Masson pine, the R&D costs are too high and do not have wide applicability.
In addition, the associated markers obtained by using the GWAS method usually need to be verified by the population PCR method, and the false positive rate of the associated markers obtained is high
Method used
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Embodiment 1
[0074] (1) Technical process
[0075] The present invention mainly uses multi-omics to screen candidate genes, then uses PCR to develop SNP sites for candidate genes, and obtains functional markers through population verification. The technical flow chart is shown in figure 1 .
[0076] (2) Select materials based on phenotype
[0077] Using 150 clone germplasm resources of Pine massoniana collected and preserved in the Pine massoniana seed orchard (150 clones in total, 5-20 individual plants for each clone, these single plants are equivalent to biological repetitions; these experimental materials were all Resin-yielding capacity (RYC) calculations can be carried out for the calculation of resin-yielding capacity (RYC), and resin-yielding capacity (RYC) can be calculated for each tree age (generally Masson pine is 10 years old or the diameter at breast height reaches 16cm or more). , what this experiment used was the data measured by the 29-year-old Pinus massoniana. Although...
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The invention discloses a screening method and application of a masson pine high-fat-yield functional SNP (Single Nucleotide Polymorphism) marker. The method comprises the following steps: collecting turpentine of clone pinus massoniana, respectively measuring the fat production capacity of the turpentine, collecting tissue materials from clone pinus massoniana samples with high, medium and low fat production capacities, extracting RNA (Ribonucleic Acid), then carrying out PacBio full-length transcriptome sequencing, simultaneously carrying out second-generation transcriptome, metabolome and proteome sequencing, carrying out differential expression analysis and screening candidate genes, and carrying out high-yield detection on the turpentine of the clone pinus massoniana. Therefore, the SNP functional marker is developed. According to the method, only transcriptome sequencing and other multi-omics sequencing need to be carried out on a small number of samples, high SNP site development efficiency is achieved, development cost is low, and the researched and developed SNP marker is beneficial to early selection of the pinus massoniana fat production capacity character and can be used in the field of molecular assisted selection breeding.
Description
technical field [0001] The invention belongs to the technical field of forestry, and in particular relates to a screening method and application of massoniana pine high lipid-yielding functional SNP markers. Background technique [0002] The development of functional markers for target traits is a common method for plant molecular assisted breeding, and it is also an important means to achieve early selection of superior individuals. Now commonly used SNP development techniques include: enzyme-based simplified genome sequencing (SLAF-seq, GBS, RAD, etc.), whole genome resequencing (genome re-sequencing), etc. These methods usually need to collect a large amount of population genetic resources first, and after extracting genomic DNA, sequence all individual DNA of the population by enzyme digestion or random fragmentation, and develop population SNP molecular markers through sequence comparison. Subsequently, the use of GWAS methods to develop target trait-associated markers...
Claims
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Login to View More IPC IPC(8): G16B20/20G16B20/30G16B30/10C12Q1/6895C12N15/11
CPCG16B20/20G16B20/30G16B30/10C12Q1/6895C12Q2600/156C12Q2600/13
Inventor白青松何波祥汪迎利连辉明陈杰连
OwnerGUANGDONG ACAD OF FORESTRY