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Construction method for forest-tree genome DNA (deoxyribonucleic acid) methylation inheritance linkage maps

A technology of genetic linkage map and construction method, which is applied in the fields of biochemical equipment and methods, determination/inspection of microorganisms, etc., to achieve the effect of stable construction and high accuracy

Inactive Publication Date: 2017-05-31
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

The construction of epigenetic linkage maps in forest trees has not been reported

Method used

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  • Construction method for forest-tree genome DNA (deoxyribonucleic acid) methylation inheritance linkage maps
  • Construction method for forest-tree genome DNA (deoxyribonucleic acid) methylation inheritance linkage maps
  • Construction method for forest-tree genome DNA (deoxyribonucleic acid) methylation inheritance linkage maps

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Embodiment 1

[0042] 1. The population of hybrid offspring used in this example is Populus alba "YX01" (Populus alba×P.glandulosa) as the female parent, and Populus tomentosa male elite clone "LM50" (P.tomentosa Carr.) as the male parent From the F1 generation families obtained by crossing, 552 plants were randomly selected as experimental materials. The bark (approximately 5 cm × 5 cm) at the DBH of the plant was peeled off with a sharp blade, and small pieces of xylem material were taken out and put into liquid nitrogen for freezing and preservation for the extraction of genomic DNA.

[0043] 2. The modified CTAB method was used to extract the mature xylem genomic DNA of 552 individuals of different genotypes in F1 population of poplar hybrids. Specifically include the following steps:

[0044] 1) Preheating the CTAB extraction buffer containing β-mercaptoethanol in a water bath at 65°C;

[0045] 2) Grinding the xylem material into powder in liquid nitrogen, taking 0.5 g of the powder a...

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Abstract

The invention relates to a construction method for forest-tree genome DNA (deoxyribonucleic acid) methylation inheritance linkage maps, and belongs to the technical field of molecular genetics. The construction method includes the steps: 1) extracting genome DNA of xylem tissues; 2) respectively performing dual-enzyme digestion for the genome DNA by the aid of first group of EcoRI and HpaII enzyme and second group of EcoRI and MspI enzyme, connecting connectors on two groups of dual-enzyme digestion products to obtain two groups of enzyme digestion connecting products; sequentially performing pre-amplification and selective amplification by the aid of connecting amplification products, separating selective amplification products by the aid of capillary electrophoresis to obtain population genome methylation marker sites; 4) selecting population methylation polymorphic sites meeting Mendelian inheritance separation from the population genome methylation marker sites to construct the methylation inheritance linkage maps. The construction method is efficient, stable and easy to operate, so that forest-tree population epigenetics research progress can be accelerated.

Description

technical field [0001] The invention relates to the technical field of molecular genetics, in particular to a method for constructing a forest tree genome DNA methylation genetic linkage map. Background technique [0002] Among the many forms of epigenetic modification, the most common and most studied is DNA methylation (Jaenisch and Bird, 2003), which refers to the use of S-adenosylmethionine as a donor under the catalysis of methyltransferases, A modification in which a methyl group is covalently added to a DNA base (Santi et al., 1983). DNA methylation plays an important role in many biological processes, including regulation of gene expression, gene imprinting, and transposon silencing. Numerous studies focusing on DNA methylation have shown that DNA methylation polymorphisms can also affect phenotypes (Katherine et al., 2013). As early as the 1980s, many studies reported that the main function of cytosine methylation is to act as a repressor of gene expression (Razin...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6806C12Q1/6895C12Q2600/156C12Q2565/125C12Q2525/131C12Q2525/191C12Q2563/107
Inventor 张德强王情世杜庆章
Owner BEIJING FORESTRY UNIVERSITY
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