Method and kit for constructing poplar core collection through micro-satellite DNA molecular marking technology

A technology of DNA molecules and core germplasm, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of not being suitable for the construction of poplar core germplasm, so as to avoid the omission of precious germplasm resources, multiple The effect of rich behavior and high selection accuracy

Inactive Publication Date: 2015-01-21
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

[0006] The present invention aims to provide a method and kit for constructing poplar core germplasm using microsatellite DNA molecular marker technolog

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  • Method and kit for constructing poplar core collection through micro-satellite DNA molecular marking technology
  • Method and kit for constructing poplar core collection through micro-satellite DNA molecular marking technology
  • Method and kit for constructing poplar core collection through micro-satellite DNA molecular marking technology

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Embodiment approach

[0027] According to a typical embodiment of the present invention, the developed SSR markers are used for PCR amplification to establish the genotype database of the original population, and then the UPGMA-clustering diagram is established using PowerMarker software, and the sub-branches containing 2 genotype individuals are deleted Any one of the genotype individuals will be reserved for other sub-branch types (including 1 or no less than 3 genotype individuals) and enter the next round of clustering; according to the above criteria, multiple clustering selections will be made to obtain different proportions of one level core group. For these groups, we use STRUCTURE group structure detection to detect all core groups within the range of 8% to 30%, and finally obtain 8%, 10%, 15%, 20%, 25%, and 5 groups without subgroup structure. For the candidate core germplasm population, further, carry out genetic diversity detection of population molecules and phenotypes, select a candid...

Embodiment 1

[0044] Using microsatellite DNA molecular marker technology "UPGMA-cluster analysis sequential random sampling" and "STRUCTURE model re-testing" joint analysis method to construct the core germplasm population of Populus tomentosa.

[0045] The sample object is the Populus tomentosa germplasm resource gene bank (National Populus tomentosa germplasm resource bank) planted in the state-owned nursery in Guanxian County, Shandong Province in 1984. The gene bank is composed of 1047 excellent clones or wild ecotype individuals of Populus tomentosa collected by the National Populus tomentosa Cooperative Group in the distribution area of ​​1 million square kilometers of Populus tomentosa in the Yellow River Basin in northern my country in the early 1980s.

[0046] Using the above-mentioned primer development method, obtain the genome SSR primer set, and perform PCR amplification and capillary electrophoresis detection on all DNA samples, and use GeneMapper (Applied Biosystems) software...

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Abstract

The invention discloses a method and a kit for constructing a poplar core collection through a micro-satellite DNA molecular marking technology. The method includes following steps: (S1) collecting populus tomentosa germplasm resources; (S2) extracting genomic DNA of the populus tomentosa in the step (S1); (S3) performing PCR amplification on a polymorphism micro-satellite DNA marker developed in a whole genome region of the populus tomentosa; (S4) construction a candidate core collection group, having different scales being 8-30%, through UPGMA-cluster analysis gradually random sampling and an STRCUTURE model re-detection combined analytic method; and (S5) determining 15% of the core collection group through comparison between the candidate core collection group and an original group according to differences in molecular genetic diversity and quantitative character variation level. By means of the method and the kit, on one hand, phenotypic character diversity of the species can be reflected well and precious core collection resources are free from being lost; and on the other hand, a good core collection resource material can be provided for developing molecule-marking directional selective breeding in a later period.

Description

technical field [0001] The invention relates to the field of tree molecular breeding, in particular to a method and a kit for constructing poplar core germplasm using microsatellite DNA molecular marker technology. Background technique [0002] Frankel and Brown first proposed the concept of core collection in 1984. The core germplasm is considered to be a core subset of the original germplasm resources, and the genetic diversity of the morphological characteristics, geographical distribution, genes and genotypes of the entire resource population can be preserved to the greatest extent with the least amount of genetic resources. Therefore, the core collection can be used as an entry point for the research and utilization of germplasm resources, thereby improving the management and utilization of the entire gene bank. [0003] Populus tomentosa is an important timber tree species in northern my country. It is widely distributed and covers an area of ​​one million square kilo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张德强杜庆章巩琛锐
Owner BEIJING FORESTRY UNIVERSITY
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