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Identification method and identification kit for different species of Populus species

A kind of kinship and identification method technology, which is applied in the field of identification methods and identification kits for different kinship relationships among Populus sects, can solve the problems of poor stability, complex identification methods, etc., and achieves the effect of rapid method and easy automation.

Active Publication Date: 2016-10-05
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

[0005] The present invention aims to provide an identification method and an identification kit for the relationship between different species of Populus species, so as to solve the technical problems of complex and poor stability in the individual identification methods among poplar species in the prior art

Method used

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  • Identification method and identification kit for different species of Populus species
  • Identification method and identification kit for different species of Populus species
  • Identification method and identification kit for different species of Populus species

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Experimental program
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Embodiment 1

[0038] S1, using the 5 pairs of polymorphic microsatellite DNA primer combinations (as shown in Table 1) of the present invention to determine the genotype individuals of Populus tomentosa (taken from the National Populus tomentosa germplasm resource bank) of 25 common species of the five poplar schools in my country Perform genotyping and construct microsatellite DNA fingerprints of interspecies individuals, such as figure 2 shown. After the fingerprint is made, it can be used for follow-up experiments without having to make it again every time.

[0039] in, figure 1 Shown is the use of Multiplex Manager software to design primer combinations based on fragment length; figure 2 The fragment lengths of the amplified products are shown to construct the microsatellite DNA fingerprints of different species of Populus genus, and the sizes of the amplified fragments of the five pairs of primers in the primer set correspond in sequence from low to high figure 1 Populus SSR Primer...

Embodiment 2

[0048] Using microsatellite DNA molecular marker technology to identify the male parent of the hybrid progeny of Populus silver glandularis.

[0049] Three adult female plants of Populus silver gland were selected from the poplar mixed forest, and the capsules were collected, and all 10 adult male plants near the female plants were collected at the same time. The seeds in the capsules were propagated by tissue culture. After 3 months, the tissue cultured seedlings were transplanted into nutrition cups, hardened in the artificial climate chamber, and then transferred to the greenhouse seedbed for unified maintenance. Collect 2-3 pieces of young leaves from a single plant.

[0050] The DNA of all parents was extracted by CTAB method, and 20 progeny were selected from each female plant to extract DNA, totaling 73 plants. The DNA was diluted to 10 ng / μl as a template for multiplex PCR.

[0051] All DNA samples were amplified using the previously described microsatellite DNA mark...

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Abstract

An identification method and an identification kit for genetic relationships among different species among sections of populus are disclosed. The method includes following steps of: S1) subjecting populus to genetic typing by adoption of polymorphic microsatellite DNA primer pairs, and constructing microsatellite DNA finger-prints of individuals in the species; and S2) subjecting populus to be identified to genotype amplification by adoption of the polymorphic microsatellite DNA primer pairs, and determining the genetic relationships of the populus to be identified according to the microsatellite DNA finger-prints, wherein the polymorphic microsatellite DNA primer pairs comprise a primer pair 1, a primer pair 2, a primer pair 3, a primer pair 4 and a primer pair 5. By application of the technical scheme, the microsatellite DNA finger-prints are constructed by utilization of the five pairs of polymorphic microsatellite DNA primers which are high in polymorphism and free of interaction between each two, and then genetic relationships of the populus among sections are determined through the microsatellite DNA finger-prints. The method is rapid, efficient, stable and suitable for different growth periods of the populus.

Description

technical field [0001] The invention relates to the technical field of molecular genetics, in particular to an identification method and an identification kit for the genetic relationship among different species of Populus sects. Background technique [0002] Poplar (Populus) is an important forest tree species in the northern hemisphere. It is rich in species, widely distributed, fast-growing and high-yield, and plays a pivotal role in industrial production and environmental protection. my country is located in the central distribution area of ​​poplar in the world. According to the statistics of Xu Weiying (1988), there are as many as 53 unique native poplar species in my country. Therefore, rich genetic variation resources lay a solid foundation for poplar genetic improvement and population evolutionary genetics research. It has established the foundation, and it also poses a challenge for us to better protect and utilize it. However, the phenomenon of homonym, synonym, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 张德强杜庆章潘炜
Owner BEIJING FORESTRY UNIVERSITY
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