SSR marker primer set for identifying germplasm resource genetic relationship of halogeton glomeratus and application of SSR marker primer set

A technology for labeling primers and kinship relationships, applied in the field of plant biology, can solve the problems of germination characteristics, growth characteristics, salt enrichment characteristics of plants, differences in growth periods, and it is difficult to effectively distinguish the ecotypes of halophytes in different regions, and achieve stability. Good, abundant, high polymorphism effect

Active Publication Date: 2020-09-11
GANSU AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Problems existing in the prior art: Due to factors such as the genetic basis of various germplasm materials of halophytic grass, differences in growth environments, and geographical isolation, the germplasm characteristics, growth characteristics, and plant richness of halophytic grass germplasms in different regions are under salt stress. There are different degrees of differences in characteristics such as salt content and growth period
It is difficult to effectively distinguish the ecotypes of halophytic grass in different regions only based on the morphological characteristics of halophytic grass germplasm, which has become an urgent problem to be solved in the process of germplasm screening, introduction and germplasm purification of halophytic grass

Method used

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  • SSR marker primer set for identifying germplasm resource genetic relationship of halogeton glomeratus and application of SSR marker primer set
  • SSR marker primer set for identifying germplasm resource genetic relationship of halogeton glomeratus and application of SSR marker primer set
  • SSR marker primer set for identifying germplasm resource genetic relationship of halogeton glomeratus and application of SSR marker primer set

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Experimental program
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Effect test

Embodiment 1

[0043] This embodiment provides a set of SSR marker primers for identifying the genetic relationship of halophyte germplasm resources, including 33 pairs of primers with polymorphisms, and the primer sequences are shown in the sequence table SEQ ID 1 to SEQ ID 66 and the following table , the annealing temperature is 58-60 °C, and the number of repetitions is between 12-30.

[0044]

[0045]

Embodiment 2

[0047] This embodiment provides a method for identifying the kinship of halophyte germplasm resources using the SSR marker primer set described in Example 1, including the following main steps:

[0048] 1. DNA extraction. Take the sample to be identified, and use the seeds or seedlings to extract the total plant DNA.

[0049] 2. PCR amplification. Using the total DNA obtained in step (1) as a template, PCR amplification was carried out with the above-mentioned 33 pairs of SSR marker primer sets for identifying the genetic relationship of halophyte germplasm resources.

[0050] 3. Electrophoresis detection. The PCR amplification products were detected by denaturing polyacrylamide gel electrophoresis, and stable and clear PCR amplification products were obtained.

[0051] 4. Data statistics. The PCR amplification products obtained in step (3) are counted, and the SSR analysis results are recorded in binary and genotype. , 2, 3, etc. represent different genotypes.

[0052] ...

Embodiment 3

[0060] This example provides the application of the SSR marker primer set design method described in Example 1 and the identification of the affinity of halophyte germplasm resources, including:

[0061] 1. Sample collection and DNA extraction

[0062] Before and after December in winter, the seeds of different halophyte ecological types were collected from 18 sites in the arid area of ​​central Gansu and Hexi Corridor (attached). figure 1 ), using the seeds or seedlings cultivated in flowerpots for 1 month as materials, using the CTAB method to extract the genomic DNA of halophyte, and using a micro-ultraviolet spectrophotometer to detect the DNA concentration and purity, the concentration is 300-500ug / mL, The absorbance value OD260 / OD280 is between 1.7-1.9.

[0063] 2. Screening of primers for SSR polymorphism

[0064] The SSR primers in the halophyte have not yet been developed, so the halophyte three-generation full-length transcriptome was sequenced (https: / / www.ncbi.nl...

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Abstract

The invention discloses an SSR marker based primer set for identifying germplasm resource genetic relationship of halogeton glomeratus. A total of 33 pairs of SSR polymorphic molecular markers are identified, an accurate and efficient method for identifying the genetic structure of the halophyte germplasm from different sources is established, and reference can be provided for identifying the halophyte germplasm from different sources. The method for identifying the halophyte germplasm resources comprises the steps of halogeton glomeratus DNA extraction, SSR primer amplification, polymorphismscreening, classification of halogeton glomeratus materials according to amplification results and the like. The germplasm resource genetic relationship of the halogeton glomeratus can be identified accurately and efficiently.

Description

technical field [0001] The invention relates to an SSR marker primer set for identifying the genetic relationship of halophyte germplasm resources and its application, and belongs to the field of plant biotechnology. Background technique [0002] Halogeton glomeratus is an annual herbaceous salt-tolerant plant of Chenopodiaceae. It has excellent characteristics of drought resistance and salt tolerance. There is growth, and the ecological types of germplasm resources are rich. From the morphological point of view, the most notable feature of halophyte plants is that the leaves are succulent, which not only has the characteristics of enriching salt, but also has the characteristics of enriching heavy metal ions. Halophyte Halogeton glomeratus, a promising candidate for phytoremediation of heavy metal-contaminated saline soils [J]. Plant and Soil, 2019: 323-331). [0003] Problems existing in the prior art: due to factors such as the genetic basis of various halophyte materia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12Q1/6806C12N15/11G16B40/10G16B40/00
CPCC12Q1/6895C12Q1/6858C12Q1/6806G16B40/10G16B40/00C12Q2600/156C12Q2531/113C12Q2565/125Y02A40/22
Inventor 王化俊汪军成姚立蓉何建军孟亚雄李葆春马小乐司二静杨轲马增科黄志磊
Owner GANSU AGRI UNIV
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