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Method for constructing cynodon dactylon core germplasm by utilizing SSR markers

A technology of core germplasm and bermudagrass, applied in the field of plant germplasm resource evaluation

Pending Publication Date: 2019-12-24
TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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  • Abstract
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  • Application Information

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Problems solved by technology

[0003] The Institute of Tropical Crop Variety Resources of the Chinese Academy of Tropical Agricultural Sciences has collected a large number of bermudagrass provenances in recent years, and has studied the collected representative resources in terms of morphology, molecules, stress resistance, flat use value, and feeding value. , however, the existing research generally stays in the use of morphological markers to construct the core germplasm resources of Bermudagrass

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  • Method for constructing cynodon dactylon core germplasm by utilizing SSR markers
  • Method for constructing cynodon dactylon core germplasm by utilizing SSR markers
  • Method for constructing cynodon dactylon core germplasm by utilizing SSR markers

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Effect test

Embodiment 1

[0120] Bermudagrass DNA extraction and detection

[0121] Agarose gel electrophoresis detection was carried out on the DNA extracted from part of the wild bermudagrass (see figure 1 ). Depend on figure 1 It can be seen that the DNA bands extracted in this test are bright and clear, without bands, and the brightness of the sample wells is low, indicating that the DNA content is high, there is no degradation phenomenon, and there are few impurities such as proteins. In addition, the detection results of the nucleic acid protein content analyzer showed that the OD of DNA 260 / OD 280 All between 1.8 and 2.0, the concentration ρ≥300ng / μL. The results of these two tests showed that the quality and concentration of the DNA used met the requirements.

Embodiment 2

[0123] According to the bermudagrass samples, 63 pairs of amplification primers were designed, and by using the SSR-PCR reaction system, 22 pairs of primers with good polymorphism, clear bands and obvious main bands were screened out from the 63 pairs of SSR primers. Some SSR primers (SC01, Screening of SC02, SC03, SC04, SC05, SC06, SC07, SC08, SC09, SC10, SC11, SC12) see appendix figure 2 .

Embodiment 3

[0125] Bermudagrass SSR amplification band statistics

[0126] Table 3 lists the amplification diversity results of 22 pairs of SSR primers for 544 copies of Bermudagrass. A total of 135 sites were amplified by 22 pairs of primers, of which 134 were polymorphic sites. The percentage of polymorphic sites It was 99.26%, and each primer amplified 6.14 loci on average. The primers with the largest number of amplified sites were SC18 and SC12, which amplified 13 and 12 sites, respectively; the primers with the least number of amplified sites were SC51, SC56, and SC58, which amplified 3 sites. The average percentage of polymorphic loci, the average number of observed alleles, the average number of effective alleles, the average Nei's genetic diversity index and the average shannon-weaver index of 22 pairs of primers were 99.26%, 1.99, 1.32, 0.20, and 0.32, respectively . Different primers have different values ​​on each parameter, among which SC51 is the lowest in these parameters...

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Abstract

The invention discloses a method for constructing cynodon dactylon core germplasm by utilizing SSR markers, and belongs to the technical field of plant germplasm resource evaluation. The method comprises the following steps: 1) extracting genome DNA of cynodon dactylon samples of an original group, and performing SSR-PCR amplification to obtain cynodon dactylon SSR molecular marker data; 2) underthe condition of grouping cynodon dactylon samples of an original group, sampling by adopting a multiple clustering priority sampling method based on SSR molecular marker data, wherein the total sampling proportion is 20%, the intra-group sampling proportion is square root proportion, the genetic distance is Nei & Li genetic distance, and the clustering method is used for constructing a cynodon dactylon core germplasm by adopting a gravity center method. The cynodon dactylon core germplasm constructed by the method can effectively retain the genetic structure of the original germplasm, effectively removes redundant materials, and can be used as a representative sample of overall resources.

Description

technical field [0001] The invention relates to the technical field of plant germplasm resources evaluation, and more specifically relates to a method for constructing bermudagrass core germplasm using SSR markers. Background technique [0002] Bermudagrass belongs to the C4 perennial herb of the Poaceae Teff subfamily Sansevieria family. It is one of the three warm-season turfgrasses in the world and is also an excellent pasture grass. Scholars at home and abroad have made some progress in the collection of bermudagrass germplasm resources and the evaluation of genetic diversity. These studies have laid a good foundation for the construction of the core collection of bermudagrass provenance. [0003] The Institute of Tropical Crop Variety Resources of the Chinese Academy of Tropical Agricultural Sciences has collected a large number of bermudagrass provenances in recent years, and has studied the collected representative resources in terms of morphology, molecules, stress r...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11G16B50/30
CPCC12Q1/6895G16B50/30C12Q2600/156
Inventor 黄春琼龙萄王文强张瑜刘国道
Owner TROPICAL CORP STRAIN RESOURCE INST CHINESE ACAD OF TROPICAL AGRI SCI
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