A method and application of genome ssr molecular marker development of sugarcane cultivars

A cultivar and genome technology, applied in biochemical equipment and methods, recombinant DNA technology, DNA/RNA fragments, etc., can solve the problems of backward construction of sugarcane SSR molecular marker genetic linkage map, limited number of SSR molecular markers, and difficult development, etc. To achieve the effect of promoting the improvement of sugarcane genetics and breeding level, enriching polymorphism information, and protecting legitimate rights and interests

Inactive Publication Date: 2020-11-17
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

However, compared with model crops such as other grass crops, the development of sugarcane SSR molecular markers and the construction of genetic linkage maps are relatively backward, and there are relatively few related reports at home and abroad.
Compared with other grass crops, sugarcane has developed SSR markers with a small number of markers and low polymorphism, which cannot meet the requirements of sugarcane molecular marker-assisted breeding and genetic mapping.
In recent years, the application of SSR molecular markers in sugarcane cultivars has also been gradually carried out, but the number of SSR molecular markers publicly available for sugarcane cultivars is currently limited. Molecular markers are not available
However, the traditional SSR marker development method has many shortcomings, which consumes manpower, material resources, and low efficiency, especially for polyploid sugarcane, the development is more difficult

Method used

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  • A method and application of genome ssr molecular marker development of sugarcane cultivars
  • A method and application of genome ssr molecular marker development of sugarcane cultivars
  • A method and application of genome ssr molecular marker development of sugarcane cultivars

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Experimental program
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Effect test

Embodiment 1

[0022]A total of 4,460 sequences were downloaded from the Sugarcane Genome Center of the French Agricultural Research Institute (http: / / sugarcane-genome.cirad.fr) using the above method, 27,241 SSR sites were found, and 22,932 pairs of SSR primers were successfully designed. The specific operations are as follows :

[0023] From 22,932 pairs of designed primers, search for SSR primers with TG and CA motif types, randomly select 50 pairs, and use 4 Saccharum materials (cultivar R570, cultivar ROC1, tropical species LA purple and cut hands) species SES208) for amplification efficiency verification (see Table 1). The amplification results showed that: a total of 45 pairs of primers could amplify clear amplification bands, the remaining 5 pairs of primers had no amplification bands or the amount of amplified products was weak, and another 35 pairs of primers showed multiple The polymorphic rate was 70% (35 / 50), among which there were 28 pairs of primers of TG repeat type and 7 pa...

Embodiment 2

[0027] In order to further verify the polymorphism of the SSR primer pairs identified in this study, 20 pairs of SSR primers with high polymorphism were selected from the 35 pairs of primers screened above, and the 18 backbone parents in my country (whose blood relationship comes from Tropical species, 2-4 species of sarcasm, large-stem wild and Indian species) (see Table 2), 2 sugarcane ancestors (sarcasm SES 208 and tropical species LA purple) and 4 world Genetic diversity analysis and SSR primer polymorphism evaluation of important sugarcane cultivars (LCP85-384, R570, ROC 16 and ROC 22). The results showed that: 20 pairs of primers showed obvious polymorphisms on 24 sugarcane experimental materials, 95 alleles were amplified in total, 1-7 alleles were amplified by each pair, and each pair of primers amplified on average 4.75 alleles were identified. figure 2 The PCR amplified electrophoresis patterns of 2 SSR primers on 24 sugarcane materials tested were displayed, and the...

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Abstract

The invention aims to provide saccharum cultivar R570 haploid whole genome data formed by 4460 BAC library fragments. A primer developed by an SSR marker primer which is designed, synthesized and verified according to a primitive type with high polymorphism has the advantages of stable amplification results, clear and distinguishable electrophoretic bands and high polymorphic sites, and can be widely applied to genetic diversity analysis of saccharum cultivars, identification and protection of new cultivars and construction of DNA finger-prints and genetic linkage maps.

Description

technical field [0001] The invention belongs to the technical field of sugarcane molecular breeding, and relates to the development and application of SSR markers based on sugarcane whole genome data. Background technique [0002] Sugarcane (Sacchrum spp. Hybrid) is an important C4 plant. It has strong adaptability, high biomass, high photosynthetic efficiency, low-carbon crops that can be planted continuously for many years and fixes more CO2. It is an important sugar crop in the world. (accounting for 80% of the world's total sugar) and one of the bioenergy crops (accounting for 40% of the world's bioethanol). In 1887, Soltwedel, J B Harrison, and J R Bovell discovered that sugarcane seeds could produce seedlings in Java and Barbados, West India, respectively, which opened the history of sexual hybridization in sugarcane (Luo 1992). Modern sugarcane cultivars are interspecific crosses between tropical species (Saccharum officinarum L., 2n=80, x=10) and Saccharum spontaneu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 王恒波陈姝琦祁舒婷张华郭晋隆
Owner FUJIAN AGRI & FORESTRY UNIV
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