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Development method and application of saccharum cultivar genome SSR (simple sequence repeat) molecular marker

A technology of molecular markers and cultivars, applied in biochemical equipment and methods, recombinant DNA technology, DNA/RNA fragments, etc., can solve the problem of backward construction of genetic linkage map of sugarcane SSR molecular markers, limited number of SSR molecular markers, and great difficulty in development To achieve the effect of improving the level of sugarcane genetics and breeding, enriching polymorphism information, and protecting legitimate rights and interests

Inactive Publication Date: 2019-12-10
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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  • Development method and application of saccharum cultivar genome SSR (simple sequence repeat) molecular marker
  • Development method and application of saccharum cultivar genome SSR (simple sequence repeat) molecular marker
  • Development method and application of saccharum cultivar genome SSR (simple sequence repeat) molecular marker

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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) SES 208) 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 pairs of p...

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 sarcophagus, large-stem wild species and Indian species) (see Table 2), 2 sugarcane ancestor species (saccharomyces SES 208 and tropical species LApurple) and 4 worldwide Important sugarcane cultivars (LCP85-384, R570, ROC 16 and ROC 22) were analyzed for genetic diversity and polymorphism evaluation of SSR primers. 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 amplification electrophoresis patterns of 2 pairs of SSR primers on 24...

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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 with strong adaptability, high biomass, high photosynthetic efficiency, continuous planting for many years and CO fixation 2 It is one of the most important sugar crops (accounting for 80% of the world's total sugar) and bioenergy crops (accounting for 40% of the world's bioethanol) in the world. 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). While modern sugarcane cultivars were derived from the tropical ancestors of sugarcane ( Saccharum officinarum L., 2n=80, x=10) and cut hand secret ( Saccharum spontane...

Claims

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Application Information

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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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