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EST-SSR primer group developed on basis of transcriptome sequences of hemarthria compressa and hemarthria altissima and application of EST-SSR primer group

A technology of bougainvillea and transcriptome sequence, which is applied in the field of SSR primer sets, can solve the problems of no molecular marker development and application report, no transcriptome sequencing technology, etc., and achieves rich polymorphism, rich quantity and repeatability good effect

Inactive Publication Date: 2016-05-18
SICHUAN AGRI UNIV +1
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Problems solved by technology

So far, no transcriptome sequencing technology has been applied to verbena plants, and there is no report on the development and application of EST-SSR molecular markers in a certain species of verbena

Method used

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  • EST-SSR primer group developed on basis of transcriptome sequences of hemarthria compressa and hemarthria altissima and application of EST-SSR primer group
  • EST-SSR primer group developed on basis of transcriptome sequences of hemarthria compressa and hemarthria altissima and application of EST-SSR primer group
  • EST-SSR primer group developed on basis of transcriptome sequences of hemarthria compressa and hemarthria altissima and application of EST-SSR primer group

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Embodiment Construction

[0026] The present invention will be further described below in conjunction with embodiment.

[0027] Transcriptome sequencing was carried out on the young leaves and roots of Verbena flat tachycarpus and Verbena genus, and the spliced ​​Unigenes larger than 1kb obtained were merged and then used the software MISA (http: / / pgrc.ipk-gatersleben.de / misa / ) Scanning of EST-SSR loci was performed. The screening criteria were that 2 bases were repeated at least six times, and 3, 4, 5, and 6 bases were repeated at least five times before they were identified as SSR sites. Finally, 10,888 EST-SSR sites were obtained, and then Primer3v2.23 ( http: / / primer3.sourceforge.net ) designed primers for these candidate sites, and the design parameters were set as follows: the primer length range was between 18-23nt, and the optimum length was 21nt; the length of the PCR product was within the range of 100bp and 300bp; the annealing temperature of the primers was 52-58 °C, of ​​which 55 °C ...

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Abstract

The invention discloses an EST-SSR primer group developed on the basis of transcriptome sequences of hemarthria compressa and hemarthria altissima. The EST-SSR primer group comprises 52 pairs of primers, and nucleotide sequences of the primers are represented as sequence tables SEQUENCE ID NO.1-104. The invention further relates to an application of the EST-SSR labeled primers in the aspects of genetic diversity analysis of hemarthria sibirica and transferability of the hemarthria sibirica in gramineae species. The 52 pairs of primers in the EST-SSR primer group developed on the basis of the transcriptome sequences of hemarthria compressa and hemarthria altissima have the advantages of abundant polymorphism, stable amplification, good repeatability, convenience in counting and the like, can be effectively applied to research fields such as hemarthria sibirica germ resource genetic diversity analysis, variety identification, fingerprinting construction, molecular marker assisted breeding and the like and are applied to correlational research of other gramineae forage species.

Description

technical field [0001] The invention relates to the field of SSR primer sets, in particular to an EST-SSR primer set developed based on the transcriptome sequences of Verbena flat tachycarpa and Verbena sylvatica and its application. Background technique [0002] Verbena is a general term for plants of the genus Hemarthria R.Br. in the grass family (Gramineae). It is also known as cowboy grass and iron verbena. It is also distributed in warm temperate and tropical regions such as Southeast Asia, Indonesia, India, the United States, and Brazil. It has many species, and there are nearly 20 species in the world, among which Hemarthriacompressa and its close relative Hemarthria altissima are two particularly important perennial warm-season types in the Verbena genus. Forage grass species are mainly distributed in my country and Africa. Due to the advantages of fast growth, high yield and high quality, rich nutrition, good palatability, tolerance to poor soil, high resistance an...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2600/156C12Q2565/125
Inventor 黄琳凯张健黄秀张新全杨盛婷黄德均严海东周思凡王登平闫艳红马啸彭燕
Owner SICHUAN AGRI UNIV
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