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The est-ssr genetic marker locus and the corresponding marker primer sequence and its application

A technology of genetic markers, lentils, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of low polymorphism

Active Publication Date: 2022-06-21
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, compared with the intergenic region with large variation in the genome, the nucleotide sequence of the transcribed region is highly conserved among species, and there are many transferable EST-SSR markers among species with distant relatives within the genus. attitude tends to be low

Method used

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  • The est-ssr genetic marker locus and the corresponding marker primer sequence and its application
  • The est-ssr genetic marker locus and the corresponding marker primer sequence and its application
  • The est-ssr genetic marker locus and the corresponding marker primer sequence and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Qinghai-Tibetan lentil bean EST-SSR genetic marker site, the EST-SSR genetic marker site refers to the transcript obtained by sequencing the Qinghai-Tibetan lentil bean seedling transcriptome. After detecting the SSR site in the transcript, according to the transcription The nucleotide sequences of the primers designed by the flanking sequences of the SSR site are shown in SEQ ID NO. 1~700 in the sequence listing.

[0023] The lengths of the amplification products of the primer sequences in Qinghai-Tibet lucerne bean and lentil bean genomic DNA are the same as and close to the base sequence lengths of the expected amplification products of the primers shown in SEQ ID NOs. 1 to 700 in the sequence table.

[0024] 1. Obtaining the SSR locus of Qinghai-Tibet lentils.

[0025] 1. Low temperature stress treatment and transcriptome sequencing of Qinghai-Tibet lentil plants:

[0026] (1) Lentil bean plant culture and low temperature stress treatment:

[0027] Qingh...

Embodiment 2

[0072] Example 2 Qinghai-Tibetan lentil bean EST-SSR genetic marker site and its primer sequence, derived from Qinghai-Tibetan lentil bean, it has the marker site characteristics and nucleotide sequence listed in the attached table 1, and is applied to lentil bean and Detection and analysis of population genetic diversity, population genetic structure and genetic differentiation of Qinghai-Xizang lentils.

[0073] 1. STR genotyping of individuals in Qinghai-Tibet lentils and lentils wild populations

[0074] Listed in table 2 from 3 Qinghai-Xizang lentils and 3 lentils wild populations of 94 individuals in total to extract DNA as templates, take from the polymorphic primers of the verification obtained in Example 1 as shown in Table 4 The 75 pairs of primers with the listed numbers corresponding to the primer sequences in Table 1 were labeled with HEX, 6-FAM or TAMARA fluorophore at the 5' end of the reverse primer, and PCR amplification was performed. The amplification condit...

Embodiment 3

[0093] Example 3 Application of Qinghai-Tibetan lentil bean EST-SSR genetic marker locus in the breeding of Qinghai-Tibetan lentil bean and lentil bean varieties.

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Abstract

The present invention relates to an EST-SSR genetic marker site of the Qinghai-Tibetan fenugreek bean. The EST-SSR genetic marker site refers to a transcript obtained by sequencing the transcriptome of the Qinghai-Tibetan fenugreek bean seedling. After detecting the SSR site in the transcript, According to the flanking sequence of the SSR site on the transcript, the nucleotide sequence of the primer shown in SEQ ID NO.1-700 in the sequence listing is designed. The length of the amplification product of the primer sequence in the Qinghai-Tibetan fenugreek and the lentil genome DNA is the same and close to the base sequence length of the expected amplification product of the primer shown in SEQ ID NO.1~700 in the sequence table . The EST-SSR genetic marker locus of the Qinghai-Tibetan fenugreek of the present invention can be applied to the evaluation of the commercial genetic diversity of the Qinghai-Tibetan fenugreek and the germplasm resources of the Qinghai-Tibetan fenugreek and the commercial varieties of the Qinghai-Tibetan fenugreek and the lentil-bean, and The commercial identification and development of Qinghai-Tibetan fenugreek and functional genes of lentil bean.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the EST-SSR genetic marker site of Qinghai-Tibetan lentil bean and the corresponding marker primer sequence and application thereof. Background technique [0002] Tibetan lentils (Medicago archiducis-nicolai Sirjaev) and lentils (Medicagoruthenica (L.) Ledebour) belong to the genus Alfalfa ( Medicago ) moment pod alfalfa group ( Platycarpae ), which has strong adaptability to extreme environments such as drought, cold and salinity. Among them, the Qinghai-Tibet lentils are distributed in the Qinghai-Tibet Plateau, and are the only species in the genus Alfalfa that can survive the winter in the alpine natural grasslands of the Qinghai-Tibet Plateau. Lentils are widely distributed in the high latitudes and alpine regions of the northern hemisphere. Due to its good adaptability to harsh natural environment, lentil is considered to be an excellent legume genetic resource of alfalfa w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 王海庆王英芳窦全文陈志国
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST