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EST-SSR (expressed sequence tag-single sequence repeat) genetic marker site of medicago archiducis-nicolai sirjaev and application thereof

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

Active Publication Date: 2019-05-28
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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  • EST-SSR (expressed sequence tag-single sequence repeat) genetic marker site of medicago archiducis-nicolai sirjaev and application thereof
  • EST-SSR (expressed sequence tag-single sequence repeat) genetic marker site of medicago archiducis-nicolai sirjaev and application thereof
  • EST-SSR (expressed sequence tag-single sequence repeat) genetic marker site of medicago archiducis-nicolai sirjaev and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 The ESR-SSR genetic marker site of fenugreek bean, the ESR-SSR genetic marker site refers to the transcripts obtained by sequencing the transcriptome of fengal bean seedlings, after detecting the SSR site in the transcript, according to the transcript The nucleotide sequences of the primers designed for the flanking sequences of the SSR sites above are shown in Man1~Man350 in the sequence listing.

[0023] The length of the amplification product of the primer sequence in the Qinghai-Tibet fenugreek and the lentil genome DNA is the same and close to the base sequence length of the expected amplification product of the primers shown as Man1~Man350 in the sequence table.

[0024] 1. Acquisition of the SSR loci of Alfalfa sativa.

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

[0026] ⑴Plant cultivation and low temperature stress treatment:

[0027] The Qinghai-Tibetan sativa plants were collected from the natu...

Embodiment 2

[0072] Example 2 The EST-SSR genetic marker locus and primer sequence of A. chinensis, derived from A. sativa. It has the characteristics and nucleotide sequences of the marker sites listed in Table 1, and is applied to A. sativa and A. sativa Detection and Analysis of Genetic Diversity, Population Genetic Structure and Genetic Differentiation of A.

[0073] 1. STR genetic typing of individuals in Qinghai-Tibet fenugreek and lentil wild populations

[0074] A total of 94 individuals listed in Table 2 from 3 Qinghai-Tibetan lentils and 3 wild populations of lentils were extracted as templates respectively, and the verified polymorphic primers obtained in Example 1 were taken as shown in Table 4 Listed 75 pairs of primers corresponding to the primer sequences in Table 1, the 5' end of the reverse primer is labeled with HEX, 6-FAM or TAMARA fluorescent group, and PCR amplification is carried out. The amplification conditions are as described in Example 1 . The amplified product...

Embodiment 3

[0093] Example 3 The application of the ESR-SSR genetic marker loci in the Qinghai-Tibetan fenugreek and the commercial varieties of the Qinghai-Tibetan fenugreek.

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Abstract

The invention relates to an EST-SSR (expressed sequence tag-single sequence repeat) genetic marker site of medicago archiducis-nicolai sirjaev. The EST-SSR genetic marker site means that transcriptomesequencing on the medicago archiducis-nicolai sirjaev is carried out to obtain a transcript; after the SSR site in the transcript in the detected, a primer nucleotide sequence which is designed by aflanking sequence arranged on the SSR site of the transcript and is shown in Man1 to Man350 in a sequence list is acquired; the length of an amplification product in the medicago archiducis-nicolai sirjaev and a Medicagoruthenica (L.) Ledebour gene group DNA is the base sequence length of a predicted amplification product which is identical and close to the primer shown in the sequence lift Man1 to Man350. The EST-SSR genetic marker site of the medicago archiducis-nicolai sirjaev can be applied to evaluation of commercial genetic diversity of germplasm resources of the medicago archiducis-nicolai sirjaev and the Medicagoruthenica (L.) Ledebour, selective breeding of the commercial variety of the medicago archiducis-nicolai sirjaev and the Medicagoruthenica (L.) Ledebour and commercial identification and development of functional genes of the medicago archiducis-nicolai sirjaev and the Medicagoruthenica (L.) Ledebour.

Description

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

Claims

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Application Information

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Patent Type & Authority Applications(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