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The ssr marker sp_ssr04 tightly linked to spinach male and its application

A spinach and marker technology, applied in the field of molecular biology, can solve problems such as protection of unfavorable intellectual property rights of spinach varieties, less sex-linked molecular markers, etc., and achieve the effect of improving breeding efficiency

Inactive Publication Date: 2021-03-16
HENAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Except for T11A and V20A, the above markers are often not universal and can only be applied to certain varieties, and there are very few gender-linked molecular markers developed by domestic scientific research institutions, which are not suitable for the protection of intellectual property rights of spinach varieties

Method used

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  • The ssr marker sp_ssr04 tightly linked to spinach male and its application
  • The ssr marker sp_ssr04 tightly linked to spinach male and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Acquisition and Verification of SSR Marker SP_SSR04 Closely Linked to Spinach Male

[0016] 1. Whole-genome sequencing of spinach XX and YY genomes using PacBio sequencing technology

[0017] Screening the germplasm resources of spinach, the population of spinach cultivar Cornell #9 has both female plants, male plants and monoecious plants. The female plant is XX plant, and YY plant is obtained by hybridization between monoecious plants (XY x XY→YY:XY:XX), and the whole genome of spinach XX and YY genomes is sequenced by PacBio sequencing technology.

[0018] 2. Perform comparative genome analysis on the obtained whole genome sequence to obtain male-specific sequence

[0019] Comparative genome analysis was performed using bioinformatics analysis software to obtain male-specific sequences.

[0020] 3. Use SOAPdenovo2 software to assemble the obtained male-specific sequences to obtain male-specific contigs

[0021] 4. Use the RepeatMasker software to analyze the obtai...

Embodiment 2

[0032] Application of SSR Marker SP_SSR04 Closely Linked to Spinach Male in Sex Identification of Cultivated and Wild Spinach Species

[0033] 1. Extraction of single plant genomic DNA of spinach cultivars and wild species

[0034] Genomic DNA of spinach strains to be tested was extracted by CTAB method.

[0035] 2. Application of SSR marker SP_SSR04 in sex identification of spinach cultivars and wild species

[0036] Utilize the designed specific primer pair:

[0037] Upstream primer: 5'-TTGATTGCCTCAAGGTGTTG-3'

[0038] Downstream primer: 5'-TGGGTTCTTAGCCACTTTTTG-3'

[0039] The genomic DNA of the tested spinach cultivars and wild species were respectively amplified. The results of agarose gel electrophoresis showed a 176bp specific band as the male plant, and the one without the amplification product was the female plant.

[0040] The PCR amplification reaction system is: 10×PCR Buffer (2 μL), 2.5 mM dNTP Mixture (1 μL), 10 μM / μL upstream primer (1.25 μL), 10 μM / μL downs...

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Abstract

The invention discloses an SSR marker SP_SSR04 closely linked with male spinach and application thereof to sex determination of spinach, and belongs to the technical field of molecular biology. The technical scheme is characterized in that the SSR marker SP_SSR04 closely linked with the male spinach is obtained through amplification of a specific primer pair, wherein the nucleotide sequence of theSSR marker SP_SSR04 is shown as SEQ ID NO.1; a specific stripe of 176bp is amplified in the male spinach; no amplification product exists in the male spinach; the specific primer pair consists of anupstream primer: 5'-TTGATTGCCTCAAGGTGTTG-3' and a downstream primer 5'-TGGGTTCTTAGCCACTTTTTG-3'. The molecular marker provided by this invention can not only be applied to early sex determination of spinach cultivated species, but also can be applied to early sex determination of spinach wild species, can realize early screening of female and male spinach plants, greatly improves the breeding efficiency, and has an important value in the production practice and breeding of the spinach.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to an SSR marker SP_SSR04 closely linked with spinach male and its application in spinach sex identification. Background technique [0002] spinach( Spinacia oleracea L.) Also known as Persian grass and red root vegetable, it is a one or two-year-old herbaceous plant in the genus Spinach of Rico with green leaves as the main product organ. Spinach is rich in nutrition, contains a variety of vitamins, has strong cold resistance, short production cycle, high multiple cropping index, high yield and output value, and is deeply loved by producers and consumers. At present, it is widely cultivated in the north and south of my country. It is one of the main green leafy vegetables in spring, autumn and winter, and it is also one of the main export vegetables in my country. According to the statistics of the United Nations Food and Agriculture Organization (FAO) in 2...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6879C12N15/11
CPCC12Q1/6879C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 秦瑞云邓传良李晓月李书粉高武军
Owner HENAN NORMAL UNIV