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An ssr marker sp_ssr19 tightly linked to spinach males and its application in spinach sexing

A spinach and marker technology, applied in the field of molecular biology, can solve problems such as the protection of intellectual property rights of unfavorable spinach varieties, the lack of 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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  • An ssr marker sp_ssr19 tightly linked to spinach males and its application in spinach sexing
  • An ssr marker sp_ssr19 tightly linked to spinach males and its application in spinach sexing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Acquisition and Validation of SSR Marker SP_SSR19 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 obtaine...

Embodiment 2

[0032] Application of SSR Marker SP_SSR19 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_SSR19 in sex identification of spinach cultivars and wild species

[0036] Utilize the designed specific primer pair:

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

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

[0039] The genomic DNA of the tested spinach cultivars and wild species were respectively amplified. The results of agarose gel electrophoresis showed a 178bp 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 downst...

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Abstract

The invention discloses an SSR (Simple Sequence Repeat) marker SP_SSR19 closely linked with male spinach and application thereof in sex determination of spinach and belongs to the technical field of molecular biology. The invention adopts the technical scheme with the following key points that the SSR marker SP_SSR19 closely linked with spinach male is obtained by amplification of the following specific primer pair and the nucleotide sequence is shown in SEQ ID NO.1 in a sequence table, 178bp specific strips are amplified in male spinach plants, and no amplification products exist in female spinach; the specific primer pair includes an upstream primer 5'-GGTACTGGTTAGGGCTCTGG-3' and a downstream primer 5'-CATCTGGTCATCCGATGTGA-3'. The molecular marker provided by the invention not only can be used for early sex determination of spinach cultivation seeds, but also can be applied in early sex determination work of wild spinach seeds; the female spinach plants and the male spinach plants can be screened in advance, the breeding efficiency can be greatly improved, so that the molecular marker has an important value in 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_SSR19 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 邓传良李晓月秦瑞云李书粉高武军
Owner HENAN NORMAL UNIV