Preparation method and application for Miscanthus Genic-SSR mark
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A Miscanthus and plant technology, applied in the field of molecular biology, can solve the problems of unable to meet the research needs of Miscanthus plants, difficult to meet the development of EST-SSR markers, time-consuming and labor-intensive, etc.
Inactive Publication Date: 2012-04-25
湖北光芒能源植物有限公司
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Problems solved by technology
Among them, the first two methods are time-consuming, laborious and costly to develop, and the third method relies on the existing EST data in NCBI, and its application to species with few EST sequencing is very limited.
[0004] As of November 21, 2011, there are only 5 EST sequences of Miscanthus that can be retrieved in the NCBI database, which is difficult to meet the needs of developing EST-SSR marke
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Embodiment 1
[0050] This embodiment is the preparation and screening method of Miscanthus Genic-SSR marker, which mainly includes the following main steps:
[0051] 1. Acquisition of EST sequence of Miscanthus:
[0052] Select the young leaves of five species of Miscanthus plants (Awn, Wujiemans, Di, Nandi, and Qigang) to extract total RNA, use the standard Solexa sequencing method to obtain transcriptome Reads, and use Repeat Masker to remove carrier and other RNA Sequence pollution, using SOAP splicing software to complete sequence splicing, and finally obtain the non-redundant Unigene-EST sequence of Miscanthus, see Table 1 for details.
[0053] Table 1 Unigene-EST statistical information generated by sequencing
[0054] species name (abbreviation)
Unigene number / article
Total number of designed primers / article
Total number of SSRs / piece
Nandi (LT)
64663
2347
3381
Hagi(TS)
103114
2916
4463
Mount (MS)
97...
Embodiment 2
[0073] A kind of application of Miscanthus Genic-SSR marker in Miscanthus genetic diversity analysis, its steps are:
[0074] 1. Experimental materials:
[0075] A total of 17 materials from Miscanthus and Miscanthus were selected to test the effectiveness of Genic-SSR markers of Miscanthus in the analysis of genetic diversity of Miscanthus. See Table 3 for details of the materials.
[0076] Table 3 is used for carrying out the material information of primer screening and genetic diversity analysis in the present invention
[0077] material code
Genus-species / variety name
source of material
Type information
1
Miscanthus - Miscanthus
Changling Town, Ezhou City, Hubei Province
wild species
2
Miscanthus - Miscanthus
Changling Town, Ezhou City, Hubei Province
wild species
3
Miscanthus - Miscanthus
Germplasm Resource Nursery of Wuhan University
cultivar
4
Miscanthus - Miscanthus
Ge...
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Abstract
The invention discloses a preparation method and application for a Miscanthus Genic-SSR mark. The method comprises the following steps: A, acquiring Miscanthus transcriptomic sequence, that is, selecting young and tender Miscanthus leaves, extracting total RNA respectively so as to obtain transcriptome Reads, removing the pollution of carrier RNA sequence and carrying out splicing so as to obtainthe Miscanthus transcriptomic sequence; B, identifying SSR sites in Miscanthus ESR sequence, that is, searching SSR sites in obtained non-redundant Unigene-EST sequence by using SSR retrieval software, with repeats being 2 to 6 bases, so as to obtain a series of Unigene sequences containing information about the SSR sites; C, designing Miscanthus Genic-SSR primers, that is, using primer design software to design primers according to the obtained Unigene sequences containing the SSR sites so as to obtain a series of the Miscanthus Genic-SSR primers. According to the invention, the method is simple, has high efficiency and is simple to operate; a huge amount of marks are obtained; EST sequence obtained through transcriptomic sequencing lays a foundation for genetic research, molecular improvement and the like of the critical energy plant Chinese silvergrass.
Description
technical field [0001] The invention belongs to the technical field of molecular biology, and more specifically relates to a preparation method of a Miscanthus Genic-SSR marker, and also relates to an application of a Miscanth Genic-SSR marker. technical background [0002] Miscanthus belongs to the C4 perennial herbaceous plants of Poaceae, mainly including Miscanthus, Wujiemans, Di, Nandi, and Qigang, which are concentrated in the east and southeast of Asia. Miscanth plants have the characteristics of strong vitality, high photosynthetic utilization efficiency, and strong stress resistance, and can be used in pharmaceuticals, papermaking, water and soil conservation, and ecological restoration. With the continuous improvement of human energy demand and the continuous reduction of global energy reserves, Miscanthus has attracted more and more attention as a potential energy plant. At present, Qigang, which has been widely planted in Europe and the United States, has an ann...
Claims
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