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Universal SSR (simple sequence repeat) molecular marker CSSR1 for chrysanthemum and related plants and application thereof

A technology of molecular markers and related genera, applied in the field of molecular biology, can solve the problems of no SSR molecular markers, no SSR markers have been found, etc., and achieve the effect of broad application prospects.

Active Publication Date: 2014-05-07
BEIJING FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the development of SSR markers in chrysanthemum is currently in its infancy, and there is no report on the development of SSR markers in Ajania and Opisthopappus due to the lack of gene sequence information; therefore, currently Ajania and Opisthopappus No SSR molecular markers have been developed for Opisthopappus, and there is no SSR marker that can be used for general use between Chrysanthemum and related genera; For the development and utilization of wild resources of Chrysanthemum, Chrysanthemum and Taihang Chrysanthemum, it is of great significance to carry out intergeneric distant hybrid breeding and molecular level research

Method used

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  • Universal SSR (simple sequence repeat) molecular marker CSSR1 for chrysanthemum and related plants and application thereof
  • Universal SSR (simple sequence repeat) molecular marker CSSR1 for chrysanthemum and related plants and application thereof
  • Universal SSR (simple sequence repeat) molecular marker CSSR1 for chrysanthemum and related plants and application thereof

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Experimental program
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Embodiment 1

[0024] Example 1 Acquisition of General SSR Molecular Marker CSSR1 of Chrysanthemum and Related Genus Plants

[0025] (1) Download the 7180 ESTs of all Chrysanthemum plants published on the website of the National Center for Biotechnology Information (NCBI) (http: / / www.ncbi.nlm.nih.gov / Genbank / ) as of April 2012 Sequences were spliced ​​using CAP3 software to obtain spliced ​​gene sequences. Then use SSRIT software (http: / / www.gramene.org / db / markers / ssrtool) to search for SSR sites, and then use Primer Premier5.0 software to design upstream and downstream primers for SSR sites.

[0026] Wherein the primers for amplifying the molecular marker CSSR1 are the following primer pairs:

[0027] CSSR1 upstream primer F: 5'-TTCAAGGGCTGGATGCCTTATT-3'

[0028] CSSR1 downstream primer R: 5'-GTGTTTGCGATGTTCCATTCTC-3'

[0029] (2) DNA samples of common plant materials of Chrysanthemum, Ajania and Opisthopappus were extracted (see Table 1).

[0030] Table 1 Plant materials of the genera ...

Embodiment 2

[0036]Example 2 Using the SSR molecular marker CSSR1 to carry out hybridization (AP1, AP2, AP3) between the ground cover chrysanthemum variety "Pudifendai" (C.Morifolium "Pudifendai") and A.Acificum identification

[0037] (1) Extract the genomic DNA samples of the ground cover chrysanthemum variety "Pudi Danfen", the chrysanthemum of the genus Chrysanthemum and the hybrid offspring AP1, AP2, and AP3.

[0038] (2) Using the PCR reaction system for amplifying the SSR molecular marker in Example 1, using the upstream and downstream primers of the CSSR1 site and using the DNA of the hybrid parents and hybrid offspring as templates, PCR reactions were performed.

[0039] (3) STR typing analysis of parents and hybrid offspring ( figure 1 ), the hybrid offspring all showed the characteristic bands of the male and female parents, proving that AP1, AP2, and AP3 are true hybrids of the ground cover chrysanthemum variety "Pudi Danfen" and the chrysanthemum ( figure 2 ).

[0040] Tab...

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Abstract

The invention provides a universal SSR (simple sequence repeat) molecular marker CSSR1 for chrysanthemum and related plants, which is used for PCR (polymerase chain reaction) amplification of the marker CSSR1 primer sequences disclosed as SEQ ID NO. 1 and 2. The invention also provides application of the SSR marker in identification of intergeneric cross hybrids of chrysanthemum and Ajania. Besides, the invention also provides a method for amplifying the SSR molecular marker, which establishes a universal SSR molecular marker technical system among chrysanthemum, Ajania and Opisthopappus plant materials. The marker can be used for intrageneric and intergeneric hybrid identification of chrysanthemum, Ajania and Opisthopappus, plant genetic diversity analysis, fingerprint construction, molecular marker assisted breeding and the like, and has very wide application prospects.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a universal SSR molecular marker CSSR1 for plants of Chrysanthemum and related genera and its application. Background technique [0002] Chrysanthemum (Chrysanthemum) belongs to the dicotyledonous class Asteraceae. The Chrysanthemum plants native to my country mainly include: Wild Chrysanthemum (C.indicum), Shennongxiangju (C.indicum var.aromaticum), Maohuaju (C.vestitum), Chamomile (C.lavandulifolium), Fever Chrysanthemum (C.chanetii), Purple Wild Chrysanthemum (C.zawadskii), Chrysanthemum Brain (C.mankingense), etc. Among them, chrysanthemum (Chrysanthemum morrofolium) is the species with the highest ornamental value. Chrysanthemum is one of the top ten famous flowers in China and one of the four major cut flowers in the world. It has a cultivation history of more than 3,000 years in China and has formed a variety group with rich flower patterns and various colors. . In additi...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 张启翔刘华孙明高亦珂潘会堂程堂仁王佳杨炜茹
Owner BEIJING FORESTRY UNIVERSITY
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