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Fingerprint spectrum of butterfly-leaf platycladus orientalis SSR marker and construction method and application thereof

A technology of oriental orientalis and butterfly leaves is applied in the field of oriental orientalis germplasm resource detection to achieve the effects of high identification accuracy, short identification time and good stability.

Active Publication Date: 2020-12-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, some SSR primers have been developed in the plant materials of Arborvitae, and have been applied in the analysis of genetic diversity and pedigree reconstruction, but the use of SSR markers to construct fingerprints of Arborvitae species has not been reported.

Method used

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  • Fingerprint spectrum of butterfly-leaf platycladus orientalis SSR marker and construction method and application thereof
  • Fingerprint spectrum of butterfly-leaf platycladus orientalis SSR marker and construction method and application thereof
  • Fingerprint spectrum of butterfly-leaf platycladus orientalis SSR marker and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Example 1. Identification of the screening of SSR primers for 'Orientalis arborvitae' and the establishment of a PCR detection method

[0095] 1. Screening of SSR primers for identification of ‘Arborvitae arborvitae’

[0096] Based on the amplified product of simple repeat sequence (SSR) developed based on the sequencing of the Thuja arborvitae transcriptome, the amplified band type, rich polymorphism, stability and identification obtained after PCR amplification of different Thuja arborvitae germplasm resources High-efficiency SSR primers. Using the established Thuja SSR reaction technology system, agarose gel electrophoresis and polyacrylamide gel electrophoresis, 4 arborvitae materials with large phenotype differences were selected, and 182 pairs of primers were screened for the first time. The primers of the extended bands were retained, and polymorphism re-screened the primers amplified for the first time by polyacrylamide gel electrophoresis. After the second amp...

Embodiment 2

[0130] Embodiment 2, the application of identifying whether the original plant of Arborvitae to be tested or its asexually reproduced progeny is Arborvitae phalangifolia

[0131] 1. Genomic DNA extraction:

[0132] Genomic DNA of 81 leaves of Arborvitae were extracted; among them, DY-1 # Subsequent cutting seedlings for ‘Butterfly arborvitae’, DY-2 # It is the original seedling of ‘Thuja arborvitae’;

[0133] 2. PCR amplification of SSR markers:

[0134] Using the above genomic DNA as a template, perform PCR amplification with 8 pairs of SSR primers shown in Table 1;

[0135] SSR primer system for the above PCR amplification (20 μl in total): ddH 2 O 14.8 μl, dNTP 0.4 μl, Buffer 2 μl, primer F 0.3 μl (20 μM), primer R 0.3 μl (20 μM), DNA template 2 μl, Taq 0.2 μl.

[0136] The reaction of the above-mentioned PCR amplification adopts the following cycle parameters:

[0137] Pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30 seconds, renaturation at 54°C f...

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Abstract

The invention discloses a fingerprint spectrum of a butterfly-leaf platycladus orientalis SSR marker and a construction method and application thereof. The invention provides a set of primers, which are composed of a primer pair 1 to a primer pair 8; and the nucleotide sequences of the primer pair 1 to the primer pair 8 are respectively sequence 1 to sequence 16 in a sequence table. The SSR markersuitable for butterfly-leaf platycladus orientalis is developed based on platycladus orientalis transcriptome sequencing, and the construction method and application of the fingerprint spectrum of the butterfly-leaf platycladus orientalis SSR marker are established. Compared with morphological detection, the technology has the advantages that the identification result is not affected by climate,environment, human factors and tree age, identification and differentiation can be effectively carried out in the seedling stage, the identification time is short, the identification accuracy is high,the detection efficiency is high, the stability is good, and the technology has important significance and good application effect on genetic specificity identification of butterfly-leaf platycladusorientalis and other platycladus orientalis germplasm.

Description

technical field [0001] The invention belongs to the field of detection of germplasm resources of arborvitae, and in particular relates to a fingerprint of arborvitae SSR markers and a construction method and application thereof. Background technique [0002] Platycladus orientalis (L.) Franco is an evergreen tree belonging to the genus Arborvitae of the Cupressaceae subfamily, and it is a unique tree species in my country. Arborvitae is a widely distributed tree species that is distributed in most parts of the country. It is a good tree species for afforestation and greening. It has the functions of water and soil conservation, windbreak and sand fixation, and landscaping. Moreover, Arborvitae has a long life span, is resistant to cold and drought, and its seeds have extremely medicinal value, and it is also one of the food sources for birds. Economic and social benefits. As early as 1987, Arborvitae was identified as the city tree of Beijing together with Chinese pagoda t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156Y02A40/22Y02P60/40
Inventor 刘国彬曹均张玉平廖婷王烨郭丽琴姚砚武
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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