Method for identifying new variety Qingbi of fraxinus velutina by using capillary electrophoresis fluorescence SSR (Simple Sequence Repeat) fingerprint spectrum

A technology of capillary electrophoresis and fingerprinting, which is applied in the field of identification of a new species of white wax 'Qingbi', achieving high resolution, stable specificity, and easy operation

Active Publication Date: 2019-04-09
SHANDONG FOREST SCI RES INST +1
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, the search found that there is no report on the development of a method for identifying the new white wax variety 'Qingbi' based on capillary electrophoresis fluorescent SSR molecular labeling technology using capillary electrophoresis fluorescent SSR fingerprints.

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  • Method for identifying new variety Qingbi of fraxinus velutina by using capillary electrophoresis fluorescence SSR (Simple Sequence Repeat) fingerprint spectrum
  • Method for identifying new variety Qingbi of fraxinus velutina by using capillary electrophoresis fluorescence SSR (Simple Sequence Repeat) fingerprint spectrum
  • Method for identifying new variety Qingbi of fraxinus velutina by using capillary electrophoresis fluorescence SSR (Simple Sequence Repeat) fingerprint spectrum

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Embodiment 1

[0035] 1. Identification of materials

[0036] Genomic DNA was extracted from young leaves of ‘Qingbi’.

[0037] 2. Extraction of DNA

[0038] 2.1 Preheat the prepared CTAB extraction buffer in a water bath at 65°C for 30 minutes, put chloroform-isoamyl alcohol (24: 1), absolute ethanol, and 70% ethanol in a -20°C refrigerator for pre-cooling for later use. The bowl needs to be pre-cooled in advance, and then pre-cooled with a small amount of liquid nitrogen before use;

[0039] 2.2 Take 0.4-0.5g young white wax leaves, add enough liquid nitrogen to grind to powder, quickly transfer to a 2mL centrifuge tube, add 600 μL 2% CTAB extract (containing 2% β-mercaptoethanol), and mix well. Bathe in a water bath at 65°C for 30 minutes, during which time gently invert 3-4 times;

[0040] 2.3 After cooling to room temperature, add 600 μL of chloroform: isoamyl alcohol (24:1) to extract for 10 minutes, shake gently constantly, cool to room temperature, and centrifuge at 12000 r / min fo...

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Abstract

The invention discloses a method for identifying a new variety Qingbi of fraxinus velutina by using a capillary electrophoresis fluorescence SSR (Simple Sequence Repeat) fingerprint spectrum, and thenew variety Qingbi of the fraxinus velutina is identified by extracting a genomic DNA of the fraxinus velutina from a tender leaf of the fraxinus velutina, taking the extracted DNA as a template, using screened SSR primers to implement PCR (Polymerase Chain Reaction) amplification, implementing capillary electrophoresis on a PCR amplification product, and combining to form the SSR fingerprint spectrum for the new variety Qingbi of the fraxinus velutina according to the peak value of each pair of SSR primers in an electrophoresis result. The six pairs of SSR specific primers stable in specificity and good in polymorphism are screened by using the method, the fingerprint spectrum for the new variety Qingbi of the fraxinus velutina is constructed, and the new variety Qingbi of the fraxinus velutina is identified conveniently, rapidly and accurately.

Description

technical field [0001] The invention belongs to the technical field of molecular identification of plant varieties, and in particular relates to a method for identifying a new variety of white wax 'Qingbi' using capillary electrophoresis fluorescence SSR fingerprints. Background technique [0002] Ash plants are wind-pollinated flowers, and natural pollination hybridization is easy to occur when mixed planting, especially F. velutina, F. pennsylvanica (F. pennsylvanica), and F. americana are equivalent to the sub-genus group , their morphological characteristics are very similar, the kinship is very close, and the traditional methods of hybrid offspring are often indistinguishable. ‘Qingbi’ is a new female species of Fraxinus Velutina ‘qingbi’. The trunk of this variety is yellowish brown, the branches of the current year are slightly bent and drooping, and it is resistant to salt and alkali. It has a wide range of applications and is suitable for landscaping and afforestat...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2525/151C12Q2565/125
Inventor 吴德军燕丽萍王因花姚俊修任飞王开芳刘翠兰束德峰张子通
Owner SHANDONG FOREST SCI RES INST
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