A kind of Cattelan ssr molecular marker primer composition and application thereof

A primer composition and molecular marker technology, applied in the field of molecular biology, can solve problems such as the adverse effects of Cattleya hybrid breeding and the distribution of undiscovered native species, and achieve the effects of early selection at seedling stage, accelerated breeding process, and stable identification

Active Publication Date: 2021-08-27
ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Cattleya has not been found in the distribution of native species in my country, and it needs to be introduced and cultivated from other countries. However, there are many homonyms and homonyms in the introduced Cattleya, which has a negative impact on the hybrid breeding of Cattleya

Method used

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  • A kind of Cattelan ssr molecular marker primer composition and application thereof
  • A kind of Cattelan ssr molecular marker primer composition and application thereof
  • A kind of Cattelan ssr molecular marker primer composition and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Embodiment 1 Cattleya Germplasm Resources

[0087] The present invention adopts totally 26 Cattleya germplasm resources of different species collected and preserved in the Orchid Resource Garden of the Institute of Environmental Horticulture, Guangdong Academy of Agricultural Sciences, and the specific information is shown in Table 3 below. All experimental materials were planted in the greenhouse of the Institute of Environmental Horticulture, Guangdong Academy of Agricultural Sciences, Guangdong Agricultural Modernization Technology Demonstration Zone.

[0088] Table 3 Basic information of 26 germplasm resources

[0089]

[0090]

Embodiment 2

[0091] Embodiment 2 Cattleya Plantation Resource Identification

[0092] 1. Total DNA Extraction

[0093] Extract the total DNA of Cattleya plants according to the operation steps of the new plant genome DNA extraction kit of Tiangen Biotechnology (Beijing) Co., Ltd. The specific steps are as follows:

[0094] (1) Take Cattleya plant young root tip or young leaf tissue and add liquid nitrogen to grind thoroughly, and weigh about 100 mg of fresh plant tissue.

[0095] (2) Quickly add 400 μL buffer solution GPS and 10 μL RNase A (10 mg / mL) to the ground powder, vortex quickly and mix well, then place the centrifuge tube in a 65°C water bath for 15 minutes, invert the centrifuge tube during the water bath to mix Sample several times.

[0096] (3) Add 100μL buffer GPA, vortex for 1min, centrifuge at 12000rpm for 5min, transfer the supernatant to the filter column CS (the filter column CS is placed in the collection tube), then centrifuge at 12000rpm for 1min, transfer the filtra...

Embodiment 3

[0118] Embodiment 3 primer amplification result

[0119] The amplification results of the SSR molecular marker primers described in the present invention are detailed in Table 5 below. Because there are many kinds of SSR molecular marker primers and Cattleya, the peak map of the amplification result of 1-N1006714 is shown as an example. The peak map of the amplification result of 1-N1006714 is as follows figure 1 shown. From Table 5 and figure 1 It can be seen that the SSR molecular marker primers of the present invention have good amplification effect, high detection rate, and can amplify stable DNA bands.

[0120] Table 5 Primer Amplification Results

[0121]

[0122]

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Abstract

The invention provides a Cattleya SSR molecular marker primer composition, a fingerprint code and its application in the identification of Cattleya germplasm resources, belonging to the technical field of molecular biology. The present invention provides 15 SSR molecular markers of Cattleya and their amplification primer compositions, and utilizes TP-M13-SSR technology to analyze the genetic diversity of 26 different germplasm resources within the genus and construct molecular ID cards , can accurately, efficiently and stably identify the germplasm resources of Cattleya, and lay a solid foundation for the identification of germplasm resources of Cattleya, analysis of kinship, mapping of trait genes, molecular marker-assisted breeding and related molecular research of Cattleya. Base.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a Cattleya SSR molecular marker primer composition, a fingerprint code and its application in the identification of Cattleya germplasm resources. Background technique [0002] Cattleya, also known as Cattleya orchid, Cattleya orchid, Cattleya orchid, etc., is the collective name for the species of the genus Cattleya (Orchidaceae) and its close relatives, native to the Americas Tropical and subtropical, distributed from Mexico to Brazil, all are epiphytic orchids, because of their large and beautiful flowers, bright colors, and different varieties bloom in different seasons, they are very popular among the people, and they are also popular in Brazil, Colombia, Costa Rica The national flower of other countries has a good international recognition, so it enjoys the reputation of "King of Yanglan" in the world. [0003] Generalized Cattleya is a general term for a collecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6809C12Q1/6858C12N15/11
CPCC12Q1/6809C12Q1/6858C12Q1/6895C12Q2600/156C12Q2535/122C12Q2537/165C12Q2531/113C12Q2565/125
Inventor 李佐肖文芳吕复兵陈和明朱根发
Owner ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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