A kind of ssr primer set and application thereof for distinguishing litchi varieties

A primer set and variety technology, applied in the field of molecular biology, can solve the problems of poor repeatability of RAPD, complex operation of AFLP, low efficiency, etc., and achieve the effect of promoting healthy development

CN112176088BActive Publication Date: 2022-04-15特稳植物检测技术服务(广州)有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Publication Date
2022-04-15

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Abstract

The invention belongs to the technical field of molecular biology, and in particular relates to an SSR primer set for distinguishing litchi varieties and its application. The SSR primer set includes 6 sets of litchi SSR primer pairs, and the nucleotide sequences of the forward primer and the primer in the 6 sets of litchi SSR primer pairs are respectively shown in SEQ ID NO.1~SEQ ID NO.12 in the sequence list Show. The band patterns amplified by the obtained SSR primers in litchi are clear and easy to interpret, and are suitable for detection on common denaturing polyacrylamide gel electrophoresis detection platforms and capillary fluorescence detection platforms. The SSR primers have high polymorphism and good amplification repeatability; The SSR primers of the present invention are applied in the fields of litchi genetic diversity analysis, variety identification, DNA fingerprinting and the like, which is beneficial to protect the legitimate rights and interests of breeders, producers and consumers, and promote the sound development of the litchi industry.
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Description

technical field

[0001] The invention belongs to the technical field of molecular biology, and in particular relates to an SSR primer set for distinguishing litchi varieties and its application. Background technique

[0002] Litchi is native to China and has been cultivated for more than 2,000 years. There are more than 500 wild germplasm resources and cultivated varieties. According to statistics, in 2018, the domestic litchi cultivation area reached 8.6 million mu, with an annual output of more than 600,000 tons, ranking first in the world in both cultivation area and annual output.

[0003] As a unique tropical and subtropical fruit tree in South my country, litchi has high economic value. Litchi is an evergreen tree that can be used to improve the ecological environment. Moreover, the trunk wood is solid and resistant to decay, and can be used for furniture making; some lychee roots that have been cultivated for a long time have good shape, and are even more suitable fo...

Examples

Embodiment 1

[0031] A method for distinguishing litchi varieties based on SSR genotype

[0032] 1) Extraction of Litchi Genomic DNA

[0033] a) 74 parts of lychee variety materials

[0034] Table 1 74 varieties of litchi

[0035]

[0036]

[0037] b) Genomic DNA extraction by CTAB method

[0038] (1) First heat the water bath to 65°C in advance, add 1.5ml 3% CTAB extract and 48ul β-mercaptoethanol, mix well and put it in the water bath to preheat;

[0039] (2) Weigh 0.5g of tender green leaves, put them into a mortar pre-cooled with liquid nitrogen, quickly add a small amount of PVP, and then add liquid nitrogen to quickly grind until the sample is powdery;

[0040] (3) Quickly pour the ground sample into the preheated CTAB extract, shake it upside down, put it in a water bath at 65°C for 1 hour, and shake it gently every 5 minutes. Centrifuge at 8000 rpm for 6 minutes at 4°C, and perform the remaining steps on ice;

[0041] (4) Transfer 1ml of the supernatant carefully into a n...