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Group of cannabis sativa SSR molecular markers and application thereof

A molecular marker and molecular marker-assisted technology, applied in the field of molecular biology of crops, can solve the problems of low accuracy, large workload, occupying land, etc., and achieve the effects of convenient operation, improved efficiency and cost saving.

Active Publication Date: 2021-11-02
黑龙江省农业科学院农产品质量安全研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these identification methods have application limitations such as long time, heavy workload, low accuracy, and land occupation. At the same time, this method is only suitable for the identification of samples with obvious differences between varieties.

Method used

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  • Group of cannabis sativa SSR molecular markers and application thereof
  • Group of cannabis sativa SSR molecular markers and application thereof
  • Group of cannabis sativa SSR molecular markers and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1 Determination of cannabis SSR molecular markers and primers

[0050] According to 22 cannabis varieties (Yousha-31, Longma No. 1, Longmai No. 2, Huoma No. 1, Qingmai No. 1, Longmai No. 3, Longma No. 1, Hanma No. 2, Hanma No. 1 , Longma No. 2, Grecia, Hanma No. 5, Hanma No. 4, Muma No. 1, Qingma No. 2, Xianma No. 1, Longma No. 4, Qingmai No. 3, Qingmai No. 4, Hanma No. 6, Hanma No. 7, Longmai No. 5) gene sequences, search for SSR sites, and design 300 pairs of SSR primers; extract Qingmai No. 3, Hanma No. 7, Longmai No. 5, Huomayi The DNA of No. 1, Longma 1 and Yousha-31 were used to carry out PCR and polyacrylamide gel electrophoresis for these six varieties with these 300 pairs of primers, and analyzed and compared the annealing temperature, amplifiability, and stripping of each pair of primers. According to parameters such as band specificity, clarity and whether a stable single band can be obtained in the six varieties, 230 pairs of primers were prelimi...

Embodiment 2

[0053] Embodiment 2 Utilizes the SSR molecular marker provided by the present invention to distinguish cannabis varieties

[0054] (1) Rapid extraction of DNA

[0055] DNA extraction from hemp seeds was carried out by CTAB method. This method is quick and easy to operate, and the quality of the extracted DNA is high. It is suitable for DNA preparation in the field of plant molecular biology, and it is of great significance for greatly shortening the time of seed purity inspection and transgenic detection, improving detection efficiency, and reducing detection costs. The specific operation steps are as follows: take a few hemp seeds and grind them into powder, put them in a 1.5mL centrifuge tube; add 700 μL of CTAB extract to the centrifuge tube, and incubate at 65°C for 30 minutes; add 500 μL of chloroform-isoamyl alcohol ( 24:1), shake vigorously, centrifuge at 12000rpm for 10min; take the supernatant, add 0.7 times the volume of isopropanol, and centrifuge at 12000rpm for 5...

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Abstract

The invention relates to the technical field of molecular biology of crops, in particular to a group of cannabis sativa SSR molecular markers and application thereof. The invention provides a group of cannabis sativa SSR molecular markers, totally 11 SSR molecular markers, and also provides a set of primer combination suitable for detecting the SSR molecular markers on a capillary electrophoresis platform. The 11 SSR molecular markers can be used for identifying the variety of cannabis sativa and analyzing the genetic diversity; the varieties such as qing hemp No. 3 and China hemp No. 7, dragon hemp No. 5 and hemp No. 1, dragon hemp No. 1 and Euexin-31 are distinguished. Through the application of the cannabis sativa SSR molecular marker, the range of available marker sites of cannabis sativa can be expanded on the genome level; and a new tool is provided for researches on cannabis sativa varieties, germplasm resource identification, genetic relationship evaluation, cytoplasm hereditary characteristics and the like, and the application prospect is good.

Description

technical field [0001] The invention relates to the technical field of crop molecular biology, in particular to a group of marijuana SSR molecular markers and applications thereof. Background technique [0002] Hemp (Cannabis sativa L.) is a traditional economic crop, mainly used for textiles and oil extraction, and is an annual herb belonging to the genus Cannabis of the family Cannabinaceae. Hemp is an economic crop with special effects, and its bark, stalks, seeds, roots, leaves, and flowers are all valuable, and are widely used in textiles, food, papermaking, construction, medicine, transportation, and national defense. Especially cannabidiol (CBD, cannabidiol) in cannabis, which belongs to terpene phenolic compounds, has abundant pharmacological effects. Since the hallucinogenic component tetrahydrocannabinol (THC) in marijuana is easily used by criminals, reducing the THC content has become one of the important indicators for marijuana breeding. In order to distingui...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125C12Q2563/107
Inventor 张晓磊关海涛黄盈莹张瑞英温洪涛兰青阔
Owner 黑龙江省农业科学院农产品质量安全研究所