Cypress microsatellite molecular marker combination, primer screening method and application of cypress microsatellite molecular marker combination

Abstract

The invention discloses a cypress microsatellite molecular marker combination, and a primer combination and screening method and application thereof, and belongs to the technical field of forest molecular biology. In the application, a method for identifying the polymorphism of cypress chloroplast microsatellite molecular marker primers comprises the steps that an M13 tail sequence is added at the 5' terminal of a forward primer of a designed primer pair, then the designed primer pair is mixed with a specific reverse primer, a general M13 primer with a fluorescence marker and a genome DNA, PCR amplification and electrophoretic separation sequencing are carried out, and polymorphic primers of cypress microsatellite molecular markers are obtained. A method for identifying the genetic relationship of cypress comprises the steps that the genome DNA of cypress is extracted, PCR amplification and electrophoretic separation are carried out with the polymorphic primers of the cypress microsatellite molecular markers, capillary electrophoresis sequencing is carried out, STR polymorphism is counted, the polymorphic primers can reflect the genetic information of cypress plasmon and can be used for researches such as genetic resource classification, genetic diversity analysis and the like.

Description

technical field [0001] The present invention relates to the field of forestry molecular biotechnology, in particular to cypress molecular marker technology, more specifically to a cypress microsatellite molecular marker and its screening method and its application in the fields of identification of cypress parentage relationship, variety identification and assisted breeding. Background technique [0002] Many species of Cupressaceae are the main wood raw materials, and are widely used in fields such as horticulture and medicine. The whole body of cypress is a treasure, and its resin, tree oil, fruit, branches and leaves can all be used as medicine. The most widely used clinically are the seeds, cypress kernels and leaves of orientalis arborvitae. Seed kernel: sweet, flat, can nourish the heart and calm the nerves, stop sweating, and moisten the intestines. Branches and leaves: bitter, slightly cold, cools blood and stops bleeding, grows hair and black hair, can cure hemate...

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Problems solved by technology

[0008] In order to solve the problem of the lack of effective means for identifying cypress germplasm resources in the prior art, the purpose of the invention is to provide a group of high-efficiency, operational methods that can be directly applied to intraspecific and interspecific cypress, and have good versatility and practicability. Simple, high detection accuracy, stable and efficient chloroplast polymorphism microsatellite molecular marker, its screening method and amplification primers, variety identification method and parent-origin relationship identification method

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