Efficient akebia trifoliate ploidy detection method

Abstract

The invention discloses an efficient akebia trifoliate ploidy detection method. The method comprises the steps of preparing a sample pretreatment agent; to-be-detected material sampling: selecting materials such as calluses, hypocotyls, cotyledons, suspension culture cells, leaves and the like, repeatedly washing and cleaning by using sterilized double distilled water when the experimental materials are taken, selecting tender parts, taking 480 to 510mg of samples, and avoiding tissues such as plant veins, vascular bundles and the like with high metabolite content during sampling; preparing a cell nucleus suspension; performing instrument detection and quality control of a corresponding fluorescence channel; performing contrast and standard flow cytometry detection; and performing data comparison and ploidy determination. The method has the advantages that pretreatment steps are simplified, the cell nucleus suspension required by detection is efficiently prepared, RNA interference is eliminated, flow cytometry sample loading parameters are standardized, identification results are unified and standard, multiple methods are combined to guarantee accurate results, large-scale detection is achieved, results are visualized, and data transmission is convenient and fast.

Description

technical field [0001] The invention relates to the technical field of plant ploidy detection, in particular to an efficient method for detecting ploidy of Akebia trifoliate. Background technique [0002] Akebia trifoliata is a perennial woody vine of the genus Akebia in the family Akebia. The related research in China, Japan and Korea has developed rapidly, and the research fields involve cultivation technology, breeding technology and molecular basic research. As early as 1986, Chinese scholar Jiang Like analyzed the amino acids of its ripe fruit, and found in subsequent research that it is rich in free amino acids, oleanolic acid, flavonoids and saponins, etc., and is a new type of therapeutic fruit. In 2018, my country's Sichuan Province promulgated the "Sichuan Provincial August Melon Food Safety Local Standards" to boost the development of the Sanye Akebia industry. Its artificial planting area has accumulated more than 70,000 mu in Sichuan Province. Related deep-proce...

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Problems solved by technology

So far, there is no documented successful case of flow cytometry ploidy identification related to Akebia trifoliate in my country. Only Shi Xiaobing of Guizhou University used this technology in 2015, but due to the toxic effect of colchicum and the polyphenol content of Akebia trifoliata Many characteristics lead to strong interference peaks in the identification results and inaccurate results

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