Quick extraction method of high-activity mitochondria in plant organs and special extracting solution thereof

Abstract

The invention discloses a quick extraction method of high-activity mitochondria in plant organs and special extracting solution thereof. The quick extraction method comprises the following steps: mixing the plant organs, the extracting solution and grinding medium, homogenizing, collecting filtrate, carrying out differential centrifugal separation on the mitochondria in the plant organs, and thenadopting mitochondrial respiratory buffer solution for storage. The extracting solution is prepared from sucrose, sodium pyrophosphate, EDTA (Ethylene Diamine Tetraacetic Acid), potassium dihydrogen phosphate, BSA (Bovine Serum Albumin), cysteine, ascorbic acid and pvp-40. Proved by the experiment, the quick extraction method disclosed by the invention has the following advantages that the efficiency is high, i.e., the activity of the mitochondria is high; the applicability is wide, so that the quick extraction method can be applicable to extraction of the mitochondria of flower organs of multiple angiosperms; the storage time is long, and the activity can be maintained for 5 hours on an ice box, so that a great number of studies on the energy metabolism and the respiration of the mitochondria can be met; the operation is simple and the consumed time is short; the method provided by the invention lays a solid foundation for study on the respiratory chain and the energy metabolism of the mitochondria and has important practical application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a rapid extraction method of highly active mitochondria in plant organs and a special extraction solution thereof. Background technique [0002] As one of the important characteristics of angiosperms, flowers are composed of sepals, petals, stamens (stamen group) and pistils (gynosperm group), which are closely related to the reproductive process of plants and are important for ensuring the pollination, fertilization and smooth fruiting of angiosperms. organ. Therefore, floral organs have attracted much attention as the key object of plant reproductive biology research. Moreover, as the flower organ is an important place for plant reproduction and reproduction, its life activities are vigorous, and mitochondria, as an important organelle of energy metabolism, can perform physiological activities such as oxidative phosphorylation and electron transfer in response to diffe...

AI Technical Summary

Problems solved by technology

[0003] At present, the research systems for extracting highly active mitochondria for respiration, energy metabolism and superoxide levels are mostly reported in animal studies, and the extraction technology in cardiomyocytes and skeletal muscle cells is more mature; however, in plants, extracting mitochondria The research objects mainly focus on the model plants Arabidopsis thaliana, rice and their single-cell pollen tubes, etc., and the extraction method is too cumbersome

Most of the plant species in the current research are non-model plants, and the research objects are mostly tissues and organs with active plant physiological states. So far, there is no rapid extraction of non-model plants, especially for flower organs with important reproductive functions. The technical method of highly active mitochondria has led to a relatively lagging research on the mitochondrial respiration of floral organs in plant reproductive development and even plant energy metabolism. Therefore, the extraction technology of high-efficiency and active plant mitochondria is urgent and crucial.

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