A method for isolating monocot guard cells

Abstract

The invention provides a method for isolating guard cells of monocotyledonous plants, comprising the following steps: (1) taking wheat seeds and inducing germination; (2) growing the wheat seeds in step (1) until the first leaf is not unfolded, Cutting and peeling the coleoptile from the seed embryo, longitudinally cutting the coleoptile into two halves, and tearing off the epidermal strip; (3) placing the epidermal strip prepared in step (2) in an enzymatic hydrolysis solution for enzymolysis; ( 4) Washing the cuticle strip prepared in step (3); (5) placing the washed cuticle strip under a microscope for microscopic examination. The method of the present invention uses wheat coleoptiles containing fewer types of cells, tears off epidermal cells, and then undergoes enzymolysis treatment with enzymolysis liquid, and the coleoptiles after enzymolysis treatment are cleaned to finally obtain high-purity, Highly active dumbbell-shaped wheat guard cells, isolated guard cells can be used for transcriptomic, proteomic and metabolomic studies.

Description

technical field [0001] The invention belongs to the field of plant physiology, and relates to a method for obtaining monocotyledonous plant single-type cells, and more specifically, relates to a method for isolating monocotyledonous plant guard cells. Background technique [0002] The stomata on the plant surface are surrounded by two guard cells that control CO 2 The absorption and loss of water, which in turn affects the photosynthesis and transpiration of plants. Guard cells can sense changes in environmental factors and respond to environmental changes by adjusting stomatal openings. The opening of the stomata is determined by the turgor pressure of the guard cells. When the concentration of osmotic adjustment substances in the guard cells increases, the water potential drops, and water flows into the guard cells from the surrounding cells, which increases the turgor pressure and the stomata open. On the contrary, the stomata will open. closure. Under the conditions o...

AI Technical Summary

Problems solved by technology

Micromanipulated dissection methods include laser capture microdissection and manual microdissection, but the application of these two methods in the study of guard cells is relatively limited because of the different requirements for material preparation, equipment and manipulation technique of these two methods. Relatively high, and the amount of isolated cells is limited, which is not conducive to the promotion of later research and technical methods

The free protoplast method requires less equipment and can obtain a large number of cells, but it will cause greater damage to the cells during the separation process

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