In vivo observation of the microfilament skeleton of plant cells
In a broad sense, the cytoskeleton is a type of organelle that contains three different components: microtubules, microfilaments, and intermediate fibers. So far, only two types of cytoskeleton, microtubules and microfilaments, have been identified in plant cells. They are highly dynamic within the cell and are involved in many important physiological processes: cell morphogenesis, cell division, cytoplasmic transport, and organelle movement. Intracellular skeleton binding proteins are involved in the dynamic regulation of the cytoskeleton.
Operation method
In vivo observation of the microfilament skeleton of plant cells
Materials and Instruments
Material: Move The basic process of in vivo observation of the microfilament skeleton in plant cells can be divided into the following steps: In order to adapt to the needs of function, different types of plant cells form their unique morphology. Young roots and leaves of Arabidopsis thaliana transfected with GFP-Lifeact gene were taken to make clinical slices, and the epidermal cells and leaves of young leaves were carefully observed under the bright field of a microscope. The morphology of epidermal cells on young leaves and root hair cells on young roots was carefully observed in the bright field of a microscope. Then we observed the arrangement of microfilament skeleton in these two types of cells under fluorescence (excitation wavelength 488 nm). 1. Root hair cells in the elongation zone of young roots The shape of the cells was long and tubular, and the arrangement of the microfilament skeleton in most of the root hair cells was roughly parallel to the growth direction of the root hairs. 2. Leaf epidermal cells of young leaves Most of the epidermal cells of mature dicotyledonous plants are tightly arranged and irregularly polygonal. The epidermal cells of young leaves are still growing and developing, and the microfilament skeleton within them is densely packed at certain parts of the cell and plays a role in the further extension of that part.
Arabidopsis thaliana transferred with GFP-Lifeact gene.
Apparatus:
① Fluorescence microscope
② Tweezers
① Fluorescence microscope ② Tweezers ③ Slide
① Fluorescence microscope ② Tweezers ③ Slides ④ Coverslips
Reagents:
① PBS solution
① PBS solution ② NaCl solution (0.5 mol/L)
The microfilament skeleton within plant cells is highly dynamic, and the arrangement of the microfilament skeleton can change significantly depending on the growth state of the plant cell itself, as well as the different stimuli of external environmental signals.
The microfilament skeletons in leaf epidermal cells and root hair cells were observed under a fluorescence microscope (excitation wavelength 488 nm), and the microfilament skeleton arrays in most of the leaf epidermal cells and root hair cells were plotted respectively. The mount was then removed, and NaCl solution (0.5 mol/L) was added dropwise on one end of the coverslip, and absorbent paper was placed on the other end of the coverslip to gently infiltrate the specimen with NaCl solution, and then the specimen was left to stand for 2 min to observe the changes of the microfilament skeleton array under the fluorescence microscope.
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