Vegetative microspores

Vegetative microspores of Cryptogams are unicellular objects with hard cover, blue-fluorescence in ultra-violet light and elaters [serve as anchor to a substrate (soil)]. The cells are diploid and have autotrophic nutrition due to the presence of chloroplasts, where photosynthesis occurs. They germinate well in artificial nutrient medium or in water and ultra-violet light induces significant autofluorescence. 

Materials required Vegetative microspores of Equisetum arvense L. [collected from the meadows in end April or in May and stored in dry conditions] ordinary and luminescent microscopes with photocamera (one microscope with luminescent and transparent regimes), and if possible,... 

Fig. 1 Vegetative microspores of horsetail Equisetum arvense. Left - Dry spore with elaters fluoresce under UV light (360-380 nm) Right - Germinated spore without elaters. The blue-fluorescing cover is missed.

Experiment 1. Effects of allelochemicals on the germination of horsetail vegetative microspores. 

Table 1 The effects of volatile plant excretions and water extracts (1 1000) w/v) from allelopathic species with pesticidal features on the germination of horsetaiI vegetative microspores.

Experiment 2. Effects of liquid allelochemicals on the germination of vegetative microspores. 

Fig. 2 The germination of Equisetum arvense vegetative microspores in the presence of anticontractile agents (left) and energetic inhibitors 10"5M (right). 1- colchicine 2-cytochalasin B oua - ouabain, ant - antimycin A diu - diuron, val - valinomycin.

Fig. 5 The effects of alkaloids -allelochemicals on the germination of vegetative microspores from Equisetum arvense and generative microspores (pollen) from Hippeastrum hybridum.

Materials required Vegetative microspores of Equisetum aroense or pollen from Hippeastrum hybridum microspores, ordinary light microscope, luminescent microscope... 

Fig. 2 LSCM view of dry and moistened (15 min) fluorescent vegetative microspore of Equisetum arvense under three laser excitation. 1 - channel 488 nm 2 - channel 533 nm 3- channel 633 nm 4-summed image with mixed (in a superposition pseudocolours. 1 bar = 20 pm.

Experiment 2. Visual slices of the vegetative microspore structure and computer modelling... 

Fig. 3 Optical slices (through 1 pm) of vegetative microspore of Equisetum arvense. Excitement with laser beams 488 (emission > 520 nm, green pseudocolor), 543 and 633 nm (fluorescence is observed at 650-750 nm, red pseudocolor).

Fig. 6 Microphotograph of fluorescent developing vegetative microspore of Equisetum arvense under laser beam 633 nm, (emission >670 nm).

Green-yellow fluorescence of the alkaloids is seen on the cell surface and in elaters of vegetative microspores from Equsetrum arvense (Fig. 9) shows the presence of tubulin in the parts. The middle of the spore lights in red due to... 

Procedure Cholinesterase activity was measured according to the modified biochemical methods developed for crude preparations (Gorunef ah, 1978), using Ellman reagent 5,5"-dithio-bis(p-nitrobenzoic acid) or its red analogue 2,2-dithio-bis-(p-phenyleneazo)-bis-(l-oxy-8-chlorine-3,6) -disulfur acid in the form of sodium salt, which interact with thiocholine salt (Roshchina 2001). Water extracts of vegetative microspores of horsetail (Equisetum arvense) or Hippeastrum hybridum microspores (150 mg of microspores in 30 ml for 1 h) were used. 

Observations The reaction lasts 1 h. The results are expressed in pM s 1kg" 1 of fresh mass. As shown in our experiments, the rates of hydrolysis of acetylthiocholine of studied plant cells varied from 0.4 0.05 pM s 1kg 1 of fresh mass for pollen of knights star up to 0.105 0.01 pM s 1kg 1 of fresh mass for horsetail vegetative microspores. Table 1 shows I50 for inhibition of cholinesterase from microspores by used inhibitors. 

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