Samples from synchronized cultures of Blastocladiella emersonii were examined by electron microscopy from the late log phase to the completion of zoospore differentiation. Log‐phase plants contain the usual cytoplasmic organelles but also have an unusual system of large tubules ca. 45 mμ diam that ramify in organized bundles throughout the protoplast. After induction, zoosporangium differentiation requires a 2‐hr period in which the nuclei divide, a cross wall forms to separate the basal rhizoid region, and an apical papilla is produced. Nuclear division in B. emersonii is intranuclear with a typical microtubular spindle apparatus and paired, unequal, extranuclear centrioles at each pole. The papilla is formed by a process of localized cell wall breakdown and deposition of the papilla material by secretory granules. Differentiation of zoospores begins when one of the two centrioles associated with each nucleus elongates to form a basal body. The flagella fibers arise from the basal body and elongate into an expanding vesicle formed by the fusion of small secondary vesicles. The cleavage planes are formed by fusion of vesicles similar to those associated with flagellum initiation. When cleavage is complete, each sporangium contains ca. 250–260 uninucleate spore units with their flagella lying in the cleavage planes. Probable fusion of mitochondria to produce the single mitochondrion of the zoospore occurs after cleavage; the mitochondrion does not take its position around the basal body and rootlets until just before zoospore release. The ribosomal nuclear cap is organized and enclosed by a membrane formed through fusion of many small vesicles during a short period near the end of differentiation.
Freshly harvested zoospores of Blastocladiella emersonii begin to germinate about 15 min after inoculation into a defined growth medium at a density of 106 zoospores per ml. Flagellum retraction accompanies encystment, and dispersal of the ribosomal nuclear cap takes place shortly thereafter. The primary rhizoid begins to emerge at 25 to 30 min and starts to branch at ca. 60 min. The first nuclear division occurs between 120 and 190 min. The dry weight per cell increases linearly after 60 min, whereas the deoxyribonucleic acid per cell doubles between 120 and 240 min. A linear increase in total ribonucleic acid (RNA) is detectable beginning at 40 to 45 min, and in total protein beginning at 80 min; neither process is interrupted during nuclear division. Encystment and nuclear cap disorganization are associated with a sharp rise in the rates of precursor incorporation into RNA and protein. Cycloheximide at 20 ,ug/ml prevents leucine incorporation at all stages and inhibits development beyond the earliest encystment stage. Actinomycin D at 25 ,4g to 50 ,ug/ml prevents uracil incorporation, but it has no effect on leucine incorporation or development until 40 to 45 min. At the latter stage, actinomycin D causes a sharp developmental arrest and begins to inhibit leucine incorporation. It is concluded that early protein synthesis must occur on the ribosomes formed during the prior growth phase and conserved through the zoospore stage in the nuclear cap. The results further indicate that this synthesis is dependent upon messenger RNA already present in the zoospore before germination.
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