Sucrose density gradient analysis was used to show that polysomes were present in the mitospores ofAllomyces macrogynus. Fifty percent of the spore monosomes were shown to be resistant to dissociation by 0.8 M KCl, indicating that messenger ribonucleic acid (mRNA) was bound to them. These polysomes and all the spore ribosomes were contained in the nuclear cap. Only 4S RNA could be demonstrated in the extra-cap fraction. Hybridization studies using 3H-labeled polydeoxythymidylic acid indicated that polyadenylate was present to the extent of 0.08% of the total spore RNA. Sixty-eight percent of the polyadenylic acid is found in the nuclear cap, and 32% is found in the extra-cap fraction. It was demonstrated that [3H]uridine was taken up by the spores and converted to uridine triphosphate. Lack of incorporation of 3H into RNA indicated that the spores do not synthesize RNA. Thus, the mRNA found in spores is synthesized prior to spore formation.

The relationship between ribonucleic acid (RNA) synthesis and germination of Allomyces macrogynus mitospores was investigated. It was determined that the synthesis of all classes of RNA was initiated during the first 10 min of germination, around the time of encystment. It is during this stage that the membrane of the nuclear cap structure begins to break down, dispersing the cell complement of ribosomes throughout the cytoplasm. After encystment, there was an increase in the rate of synthesis of the four stable RNA species (4S, 5S, 19S, and 27S) which leveled off as the germ tube emerged. Data suggested that messenger RNA was synthesized at an increasing rate during the course of germination. Studies of RNA precursor pool behavior and RNA synthesis in the presence and absence of actinomycin D indicated that no species of RNA, including messenger RNA, was synthesized in the presence of actinomycin D. Further, precursor pool measurement indicated that the apparent increase in the rate of RNA synthesis during germination was largely due to increased specific activity of the RNA precursor pool.

Protein synthesis in Allomyces macrogynus mitospores began at the time of encystment and was required for further germination. The rate of protein synthesis in germinating spores decreased in the absence of ribonucleic acid synthesis relative to the uninhibited control. There was a concomitant decrease in polysome content, suggesting that some of the messenger ribonucleic acid synthesized during early germination is translated concurrently.

SUMMARYThe ability of 18 compounds to inhibit morphogenesis and/or growth of the diploid stage of Allomyces macrogynus was assayed. Of the three stages of development -germination, vegetative growth, and sporogenesis -germination appeared to be the most sensitive to inhibition and vegetative growth the least sensitive. Sporogenesis was sensitive to most of the inhibitors tested, but high concentrations were required for effective activity. Inhibitors could be grouped according to the stage of germination (cyst or rhizoid) at which they blocked development; those effective at the rhizoid stage, could be divided further on the basis of the resultant morphology of the germling.

Mitospores or cysts of Allomyces macrogynus do not take up the glucose analogue 2-deoxyglucose. Uptake of 2-deoxyglucose by germlings begins at 25 min into germination, the start of the rhizoid stage, and increases in rate by approximately 50-fold until 100 min into germination. The rate remains constant from 100 to 200 min, at which time germination is completed and hyphal formation begins. The presence of glucose in the germination medium blocks the uptake of 2-deoxyglucose. Of the other sugars tested, only galactose had any effect on 2-deoxyglucose uptake. Actinomycin D treatment during germination in a glucose-containing medium prevented the appearance of the uptake system, but actinomycin D was not effective after the transfer to a glucose-free medium. Cycloheximide treatment prevented the appearance of the uptake system if it was added at the time of the transfer to the glucose-free medium; it inhibited uptake only partially if the germlings were starved of glucose before its addition. It appears, therefore, that both ribonucleic acid synthesis during germination and protein synthesis after the removal of glucose are required for the uptake of 2-deoxyglucose.