Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture I. Some characteristics of the cell cycle

Cells of Chlamydomonas reinhardi Dangeard were grown synchronouslyunder a 12 hr light-12 hr dark regime. Time courses of nucleardivision, chloroplast division, "apparent cytokinesis" and zoosporeliberation were followed during the vegetative cell cycle inthe synchronous culture. Liberation of zoospores occurred atabout 23–24 hr after the beginning of the light periodat 25°C. Four zoospores were produced per mother cell underthe conditions used. At lower temperatures, the process of zoosporeliberation as well as length of the cell cycle was markedlyprolonged, but the number of zoospores produced per mother cellwas approximately the same. At different light intensities,lengths of the cell cycle were virtually the same, while thenumber of zoospores liberated was larger at higher rather thanat lower light intensities. During the dark period, nuclear division, chloroplast divisionand apparent cytokinesis took place, in diis order, and proceededless synchronously than did the process of zoospore liberation.When the 12 hr dark period was replaced with a 12 hr light periodduring one cycle, the time of initiation as well as the durationof zoospore liberation was litde affected in most cases, whereasnuclear division, chloroplast division and apparent cytokinesiswere considerably accelerated by extended illumination. Whenalgal cells which had been exposed to light for 24 hr were furtherincubated in the light, zoospore liberation started much earlierand proceeded far less synchronously, compared with that under12 hr light-12 hr dark alternation. (Received October 12, 1970; )     

FURTHER NOTES ON THE GROWTH AND CHLOROPHYLL FORMATION OF CHLORELLA PROTOTHECOIDES

1. Using area of a fixed concentration as a nitrogen source,cells of Chlorella protothecoides were grown in the presenceof various carbon compounds. Magnitudes of growth of the cellswere widely different depending on the carbon sources used;glucose and fructose being most favourable substrates and galactose,glycerol and acetate coming next. But the amounts of chlorophyllformed in the cells during the experimental period were almost,the same irrespective of the different carbon sources, withsome exceptions. The similarity of the chlorophyll level observedin these experiments seemed to indicate that the formation ofchlorophyll was limited largely by the nitrogen source but notby the carbon source. 2. Strong bleaching effect was recognized with glucose and fructoseat their high concentration, which produced totally chlorophyll-lesscells. On the other hand, a stimulating effect on chlorophyllformation was observed with galactose at the different concentrationsexamined. 3. Effects of glycine and ammonium carbonate as the nitrogensource on the algal growth and pigmentation were studied insome details. The results were similar to those previously obtainedwith urea, confirming our previous conclusion that the algalpigmentation is profoundly affected by the concentration balancebetween glucose and nitrogen source. ¹Present address: Tokyo Research Laboratories, Tanabe SeiyakuCo., Toda-machi, Saitama.     

DE- AND RE-GENERATION OF CHLOROPLASTS IN THE CELLS OF CHLORELLA PROTOTHECOIDES: II. EFFECTS OF ACTINOMYCIN ON GREENING OF "GLUCOSE-BLEACHED" AND "ETIOLATED" ALGAL CELLS

The "glucose-bleached" and "etiolated" cells of Chlorella protothecoideshaving plastids of different degrees of degeneration were preparedby the methods previously reported, and the effects of actinomycin(C complex) upon the processes of greening of these cells wereinvestigated under various experimental conditions. As has beenshown previously, these cells formed normal chloroplasts onbeing incubated in the light with provision of nitrogen source(urea), but without glucose. The greening process of the glucose-bleachedcells has been found to differ from that of the etiolated cellsin the point that it involves a light-independent phase precedinga light-requiring phase. It was revealed that the greening ofglucose-bleached cells is inhibited by actinomycin much morestrongly than that of etiolated cells. On applying the antibioticat different times during the chloroplast development in glucose-bleachedcells, it was found that the inhibitory effect was remarkablyreduced with the progress of the developmental process. Thisindicated that the antibiotic attacked more strongly the light-independentphase than the light-requiring phase in question. Based on theseobservations it was inferred that, in the process of chloroplastdevelopment in glucose-bleached cells, DNA and RNA are playingimportant roles, especially during the early light-independentphase of chloroplast development. (Received September 18, 1964; )     

Reproductive and subsocial behaviour in the ovoviviparous leaf beetle Gonioctena sibirica (Coleoptera: Chrysomelidae)

Abstract.

• 1 In the spring, females of the leaf beetle Gonioctena sibirica deposited larvae on the ventral surface of growing young leaves situated on the apical position of shoots of the willow Salix bakko.
• 2 The parent females remained with the larvae usually on the underside of the basal part of leaves, facing toward the base of shoots. When other arthropods approached, the females temporarily moved towards these intruders, showing aggressive behaviour such as swinging the body or stamping the legs. Many females remained with their larvae until the larvae grew into the final (fourth) instar. No female produced an additional brood in the field.
• 3 Broods from which parent females were experimentally removed suffered higher mortality than those in which females were left intact. Arthropods such as spiders and ants were observed preying on the larvae. In contrast, the survivorship of broods from which females were removed and intruders were excluded with a sticky substance applied to the base of twigs was not different from that of control broods. These results demonstrate that the main mortality factor of offspring is pedestrian arthropod predators and females physically repel the predators.
• 4 Potentially alternative reproductive strategies, such as producing a large number of offspring by iteroparity and/or larger brood(s) with less or no care, seem to be inhibited in G.sibirica by larval dependence on growing young leaves which are temporally limited and by ovoviviparity which may have limited brood size.

     

BIOSYNTHESIS OF ECDYSONE IN THE ISOLATED ABDOMEN OF THE SILKWORM,BOMBYX MORI*

During pupal-adult development of the silkworm, Bombyx mori, the ecdysone titer changes, exhibiting two maxima in the females: one on the second day of pupal development and the other just before adult emergence. During the second maximum, ecdysone accumulates in the ovaries. It also accumulates in isolated abdomens, which were prepared just after pupal ecdysis and induced to initiate adult development by injection with β-ecdysone. Several lines of evidence suggest that ecdysone is synthesized in the ovary itself.     

POLYOL ACCUMULATION IN IN VITRO SYSTEMS OF BOMBYX EGGS

The mechanism of polyol accumulation in diapausing Bombyx eggs, conversion of [6-¹⁴C] glucose-6-phosphate into polyols and other neutral sugars was investigated in in vitro reaction systems. When a crude homogenate or a press juice of the eggs was incubated with [6-¹⁴C]glucose-6-P, the labelled trehalose, sorbitol and glycerol accumulated in the reaction mixture. In the press juice incubation system of developing eggs at day 1, ¹⁴C-sorbitol was detected in appreciable amounts, but it decreased rapidly with the development of the embryos. When the press juice was prepared from eggs in diapause, the formation of ¹⁴C-sorbitol was 3–5 times greater in eggs at early stages (day 2 to day 4) than in developing eggs.     

CYCLIC NUCLEOTIDE-DEPENDENT PROTEIN KINASES FROM SILKWORM EGGS: OCCURRENCE OF TWO CYCLIC NUCLEOTIDE-DEPENDENT PROTEIN KINASES, CHANGES IN THEIR ACTIVITIES AND LEVELS OF CYCLIC NUCLEOTIDES DURING DEVELOPMENT OF THE EGGS

Two cyclic nucleotide-dependent protein kinases, designated as protein kinase-I and -II, were obtained from the eggs of the silkworm, Bombyx mori . Protein kinase-I is highly dependent on cGMP, whereas protein kinase-II is dependent on cAMP. In developing non-diapause eggs, the level of cyclic nucleotide-dependent protein kinase activity is quite high but that in the diapause eggs is not. The developmental changes in the two protein kinases and the level of cyclic nucleotides were also studied during the development of the eggs.     

Effects of temperature of adults and eggs on the induction of embryonic diapause in the band-legged ground cricket, Dianemobius nigrofasciatus

Abstract.  Eggs laid by adult female Dianemobius nigrofasciatus , reared under long-day (LD 16 : 8 h, 25 °C) or short-day (LD 12 : 12 h, 25 °C) conditions from the nymphal stage, are kept at several constant temperatures. At 22.5–30.0 °C, eggs laid by long-day adults show lower incidences of diapause than those laid by short-day adults. In both eggs laid by adults under long-day conditions and those under short-day conditions, the higher the temperature at which the eggs are kept, the lower the incidence of diapause. When eggs of long-day adults are exposed to a low-temperature pulse (10 °C, 24 h) on the day of deposition (day 0), the incidence of diapause increases. The low-temperature pulse on day 1 does not increase the incidence of diapause. By contrast, when the eggs of short-day adults are exposed to a high-temperature pulse (35 °C, 24 h) on day 0 or day 1, the incidence of diapause decreases. The temperature pulses on day 0 are more effective at diapause prevention. Staining of diapause eggs by the Feulgen–Rossenbeck method shows that the eggs enter diapause at the blastoderm stage, which is on day 1 or day 2 at 25 °C. The exposure of adults to long days and higher temperatures prevents the eggs from entering diapause. In D. nigrofasciatus , embryonic diapause is controlled by maternal effects, adult photoperiod and temperature, and egg temperature before or at diapause.     

Changes during Development in the Activities and Intracellular Locations of Glycerol Kinase and Hexokinase in Rat Brain and Liver

The changes in activities and intracellular locations of glycerol kinase in rat brain and liver during development were compared. Glycerol kinase activity was consistently much higher in the liver than in the brain from just before the birth to the adult stage. Most of the activity was bound to mitochondria in the brain, but the intracellular distribution of the activity in the liver changed during development, the amount of activity bound to mitochondria being high just after birth and then decreasing gradually. These changes of glycerol kinase during development were compared with those of hexokinase, and the significance of the changes in the two enzymes is discussed in relation to dietary changes during development.