THE LOSS OF KINETOPLASTIC DNA IN TWO SPECIES OF TRYPANOSOMATIDAE TREATED WITH ACRIFLAVINE

The effects of acriflavine on two species of Trypanosomatidae, Crithidia luciliae and Trypanosoma mega, have been investigated. It has been observed that kinetoplastic (i.e. mitochondrial) DNA is lost in a high percentage of acriflavine-treated cells. Resting flagellates, from stationary-phase or hemin-deficient cultures, are considerably more resistant to the acridine than are flagellates from a log-phase culture. When the kinetoplast has retained some DNA and still remains visible in stained smears, it appears reduced in size, and its ultrastructure is extremely abnormal: the DNA fibrils, clearly visible in normal kinetoplasts, are condensed; they appear as an electron-opaque, apparently homogeneous mass, separated from the membranes by a space of low electron-opacity. Analyses of DNA extracts, with high speed centrifugation in CsCl density gradients, revealed that the satellite band, presumably kinetoplastic DNA, is lost by trypanosomes grown for 5 days in the presence of acriflavine. Radioautography was used to study the effects of acriflavine on thymidine-³H incorporation in C. luciliae. At the concentration which affects the kinetoplast specifically, the dye produces an 87% inhibition of thymidine incorporation in this organelle. The kinetics of this inhibition suggest a direct effect on replication. No decrease in incorporation occurs in the nucleus. These results lead to the conclusion that loss of kinetoplastic DNA is due to continued growth and cell division in the absence of kinetoplastic DNA replication. Several hypotheses are discussed concerning the specificity of the dye's action upon the replication of extrachromosomal DNA.     

Effects of Ethidium Bromide and Several Acridine Dyes on the Kinetoplast DNA of Leishmania tropica*†

Whole cell DNA from Leishmania tropica has 2 peaks when banded by CsCl equilibrium density centrifugation. The main band has a buoyant density of 1.721 and the satellite band a buoyant density of 1.705, with Clostridium perfringens DNA (ρ= 1.6915) used as a reference. The satellite band has been identified as the kinetoplast DNA by purifying DNA from isolated kinetoplasts. L. tropica has the highest G + C content of both nuclear and kinetoplastic DNA thus far reported for trypanosomatids. The effects of ethidium bromide, acriflavin, proflavin, and 5-aminoacridine on the kinetoplast of L. tropica have been compared. Ethidium bromide and acriflavin, but not proflavin or 5-aminoacridine, induce dyskinetoplasty. L. tropica is one of the most sensitive trypanosomatids to ethidium bromide and acriflavin. Examination of the DNA from drug-treated cells in CsCl gradients revealed a loss of the satellite band after ethidium bromide or acriflavin treatment, but not after proflavin or 5-aminoacridine treatment. Cell division was required to produce these effects on the kinetoplast.     

The Morphology of Ovine Trypanosoma melophagium (Zoomastigophorea: Kinetoplastida)1

Morphologic and biometric data on bloodstream stages of Trypanosoma melophagium are presented. An increasing parasitemia with 111 trypomastigote stages of T. melophagium were found in Giemsa-stained thin blood smears taken from a splenectomized, cortisone-treated sheep recently infested with Melophagus ovinus infected with T. melophagium. The arithmetic mean and standard deviation in μm of the distances between posterior end and kinetoplast were 14.7 and 2.9, from the kinetoplast to the center of the nucleus 5.1 and 1.1, and from there to the anterior end 19.5 and 1.9. The free flagellum measured 6.0 μm ± 1.6 μm. The median and the range of the central 70% of values (median ± 35%) of the nuclear index were 1.1 and 0.9–1.2 and of the kinetoplastic index 3.8 and 3.3–4.9. The same data in μm for the maximal width were 3.1 and 2.1–4.6, and for the width at the level of the nucleus 2.9 and 2.2–4.6. The larger and smaller diameters of the nucleus measured 2.6 (2.2–3.7) μm and 1.7 (1.3–1.7) μm, respectively. The corresponding kinetoplast diameters were 1.1 (0.9–1.3) μm and 0.9 (0.6–0.9) μm, respectively.     

The morphology of ovine Trypanosoma melophagium (zoomastigophorea: kinetoplastida)

Morphologic and biometric data on bloodstream stages of Trypanosoma melophagium are presented. An increasing parasitemia with 111 trypomastigote stages of T. melophagium were found in Giemsa-stained thin blood smears taken from a splenectomized, cortisone-treated sheep recently infested with Melophagus ovinus infected with T. melophagium . The arithmetic mean and standard deviation in micron of the distances between posterior end and kinetoplast were 14.7 and 2.9, from the kinetoplastic to the center of the nucleus 5.1 and 1.1, and from there to the anterior end 19.5 and 1.9. The free flagellum measured 6.0 microns +/- 1.6 microns. The median and the range of the central 70% of values (median +/- 35%) of the nuclear index were 1.1 and 0.9-1.2 and of the kinetoplastic index 3.8 and 3.3-4.9. The same data in microns for the maximal width were 3.1 and 2.1-4.6, and for the width at the level of the nucleus 2.9 and 2.2-4.6. The larger and smaller diameters of the nucleus measured 2.6 (2.2-3.7) micron and 1.7 (1.3-1.7) micron, respectively. The corresponding kinetoplast diameters were 1.1 (0.9-1.3) microns and 0.9 (0.6-0.9) micron, respectively.     

Culture and morphological observations on Trypanosoma melphagium.

The cultural characteristics of Trypanosoma melophagium of sheep were studied. Aspects investigated were size of the inoculum and population growth in Modified Monophasic Medium for Trypanosomes (MMMT), population growth in Medium 199 with 10% inactivated calf serum containing 5, 10, and 15% hemolyzed defibrinated rabbit blood (199-CS-5, 199--CS-10, 199-CS-15) at 27 degrees C, effects on population growth of temperature and hydrogen ion concentration in MMMT, and morphology and morphometrics of the developmental stages found under different experimental conditions. The best growth occurred in medium MMMT at 30 degrees, C, pH 7.25. Temperature seemed to be a critical factor for differentiation of epimastigotes to trypomastigotes. Statistically significant differences were found between the trypomastigotes in MMMT and 199--cs-5 at 37 degrees C on day 4 of incubation for the following measurements: PK (distance from posterior end to kinetoplast), KN (from kinetoplast to middle of nucleus), PN (from posterior end to middle of nucleus), and nuclear and kinetoplastic indices. The trypomastigotes formed in both media were much smaller in size than the blood forms reported by Hoare (1972).     

FINE STRUCTURE AND REPLICATION OF THE KINETOPLAST OF TRYPANOSOMA LEWISI

The kinetoplastic DNA of Trypanosoma lewisi is described as a filamentous body lying within a mitochondrion, with the filaments oriented parallel to the long axis of the cell. The manner of fixation, the replicative state, and perhaps the physiological state of the cell, may result in slight morphological differences among such bodies. The kinetoplastic DNA replicates to form "left" and "right" rather than "upper" and "lower" members, and both the kinetoplast and nucleus incorporate radiothymidine as shown by radioautography. Radioautographic analyses suggest a random incorporation of radiothymidine by kinetoplasts. Silver grains were occasionally observed over centriolar elements. Finally, the observations are discussed with respect to the sequential replication of the aforementioned organelles by T. lewisi.     

Isolation and characterization of circular DNA molecules heterogeneous in size from a dyskinetoplastic strain of Trypanosoma equiperdum

In a naturally occuring dyskinetoplastic mutant strain of $\text{T. equiperdum}$, covalently closed circular DNA molecules of assumed mitochondrial origin were isolated. These molecules, heterogeneous in size, represent 6–9 % of total DNA and are essentially organized in catenated oligomers composed of molecules of different length. The typical molecular organization of the kinetoplast DNA from kinetoplastic trypanosomes, the network, was not observed.     

Ultrastructure of Trypanosoma conorhini in the Crithidial Phase*

SYNOPSIS. The ultrastructure of the crithidial phase of T. conorhini culture has been studied. The structure of the plasma membrane is not easy to make out; only at a few points in highly magnified picit is seen as a double osmiophilic membrane with an intermedilayer of low density. Sub-pellicular tubules are present also around the flagellar pocket. The nucleus usually has a single large nucleolus, but sometimes this may be double. The flagellum has the sual 2 central and 9 double peripheral fibers, the latter having lateral arms. The structure of the kinetoplast is similar to that of other rypanosomatids but the division of the organelle seems to be more complex than has been described: the central, DNA-bearing lamellae duplicate, forming a double transverse band; 1 of these bands probably migrates toward the side before invagination of the membrane completes the division. Mitochondria are long tubular structures with few cristae, disposed chiefly along the periphery of the cell. A communication between the kinetoplast and tubular mitochondria is very frequent. The endoplasmic reticulum is poorly developed, represented chiefly by smooth membranes surrounding vesicles, rough-surfaced membranes being scanty in most cells. Several inclusions may be found, some probably lipids, others being of unknown nature.     

Cytochemical study of nucleic acids in relation to the polymorphism of the trypomastigote forms of Trypanosoma cruzi

Four morphological types of T. cruzi trypomastigotes are distinguished in mouse blood. These differ in RNA contents, in the distribution pattern of RNA in the cytoplasm, in the intensity of the Feulgen reaction and the topography of DNA in the nucleus, and in the contents and distribution of both the nucleic acids in the kinetoplast. Among the trypomastigotes examined, forms C and S differ at a lesser degree, than their slender and middle variants differing much stronger. The early slender trypomastigotes are characterized by poor and diffuse RNA in the cytoplasm, by a homogeneous distribution of DNA in the nucleus and by a low content of DNA (sometimes RNA) in the kinetoplast. The middle trypomastigotes, dominating at the final step of the infection, are rich in granular RNA, differ (despite their inability to divide) in their nuclear organization mostly characterized by a large karyosome and uneven distribution of chromatin at the periphery; the kinetoplast is rich in DNA and often contains RNA. The peak of trypomastigotes with the kinetoplast deprived of obviously stained RNA precedes the impetuous increase of parasitemia. It coincides with the decrease in the number of destroyed parasites, and with the active substitution of slender variants by the middle ones within both C- and S-forms. Thus, changes in the nucleus and kinetoplast are involved in the trypomastigote transformation and in the development of infection.     

In Vitro Cellular Division of Trypanosoma abeli Reveals Two Pathways for Organelle Replication

Since the observation of the great pleomorphism of fish trypanosomes, in vitro culture has become an important tool to support taxonomic studies investigating the biology of cultured parasites, such as their structure, growth dynamics, and cellular cycle. Relative to their biology, ex vivo and in vitro studies have shown that these parasites, during the multiplication process, duplicate and segregate the kinetoplast before nucleus replication and division. However, the inverse sequence (the nucleus divides before the kinetoplast) has only been documented for a species of marine fish trypanosomes on a single occasion. Now, this previously rare event was observed in Trypanosoma abeli, a freshwater fish trypanosome. Specifically, from 376 cultured parasites in the multiplication process, we determined the sequence of organelle division for 111 forms; 39% exhibited nucleus duplication prior to kinetoplast replication. Thus, our results suggest that nucleus division before the kinetoplast may not represent an accidental or erroneous event occurring in the main pathway of parasite reproduction, but instead could be a species‐specific process of cell biology in trypanosomes, such as previously noticed for Leishmania. This “alternative” pathway for organelle replication is a new field to be explored concerning the biology of marine and freshwater fish trypanosomes.     

Development of hamster circadian rhythms: Role of the maternal suprachiasmatic nucleus

Summary During development, the circadian rhythms of rodents become entrained to rhythmicity of the mother. Rhythms in behavior and in neuroendocrine function are regulated by a circadian pacemaker thought to be located within the suprachiasmatic nucleus (SCN) of the hypothalamus. Evidence indicates that this pacemaker begins to function and to be entrained by maternal rhythms before birth. Although the maternal rhythms which mediate prenatal entrainment of the fetal circadian pacemaker have not been identified, it is likely that they are regulated by the maternal SCN.The role of the maternal SCN in entrainment of the offspring was examined in Syrian hamsters (Mesocricetus auratus) by measuring the activity/rest rhythms of pups. Using the synchrony among the rhythms of pups within a litter as an indication that the pups had been entrained, the effect on entrainment of ablating the maternal SCN was determined. Lesions of the maternal SCN which were performed early in gestation (day 7) and which destroyed at least 75% of the SCN were found to disrupt the normal within litter synchrony among pups, indicating interference with the normal mechanism of entrainment.The effect of lesions on day 7 of gestation could mean that the maternal SCN is important for entrainment of the pups before birth, after birth, or during both of these times. To determine if the maternal SCN is specifically important for prenatal entrainment, lesions were performed two days before birth on day 14 of gestation. Lesions of the maternal SCN on day 14 were not as disruptive as were lesions on day 7. This suggests that the maternal SCN is important between days 7 and 14 of gestation and that the synchrony normally observed at weaning is already established, in part, on or before day 14 of gestation. This further suggests that an entrainable circadian pacemaker is present in the fetus only two weeks after fertilization.Abbreviations SCN suprachiasmatic nucleus - L:D light:dark - LL constant light - r mean vector length - 2DG 2-deoxyglucose - NAT N-acetyltransferase     

The kinetoplast DNA of Trypanosoma equiperdum

We have analyzed the kinetoplast DNA for Trypanosoma equiperdum (American Type Culture Collection 30019) and two dyskinetoplastic strains derived from it. The DNA networks from the kinetoplastic strain are made up of catenated mini-circles and maxi-circles, like the networks from the closely-related Trypanosoma brucei. The mini-circles of T. equiperdum lack the pronounced sequence heterogeneity of T. brucei mini-circles, as shown by the fragment distribution of restriction digests and by the predominance of well-matched duplexes in electron micrographs of renatured DNA. The electrophoretic analysis of kinetoplast DNA digested with various restriction endonucleases shows the maxi-circle of T. equiperdum to consist of circular DNA molecules of 8.4 x 10(6) daltons, without size or sequence heterogeneity or repetitious segments. A comparison of the sequence by restriction endonuclease fragmentation and hybridization shows extensive sequence homology. The size difference between both maxi-circles is due to the deletion of one continuous segment of 5.10(6) daltons. In the two dyskinetoplastic strains, we cannot detect DNA sequences that hybridize with kinetoplast DNA from T. brucei or from the kinetoplastic strain of T. equiperdum. In one of these strains, a 'low-density' DNA fraction contained a simple sequence DNA, cleaved by restriction endonuclease HindIII into fragments of 180 base-pairs and multimers of this. The relation of this DNA to kinetoplast DNA, if any, is unknown.     

Mitosis in the Hemoflagellate Trypanosoma cyclops*

SYNOPSIS. The ultrastructure of interphase and mitotic nuclei of the epimastigote form of Trypanosoma cyclops Weinman is described. In the interphase nucleus the nucleolus is located centrally while at the periphery of the nucleus condensed chromatin is in contact with the nuclear envelope. The nucleolus fragments at the onset of mitosis, but granular material of presumptive nucleolar origin is often recognizable in the mitotic nucleus. Peripheral chromatin is in contact with the nuclear envelope throughout mitosis, and it seems reasonable to assume that the nuclear envelope is involved in its segregation to the daughter nuclei. Spindle microtubules extend between the poles of the dividing nucleus and terminate close to the nuclear envelope. The basal body and kinetoplast divide before the onset of mitosis and do not appear to have any morphologic involvement in that process. Spindle pole bodies, kinetochores, and chromosomal microtubules have not been observed.     

Effects of temperature on phenological synchrony and altitudinal distribution of jumping plant lice (Hemiptera: Psylloidea) on dwarf willow (Salix lapponum) in Norway

Summary.

• 1 The geographical distributions of three species of jumping plant lice (psyllids) along an altitudinal transect (988–1300 m a.s.l.) in southern Norway were restricted within the range of their host plant Salix lapponum. One species, Cacopsylla propinqua, occurred at all sampling locations between 988 and 1222 m, whereas C.palmeni was confined to higher altitudes (1153–1222 m) and C.brunneipennis was more abundant at lower altitudes (988–1101 m).
• 2 C.brunneipennis and C.palmeni developed only on female catkins. Development times of catkins and psyllids were similar (approximately 50 days) and successful psyllid development depended on close phenological synchrony with catkins.
• 3 Thermal requirements for development of female catkins were greater at low altitude (988 m) compared with higher altitude (1222 m), showing local adaptation of S.lapponum to altitude. In general, thermal requirements of psyllids were less than those of catkins at the same location. C.brunneipennis had higher thermal requirements than C.palmeni.
• 4 Field experiments, using polythene enclosures to elevate temperatures at two sites at different altitudes (by 0.6–1.4 deg. C), showed that insects had an enhanced relative rate of development under elevated temperatures compared with their host plants.
• 5 Indices of phenological synchrony were calculated from thermal requirements of psyllids and catkins. Under elevated temperatures, phenological synchrony decreased at both sites. This resulted in the subsequent development of smaller adult insects at low altitude, although at higher altitude, insects developing under elevated temperatures were larger and had a higher survival rate compared with controls.
• 6 Effects of temperature on phenological synchrony may explain the limits to the geographical range of psyllids. The consequences of climate change on psyllid populations will depend on the effects of decreased phenological synchrony on insect development and this may differ within the insect's geographical range.

     

Variations in morphological and life-history traits under extreme temperatures in <Emphasis Type="Italic">Drosophila ananassae</Emphasis>

Using half-sib analysis, we analysed the consequences of extreme rearing temperatures on genetic and phenotypic variations in the morphological and life-history traits of Drosophila ananassae. Paternal half-sib covariance contains a relatively small proportion of the epistatic variance and lacks the dominance variance and variance due to maternal effect, which provides more reliable estimates of additive genetic variance. Experiments were performed on a mass culture population of D. ananassae collected from Kanniyakumari (India). Two extremely stressful temperatures (18°C and 32°C) and one standard temperature (25°C) were used to examine the effect of stressful and non-stressful environments on the morphological and life-history traits in males and females. Mean values of various morphological traits differed significantly among different temperature regimens in both males and females. Rearing at 18°C and 32°C resulted in decreased thorax length, wing-to-thorax (w/t) ratio, sternopleural bristle number, ovariole number, sex comb-tooth number and testis length. Phenotypic variances increased under stressful temperatures in comparison with non-stressful temperatures. Heritability and evolvability based on among-sires (males), among-dams (females), and the sum of the two components (sire + dam) showed higher values at both the stressful temperatures than at the non-stressful temperature. These differences reflect changes in additive genetic variance. Viability was greater at the high than the low extreme temperature. As viability is an indicator of stress, we can assume that stress was greater at 18°C than at 32°C in D. ananassae. The genetic variations for all the quantitative and life-history traits were higher at low temperature. Variation in sexual traits was more pronounced as compared with other morphometric traits, which shows that sexual traits are more prone to thermal stress. Our results agree with the hypothesis that genetic variation is increased in stressful environments.     

Fine Structure and Cytochemistry of the Nucleus and the Kinetoplast of Epimastigotes of Trypanosoma cruzi1

ABSTRACT. Ultrastructural cytochemical techniques were used to analyze the nucleus and the kinetoplast of epimastigotes of Trypanosoma cruzi. With the use of ethanolic phosphotungstic acid, which detects basic proteins, reaction product was seen in the chromatin and at the periphery of the kinetoplast. Thallium alcoholate, which interacts with DNA, stained strongly the whole kinetoplast and the chromatin. With the use of a silver impregnation method that detects acidic nucleolar proteins, silver granules were seen preferentially located in the central region of the nucleolus. With the EDTA method, which reveals the presence of ribonucleoproteins, staining was observed in the nuclear pores. Also 6–8 nm fibrils, 25 nm and 40 nm granules, which correspond to the perichromatin fibers, interchromatin granules and the perichromatin granules, respectively, were identified in the nucleus. The EDTA method also revealed the presence of 40 nm granules in the kinetoplast. These granules were seen mainly at the two extremities of the kinetoplast. Freeze-fracture images indicate that the nuclear membrane contains ca. 9 pores/μm² of nuclear surface area. The mean diameter of the pores was 80 nm. All these results suggest that epimastigotes of T. cruzi have a very active nucleus and a high rate of nucleocytoplasmic interchange.     

In vitro Transcription of Kinetoplast and Nuclear DNA in Kinetoplastida*†

SYNOPSIS. DNA-dependent RNA polymerases have been solubilized from homogenates of Crithidia fasciculata using gentle extraction procedures. RNA polymerase I and II are separated on DEAE cellulose at 0.07M (NH₄)₂SO₄ and 0.13M (NH₄)₂SO₄ respectively. RNA polymerase II is inhibited 80% by α-amanitin (25 μg/ml). Both RNA polymerases require DNA as a template, ribonucleoside triphosphates and Mn²⁺. The synthesis of RNA as a product is inhibited by DNase. RNase, pronase and actinomycin D. Purified kinetoplast and nuclear DNA can serve as templates for the RNA polymerases. Denatured DNA templates are preferred. The synthesis of RNA continues for at least an hour and is inhibited by trypanocidal drugs including suramin. antrycide, acriflavine, ethidium bromide and berenil. Complementary RNA synthesized in vitro from C. fasciculata kinetoplast DNA hybridizes with C. fasciculata kinetoplast DNA but not with C. fasciculata nuclear DNA or Blastocrithidia culicis kinetoplast DNA, Escherichia coli, T4 or calf thymus DNAs. The complementary RNA synthesized in vitro from C.fasciculata kinetoplast DNA sediments at 4–5S.     

The Fine Structure of the Flagellum and Kinetoplast-Chondriome of Trypanosoma (Schizotrypanum) cruzi in Tissue Culture*

SYNOPSIS. Trypanosoma cruzi in tissue cultures was studied with the electron microscope after double fixation in glutaraldehyde and osmium tetroxide, and embedding in Epon. Previous findings on its fine structure were confirmed, and some new structures were found in the flagellum and kinetoplast-chondriome. In the flagellum, an intraflagellar body was found, similar to that observed in other trypanosomes, beginning at the base of the flagellum and running along the axial fibre bundle thruout its length. The axial fibre bundle is formed by interconnecting tubules, the outer ones apparently smooth, the inner ones with a helical substructure. Lateral extensions from the outer tubules in the flagellar bundle seem to enter the intraflagellar body. The kinetopiast in the leishmania bodies has the same electrondense structure described before. In the trypanosome form it has assumed a large spherical shape, in which the formerly short, compressed fibres have grown in length, are more dispersed and have an irregular shape. They are oriented in the direction of the body's length in parallel array. The whole formation is continuous with a long mitochondrion which begins in the region of the nucleus and extends up almost to the tip of the trypanosome. The matrix of the kinetoplast in these forms is electron-transparent; the matrix of the mitochondria is rather dense. In a few extracellular trypanosomes, a special structure was found in which the kinetoplast is composed of electron-transparent formations, arranged in orderly horizontal lines quite similar to the mitochondrial cristae of the parasite. The significance of this structure is uncertain.     

Multiplication of Trypanosoma pacifica (Euglenozoa: Kinetoplastea) in English sole, Parophrys vetulus, from Oregon coastal waters

Multiplication of Trypanosoma pacifica was common in the fish host from observations of live flagellates and Giemsa-stained blood smears. Multiplication began with the elongation of the kinetoplast, thickening of the posterior portion of the body, and appearance of a new flagellum near the kinetoplast. The new flagellum was very rigid when less than 3 microm in length, but it became flexible as it elongated. When the new flagellum was approximately 12 microm in length, cell division began and the kinetoplast also began to divide. The timing of nuclear division was variable. Generally, it did not occur until division of the kinetoplast had begun, but occasionally binucleate individuals were observed before cell or kinetoplast division was apparent. As division continued, 1 nucleus migrated past the dividing kinetoplast into the future daughter trypanosome. Finally, the kinetoplast completed division and the trypanosomes separated. Cell division was unequal, with the daughter trypanosome being smaller than the parent and with a more weakly developed undulating membrane.     

Two alleles of a developmentally regulated alpha-tubulin locus in Physarum polycephalum replicate on different schedules.

The replication timing of a pair of natural alleles was compared at two alpha-tubulin loci of the Physarum plasmodium. Taking advantage of the naturally synchronous cell cycle of nuclei within the syncytial plasmodium, we analyzed the replication schedule of specific DNA fragments to a resolution of 10-min intervals within a 3-h S phase. At this level of resolution, differences in replication timing between polymorphic alleles at the same locus can be detected in a heterozygote. Specifically, the 3' region of the altA1 allele completes replication at between 20 and 40 min of S phase. The same region of the altA2 allele completes replication at between 40 and 80 min of S phase. In contrast, both alleles at the altB locus replicate concurrently within the first 10 to 15 min of S phase. Previous studies showed that both altA and altB are expressed in the plasmodium, their message levels peaking at mitosis, just minutes before the onset of S phase. However, altB message is detected at substantially higher levels than altA message on Northern (RNA) blots. The temporal windows over which the altA alleles each replicate are very broad in comparison with the levels of mitotic synchrony and altB replication synchrony in a single plasmodium. The allele-specific replication schedule of the altA locus demonstrates that the temporal organization of replicons is not strictly conserved between homologous chromosomes.