Genetic divergence in Northamerican freshwater planarians of the Dugesia dorotocephala group (Turbellaria, Tricladida, Paludicola)

The genetic differentiation between the members of the Dugesia (Girardia) dorotocephala group was analyzed by means of multilocus electrophoresis, and comared to that of another planarian secies, D. tahitiensis, also belonging to the subgenus Girardia. The species examined were: D. dorotocephala s.s (2n = 16), D. arizonensis (2n = 8), D. jenkinsae (2n = 8), and the above mentioned D. tahitiensis (2n = 16). The former three species inhabit North America, and show different proportion of fissiparous and sexual individuals; the latter species inhabits Polynesia and is fully asexual. A total of 11 enzyme loci were genetically analyzed: Mdh-1, Mdh-2, Zdh-1, Idh-2, G3pdh, Got-1, Ck, Pgm-2, Ada, Mpi, and Gpi. Low values of observed mean heterozygosity per locus (Ho) were found in the populations studied, ranging from 0 to 0.18 (average 0.08. In asexual populations (except that of D. tahitiensis) fixed heterozygosity was observered in all the individuals for 1 or 2 loci. The genetic divergence between the species examined is very high, with many loci showing discriminating alleles in different taxa (Nei's genetic distance varies from 0.871 to 1.759). The populations of D. dorotocehala s.s., on the contrary, appear to be genetically quite homogenous average D= 0.019), and the genetic distance values are apparently unrelated to their geographic location and to their way of reproduction. The genetic distance between D. tahitiensis, a species not included in the D. dorotocephala group and D. dorotocephala s.s. is 1.314 and hence similar to the D value between two members of;he dorotocephala group: D. dorotocephala and D. jenkinsae (D = 1.303). The genetic relationships among the populations studied were established by UPGMA cluster analysis and multidimensional scaling. The descendence of the North American species with 2n = 8 from a dorotocephala-like ancestor with 2n = 16 is considered. It is suggested that the latter, as well as a tahitiensis-like line, also having 2n = 16, have originated from a common ancestor by geographic isolation.     

Synchronous and early activation of planarian Hox genes and the re-specification of body axes during regeneration in Dugesia (G.) tigrina (Turbellaria; Tricladida)

Seven Hox cluster-related genes (Dthox-A to -G) have been isolated from the freshwater triclad Dugesia (G.) tigrina, their sequence compared to other Hox genes and their expression in intact and regenerating organisms analyzed by whole mount in situ hybridization. Sequence comparison analyses show high similarities of D. tigrina Hox genes to anterior and medial groups of coelomate Hox genes. Expression analyses show very early, synchronous, and overlapping expression of Dthox -A, -E, -G and -F in anterior, posterior and lateral regenerative tissues. At one hour of regeneration all Dthox genes studied showed a neat, clear expression at the wound boundary. Later, as the blastema grows, the expression area expands to more proximal regions covering the blastema and the distal postblastema regions. Blastemas formed by intercalary regeneration also show a synchronous expression of the same Hox genes though the onset of activation is much delayed. The finding that the same set of Hox genes is synchronously activated in anterior, posterior, intercalary and lateral regeneration is in sharp contrast to its well established role in specifying antero-posterior pattern during embryonic development. The implications of these results as regards ancestral versus co-opted roles of Hox genes in development and regeneration are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

The karyotypes of Dugesia species from Spain (Turbellaria,Tricladida)

Some species of Planarians, new to Spain, are recorded. Dugesia polychroa, D. sicula, D. iberica and D. gonocephala s. 1. have been investigated karyologically. The former possesses a diploid complement characteristic of the biotype A (2n = 8); the second is diploid with 2n = 18; diploidy and triploidy were found in sexual populations of D. iberica with n = 8. Triploidy occurred in all the asexual strains of the D. gonocephala group with a basic number of either 8 or 9. In this latter case B-chromosomes were occasionally found.     

Phylogenetic relationships of the genus Dugesia (Platyhelminthes, Tricladida, Paludicola)

A phylogenetic analysis was performed on the genera and subgenera within the freshwater triclad family Dugesiidae, based on 19 terminal taxa and 17 morphological characters. The phylogenetic tree proposed has length of 27 steps and consistency index of 0.66. This phylogenetic hypothesis implies that the current genus Dugesia is paraphyletic and that its subgenera Girardia, Schmidiea and Dugesia S.S. should be elevated to the rank of genus. The genera Cura, Spathula and Neppia are presumed monophyletic by default because the database was unable to provide autapomorphies for any of these genera. The genera Dugesia S.S. and Neppia share sistergroup relationship. Several characters are discussed which were previously considered to be of phylogenetic importance but were not included in the present analysis. It is emphasized that sensory organs form potentially useful set of phylogenetic characters for the Dugesiidae.     

Chromosomal polymorphism in planarians (Turbellaria,Tricladida) and the plate tectonics of the western Mediterranean

In two populations of Dugesia (Schmidtea) mediterranea the third chromosome pair consists of two heteromorphous elements. This chromosomal polymorphism, which is common in other animal groups, was hitherto unknown in planarians.The evolutionary and biogeographical significance of this chromosomal polymorphism is discussed, for the geographical distribution of the two aberrant populations may have relevance for the theories of microplate dispersal involving the western Mediterrancan.     

Unusual Pre-copulatory Behaviour in the Hermaphroditic Planarian Flatworm,Dugesia gonocephala (Tricladida,Paludicola)

Hermaphrodite mating behaviour is poorly understood, particularly in species with internal fertilization and mutual penis insertion. We collected detailed recordings on the frequency, timing and outcome of the mating sequence of the simultaneous hermaphroditic flatworm Dugesia gonocephala (Tricladida, Paludicola). Mating in this species differs from that in other planarians in that receptive partners show a stereotypic pre-copulatory behaviour which results in copulation in only 17% (n = 342) of all attempts. It includes an unusual phase during which one partner glides on top of the other and first the lower, then both individuals, spread out and flatten. Genital intromission is not possible in this position. All observed copulations (n = 57) were preceded by this behaviour and most mating sequences were interrupted at this stage. Pairs were seen starting new mating sequences up to 11 times. Which animal lay on top varied randomly within a series of attempts. Cocoon production was a good predictor of mating activity, and copulations mainly occurred during the night. Copulations lasted between 24 min and 9.3 h (mean = 4.7 ± 2.3 h) and penises were mutually inserted. In the field, 99% (n = 70) of all matings involved reciprocal spermatophore exchange, compared with only 50% in the laboratory (n = 26). The likelihood of reciprocal spermatophore transfer increased with increasing duration of copulation. Duration was independent of the events that took place during courtship. We discuss the possibility that partners interrupt a copulation when there is no guarantee of reciprocal spermatophore transfer.     

Observations on Fission, Maturation and Reproduction in Dugesia biblica (Turbellaria, Tricladida) from Israel

Laboratory studies on the normally fissiparous Dugesia biblica from Israel (3 n =27+ x supernumeraries) have been made in order to investigate fission rates and spontaneous maturation of four fissioning populations and one clone. Laboratory induced maturation in a sixth population of D. biblica resulted in the laying of fertile cocoons, the offspring of which were either 3 n =27+ x , 2 n = 18+ x or 2 n = 18+ x It is suggested that triploidy is primarily responsible for the lack of sexual reproduction in the field, and that unequal inheritance of supernumerary chromosomes during mitosis may explain the differing tendencies to undergo spontaneous maturation by individuals of a clone of D. biblica. The karyotype, habitat and reproduction of D. biblica is compared with a closely related but diploid, normally sexually reproducing Dugesia form and possible relationships between these two are discussed.     

Distribution,abundance, and size of rhabdoids in Dugesia polychroa. (Turbellaria: Tricladida)

The ontogenetic changes in the distribution, abundance, and size of rhabdoids were examined in an undisturbed laboratory population of Dugesia polychroa. Irrespective of triclad age, rhabdoids in the epidermis and parenchyma were more abundant on the dorsal than the ventral side of the body. No significant differences were found in the abundance of epidermal or parenchymal rhabdoids among the anterior, medial, and posterior regions of the body. Rhabdoid number and size changed significantly with triclad age, with a marked depression coinciding with the onset of cocoon production. Rhabdoid discharge was correlated with physical and/or physiological disturbance and occurred in the absence of any overt environmental disturbance. Simple allometric relationships were observed between rhabdoid size and number on one hand and body plan area on the other. Different allometric trends were observed from field-collected individuals compared to the undisturbed laboratory population. The potential function of rhabdoids in the Tricladida is discussed in light of these findings.     

Asexual reproduction,regeneration, and somatic embryogenesis in the planarian Dugesia tigrina (Turbellaria)

In reviewing recent research published in Russian on regeneration and asexual reproduction, the following morphogenetic processes in the planarian Dugesia tigrina are considered: 1) regeneration of lost parts of the body; 2) regeneration of the whole worm from fragments of the body, either by normal regeneration when the inital polarity of the fragment is retained or by somatic embryogenesis when one or more new axes of polarity arise; 3) somatic embryogenesis, or development of individuals from somatic cells; 4) hypermorphosis, or the presence of more than the usual number of organs or body parts, a process that can be interpreted in terms of somatic embryogenesis; and 5) asexual reproduction. Some morphological, biochemical, and physiological studies of the division zone in D. tigrina demonstrate peculiarities of a local breakdown of integrative functions, a breakdown which in turn causes division of the individual to take place at this zone; timing of division is controlled by the organism as an integrated whole.     

Morpho-karyological Types of Dugesia (Turbellaria, Tricladida) in Israel and Their Distribution Patterns

Populations of Dugesia in Israel have been classified on morphological and karyological criteria into 5 types. Two of these are morphologically similar to the Dugesia lugubris group (n = 4) but differ karyologically ( Types 1 and 2 : n=8), and three types are morphologically similar to the Dugesia gonocephala group ( n = 8) but two differ karyologically (Types 4 and 5: n = 9). The three ' gonocephala ' types are distributed in three distinct areas of Israel with little overlap, while the two ' lugubris ' types are found in springs of one of these areas, The Rift Valley, where special localised conditions prevail. Possible relationships between Types 3, 4 and 5 are discussed with particular reference to their karyotypes and the presence of supernumerary chromosomes in Type 5.     

Bursa,bursa canal,and female antrum of Dugesia tigrina (Plathelminthes,Tricladida)

Summary The bursa of Dugesia tigrina is located between pharynx and penis and is suspended in a network of muscle fibers. Two cell types are present in the bursa: small outer cells which likely represent replacement cells and tall inner cells. During copulation seminal material, consisting of secretion products, fibrillar and tubular material, and small clusters of spermatozoa, is injected through the bursa canal into the bursa. During the 48 h post-copulatory period the seminal material is absorbed by the inner cells. Phagocytosis of the seminal material is facilitated by broad apical cytoplasm and modified distal cell membranes. Phagocytosed spermatozoa are enclosed in vaculoes and morphologic breakdown occurs as early as 4 h after copulation. Fibrillar and tubular material is phagocytosed directly into the apical, organelle-free cytoplasm without vacuole formation.The bursa canal is ciliated and the distal cell ends are studded with ultrarhabdites. These are also present in the epithelial cells of the female antrum. Antrum epithelial cells furthermore display apical organelle-free cell areas. Numerous glands penetrate through the antrum wall.     

On the taxonomic status of a Dugesia species from Buenos Aires (Platyhelminthes: Turbellaria: Tricladida)

The taxonomic and distributional statuses of the freshwater planarians of South America are imperfectly understood, and a comprehensive review is sorely needed. The species known from the vicinity of Buenos Aires, Argentina, was first described by Böhmig (1902) under the name Planaria dubia Borelli, 1895. Subsequent authors (Kenk, 1930, 1974; Cazzaniga & Curino, 1987) have cast doubt on this assignation. Some have assigned it to Dugesia anceps Kenk; Hyman (1959) was inclined to regard the Buenos Aires planarian as a geographical variant of Dugesia sanchezi Hyman. A comparison of D. sanchezi from the mountainous region of central Chile with specimens from Buenos Aires confirms that the Argentinian material is a distinct species.The differences between the two species are subtle. The Argentinian specimens are distinctive in their possession of infranucleate epithelia of the atria, penis, and bursal canal, as well as in their penial structure. In both species, the epithelia of the atria and bursal canal are glandular. Those of D. sanchezi are nucleate and their secretions are cyanophilous. In the Argentinian specimens, these same epithelia are infranucleate; but while the secretion of the bursal canal is indeed cyanophilous, that of the atrium is erythrophilous. The presence, in these same specimens, of cyanophilous glands opening into the penial vesicles, and of erythrophilous glands whose secretions contribute to a viscous plug in the ejaculatory duct, suggests a difference in the physiology of copulation.It is apparent that the Argentinian specimens are distinct from D. sanchezi Hyman, and probably they represent a new species, one to be described in detail elsewhere. In any case, Hyman's (1959) suggestion that D. sanchezi occurs near Buenos Aires is mistaken; so also is Cazzaniga's & Curino's (1987) identification of specimens from Buenos Aires as D. anceps.     

The influence of body size on immediate reproductive success in Dugesia gonocephala (Tricladida,Paludicola)

The individual reproductive output of the stream-dwelling flatworm Dugesia gonocephala was investigated. Various measures of reproductive success were related to body size. (I) For the first 30 days in the laboratory small individuals produced no cocoons, individuals of intermediate size produced unfertilized cocoons and large individuals usually produced fertilized cocoons. (II) In individuals that produced a cocoon, no correlation was found between the number of cocoons produced in one month and body size. (III) Large individuals, however, produced larger cocoons. This was not due to the fact that unfertilized cocoons were smaller. (IV) Large cocoons tended to contain more young. (V) The average size of young hatching from large cocoons was larger. (VI) Large individuals produced their first cocoon soon after their arrival in the laboratory and seemed to have a higher chance of producing a fertilized first cocoon. (VII) A trade-off existed between producing many small versus few large young.     

On the karyology of Dugesia gonocephala s.l. (Turbellaria,Tricladida) from Montpellier,France

Two species belonging to the Dugesia gonocephala group are found in the area of Montpellier, France. The karyology of these two species, D. gonocephala s. str. and S. subtentaculata, and of fissiparous Dugesia races has been studied.Two populations belonging to D. gonocephala s. str. are diploids with a chromosome number of 16, whereas the specimens of a third population are sexual aneuploids; the majority of cells possess 24 chromosomes, but some cells contain 23 or 25. The specimens attributable to D. subtentaculata are triploids, the most notable karyological feature being the presence of a single unmatched acrocentric chromosome. The fissiparous Dugesia strains are all aneuploids, the most common chromosome number being 27 with up to three small B-chromosomes.     

Notes on the biology of the freshwater planarian Dugesia bengalensis (Platyhelminthes Turbellaria Tricladida)

Dugesia bengalensis was described by Kawakatsu (Kawakatsu et al., 1983) from specimens collected in West Bengal. We have been studying populations from many different localities in Santiniketan and adjoining areas of West Bengal and can provide additional biological information.The species is hermaphroditic, and its breeding season was found to occur usually between October and March when the winter temperature falls below 25 °C. Outside of the breeding season, D. bengalensis is capable of asexual reproduction by binary fission (Mahapatra et al., 1987).Development of the reproductive organs appeared to be from neoblasts and other mesenchymal cells and, therefore, to be like that typical of other triclads with the exception that some of the neoblasts used for the reproductive tissue appeared to be derived from the intestinal region (Ghosh, 1988; cf. Teshirogi, 1986).During copulation, the partners were oriented in the same direction and not in a head-to-tail position as has been reported (Hyman, 1945) for some planarians.The oval, stalked cocoons were laid in marshy places, and during the period of summer (usually from April to June) they lay dormant in the sandy soil until the onset of the monsoon rains. Then, typically three or four months after they were laid, the cocoons hatched to yield three or four young, a remarkably low number for freshwater triclads (cf Ball & Reynoldson, 1981).