Franca Scanabissi

Molecular taxonomy and phylogeny of the ‘living fossil’ lineages Triops and Lepidurus (Branchiopoda: Notostraca)

Mantovani B. Cesari M. & Scanabissi F. (2004). Molecular taxonomy and phylogeny of the ‘living fossil’ lineages Triops and Lepidurus (Branchiopoda: Notostraca). —Zoologica Scripta, 33, 367–374.

Ultrastructural observations on the oogenesis of Triops cancriformis (Crustacea, Notostraca).

SummaryEach ovarian follicle of Triops cancriformis is four-celled; these cells (one oocyte and three nurse cells) are interconnected by cytoplasmic bridges. In the course of differentiation, the nurse cells are early recognizable; they increase in size more than the oocyte and their nuclei contain many nucleoli. For the first time in Arthropoda, yolk globules are reported to be present in nurse cell cytoplasm; these globules arise from the smooth endoplasmic reticulum. The functional significance of the intercellular bridges and the trophic role of the nurse cells are discussed.

A survey of the reproductive biology in Italian branchiopods

A survey of the most important aspects of the reproductive biology of Lepidurus apus lubbocki Brauer, 1873 (Notostraca) is presented. Gametogenesis, germ cell distribution, and somatic tract structure of the male gonad are studied by means of TEM and SEM. The most important data emerging from our observations is the total and widespread degeneration of the sperms. We discuss the real functionality of male germ cells, and the implications on sexuality and reproductive strategies. On the whole, present ultrastructural observations suggest that males of Lepidurus apus lubbocki are not functional. Therefore, we suggest that this form reproduces parthenogenetically as the functional female gonad lacks any associated testicular cells. For comparison, we analyzed the male portion of the hermaphroditic gonad of Triops longicaudatus (LeConte, 1846) from California. In this case functionallity of the testis lobes and sperm were readily apparent.

ANDRODIOECY INFERRED IN THE CLAM SHRIMP EULIMNADIA AGASSIZII (SPINICAUDATA: LIMNADIIDAE)

Abstract Androdioecy (mixtures of males and hermaphrodites) is a rare mating system in both the plant and animal kingdoms. Androdioecy has been described in three branchiopod species, and is best known from the clam shrimp Eulimnadia texana Packard. Herein we describe sex ratio, genetic and histological evidence from the clam shrimp Eulimnadia agassizii Packard that suggest androdioecy is also found in this species. The E. agassizii population sampled had all-females, and when these females were isolated and allowed to produce eggs, those eggs yielded 100% female offspring in 15 out of 15 cases. Additionally, the originally isolated females proved to be completely homozygous at each of the six allozyme loci scored. The offspring from these isolated females also proved to be homozygous for the same alleles as their parent. Tissue sectioning of the gonad found that the “females” actually produced testicular tissue in the posterior portion of the gonad. Taken together, these data are entirely consistent with those of the androdioecious E. texana, and thus indicate that E. agassizii is also an androdioecious species, bringing the total number of branchiopod species with this form of reproduction to four.

FOLLICLE DUCT CELL ULTRASTRUCTURE AND EGGSHELL FORMATION IN TRIOPS CANCRIFORMIS (CRUSTACEA, NOTOSTRACA)

Electron microscopy of the cells of the follicle duct of Triops cancriformis shows that the follicular ducts are lined by a single‐layered epithelium which also produces the eggshell material. The cytoplasm is rich in rough endoplasmic reticulum that synthesizes the eggshell material which subsequently aggregates into preformed vacuoles. Newly formed spheres of eggshell material are then excreted into the lumen. At the end of vitellogenesis the oocytes descend toward the longitudinal oviduct and pass through the eggshell material which fills the follicle ducts. The production of the eggshell and its chemical composition in some Phyllopoda are compared. The paper discusses the relationship between the eggshell construction and the reproductive biology of the population.

Mitochondrial and nuclear DNA variability in the living fossil Triops cancriformis (Bosc, 1801) (Crustacea, Branchiopoda, Notostraca).

The living fossil Triops cancriformis comprises bisexual (either gonochoric or hermaphroditic) and unisexual populations. Genetic surveys have recently revealed a general trend of low differentiation of 12S and 16S mitochondrial genes. We, therefore, surveyed further mitochondrial (COI gene and control region) and nuclear markers (dinucleotide microsatellites) to assess the genetic variability and to establish any relationship with the different reproductive modes found in European populations. The mitochondrial analyses confirmed the pattern of low variability. Hence, the low mitochondrial genetic variability appears as a common feature of the genus Triops. The microsatellite analysis found that Italian populations are monomorphic or exhibit little polymorphism, while other European samples display a higher degree of polymorphism and private alleles. Spanish, Austrian and Italian populations show patterns of Hardy–Weinberg disequilibrium that could be explained by the mode of reproduction, or by a higher frequency of null alleles in these populations. The low diversity and differentiation among Italian populations lead us to question the Monopolization Hypothesis. One microsatellite locus appears to be sex-linked, with heterozygotes detected only in males and hermaphrodites.

Production of intersexes and the evolution of androdioecy in the clam shrimp Eulimnadia texana (Crustacea, Branchiopoda, Spinicaudata)

Summary The production of low numbers of offspring that exhibit a mixture of male and female traits (termed “intersexes”) is commonly reported for crustaceans. The production of intersexes has been ascribed to both genetic and non-genetic (e.g., parasitic infections and environmental pollutants) causes. Herein we report on two observed types of intersexes in the clam shrimp Eulimnadia texana: (1) a “morphological” intersex, possessing secondary male characteristics (e.g., claspers) and an eggproducing gonad, and (2) a “gonadal” intersex, possessing primarily male traits (e.g., male secondary sexual characters and male gamete production) but also producing low levels of abortive female gametes. We propose that these intersexes are likely the products of low frequencies of crossing over between the sex determining chromosomes that result in the array of observed mixed sexual phenotypes. Additionally, we suggest that the low-level production of intersexes, combined with the ephemeral nature of the habitats occupied by these shrimp, may explain the preponderance of androdioecy (mixtures of males and hermaphrodites) found in these clam shrimp, and possibly branchiopods more generally.

Genetic variability in European Leptestheria dahalacensis (Ruppel, 1837) (Crustacea, Branchiopoda, Spinicaudata)

The genetic variability of the gonochoric Leptestheria dahalacensis (Rüppel, 1837) was studied through the analysis of mitochondrial and nuclear (microsatellite loci) markers in eight Italian and two Central European populations. Mitochondrial data exhibited a low variability, as only six mitotypes were scored: five in Italy and one for both Central European samples, with a very low number of substitutions. All analysed microsatellite loci were variable, with 3–5 alleles per locus and 1–4 alleles per population. All populations were at the Hardy–Weinberg equilibrium, with the exceptions of two samples for locus ldAC-16, due to heterozygote excess, and of four populations for locus ldAC-11, probably linked to the presence of null alleles. A substantial population structuring was found between Central European and Italian samples for both utilized markers. This observation may be explained by isolation by distance and/or recent isolation events. On the other hand, the absence of a clear inter-pond variability in Italian sample comparisons may be ascribed to high dispersal ability in the short range.

Sperm transfer and occurrence of spermatophore in the Conchostraca Leptestheriidae (Crustacea, Branchiopoda)

Summary Sperm transfer in Conchostraca Leptestheriidae Leptestheria dahalacensis Rüppel, 1837, a strictly amphigonic species, occurs due to the production in males of an extremely primitive structure. This structure, analogous to spermatophores present in most Crustacea, may be observed in the lumen as thin strips of crystalline material. Shortly before mating, sperm produced in the gonads are released together with the crystalline fibres that serve as a net which helps keep the sperm compact and adherent to the males body. The surface and shape of the so-called spermatophore is described. Mating time is partly dependent on the degree of adherence of the spermatophore to the females body. The origin and conformation of this primitive structure is assessed in terms of its significance as an adaptive mechanism.

Molecular taxonomy and phylogeny of Italian Lepidurus taxa (Branchiopoda: Notostraca)

Notostraca taxonomy is controversial, mainly owing to the high intraspecific variability of morphological characters; other complexity factors are represented by the consistent rate of reproductive variation. We undertook the molecular analyses of Italian Lepidurus taxa through the characterization of 12S, 16S and COI mitochondrial genes in new L. apus demes and in the L. couesii populations recently discovered in Southern Italy. For L. apus, the analysis of a wider data set (both in terms of molecular markers and of analysed populations) confirms the previously suggested specific rank of differentiation between its so far recognized subspecies, the Italian L. apus lubbocki and the European L. apus apus. The Italian L. couesii samples pertain to a well‐defined taxonomic entity, strictly related to Canadian L. couesii. In 12S dendrograms, this taxon strongly diverges from the other North American Lepidurus species, but the relationships among nearctic taxa are unresolved. On the other hand, a high affinity between Italian/Canadian L. couesii and the circumpolar species L. arcticus is scored and a possible descent from a common ancestor is hypothesized.