Asexual Reproduction and Segmental Regeneration, but not Morphallaxis, are Inhibited by Boric Acid in Lumbriculus variegatus (Annelida: Clitellata: Lumbriculidae)

Body fragmentation, in some animal groups, is a mechanism for survival and asexual reproduction. Lumbriculus variegatus (Müller, 1774), an aquatic oligochaete worm, is capable of regenerating into complete individuals from small body fragments following injury and reproduces primarily by asexual reproduction. Few studies have determined the cellular mechanisms that underlie fragmentation, either regenerative or asexual. We utilized boric acid treatment, which blocks regeneration of segments in amputated fragments and blocks architomic fission during asexual reproduction, to investigate mechanistic relationships and differences between these two modes of development. Neural morphallaxis, involving changes in sensory fields and giant fiber conduction, was detected in amputated fragments in the absence of segmental regeneration. Furthermore, neural morphallactic changes occurred as a result of developmental mechanisms of asexual reproduction, even when architomy was prevented. These results show that fragmentation in L. variegatus, during injury or asexual reproduction, employs developmental and morphallactic processes that can be mechanistically dissociated by boric acid exposure. In regeneration following injury, compensatory morphallaxis occurred in response to fragmentation. In contrast, anticipatory morphallaxis was induced in preparation for fragmentation during asexual reproduction. Thus, various forms of regeneration in this lumbriculid worm can be activated independently and in different developmental contexts.     

Biology of Pseudopotamilla reniformis (Müller 1771) in the White Sea,with description of asexual reproduction

The tube-dwelling polychaete Pseudopotamilla reniformis (Sabellidae) forms dense and complex aggregations of flexible tubes on hard substrates in the subtidal zone of the White Sea. No sexual reproduction was observed in this study and recruitment appeared to be due to asexual reproduction by architomy in winter, from October to March. The posterior part of the abdomen undergoes spontaneous fission into from 2 to 4 fragments and depending on their position, the fragments regenerate their anterior ends or both anterior and posterior ends. Regeneration in P. reniformis takes place via a combination of epimorphosis (replacement of missing parts by cell proliferation and the growth of new tissue) and morphallaxis (the remodelling of pre-existing structures without cell proliferation). The morphogenetic events during regenerative restoration include de novo formation of branchial crown, formation of thoracic segments and restoration of the posterior end. Asexual reproduction appears to play a crucial role for formation of P. reniformis aggregations and is very important for the population in the White Sea, at the margin of the species’ range.     

The social feather duster worm Bispira brunnea (Polychaeta: Sabellidae): aggregations,morphology and reproduction

The fan worm Bispira brunnea is one of the most attractive sabellid polychaetes from Caribbean coral reef areas and it is exploited for ornamental purposes. An understanding of the structure of its aggregations, morphology and reproductive biology will provide information required to facilitate artificial propagation of this species. Ten aggregations were collected in October 2013, February and March 2014 in the Majahual reef lagoon, Mexican Caribbean. Whole aggregations were examined under light microscopy and scanning electron microscopy and the histology of oogenesis was determined. Aggregations were composed of 24–56 individuals and included juveniles and adults. The adults (5–20?mm) did not display any noticeable form of sexual dimorphism. In B. brunnea both sexual (hermaphroditism or gonochorism) and asexual reproduction occurred at the same time within the population: 92.71% reproduced sexually and 52% by architomy. The buds produced by architomy were inside the parental tube, at three regenerative stages. The sex ratio was 36.75% males, 33.11% females, 22.84% hermaphrodites and 7.28% unknown (gametes not seen). Gametes were distributed in the last thoracic segments and throughout the abdomen. Oogenesis was extra-ovarian, development followed four discrete stages and the oocytes were small and asynchronous (60.97 μm in diameter). Sperm morphology was adapted to external fertilization in the water column. Sequential (protandrous) hermaphroditism is suggested to occur in B. brunnea. The pyramidellid ectosymbiont mollusc Odostomia (Eulimastoma) caniculatum is recorded here for the first time as being associated with a sabellid worm.     

Two new DorvNea species (Polychaeta, Dorvilleidae) with obligate asexuaI reproduction

Two new species, Dorvillea bermudensis sp.n. and D. albomaculata sp.n., with obligate asexual reproduction are described. Both are closely related to sexually reproducing members of the same genus. The new species undergo repeated fission of the architomy type, with the rate of regeneration and subsequent production of segments being positively correlated with the energy content of the fragments. Regeneration and reproductive potentials vary between head and tail fragments with tail lineages producing higher rates than head lineages. Both species are very tolerant of starvation which may cause considerable degrowth and initiation of dispersal behavior.     

What role do annelid neoblasts play? A comparison of the regeneration patterns in a neoblast-bearing and a neoblast-lacking enchytraeid oligochaete

The term 'neoblast' was originally coined for a particular type of cell that had been observed during annelid regeneration, but is now used to describe the pluripotent/totipotent stem cells that are indispensable for planarian regeneration. Despite having the same name, however, planarian and annelid neoblasts are morphologically and functionally distinct, and many annelid species that lack neoblasts can nonetheless substantially regenerate. To further elucidate the functions of the annelid neoblasts, a comparison was made between the regeneration patterns of two enchytraeid oligochaetes, Enchytraeus japonensis and Enchytraeus buchholzi, which possess and lack neoblasts, respectively. In E. japonensis, which can reproduce asexually by fragmentation and subsequent regeneration, neoblasts are present in all segments except for the eight anterior-most segments including the seven head-specific segments, and all body fragments containing neoblasts can regenerate a complete head and a complete tail, irrespective of the region of the body from which they were originally derived. In E. japonensis, therefore, no antero-posterior gradient of regeneration ability exists in the trunk region. However, when amputation was carried out within the head region, where neoblasts are absent, the number of regenerated segments was found to be dependent on the level of amputation along the body axis. In E. buchholzi, which reproduces only sexually and lacks neoblasts in all segments, complete heads were never regenerated and incomplete (hypomeric) heads could be regenerated only from the anterior region of the body. Such an antero-posterior gradient of regeneration ability was observed for both the anterior and posterior regeneration in the whole body of E. buchholzi. These results indicate that the presence of neoblasts correlates with the absence of an antero-posterior gradient of regeneration ability along the body axis, and suggest that the annelid neoblasts are more essential for efficient asexual reproduction than for the regeneration of missing body parts.     

Asexual reproduction and the turbellarian archetype

The turbellarian archetype is widely believed to have been a hermaphrodite lacking asexual reproduction, and such asexual reproduction as is now seen in the Turbellaria (as paratomy and architomy) is generally assumed to have arisen secondarily several times independently. Asexual reproduction clearly prevails among the most primitive metazoans such as the placozoans, sponges, and radiates, however, and if the Platyhelminthes is indeed an early offshoot of bilaterian evolution, as some have claimed, then it is reasonable to expect asexual reproduction to be a primitive feature of the Turbellaria. Asexual reproduction by paratomy or architomy is found in all three main evolutionary lines of the Turbellaria and is most common among primitive groups such as the Catenulida and Macrostomida. The discovery of a new, apparently primitive marine genus of Macrostomida having paratomy widens the known incidence of asexual reproduction within that order. The presence of a muscle ring around the gut of several distantly related genera of the Macrostomida and similarities this ring shows with septa in the division plane of paratomizing species are evidence that paratomy was a feature of the stem species for this order — a feature only secondarily lost in most macrostomids — and suggest that asexual reproduction is a primitive feature for the Platyhelminthes as a whole.     

Subfossil chydorid (Cladocera, Chydoridae) ephippia as paleoenvironmental proxies: evidence from boreal and subarctic lakes in Finland

Chydorids (Cladocera, Chydoridae) have two reproductive strategies: asexual reproduction that prevails during favorable environmental conditions and sexual reproduction that is induced by environmental stimuli associated with seasonal or aperiodic environmental stresses. These modes of reproduction can be recognized in the subfossil sedimentary records as parthenogenetic shells of females (asexual reproduction) and by ephippia (sexual reproduction). We studied the interrelations between subfossil chydorid ephippia and environmental variables by analyzing surface sediment samples obtained from 76 Finnish lakes across a latitudinal gradient (60–70°N). The results showed that the total chydorid ephippia (TCE) increases along the climate gradient from ~2 to 3% in the south to ~25% in the north and suggested a significant dependence (r ~ −0.8, P < 0.001) with several climate factors, especially that of mean July air temperature. We used this relationship to create a model for reconstructing past mean July air temperatures. A linear regression of the log₁₀ transformed TCE as a single independent variable explained 76% (SE ± 0.76°C) of the variance of the observed mean July air temperatures. Accordingly, we propose that this novel tool may be highly suitable for reconstructing paleotemperatures in cold-temperate environments.     

Morphogenesis during asexual reproduction in Pygospio elegans Claparede (Annelida, Polychaeta)

The spionid Pygospio elegans reproduces both asexually and sexually. Using scanning electron and bright field microscopy, we examined morphogenesis following asexual reproduction to determine how "lost" body regions were regenerated after a worm spontaneously divided. Asexual reproduction occurred through transverse fission and divided the parent worm into 2 to 6 fragments (architomy). All fragments retained their original anterior-posterior polarity. Regeneration in all fragments followed a specific series of events: wound healing (day 1); extension of the blastema to generate lost body regions-specifically, the head and thorax for posterior fragments and the tail and pygidium for anterior fragments (days 2-3); segmentation (days 3-6); and differentiation of segment- or region-specific structures (days 4-8). This pattern occurred regardless of where the original division took place. Subsequent growth occurred through addition of terminal setigers anterior to the pygidium followed by differentiation of tail setigers into abdominal setigers, leaving the tail region about 6 to 10 setigers in size. Division rates were compared in worms from three populations in Nova Scotia, Canada. Worms from two populations (Conrad's Beach, Starr's Point) divided more frequently (about 1.2 and 1.3 weeks between divisions, respectively) than worms from Bon Portage Island (3.5 weeks between divisions). Fragments containing the original head (original mouth intact, generally much larger fragment) had a higher survivorship than fragments containing the original tail.     

COMPARATIVE STUDIES ON THE CELL WALLS OF SEXUAL AND ASEXUAL BANGIA ATROPURPUREA (RHODOPHYTA). II ELECTROPHORETIC PATTERNS OF POLYSACCHARIDES1

Cellulose acetate electrophoresis of the hot water soluble polysaccharide extracts from whole filaments, as well as base, mid and tip segments, of marine asexual and sexual Bangia atropurpurea (Roth) C. Ag. Yielded distinctive patterns which demonstrated that changes occur in the outer cell walls during sexual reproduction. Heterogeneity of the sulfated polysaccharide components isolated from outer cell walls was shown to be specifically related to sexual reproduction. Two components (Band I and II) were detected in extracts from tips of sexual filaments, whole only one (Band I) was present in the vegetative segments of all filaments and in asexual reproductive regions. The faster running component (Band II) was detected during the later stages of sexual development, prior to maturation.     

Morphogenetic characters of the model ciliate Euplotes vannus (Ciliophora,Spirotrichea): Notes on cortical pattern formation during conjugational and postconjugational reorganization

Ciliated protists represent a morphologically and genetically distinct group of single-celled eukaryotes which can reproduce asexually and sexually. Morphogenesis occurs in both asexual and sexual modes of reproduction which is of interest for researchers investigating cell differentiation, regeneration, systematics and evolution. However, studies of morphogenesis have concentrated almost entirely on the asexual mode. Here we use protargol staining to investigate the morphogenetic processes during sexual reproduction in the model species Euplotes vannus (Müller). The major events include: (1) two rounds of morphogenesis occur during sexual reproduction, i.e., conjugational and postconjugational reorganization; (2) in both processes the oral primordium is generated de novo in a pouch beneath the cortex; (3) the frontoventral-transverse cirri anlagen are formed de novo and fragment in a 3:3:3:3:2 pattern; (4) the leftmost cirrus and the paroral membrane do not change during conjugational morphogenesis, but reorganize de novo during postconjugational morphogenesis; (5) marginal cirral anlagen are formed de novo in both morphogenetic processes; (6) two or three caudal cirri are formed at the ends of the rightmost two or three old dorsal kineties; (7) the dorsal kineties are retained entirely. These results can serve as reference to investigate the morphogenetic events in the different stages of sexual reproduction.     

Fine‐scale asexual and sexual reproduction in Quercus crispula var. horikawae,a stunted shrub oak,on a mountain with deep snow in central Japan

Quercus crispula var. horikawae, a stunted shrub oak, occurs on mountains with deep snow on the Sea of Japan side of Japan. This oak generates patches of multiple creeping stems. It is unclear whether these patches are the result of asexual or sexual reproduction, or both. We therefore aimed to describe the clonal structure and gene flow in Q . crispula var. horikawae on Mount Nasu in central Japan by using nuclear microsatellites. Genotypes of 331 stems with no distinct connection with roots and creeping stems above the ground were determined using nine loci in two study plots, and 64 acorns from three mother genets in a plot were determined using eight loci. The results of the clonal identification indicated that the patches consisted of 51 genets; at least 85% of the stems may have been derived from asexual reproduction through sprouting and layering. The prominence of asexual reproduction may be a result of adaptation to the snowy environment. In contrast, 15% of the ramets in the study plots probably originate via sexual reproduction by seedling regeneration. Analyses of the spatial genetic structure and paternity showed that limited ability of the pollen and seeds to disperse might result in the spatial aggregations of closely related offspring at a relatively short distance (<10 m), and inbreeding, a factor that might reduce sexual reproduction, was not observed. Thus, sexual reproduction could be reduced by ecological rather than genetic factors, namely the hindrance of seedling regeneration by the dense coverage of dwarf bamboo (Sasa) on the forest floor.     

Long-term storage of sexual and asexual Trichogramma brassicae (Hymenoptera: Trichogrammatidae)

The effect of exposure duration to pre-storage (representing fall conditions) and storage (representing winter conditions) temperatures on the emergence rate in sexual and asexual Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae) was studied under controlled laboratory conditions. Asexual T. brassicae was infected by Wolbachia, an endosymbiont bacterium. We considered 0, 30, 40, 50 and 60 days as exposure duration at 10°C and 0, 30, 60, 90 followed by 180 days as exposure duration at 4°C for both T. brassicae strains. Emergence rate was significantly affected by the presence of Wolbachia, the duration of exposure to 10°C, the duration of storage at 4°C and their interactions, but not by the interaction between strain × storage duration at 4°C. Emergence rate of adults was positively correlated with exposure duration at 10°C, but it gradually decreased as the cold storage duration (4°C) increased in both strains. In conclusion, we are able to store both asexual and sexual strains for longer periods of time at 4°C, but the emergence rate of sexual T. brassicae was greater than asexual in all the treatments. The presence of Wolbachia could have a negative effect on fitness of T. brassicae and reduces its emergence rate after long-term storage.     

Stem cells in microturbellarians

Summary A combination of microscopical, immunocytochemical, and autoradiographic techniques were employed to study stem cells and their fates during asexual reproduction and regeneration in two microturbellarians,Microstomum lineare (Macrostomida) andStenostomum leucops (Catenulida). Special attention was paid to the development of the immunoreactivity (IR) to FMRF/RF-amide and 5-HT in differentiating nerve cells.Asexual reproduction inM. lineare andS. leucops occurs by paratomy, i.e., fragmentation after completed differentiation of the new organs. Regeneration, on the other hand, involves a combination of morphallactic and epimorphic processes without the formation of a regeneration blastema. The only cells incorporating tritiated thymidine ([³H]T) were the mesenchymal and gastrodermal neoblasts, which proliferate continuously replenishing the population of stem cells available for growth, asexual reproduction and regeneration. These proliferative cells occurred in two ultrastructurally different forms, differing from each other only by the presence or absence of ciliar basal bodies in the cytoplasm. Few differentiated cells were labeled in the head piece after completed regeneration. A greater amount of labeled differentiated cells were, however, observed postpharyngeally in the first zooid as well as in zooids having developed during the same time (i.e., 20–45 h after the treatment with [³H]T). Furthermore, many labeled cells were still undifferentiated at that time or just in the beginning of the differentiation process. It can therefore be concluded that neoblasts function both as reserve cells and as functional stem cells for all differentiated cell types in these worms. IR to FMRF/RF-amide neuropeptides was not observed in nerve cells differentiating from neoblasts until the occurrence of dense-core vesicles in their cytoplasm. Due to methodological difficulties only weak or no IR to 5-HT could be traced in the nervous system of the asexual and regenerating worms.Abbreviations ICC Immunocytochemical - IR immunoreactivity - [³H]T tritiated thymidine     

Temperature Effects on Shoot and Root Development From Begonia × cheimantha Petiole Segments Grown in vitro

The effect of different temperatures on the shoot and root formation in isolated petiole segments of Begonia × cheimantha was determined after 10 weeks on a modified White medium containing 0.1 mg/1 NAA and 0.5 mg/1 BA. Temperature proved to be important for the induction of shoot and root formation. At a constant temperature the best plants were obtained at 18 to21°C. If the temperature was higher, fewer cultures survived and the number of roots and shoots were lower. Lower temperatures inhibited the development of plants. A pretreatment at 15 or 18°C for two to four weeks improved the number and size of the shoots developed during a following 24°C treatment. High temperatures throughout the growing period reduced the number of shoots severely. A pretreatment of three days at 24°C or one day at 28°C reduced the shoot number by 50 %. After seven days at 28°C there was not a single shoot in any of the cultures. However, after two weeks at 15 or 18°C it was no longer possible to inhibit the shoot formation by a 24°C treatment. It is concluded that the formation of shoots in petiole segments takes place during the first two weeks after excision, and that high temperature is detrimental to the shoot initiation process.     

Agrobacterium tumefaciens-mediated transformation of Robinia pseudoacacia

Robinia pseudoacacia (black locust) plants were regenerated after co-cultivation of stem and leaf segments with Agrobacterium tumefaciens strain GV3101 (pMP90) that harbored a binary vector that included genes for β-glucuronidase (GUS) and hygromycin phosphotransferase. Successful transformation was confirmed by the ability of stem and leaf segments to produce calli in the presence of hygromycin, by histochemical and fluorometric assays of GUS activity in plant tissues, and by Southern blotting analysis. In this transformation system, about 2 months were required for regeneration of transgenic plants from stem and leaf segments. The frequency of transformation from stem segments was approximately 24%, and the morphology of regenerated plants resembled that of the original parental strain. Received: 2 September 1999 / Revision received: 30 November 1999 / Accepted: 4 December 1999     

Biology of Agistemus olivi,a new predator of eriophyid mites infesting olive trees in Egypt

Life history and prey consumption of the predatory stigmaeid mite Agistemus olivi Romeih, as affected by feeding on the motile stages of the olive bud mite Aceria oleae Nalepa and the olive rust mite Tegolophus hassani Keifer (Acari : Eriophyidae), has been studied for the first time at different temperatures and 70–75% r.h. A greater capability was shown for consuming the olive bud mite than the olive rust mite and the former prey relatively induced more fecundity. The rise of different temperatures from 20°C to 25°C and 30°C shortened development and increased reproduction and prey consumption. The maximum reproduction (6.92 and 6.08 eggs ♀ day) was recorded at the higher temperature, while the minimum reproduction (1.50 and 1.30 eggs ♀ day) was observed for A. oleae and T. hassani, respectively.     

Asexual reproduction, evisceration, and regeneration in holothurians (Holothuroidea) from Nha Trang Bay of the South China Sea

Peculiarities of asexual reproduction, evisceration, and regeneration were studied in 13 holothurian species of the orders Dendrochirotida and Aspidochirotida from Nha Trang Bay of the South China Sea. Asexual reproduction by fission has been described for the first time for Cladolabes schmeltzii. It has been shown for the first time that transected C. schmeltzii and Colochirus robustus successfully regenerate not only their anterior, but also their posterior structures. It was found that Pseudocolochirus violaceus was able to regenerate the anterior part of the body and to eject viscera through the anus. The modes of evisceration and regeneration of the gut have been elucidated for ten species of aspidochirotids. It has been shown that the intestine of Holothuria scabra forms from two anlagen rather than from a single one, as previously believed.     

Regeneration of the gut requires retinoic acid in the budding ascidian Polyandrocarpa misakiensis

The protochordate ascidian Polyandrocarpa misakiensis has a striking ability to regenerate. When the posterior half of the adult body is amputated, the anterior half completely loses the esophagus, stomach and intestine. These organs are reconstituted in a week. Histological observation revealed that the regeneration involves transdifferentiation of the atrial epithelium near the cut surface. The morphological features of the gut primordium were similar to those observed in the developing bud of this species. Inhibitors of the synthesis of retinoic acid (RA) suppressed the formation of the gut. 13‐cis RA rescued the regenerates from the inhibitor‐induced hypoplasia. These results suggest that RA is required for the regeneration of the gut. A gene encoding the RA receptor (Pm‐RAR) and its target gene, TRAMP, were expressed in and around the regenerating gut. Pm‐RAR‐specific and TRAMP‐specific double‐stranded RNA molecules inhibited the regeneration of the gut, indicating that the RA signal is mediated at least in part by Pm‐RAR and TRAMP. These results suggested that RA triggers the transdifferentiation of the atrial epithelium into the gut in regenerating animals, as it does during asexual reproduction.     

Effects of temperature, light and heterotrophy on the growth rate and budding of the temperate coral Cladocora caespitosa

Recent investigations have shown the temperate scleractinian coral Cladocora caespitosa to be a new potential climate archive for the Mediterranean Sea. Whilst earlier studies have demonstrated a seasonal variation in growth rates, they were unable to distinguish which environmental parameter (light, temperature, or food) was influencing growth. In this study, the effect of these three factors on the coral physiology and calcification rate was characterized to aid the correct interpretation of skeletal trace element variations. Two temperatures (13 and 23°C), irradiances (50 and 120 μmol m⁻² s⁻¹), and feeding regimes (unfed and fed with nauplii of Artemia salina) were tested under controlled laboratory conditions on the growth, zooxanthellae density, chlorophyll (chl) content, and asexual reproduction (budding) of C. caespitosa during a 7-week factorial experiment. Unlike irradiance, which had no effect, high temperature and food supply increased the growth rates of C. caespitosa. The effect of feeding was however higher for corals maintained at low temperature, suggesting that heterotrophy is especially important during the cold season, and that temperature is the predominant factor affecting the coral’s growth. At low temperature, fed samples had higher zooxanthellae density and chl content, possibly for maximizing photosynthetic efficiency. Sexual reproduction investment of C. caespitosa was higher during favourable conditions characterised by high temperatures and zooplankton availability.     

Reproduction and population parameters of the Nearctic predator Geocoris punctipes at constant and varying temperature regimes

The heteropteran predator Geocoris punctipes (Say) has been used in augmentative biological control since 2000 to control Lepidoptera. However, surprisingly, few data are available about the influence of temperature on its population development, which is of key importance to plan the number and moment of releases to obtain sufficient pest reduction. The objective of this study was to evaluate daily and total fecundity, longevity and life table parameters (m_x, l_x, r_m, R, λ, T and TD) of G. punctipes at constant (16.8°C, 21.5°C, 24.5°C and 28.3°C) and corresponding varying (day/night) (21/11°C, 24/18°C, 27/21°C and 30/26°C) temperatures. Pairs of adult predators aged 24 h and originating from nymphs exposed to the same temperature regimes were kept at the above‐mentioned temperature regimes in Petri dishes containing Anagasta kuehniella (Zeller) eggs and an oviposition substrate. Tests were conducted in climatic chambers at the different temperature regimes and a RH 70 ± 10% and a 14L: 10D photoperiod. Reproduction, longevity and life table parameters were significantly affected by temperature, with clear differences between treatments at low (16.8°C, 21/11°C, 21.5°C, 24/18°C) or a high (24.5°C, 27/21°C, 28.3°C, 30/26°C) temperature regimes. Highest reproduction and fastest population growth of G. punctipes took place at average temperatures ranging from 24.5°C to 30°C, and neither reproduction nor population growth was negatively influenced by varying temperatures at any of the temperature regimes.