Reproductive ecology of the deep-sea scleractinian coral <Emphasis Type="Italic">Fungiacyathus marenzelleri</Emphasis> (Vaughan, 1906) in the northeast Atlantic Ocean

The reproductive biology and its seasonality were examined in the deep-water, solitary coral Fungiacyathus marenzelleri from 2,200 m depth in the NE Atlantic, using histological techniques. A total of 186 corals were collected using either an Otter trawl (semi-balloon) or Agassiz trawl from the research vessel RRS Challenger between 1979 and 1991. All individuals were gonochoric, with all 48 mesenteries being fertile. A high fecundity was recorded (2,892 ᇀ.4 oocytes per polyp) with strong size-dependency. This species is a quasi-continuous reproducer with overlapping gametogenesis for both spermacysts and oocytes. No planulae were observed and broadcasting of gametes is inferred, with the large egg size (max. 750-µm diameter) suggesting lecithotrophic development. There is a non-significant trend towards a synchronous spawning event during June/July. Asexual fission was observed, though in low numbers.     

Mesozooplankton community structure in the Southern Ocean upstream of the Prince Edward Islands

A meso-scale oceanographic grid survey was conducted during the first cruise of the Marion Offshore Ecosystem Variability Study in the upstream region of the Prince Edward Islands in austral autumn (April/May) 2001. Mesozooplankton samples, collected using a Bongo net (fitted with 200-µm and 300-µm mesh nets), were separated into three size fractions, 200-500 µm, 500-1,000 µm, 1,000-2,000 µm, by reverse filtration. Total surface (depth<5 m) chlorophyll-a concentration during the study ranged between 0.11 and 0.34 µg l^-1 and was always dominated by picophytoplankton (0.45-2.0 µm). Total mesozooplankton abundance and biomass during the survey ranged between 49 and 1,512 ind. m^-3 and between 0.7 and 25 mg Dwt. m^-3, respectively. Throughout the survey, the 200 to 500 µm class numerically dominated the mesozooplankton community, with an average of ~69% (SD=ᆠ.3%). The dominant species in the 200- to 500-µm size fraction were the copepods, Oithona similis, Calanus simillimus and Metridia lucens, and the pteropod, Limacina retroversa. However, in terms of biomass, the 1,000- to 2,000-µm group was predominant, with dry weight values constituting an average of ~66% (SD=ᆞ.2%). The most well-represented species in this group were the carnivorous Euphausia vallentini, Thysanoessa vicina, Sagitta gazellae and Eukrohnia hamata. Three distinct groupings of stations were identified by numerical analysis. The different station groupings identified reflect changes in the relative contributions of the dominant species, as opposed to the presence/absence of species.     

Broadcasting fables: Is external fertilization really primitive? Sex,size, and larvae in sabellid polychaetes

Traditionally, broadcast spawning and planktonic larvae have been considered the plesiomorphic ‘ground plan’ for the Polychaeta and other metazoan groups. To assess whether this reproductive mode is in fact ‘primitive’, the study of monophyletic groups with various reproductive modes should be informative. A large range of body sizes would allow testing the ideas that aspects of reproductive mode may be functionally constrained. The family Sabellidac is one such group, with sexual reproductive modes ranging from broadcast spawning to intratubular brooding to ovovivi-parity, and a body size range over more than five orders of magnitude. Sabellids have previously been the subject of detailed cladistic analyses (Fitzhugh 1989, 1991); here we introduce several new characters based on morphology of reproductive structures. Larval development in four brooding sabellid species is also described with the aim of introducing new characters for future systematic analyses. Our cladistic analysis of sabellid genera suggests that gonochorism and brooding of direct-developing larvae are plesiomorphic in the Sabellidae, with external fertilization and swimming larvae limited to apomorphie clades in the subfamily Sabellinae. The presence of sperm with elongate heads may be correlated with the presence of intratubular brooding, though an adequate causal explanation for this relationship can not yet be presented. The concept that ‘modified’ sperm must be derived from ‘primitive’ sperm is shown to be false, with ‘modified’ sperm being plesiomorphic for the Sabellidae, from which ‘primitive’ sperm is derived in apomorphic Sabellinae. All sabellids have lecithotrophic development and appear to be phylogenetically constrained in this regard. Data gathered on body size and reproductive variables in the Sabellidac suggests the following (when phylogenetic effects are not controlled): (1) egg number and total egg volume are significantly correlated with body size, with small animals having fewer, larger eggs than large animals; (2) individual egg volume is not correlated with body size; (3) reproductive mode is significantly correlated with body size; intratubular brooders tend to be small-bodied, whereas broadcast spawners are large. However when the effect of body size is controlled for, then (4) egg number, egg volume and total egg volume all vary significantly with reproductive mode. Broadcast spawners expel a large number of small eggs for a high total egg volurne. Intratubular brooders have a few relatively large eggs for a small total egg volume. When statistics arc performed using phylogenetically independent contrasts there is a significant correlation between total egg volume and body size but not for egg number and body size. The effect of non-independence (due to phylogeny) of our data needs to be more fully controlled in future analyses but methods of incorporating continuous data into cladistic analyses should also be investigated. We show that some predictions can be made about reproductive mode based on body size but ad hoc patterns of reproductive character-state transformation should not be made independent of empirical hypotheses of phylogenetic relationship. Further studies of this kind throughout the Annelida are needed to determine the plesiomorphic reproductive mode for the phylum.     

Ultrastructure of dimorphic sperm and seminal receptacle in the hermaphrodites Barrimysia siphonosomae and Pseudopythina ochetostomae (Bivalvia, Galeommatoidea)

The intertidal Bivalvia Barrimysia siphonosomae (Montacutidae) and Pseudopythina ochetostomae (Kelliidae) live commensally with Siphonosoma cumanense (Sipuncula) and Ochetostoma erythrogrammon (Echiura), respectively. Both bivalves are hermaphrodites although males and females may occur. Paired, pouch-shaped seminal receptacles are located in the suprabranchial chamber near the genital opening of both species. The interior of the receptacles of sexually mature individuals contains numerous non-epithelial cells separated by narrow spaces that serve as a depot for sperm cells. Specimens of both species produce two sperm morphotypes. Euspermatozoa are oblong with elongate conical acrosomes, and a middlepiece consisting of about eight long mitochondria spirally arranged to form a sheath surrounding the basis of the flagellum. The paraspermatozoa are vermiform, about 4 µm broad and longer (up to 240 µm) in B. siphonosomae than in P. ochetostomae (ca 150 µm). The anterior, bullet-shaped, 4.5- or 3.0-µm-long acrosome is located above an elongate, 4- to 6-µm-long subcylindrical nucleus. Adjacent to the nucleus occurs a bundle of approximately 25 (B. siphonosomae) or ca 10 (P. ochetostomae) flagella. The cytoplasm of the sperm body contains spherical lipid droplets, glycogen deposits and numerous membrane-bound spindle- to rod-shaped electron-dense granules. It is presumed that both sperm types aggregate to form spermatozeugmata. The presence of a combination of almost identical and highly specialised euspermatozoa and paraspermatozoa in species of the two genera qualifies these structures as strong synapomorphies. Pseudopythina, as presently defined, is considered paraphyletic.     

Maternal brooding in the children’s python <Emphasis Type="Italic">(Antaresia childreni)</Emphasis> promotes egg water balance

Parental care provides considerable benefits to offspring and is widespread among animals, yet it is relatively uncommon among squamate reptiles (e.g., lizards and snakes). However, all pythonine snakes show extended maternal egg brooding with some species being facultatively endothermic. While facultative endothermy provides thermal benefits, the presence of brooding in non-endothermic species suggests other potential benefits of brooding. In this study we experimentally tested the functional significance of maternal brooding relative to water balance in the children’s python, Antaresia childreni, a small species that does not exhibit facultative endothermy. Clutch evaporative water loss (EWL) was positively correlated with clutch mass and was much lower than expected values based on individual eggs. The conglomerate clutch behaved as a single unit with a decreasing surface area to volume ratio as clutch size increased. Maternal brooding had a dramatic impact on evaporation from eggs, reducing and possibly eliminating clutch EWL. In a separate experiment, we found that viability of unattended eggs is highly affected by humidity level, even in the narrow range from 75 to 100% relative humidity at 30.5°C (20–33 mg m⁻³ absolute humidity). However, the presence of the brooding female ameliorated this sensitivity, as viability of brooded clutches at 75% relative humidity was higher than that of non-brooded eggs at either the same absolute humidity or at near-saturated conditions. Overall, these results demonstrate that brooding behavior strongly promotes egg water balance (and thus egg viability) in children’s pythons.     

Drei neue Polychaeten-Arten der Familien Polynoidae und Syllidae von Neu-Kaledonien,assoziiert mit einer verkalkten Hydrozoe

Three new species are described:Lagisca zibrowii (Polynoidae) andProcerastea hydrozoicola andP. parasimpliseta (Syllidae), living in association with a calcified hydrozoan, probably of the genusPseudosolanderia (Rosalindidae). The genusParaprocerastea is synonymized withProcerastea.     

Observations on population structure and reproductive features of <Emphasis Type="Italic">Laetmonice producta</Emphasis> Grube (Polychaeta,Aphroditidae) in Antarctic waters

The large scale-worm Laetmonice producta Grube 1877 is the most abundant aphroditid polychaete in Antarctic coastal waters. We investigated the demographic structure and some reproductive features of different L. producta populations from high-Antarctic (Weddell Sea) and Antarctic Peninsula (King George Island) shelf bottoms, collected in summer 1996 (ANT-XIII/3, EASIZ I cruise) and autumn 2000 (ANT-XVII/3, EASIZ III cruise). L. producta in the studied geographic areas showed a wide bathymetric range (200-850 m depth), and a different size distribution pattern with depth, characterised by a reduction of large specimens in the deepest stations. The species is gonochoric, with females more abundant in specimens of larger sizes. Eggs at different stages of maturation (ranging from 40 to 320 wm in diameter) were examined in 270 individuals from different stations and size classes. Egg size showed a slightly bimodal trend, with largely overlapping egg cohorts, suggesting a continuous reproduction, and a long-lasting gametogenesis. Significant differences (Kolmogorov-Smirnov test, P<0.05) in egg-size frequency distribution were detected only when data of the two geographic areas were compared (Weddell Sea vs King George Island), and not according to stations within each area, and females' size. The two sets of geographic samples were collected in different seasons and therefore it was not possible to assess if differences observed are due to sampling time or to geographic factors. Mature spermatozoa were recognisable only in autumn male specimens from King George Island, and showed a rounded nucleus and a short conical acrosome. Occurrence of an endosymbiont polychaete, Veneriserva pygoclava meridionalis (new sub-species of Dorvilleidae), was recorded in the coelomic cavity of 163 specimens of L. producta, 125 of which were from the deepest station of the Weddell Sea (stn. 14, 850 m depth). L. producta females with and without the endosymbiont did not show differences in egg-size distribution. The reproductive features of L. producta, together with its large size and slow growth, seem typical of a long-living predator species, and uncoupled from the typical summer environmental conditions and the pulsating system of coastal Antarctic waters.     

Nest, but Not Egg, Fidelity in a Territorial Salamander

Egg recognition and subsequent egg brooding are costly forms of parental investment in many species of vertebrates. Life history factors, such as coloniality or risk of brood parasitism, may constrain egg recognition in vertebrates. Female red-backed salamanders ( Plethodon cinereus ) from my study site are territorial and do not share nest sites with other females. They are terrestrial and neither they nor their eggs are likely to be displaced by environmental factors such as flooding. I experimentally tested, in the laboratory, the hypothesis that female red-backed salamanders can discriminate between their own eggs and the eggs of unfamiliar females. Each female was allowed to move about a test chamber containing two clutches of eggs, one clutch with which it was found in the forest and one that had been found with a distant female. Most females remained with one clutch of eggs, which they brooded during the entire observation period. However, they did not significantly prefer to brood their own eggs over the eggs of another female. In a corollary field experiment, I tested whether brooding females that were displaced 1 m from their nest sites would return to their territories and commence brooding behaviour within 3 d. All 10 displaced females returned to their own nest within this time period and were found brooding their eggs. Because female red-backed salamanders at my study site do not tend to share nest sites with other females and because their eggs remain in stationary nests, selection may not have favoured egg recognition. However, the results suggest that female salamanders indirectly recognize their own eggs by actively recognizing their territorial nest sites.     

Ichthyoplankton abundance and distribution in the Ross Sea during 1987–1996

During Italian expeditions, ichthyoplankton was collected in the Ross Sea and Terra Nova Bay, by BIONESS and Hamburg Plankton Net (250-µm and 500-µm mesh, respectively). A total of 394,453 fish larvae representing 46 species, 27 genera and 9 families were collected. Pleuragramma antarcticum dominated during three of the four cruises, whilst in 1994/1995 Trematomus lepidorhinus was also abundant. The most abundant icefish was Chionodraco myersi, which co-occurred with P. antarcticum. Macrouridae, Myctophidae and Bathylagidae were oceanic. Paralepididae occurred in all areas. Notothenioids, nearly 100% of the catch, occurred close to the coast and dominated the shelf ichthyoplankton community.     

Primary production, light and vertical mixing in Potter Cove, a shallow bay in the maritime Antarctic

Phytoplankton photosynthesis was measured during spring-summer 1991-1992 in the inner and outer part of the shallow Potter Cove, King George Island. Strong winds characterise this area. Wind-induced turbulent mixing was quantified by means of the root-mean square expected vertical displacement depth of cells in the water column, Z_t. The light attenuation coefficient was used as a measure of the influence of the large amount of terrigenous particles usually present in the water column; 1% light penetration ranged between 30 and 9 m, and between 30 and 15 m for the inner and outer cove, respectively. Obvious differences between photosynthetic capacity [P*_max; averages 2.6 and 0.6 µg C (µg chlorophyll-a)^-1 h^-1] and photosynthetic efficiency {!*; 0.073 and 0.0018 µg C (µg chlorophyll-a)^-1 h^-1 [(µmol m^-2 s^-1)^-1]} values were obtained for both sites during low mixing conditions (Z_t from 10 to 20 m), while no differences were found for high mixing situations (Z_t>20 m). This suggests different photoacclimation of phytoplankton responses, induced by modifications of the light field, which in turn are controlled by physical forcing. Our results suggest that although in experimental work P*_max can be high, wind-induced mixing and low irradiance will prevent profuse phytoplankton development in the area.     

Sexual reproduction and larval settlement of the zooxanthellate coral Alveopora japonica Eguchi at high latitudes

Sexual reproduction and larval settlement of Alveopora japonica Eguchi were studied at its northernmost distribution in Tokyo Bay (34°58'03'N, 139°46'05'E), where the annual sea temperature ranges from 13 to 27 °C. Alveopora japonica is a hermaphroditic brooding coral with oocytes and spermaries developing on separate mesenteries of the polyp. The oocytes first appeared in October, maturing in late August to early September of the following year, whereas the spermaries were first observed in May, and matured in approximately 4 months. The oocytes reached ca. 800 µm in diameter. Planulae containing zooxanthellae in their endoderm were released during the daytime in September and October. Well-developed planulae were able to settle and metamorphose within 7 h. The polyps started budding about 3 weeks after settlement, and took 3 years to grow to maturity. The population examined was sexually reproductive, indicating that A. japonica maintains local populations in Tokyo Bay by sexual reproduction.     

Brooding in Psolus patagonicus (Echinodermata: Holothuroidea) from Argentina, SW Atlantic Ocean

The mode, season, and time of brooding, egg diameter, egg number per brood, and the characteristics of newly released juveniles of Psolus patagonicus were investigated off Mar del Plata, Buenos Aires, Argentina, between October 1999 and February 2001. Individuals were attached to the Patagonian scallop, Zygochlamys patagonica. Spawning occurs between February and March. The mean egg diameter, 887 ± 26 μm, is the highest reported for the family Psolidae. Eggs are brooded under the mother’s sole until they develop into crawling juveniles within 7 months. The largest embryos reached a length of 1,941 ± 228 μm in September. During the brooding period (February–September) the number of brooded embryos decreased while their size increased. Our study confirms brooding behaviour in female P. patagonicus.     

Nesting behavior of the icefish Chaenocephalus aceratus at Bouvetøya Island, Southern Ocean

We describe in situ observations on nesting by the Scotia Sea (or blackfin) icefish Chaenocephalus aceratus (Lönnberg) that constitute the first substantive evidence of egg brooding and parental care by species of the family Channichthyidae. At Boutetoya Island six fish, all apparently male, were observed guarding egg nests at depths of 141–148 m during an ROV deployment. Eggs were laid as aggregated, round masses (~20–25 cm diameter) in shallow, circular depressions (~1-m diameter, ~20-cm depth) that were probably excavated by the parent(s) to protect the nests. The fish guardians remained tenaciously in contact with the eggs despite disturbances caused by the ROV, reacting to this threat with stress and defense behaviors. Because brooding fishes are more susceptible to the population impacts from trawl fisheries, we argue that this life history should be kept in mind in designing management schemes.     

First record of anomuran and brachyuran larvae (Crustacea: Decapoda) from Antarctic waters

Two decapod crustacean larval morphotypes belonging to the Anomura and Brachyura were found for the first time in Antarctic waters. Nine specimens were obtained from qualitative plankton hauls in Maxwell Bay (Bransfield Strait) (62°14'33S; 58°43'81W) off King George Island, Antarctic Peninsula. The anomuran morphotype belonged to the Hippidae, and apparently to the genus Emerita, whereas the brachyuran representative was assigned to the genus Pinnotheres (Pinnotheridae). At present, species determination is not possible due to lack of knowledge of larval morphology in both families. Adult forms of these reptant decapods are not known from Antarctic waters; the occurrence of the present larval forms is considered as a possible intrusion of Subantarctic water masses into the Antarctic environment. This hypothesis is supported by the additional presence of the copepod genus Acartia in the same sample material, which is exclusively known from Subantarctic waters.     

Photosynthesis, photoinhibition, and nitrogen use efficiency in native and invasive tree ferns in Hawaii

Photosynthetic gas exchange, chlorophyll fluorescence, nitrogen use efficiency, and related leaf traits of native Hawaiian tree ferns in the genus Cibotium were compared with those of the invasive Australian tree fern Sphaeropteris cooperi in an attempt to explain the higher growth rates of S. cooperi in Hawaii. Comparisons were made between mature sporophytes growing in the sun (gap or forest edge) and in shady understories at four sites at three different elevations. The invasive tree fern had 12-13 cm greater height increase per year and approximately 5 times larger total leaf surface area per plant compared to the native tree ferns. The maximum rates of photosynthesis of S. cooperi in the sun and shade were significantly higher than those of the native Cibotium spp (for example, 11.2 and 7.1 µmol m^-2 s^-1, and 5.8 and 3.6 µmol m^-2 s^-1 respectively for the invasive and natives at low elevation). The instantaneous photosynthetic nitrogen use efficiency of the invasive tree fern was significantly higher than that of the native tree ferns, but when integrated over the life span of the frond the differences were not significant. The fronds of the invasive tree fern species had a significantly shorter life span than the native tree ferns (approximately 6 months and 12 months, respectively), and significantly higher nitrogen content per unit leaf mass. The native tree ferns growing in both sun and shade exhibited greater photoinhibition than the invasive tree fern after being experimentally subjected to high light levels. The native tree ferns recovered only 78% of their dark-acclimated quantum yield (F_v/F_m), while the invasive tree fern recovered 90% and 86% of its dark-acclimated F_v/F_m when growing in sun and shade, respectively. Overall, the invasive tree fern appears to be more efficient at capturing and utilizing light than the native Cibotium species, particularly in high-light environments such as those associated with high levels of disturbance.     

Pixel by pixel: real-time observation and quantification of passive flotation speeds of three common equine endoparasite egg types

The efficacy of anthelmintic treatments against populations of endoparasites infecting livestock throughout the world is decreasing. To mitigate this, the use of fecal egg counts is recommended to determine both the necessity, and to ensure the appropriate choice, of anthelmintic treatment. Traditionally, and in order to facilitate easier identification and/or enumeration, samples are analysed after separating eggs from other fecal particulates by exposing them to a solution with a density higher than that of the eggs, but lower than the remaining fecal contents. While many parasite egg flotation protocols exist, little is known about the characteristics of these eggs with respect to their movement through a flotation solution. In this study, we have demonstrated a novel method for the observation and quantification of microscopic (65–100 µm) objects as they experience unassisted flotation. This also represents, to our knowledge for the first time, that the flotation of parasite eggs has been observed and their movement characteristics quantified as they float through solution. Particle tracking and video analysis software were utilised to automatically detect and track the movement of individual eggs as they floated. Three 30 s videos and one 2 min video of each egg type were analysed. If the first 30 s of video were discounted, the differences in mean flotation speed among all videos was statistically significant between egg types (P = 0.0004). Strongyle type eggs (n = 201) moved the fastest with a mean 51.08 µm/s (95% confidence interval: 47.54–54.62). This was followed by Parascaris spp. (n = 131) and Anoplocephala perfoliata eggs (n = 322), with mean speeds of 44.43 µm/s (95% confidence interval: 39.47–49.4) and 31.11 µm/s (95% confidence interval: 29.6–32.61), respectively. This method for evaluating the mean speed of passive flotation may represent a first step towards further optimizing fecal egg flotation and be of interest to parasitologists and veterinary practitioners.     

Optimization of in vitro micropropagation and regeneration for Populus × interamericana and Populus × euramericana hybrids (P. deltoides, P. trichocarpa, and P. nigra)

A rapid and efficient micropropagation method has been established for six European poplar cultivars of economic interest - four Populus 2 interamericana and two Populus 2 euramericana. Using a three-step procedure, we were able to regenerate plantlets from callus and acclimate them within 4 months. In the first step, callogenesis was induced when explants were cultured for 25 days on culture medium supplemented with 10 µM !-naphthaleneacetic acid and 5 µM N⁶(2-isopentenyl)adenine. Bud regeneration followed by shoot elongation was then obtained from callus tissue by combining the cytokinin-like compound thidiazuron with the surfactant Pluronic F-68 at concentrations adjusted for each cultivar. The usefulness of this procedure in the area of genetic engineering is discussed.     

Larval niche selection and evening exposure enhance adoption of a predacious social parasite, Maculinea arion (large blue butterfly), by Myrmica ants

Larvae of the butterfly genus Maculinea feed briefly on a foodplant before being adopted as social parasites into Myrmica ant nests. Each Maculinea species typically survives only with a single Myrmica species, yet the eggs are laid across the overlapping territories of 3-5 Myrmica species and several other ants. The ability of Maculinea arion - a 'predatory' species of Maculinea - to influence its adoption into host Myrmica colonies was studied for the first time in the field. Some earlier reports, involving captive non-host ants, suggested that larvae of the predatory Maculinea follow ant trails into host nests or wander some distance from their foodplant before being discovered and (after a long interaction) carried away by Myrmica foragers. No dispersal from foodplants occurred in wild Maculinea arion larvae. Instead, they increased by >100-fold their probability of being discovered and adopted by Myrmica spp., rather than by ants of other genera, by exposing themselves in the micro-niche occupied by Myrmica foragers at their time of day of peak foraging. Despite a complex, hour-long interaction with Myrmica workers before being carried to the nest, Maculinea arion did not enhance its adoption by host species of Myrmica. Eggs were laid without bias in Myrmica sabuleti (host) and Myrmica scabrinodis (non-host) territories; larval survival on Thymus was the same in both ants' territories; larvae waited to be found beneath their foodplant rather than seek their host; Myrmica sabuleti and Myrmica scabrinodis foraged in the same vertical and temporal niches, and had the same probability of discovering larvae; both ants behaved identically after finding larvae and took the same time to adopt them; and the ratio of wild larvae taken into Myrmica sabuleti or Myrmica scabrinodis nests was the same as the distribution between these ants of Thymus, eggs and pre-adoption larvae.     

Germination of synthetic seeds of pineapple (Ananas comosus L. Merr.)

. Tufts of multiple shoots were produced from dormant, axillary buds of pineapple in vitro. Tiny shoots (2-5 mm) isolated from the tuft of multiple shoots were encapsulated in 3% sodium alginate prepared using hormone-free Murashige and Skoog's basal medium, Murashige and Skoog's vitamins, 0.56 mM myo-inositol and 0.06 M sucrose. The encapsulated shoots represented synthetic seeds that germinated and formed roots in vitro after subculture onto one of the following media solidified with 0.8% agar: (1) hormone-free Murashige and Skoog's basal medium, Murashige and Skoog's vitamins, 0.56 mM myo-inositol and 0.06 M sucrose (Pin1), (2) Murashige and Skoog's basal medium, Murashige and Skoog's vitamins, 0.56 mM myo-inositol, 0.06 M sucrose, 9.67 µM 1-naphthalene acetic acid, 9.84 µM indole-3-butyric acid and 9.29 µM kinetin (Pin2), and (3) White's basal medium, White's vitamins, 0.56 mM myo-inositol, 0.03 M sucrose, 0.54 µM 1-naphthalene acetic acid and 1.97 µM indole-3-butyric acid (Pin3). Pretreatment of shoots in either liquid Pin3 or Pin4 medium (White's basal medium, White's vitamins, 0.56 mM myo-inositol, 0.03 M sucrose, 10.8 µM 1-naphthalene acetic acid and 39.4 µM indole-3-butryic acid) was required for development into plantlets with roots after culture on either Pin1, Pin2 or Pin3 media. One hundred percent germination of synthetic seeds to plantlets occurred after pretreatment of shoots in liquid Pin4 medium for 12 h followed by culture of synthetic seeds on Pin2 medium. Synthetic seeds stored at 4°C remained viable without sprouting for up to 45 days. Plantlets produced in vitro from synthetic seeds were successfully established in soil. The protocol provides an easy and novel propagation system for pineapple, an otherwise vegetatively propagated fruit crop.     

Hydrologic and behavioral constraints on oviposition of stream insects: implications for adult dispersal

The supply of recruits plays an important role in plant and animal population dynamics, and may be governed by environmental and behavioral constraints on animals. Mated females of the mayfly genus Baetis alight on rocks protruding from streams, crawl under water and deposit a single egg mass under a rock. We surveyed oviposition and emergence of a bivoltine population of B. bicaudatus in multiple stream reaches in one high-altitude watershed in western Colorado over 3 years to establish qualitative patterns at a regional scale (entire watershed), and quantitative patterns over six generations at a local scale (one stream reach). We also measured characteristics of preferred oviposition substrates, performed experiments to test hypotheses about cues used by females to select oviposition sites, and measured mortality of egg masses in the field. Our goals were to determine whether: (1) hydrologic variation necessitated dispersal of females to find suitable oviposition sites; (2) the local supply of females could provide the supply of local recruits; and (3) local recruitment determined the local production of adults. The onset of oviposition corresponded with the decline of spring run-off, which differed dramatically among years and among sites within years. However, eggs appeared before any adults had emerged in 8 of 22 site-years, and adults emerged 2-3 weeks before any eggs were oviposited in 3 site-years. Furthermore, the size distribution of egg masses differed from that predicted by the size distribution of females that emerged from seven of nine stream reaches. Protruding rocks and eggs appeared earlier each summer in smaller tributaries than in larger mainstream reaches, suggesting that hydrologic and behavioral constraints on oviposition may force females to disperse away from their natal reach to oviposit, and possibly explain the predominantly upstream flight of Baetis females reported in other studies. Local oviposition rates in one third-order stream-reach increased rapidly as soon as substrates protruded from the water surface, and females preferred large rocks that became available early in the flight season. However, females oviposited on <10% of all available rocks, and <65% of preferred rocks as determined by an empirical model. These data indicated that the timing of appearance of suitable oviposition sites determined the phenology of local recruitment, but that preferred oviposition sites were not saturated. Thus, the magnitude of local recruitment was not limited by the absolute abundance of preferred oviposition sites. Only 22% of egg masses observed in the field suffered mortality during their embryonic development, and per capita Baetis egg mass mortality was significantly lower on rocks with higher densities of egg masses. Thus, we suspect that specialized oviposition behavior may reduce the probability of egg mortality, potentially compensating for the costs of dispersal necessary to locate suitable oviposition sites. Finally, the number of adults that emerged at one stream reach was independent of the number of egg masses oviposited over six generations of Baetis; and local recruitment was not a function of the number of adults of the previous generation that emerged locally. The patterns of oviposition and emergence of Baetis found in this study are consistent with the following hypotheses. Recruitment of eggs in a stream reach is not limited by the local supply of adults, but is a function of the regional supply of dispersing adults, which are constrained by the spatial and temporal distribution of preferred oviposition habitat. Furthermore, subsequent local production of adults is not a function of the supply of recruits, arguing for post-recruitment control of local populations by processes operating in the larval stage (e.g., predation, competition, dispersal, disturbance). Processes affecting larval and adult stages of Baetis act independently and at different scales, thereby decoupling local population dynamics of successive generations.