Development of the salivary glands in embryos of Ixodes ricinus (Acari: Ixodidae)

We examined Ixodes ricinus embryos between 18 and 28 days of development with light, scanning and transmission electron microscopy. The differences in inner structure attested to establish three successive developmental stages: days 18-20, day 23, and days 26-28. Between 18 and 20 days the embryos are at early stages of organogenesis. Salivary glands cannot be identified at that stage. In 23-day-old embryos salivary glands are already outlined but the structure of alveoles is still different from that in larvae in which the embryonic development has been completed. Gland cells start to form alveoles and become active between 26 and 28 days of the development.     

Uptake and utilization of14C-glycine by embryos ofSebastes melanops

Synopsis The ability of embryos of the viviparous scorpaenidSebastes melanops to take up nutrients from an exogenous substrate was demonstrated by incubating embryos at various stages of development (18–30 days after fertilization) in¹⁴C-labeled glycine for 24 h. Uptake was highest for embryos at the latest stages (28–30 days) and increased at a linear rate during the incubation period. Nutrient uptake was not time dependent in embryos at the early stages (18–22 days). Nutrient utilization byS. melanops embryos was measured by the oxidation of¹⁴C-labeled glycine to¹⁴CO₂. The amount of respired¹⁴CO₂ by the oldest embryos increased significantly at a linear rate over the 24 h incubation period. There was no evidence of nutrient utilization by the youngest embryos. The developmental changes we observed in the uptake and utilization of exogenous glycine are supported by our previous findings that the oldest embryos have fully developed mouths and guts, and require additional nutrition from intraovarian sources at this stage of development.     

Isolation of cell lines from embryos of the cockroach,Blattella germanica

Summary Cell lines were isolated from three stages of embryos ofBlattella germanica dissociated with trypsin. The lines have been subcultured 50 to 134 times in 3 years. Line UM-BGE-1 was isolated from germ band embryos at stages of segmentation and limb-bud formation (5 days old). Line UM-BGE-2 was derived from embryos at dorsal closure (7 days old). Line UM-BGE-4 arose from embryos in the germ band and dorsal closure stages (5 and 7 days old); these cells colonize as hollow spheres or vesicles. Line UM-BGE-5, isolated during organogenesis (10 days old), developed into two distinct sublines. Subline α is composed of round cells that do not attach to the flask. Subline β grows as an attached monolayer; the cells can be removed with a saline solution containing 20mM disodium dihydrogen Versenate^?. Most of the cells of these lines have the diploid chromosome number (23 or 24) excepting line UM-BGE-1 in which the tetraploid number predominates. Presented in part at the 26th Annual Meeting of the Tissue Culture Association, at Montreal, Canada, 2–5 June 1975. This investigation was supported by U.S. Public Health Service Research Grant No. AI 09914 from the National Institute of Allergy and Infectious Diseases. This is Paper No. 9416, Scientific Journal Series, Minnesota Agricultural Experiment Station.     

Ontogeny of the sodium pump in embryos of rockfish of the genusSebastes

Synopsis The purpose of the present investigation was to determine at what stages of embryonic development the sodium pump (Na+,K+-activated ATP phosphohydrolase, EC 3.6.1.3) appears and whether the ontogeny of the sodium pump plays a role in the matrotrophic viviparity ofSebastes. Early larval stages (stages 1–14) had embryonic tissues nearly devoid of Na, K-ATPase activity. After epiboly, tissues from embryos taken from both species,S. schlegeli andS. taczanowskii, had significant levels of enzymatic activity coincident with the appearance of neuronal tissue in the head fold (stages 15–20). Maximal levels of specific activities of the enzyme were reached with the onset of the vascular circulation and the opening of the buccal cavity together with maturation of the midgut and hindgut regions of the intestinal tract (stages 26 and 28). Ouabain, a specific inhibitor of Na, K-ATPase, was used to measure survival of embryos placed in different seawater concentrations under in vitro conditions. The most sensitive stages to external ouabain were 26–30. These findings support the hypothesis that the intestinal tract is functional prior to gestation and could be transporting important nutritive material found in maternal ovarian fluid.     

Embryonic, larval and juvenile development of the roughskin sculpin,Trachidermus fasciatus (Scorpaeniformes: Cottidae)

Embryonic, larval and juvenile development of the catadromous roughskin sculpin,Trachidermus fasciatus, were described using eggs spawned in an aquarium. The eggs, measuring 1.98–2.21 mm in diameter, were light reddish-yellow and had many oil globules, 0.05–0.18 mm in diameter. Hatching occurred 30 days after spawning at 2.3–11.3°C. The newly-hatched larvae, measuring 6.9–7.3 mm BL, had a single oil globule, 9–10+25–26=34–36 myomeres and 6 or 7 large stellate melanophores dorsally along the gut. The yolk was almost resorbed, number of pectoral-fin rays attained 16–17, and two parietal, one nuchal and four preopercular spines were formed, 5 days after hatching, at 8.2–8.4 mm BL. The oil globule disappeared, and one supracleithral spine was formed, 11 days after hatching, at 8.9–9.5 mm BL. Notochord flexion began 15 days after hatching, at 9.7–10.3 mm BL. A posttemporal spine was formed 20 days after hatching, at 10.7–10.9 mm BL. The first dorsal fin spines (VII–VIII), second dorsal fin and anal fin rays (18–19, 16–18, respectively) appeared 23 days after hatching, at 12.0–13.7 mm BL. The pelvic fin spine and rays (I, 4) were formed and black bands on the head and sides of the body began to develop 27 days after hatching, at 13.8–15.8 mm BL. Newly-hatched larvae swam just below the surface in the aquaria. Preflexion larvae (8.9–9.5 mm BL), in which the oil globule had disappeared, swam in the middle layer, while juveniles (13.8–15.8 mm BL) began swimming on the bottom of the aquaria. Swimming behavior observed in the aquaria suggested that the fish started to change to a demersal existence at the juvenile stage.     

Effects of extreme changes of sea water temperature and salinity on the development of the sand dollar <Emphasis Type="Italic">Scaphechinus mirabilis</Emphasis>

The combined effects of temperatures of 14, 17, 20, 22, and 25°C and salinities of 36–12‰ on embryos and larvae of the sand dollar Scaphechinus mirabilis was studied. Embryonic development is the most sensitive stage in the early ontogenesis of S. mirabilis. It is completed at a temperature of 14–20°C in a salinity range of 36–24‰ and at temperature of 22°C to 26‰. The fertilization proceeds in wider ranges of temperature and salinity. Among the swimming larvae, blastulae showed the greatest resistance to variations of these environmental factors. All the larvae survived at a temperature of 14–22°C and a salinity of 36–20‰, and more than 70% of them at 18‰. The pluteus I is the most vulnerable stage; probably this is related to the formation of the larval skeleton and transition to phytoplankton feeding. The survival of larvae at the age of 20 days was 100% at 14–22° C and a salinity of 36–24‰, most of them survived at 14–20°C and a salinity 18‰. The temperature 25 ° C is the most damaging for early development of S. mirabilis. The duration of development of that species lasts 28.5–29 days at 20°C and a salinity of 32.2–32.6‰. At 20 and 22°C, the larvae settled and completed metamorphosis more quickly if sand from the parental habitat was present. The larvae did not settle during the experiment (14 days) at 14 ° C and in the absence of sand.     

Improvement of English walnut somatic embryo germination and conversion by desiccation treatments and plantlet development by lower medium salts

Summary Well-developed somatic embryos were selected from a repetivively somatic embryo line derived from embryonic axes of immature zygotic embryos of English walnut ‘No. 120’ (Juglans regia L.) for germination and conversion studies. In germinating dishes, somatic embryos germinated into only shoots, only roots, or both shoots and roots. Without any pretreatment, 28% somatic embryos germinated, while those treated with 2.5–5.0 mg 1⁻¹ (7.2–14.4 μmol) gibberellic acid (GA₃) germinated at 25–28% and those receiving a cold treatment of 2–3 mo. at 3–4°C germinated at 30–43%. However, only 4–19% of the germinating embryos showed both shoots and roots. Treated with desiccation, either with CaCl₂·6H₂O or Ca(NO₃)₂·4H₂O at 20°C in the dark for 3 d, somatic embryos germinated at 85–91%, 57–69% of which had both shoots and roots. Treatment with 2 mo. cold storage in combination with desiccation using Ca(NO₃)₂·4H₂O resulted in 92% of somatic embryos germinating, 70% of which showed both shoots and roots. No significant differences were observed between solid and liquid germination media. After transferring the germinating embryos to plantlet development media, 52–63% of those with both shoots and roots developed into plantlets while 11% with only shoots or 9% with only roots converted into plantlets. Plantlet development was improved by using lower medium salts and sucrose concentrations. The addition of activated charcoal enhanced root development, particularly root branching. Of 131 plants transplanted, 91 plants were acclimatized to a greenhouse.     

Optimization of <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation conditions in mature embryos of elite wheat

Immature embryos have been used frequently as target tissues in the genetical transformation of wheat. However, obtaining a large number of high quality immature embryos throughout the year is a laborious and delicate process, because of the need to cultivate the plants under controlled conditions. To circumvent this, we have employed mature embryos rather than immature ones as starter explants for Agrobacterium-mediated transformation of an elite wheat (Triticum aestivum L.) cultivar EM12. The neomycin phosphotransferase ІІ (npt ІІ) and β-glucuronidase (gus) genes were used as selectable and screenable marker genes, respectively, to assess and optimize the performance of T-DNA delivery. With the aid of an orthogonal design, the effect of four factors in combination on transfer DNA (T-DNA) delivery was studied. These factors were preculture duration, different kinds of inoculation, length of inoculation and co-culture condition. Optimal conditions for T-DNA delivery were obtained for mature embryos precultured for 14 days, followed by immersing in inoculation suspension with full strength Murashige and Skoog (MS) salts in darkness at 23–25°C for 3 h, and then co-culturing with Agrobacterium under desiccating condition in the dark at 23–24°C for 2–3 days. Complete analysis of transgene insertion demonstrated that the optimized method for Agrobacterium-mediated transformation of mature embryos of wheat was efficient and practicable.     

Gap-detecting mechanism in the seed germination ofMallotus japonicus (Thunb.) Muell. Arg., a common pioneer tree of secondary succession in temperate Japan

Germination responses ofMallotus japonicus (Thumb). Muell. Arg. seeds to temperature revealed a gap-detecting mechanism in the seed germination of the species. Among various constant and alternating temperatures examined in the range from 12–40°C, only very limited temperature regimes were found to be favourable for seed germination, specifically, alternating temperatures between 18–32°C and 28–40°C. A single several-hour higher-temperature (32–40°C) treatment could also induce the germination of seeds which had been imbibed for several days at a constant temperature in the range of 20–26°C, suggesting that there is a process requiring higher temperature among the overal germination processes. Seeds located at or near the surface of denuded soil would have a good chance of experiencing such a temperature change when several rainy days are followed by fine weather, while seeds beneath close vegetation would not. On the other hand, the pressence or absence of light or a simulated ‘canopy ligh’ had little effect on the germination. Therefore, it was concluded that the seeds ofM. japonicus have a ‘gapdetecting mechanism’ in the form of a higher-temperature requirement of a certain process involved in the overall germination processes.     

A new armored searobin <Emphasis Type="Italic">Paraheminodus longirostralis</Emphasis> (Teleostei: Peristediidae) from New Caledonia

A new species of armored searobin, Paraheminodus longirostralis, is described from five specimens collected from New Caledonia at depths of 412–467 m. It is distinguishable from its three known congeners in having 34 bony plates in the upper lateral row, a forward-directed spine on each plate between the 23rd–26th and 31st–32nd plates in the upper lateral row, 6–7 + 1 + 20–21 = 27–28 gill rakers, an elongate body posterior to the anus (49.9–52.1% standard length), an elongate rostral projection (53.0–59.3% head length), short upper jaw (42.1–43.4% head length), an elongate pectoral fin (70.6–79.4% head length), and long preopercular spine (39.2–57.7% head length).     

Doubled-haploid production in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.): role of stress treatments

This is the first report on the production of double-haploid chickpea embryos and regenerated plants through anther culture using Canadian cultivar CDC Xena (kabuli) and Australian cultivar Sonali (desi). Maximum anther induction rates were 69% for Sonali and 63% for CDC Xena. Under optimal conditions, embryo formation occurred within 15–20 days of culture initiation with 2.3 embryos produced per anther for CDC Xena and 2.0 embryos per anther for Sonali. For anther induction, the following stress treatments were used: (1) flower clusters were treated at 4°C for 4 days, (2) anthers were subjected to electric shock treatment of three exponentially decaying pulses of 50–400 V with 25 μF capacitance and 25 Ω resistance, (3) anthers were centrifuged at 168–1,509g for 2–15 min, and finally (4) anthers were cultured for 4 days in high-osmotic pressure (563 mmol) liquid medium. Anthers were then transferred to a solid embryo development medium and, 15–20 days later, embryo development was observed concomitant with a small amount of callus growth of 0.1–3 mm. Anther-derived embryos were regenerated on plant regeneration medium. Electroporation treatment of anthers enhanced root formation, which is often a major hurdle in legume regeneration protocols. Cytological studies using DAPI staining showed a wide range of ploidy levels from haploid to tetraploid in 10–30-day-old calli. Flow cytometric analysis of calli, embryos and regenerated plants showed haploid profiles and/or spontaneous doubling of the chromosomes during early regeneration stages.     

Survival of the mosquito predator,Notonecta unifasciata[Hemiptera: Notonectidae] embryos at low thermal gradients

Notonecta unifasciata Guerin eggs maintained at different stages of embryonic development in water at variable temperatures (2.2–25.6 °C) and for periods of 4–12 weeks revealed maximum viability (>80 %) at the highest temperature. However, optimum nondevelopmental viability was at 14.4 °C with eight-day-old embryos (>35 %). Short term (4 weeks) storage at 14.4 °C significantly increased egg viability. Survival was poor (<20 %) at the 2 lowest temperatures. Eggs held at 14.4 °C for 12 weeks and sustainingca. 50 % mortality, may be a practical procedure for biological control.      

Laboratory rearing of the squirrel fleaCeratophyllus sciurorum sciurorum with notes on its biology

A rearing method for the fleaCeratophyllus (Monopsyllus) sciurorum sciurorum, a pest on farmed mink in Europe, has been developed. The behaviour of egg-laying and the development period are described at 23 °C and 80% r.h. The eggs ofC. sciurorum sciurorum hatched after 4–7 days and cocoons were formed 10–12 days later. The development period from egg to newly emerged adult was 23–32 days. Females emerged, in general, earlier than males.     

Hot and salty: the temperature and salinity preferences of a temperate estuarine shrimp larva, Upogebia pusilla (Decapoda: Thalassinidea)

At a time when global climate changes are forcing life to adapt to a warming and salinity-changing environment, it is essential to understand how future changes in ocean chemistry will affect species. This study evaluates the combined effects of temperature and salinity on survival and development of Upogebia pusilla larvae. Combinations were made from three temperatures (18, 23, and 28°C) and three salinities (15, 25, and 35). Survival, larval duration and megalopa size were compared between treatments. U. pusilla larvae developed optimally in the highest salinity (35) and higher temperatures (23–28°C). Low salinities and temperatures did not support larval survival and development, with salinity being the main restricting factor for survival, while temperature affected mainly the duration of the larval stages. Larvae at higher temperatures (23–28°C) presented a higher development rate but no differences were found in megalopa size.     

Embryo rescue and plant regeneration in vitro of selfed chickpea (Cicer arietinum L.) and its wild annual relatives

The main constraint to the transfer of desired traits into cultivated chickpea from wild Cicer relatives is the presence of post-zygotic barriers which result in abortion of the immature embryo following interspecific hybridisation. Rescue of hybrid embryos in vitro and regeneration of hybrid plantlets could allow chickpea breeders to transfer desirable traits from wild relatives of chickpea. The development of embryo rescue techniques using selfed chickpea and selfed wild relatives is being used as a first step to protocols for wide hybrids. Optical microscopy studies of embryogenesis, in both selfs and hybrids, identified deleterious changes in the fertilised hybrid seed as early as 2–4 days after pollination in some crosses. These observations suggest that the appropriate time to rescue chickpea × C. bijugum hybrids is at the early globular stage of embryogenesis (2–7 days old), which requires the development of a complex tissue culture medium. In contrast hybrids between chickpea × C. pinnatifidum abort later (up to 15–20 days old) at the heart-shaped or torpedo stages, and are easier to rescue in vitro. Genotype also plays a significant role in the ability of immature selfed ovules to germinate in vitro. In this paper we report on the optimisation of␣protocols for rescueing immature embryos using selfed chickpea and its wild relatives in ovule, and subsequently to regenerate plantlets.     

A case for a free-running circannual rhythm in soldier developmental time of Formosan subterranean termites

The success of all insect societies relies on their ability to maintain optimal levels of different castes. Here we report on an apparent free-running circannual rhythm that optimizes the developmental time of the soldier caste of Coptotermes formosanus Shiraki. Over a 3 year period, bioassays were conducted each month (except June) with groups of 100 termite workers in a 28°C incubator in total darkness. The number of days needed for C. formosanus soldiers to develop varied depending on the time of the year (month). In March, just prior to the major swarming exodus for alates (April to June), 9 days were required before a worker molted to a presoldier. Longer times were required for such a molt in all other months, with an increasing trend from April to December (from 13 to 30 days) and a decreasing trend from January to February (from 25 to 12 days). Colony origin or the length of time that termites were kept in the laboratory under constant conditions (26 – 28°C, 70 – 80% RH) before testing (7 days – 1 year) did not affect this rhythm. This is the first demonstrated evidence of a free-running circannual rhythm in a social insect. Received 23 July 2007; revised 9 and 21 August 2007; accepted 23 August 2007.     

Reproductive biology of the bigeye thresher shark,Alopias superciliosus (Lowe, 1839) (Chondrichthyes: Alopiidae), in the northwestern Pacific

Reproductive aspects ofAlopias superciliosus in the northwestern Pacific were described in detail, on the basis of 629 specimens (429 females and 200 males) collected from January 1984 to October 1984 and from October 1992 to March 1994.Alopias superciliosus embryos are oophagous. Six developmental stages (3 encapsulative and 3 posthatching) based on embryonic morphology and source of nutrition were recognized. The species bears 2 embryos per litter, their size at birth being between 135 and 140 cm TL. The sex ratio of embryos was 1∶1. Total length of females at maturity was 332–341. 1. cm; of males 270.1–287.6 cm. The gestation period could not be determined because most adult females were pregnant throughout the year. The typical reproductive strategy ofA. superciliosus is the production of a few large embryos per litter, with no fixed mating or birthing season.     

Development of eggs, larvae and juveniles of laboratory-reared blue whiting,Sillago parvisquamis (Percoidei: Sillaginidae)

The embryonic, larval and juvenile development of blue whiting,Sillago parvisquamis Gill, are described from a series of laboratory-reared specimens. Mean egg diameter and mean total length (TL) of newly-hatched larvae were 0.71 mm and 1.58 mm, respectively. The eggs were non-adhesive, buoyant and spherical with an oil globule (mean diameter 0.18 mm). Hatching occurred about 20 hours after fertilization at a temperature of 24.0–25.0°C, newly-hatched larvae having 38–40 myomeres. The yolk and oil globule were completely absorbed 3 days after hatching at 2.8–3.2 (mean 3.0) mm TL. Notochord flexion was completed by 7.2–8.2 (7.7) mm TL, and pectoral and caudal fin rays fully developed by approximately 10 mm and 8.5 mm TL, respectively. Completion of fin development occurred in the following sequence: caudal, pectoral, anal and second dorsal, first dorsal and pelvic, the last-mentioned by approximately 11 mm TL. The larvae ofS. parvisquamis andS. japonica, which closely resemble each other in general morphology and pigmentation, could be distinguished as follows. Newly-hatchedS. parvisquamis larvae had more myomeres thanS. japonica (38–40 vs. 32–34) and more melanophores on the dorsal surface of the body (19–28 vs. about 40).Sillago japonica had a vertical band of melanophores on the caudal peduncle, which was lacking in postflexionS. parvisquamis larvae. In addition, juveniles ofS. parvisquamis (larger than 23 mm TL) had melanophores on the body extending anteriorly to below the lateral line to form a midlateral band, whereas no obvious band occurred on similarly-sizedS. japonica juveniles.     

Microspore embryogenesis and the development of a double haploidy protocol for cow cockle (Saponaria vaccaria)

Factors affecting microspore embryogenesis of cow cockle (Saponaria vaccaria) were evaluated including donor plant growing conditions, genotype, bud size, density, medium composition, and culture conditions. Of the two donor plant (day/night) temperature regimes evaluated (10/5°C and 20/15°C), plants grown at 20/15°C were the most embryogenic. An embryogenic frequency of greater than 350 embryos/100 buds was observed in the most embryogenic genotype, cv. ‘White Beauty’. Buds from 3–9 mm in length were evaluated for their embryogenic potential; buds that were 4–7.9 mm produced the most embryos/100 buds. Of all the media compositions evaluated, NLN medium with 15% sucrose resulted in the most embryos. Cow cockle microspores required an initial period of 32°C for 3 days for production of microspore-derived embryos (MDEs).