华重楼离体胚培养及植株再生初探

为探明华重楼离体胚培养及植株再生的基本体系,该文以华重楼离体幼胚为试验材料,以MS培养基为基本培养基,研究不同光照、不同浓度梯度组合的植物生长调节剂对华重楼离体幼胚萌发、成苗的影响。结果表明:培养到60 d时,暗培养条件下华重楼离体幼胚的生长率和萌发率分别比光培养条件下高45.25%、19.17%,故暗培养比光培养更有利于华重楼离体幼胚生长发育。当GA₃浓度相同时,离体幼胚萌发所需时间随IAA浓度增加而延长。不同浓度的GA₃都可促进离体幼胚萌发,促进作用由强到弱依次为5 mg·L^-1 GA₃>1 mg·L^-1 GA₃> 10 mg·L^-1 GA₃。在黑暗条件下,优选得到最适华重楼离体幼胚生长发育配方为1/2 MS+30 g·L^-1 蔗糖+7 g·L^-1琼脂+0.5 g·L^-1 活性炭+5 mg·L^-1GA₃+1 mg·L^-1 IAA。此配方可诱导华重楼离体幼胚在2个月左右萌发,萌发率达50%需80 d左右。在华重楼成熟胚出苗培养时发现,高浓度的植物生长调节剂会抑制华重楼成熟胚生长发育,高浓度GA₃甚至会导致华重楼成熟胚死亡。优选得到最适华重楼成熟胚出苗配方为MS+30 g·L^-1 蔗糖+6.2 mg·L^-1 硼酸+7 g·L^-1 琼脂+0.7 g·L^-1 活性炭+0.5 mg·L^-1 2,4-D+1.5 mg·L^-1 IAA + 1.5 mg·L^-1 ZT+5mg·L^-1 GA₃,诱导成熟胚出苗需43 d左右,75 d左右可形成真叶。     

A pupal lethal mutation with a paternally influenced maternal effect on embryonic development in Drosophila melanogaster

The maternal effect and zygotic phenotype of l(1)pole hole (l(1)ph) is described. l(1)ph is a zygotic lethal mutation which affects cell division of adult precursor cells in Drosophila larvae. The locus is located in 2F6 on the salivary gland chromosome map and four alleles have been characterized. Germ-line clonal analysis of amorphic alleles indicates that l(1)ph has a maternal effect lethal phenotype. Two lethal phenotypes are observed among embryos derived from female germ-line clones homozygous for amorphic alleles dependent upon the zygotic activity of l(1)ph⁺ introduced via the sperm. Class 1: If no wild-type dose of the gene is introduced, embryos form abnormal blastoderms in which nuclear migration and cell formation is disrupted leading to an ill-defined cuticular pattern. Class 2: If a wild-type copy of the gene is introduced, blastoderm cells do not form beneath the pole cells (the pole hole phenotype); subsequently such embryos are missing cuticular structures posterior to the seventh abdominal segment (the torso phenotype). When the zygotic activity l(1)ph⁺ is modulated using position effect variegation a new phenotype is observed among class 2 embryos in which torso embryos are twisted along their longitudinal axis.     

THE EFFECT OF PHOTON IRRADIANCE ON THE BEHAVIORAL ECOLOGY AND POTENTIAL NICHE SEPARATION OF FRESHWATER PHYTOPLANKTONIC FLAGELLATES1

Photoresponsive behavior in phytoplanktonic flagellates potentially controls depth regulation, vertical migration, and the accumulation of cells in preferred conditions and hence has major implications for photosynthesis, growth, and species competition. This study used microscale laboratory chambers with cell track analysis and mesoscale lake studies to investigate the responses of five phylogenetically contrasting freshwater flagellates to gradients of light. Laboratory results demonstrated that these species differed in their light preferences despite being grown at the same photon irradiance. Preferred photon irradiances were 20–120, 20, 65–120, 4–20, and>200 μmol photons·m^?2·s^?1, respectively, for Ceratium furcoides (Levander) Langhans, Chlamydomonas moewusii Gerloff, Dinobryon sertularia Ehrenberg, Euglena gracilis Klebs, and Plagioselmis nannoplanctica (Skuja) Novarino. Analysis of the response of individual cells showed that in all species, photoresponsive preference was the result of positive and negative phototaxis combined with step‐up and step‐down photophobic reactions. There was no evidence for photokinesis or cell memory. Only in C. furcoides did the preferred photon irradiance alter with growth conditions. In C. furcoides, D. sertularia, and P. nannoplanctica, irradiance preference matched the optimal irradiance for growth, whereas in the two remaining species a lower than optimal irradiance was preferred, suggesting that light may be used as an indicator of other ecological conditions. Mesoscale experiments in a lake demonstrated that the laboratory microscale measurements provide information relevant to understanding ecological distributions. Behavioral responses to light contribute to the delineation of vertical niche separation and provide a method for predicting the spatial and temporal distribution of flagellates.     

Optimization of direct somatic embryogenesis from mature zygotic embryos ofPanax ginseng C. A. Meyer

Culture conditions were optimized for somatic embryogenesis ofPanax ginseng. The highest frequency of embryo formation was obtained when tissues were excised from the middle region of the cotyledon segments of zygotic embryos. Only treatment with light could stimulate the formation of single-type somatic embryos, whereas multiple-type somatic embryos and calli were observed under dark conditions. The highest production of somatic embryos was found with an NH₄ ⁺:NO₃ ratio of 21:39. Among the tested media (MS, B5, and SH), maximum formation of somatic embryos was obtained when cotyledon expiants were cultured on an 1% agar MS medium supplemented with 5% sucrose. Regenerated ginseng plantlets were transferred to an autoclaved soil mixture in the greenhouse. These transformants showed no detectable variations in their morphology or growth characteristics compared with the donor plant.     

Adaptations of tropical marine microphytobenthic assemblages along a gradient of light and nutrient availability in Suva Lagoon,Fiji

How are microphytobenthic biofilms adapted to the high incident irradiances and temperatures, low inorganic nutrient concentrations and high desiccation stresses on intertidal flats present in tropical environments? This study investigated biofilms subject to different environmental conditions in a range of tropical sites in Suva lagoon, Fiji. PAM fluorescence was used to measure photophysiological responses to the light climate. Biofilm colloidal carbohydrate, extracellular polymeric substances (EPS) and low molecular weight (MW) carbohydrate concentrations and diel carbohydrate production patterns were measured. Average biomass (Chl a) ranged from 15 to 36?mg?m^?2, and was highest in seagrass bed sediments, but biomass was not correlated with water column or sediment porewater nutrient concentrations. Biofilm photophysiology differed significantly along a combined gradient of light and nutrient availability, with F _v/F _m, relative ETR_max and E _k of biofilms highest in mangrove and intertidal main island sites and lowest in subtidal coral reef flats. Subtidal biofilms showed photoinhibition at irradiances > 1000?µmol?m^?2. Significant correlations between Chl a and colloidal carbohydrate concentrations were present (except on intertidal sandflats), and tropical biofilms had higher ratios of colloidal carbohydrate and EPS to Chl a than temperate estuarine biofilms, probably due to a combination of high irradiance and low nutrient availability leading to the production of excess photoassimilates. The percentage of EPS present in the colloidal fraction was highest in coral sand biofilms (42%), which had the lowest nutrient concentrations, compared with other sites (25–32%). Intertidal biofilms predominantly consisted of large motile taxa and showed strong rhythms of vertical migration. During tidal emersion, high sediment temperatures (41?°C), irradiance (>2300?µmol?m^?2?s^?1) and salinity (49‰) stimulated downward migration. In silty sediments, migration resulted in a reduction in photosynthetic activity during the midday period but, in sands with high light penetration (to a depth of > 1700?µm), high production rates of EPS (18.2?µg carbo. µg Chl a^?1 h^?1) and low MW carbohydrate exudates (40.2?µg carbo. µg Chl a^?1 h^?1) occurred. Vertical migration, high E _k and high rates of photoassimilate dumping are all adaptations to living in the tropical intertidal zone. Seagrass and reef flat biofilms consisted of a diverse non-migratory flora of motile and non-motile taxa that were not subject to such extreme temperature and irradiance conditions. Low values of photosynthetic parameters and high colloidal and EPS content indicated that these biofilms were nutrient-limited.     

Effects of substrate and water velocity on migration by early‐life stages of kaluga,Huso dauricus (Georgi, 1775): an artificial stream study

The effect of two environmental factors (substrate type and water velocity regime) was studied in the ontogenetic migration intensity of kaluga, Huso dauricus, a protected species in the Amur River. Early‐life stages studied were: free embryos = days 0–8; larvae = days 9–49; and juveniles = days 50–66. Effect of treatments on fish migration intensity was observed in circular channels, allowing migration in an endless stream. Daily migration intensity of fish was characterized by counting the number of daily upstream or downstream fish passes past a specific site in the channel observed for 5 min every hour by vision or video camera. The hypothesis that substrate type (bare bottom or sand‐small cobble) affects migration intensity was accepted, depending on the life stage. For example, the substrate type had no effect on migration intensity of days 0–6 free embryos. However, the intensity in days 7–8 free embryos and days 9–29 larvae was greater over rock substrate, while intensity in days 46–49 larvae and days 50–66 juveniles was greater when the bottom was bare. Thus, the effect of the substrate on migration varied in intensity according to the life stage, and within the larva life stage, by age. The velocity regime had a positive effect on migration intensity of free embryos, but a high velocity (mean, 29.9 cm?s^?1) delayed the resting period of day 8 free embryos and day 9–15 larvae. Free embryos and larvae in low vs fast velocities showed that they may have a drive to migrate a similar distance, and moreover, that a triggering velocity may be needed to initiate or to stop migration. The hypothesis was accepted that the rearing velocity affects migration intensity, e.g. larvae reared in still water and then placed in moving water had a compensatory migration intensity response. For population enhancement stocking of H. dauricus, the results indicate culture practices must insure that: (i) same‐population individuals are mated to produce early‐life stages with the correct behaviour, (ii) migrating larvae, not juveniles, should be stocked, and (iii) larvae should be released in a river reach with a bottom velocity ≥30 cm?s^?1 containing sand and small pebbles.     

Biochemical characterization of developmental stages of cycad somatic embryos

The effect of two light intensities (25 μmol m⁻²s⁻¹ and 50 μmol m⁻²s⁻¹) on four developmental stages ofCeratozamia mexicana somatic embryos growing on semisolid plant growth medium at 25°C was measured. Growth parameters included fresh weight, morphology, and invertase and peroxidase activity. Under low light conditions, fresh weight was greater in stages 1 and 2 than in stages 3 and 4. In addition, there was a high frequency of hyperhydricity and polyembryogenesis in stages 1 and 2, whereas stages 3 and 4 were nonhyperhydric and unbranched. Stages 2–4 were green. Under high light conditions, embryos had lower fresh weights and less hyperhydricity, and stages 2–4 were green. Under low light conditions, peroxidase activity was less, although stage 1 embryos under both light conditions showed the highest activity. Stage 1 embryos required three to four months to develop to stage 2 under high light conditions and two to three months under low light conditions. Invertase activity under low light conditions was minimal in stage 2. All embryos had low invertase activity under high light intensity, and stages 2–4 had high levels of glucose. Embryo development from stage 2 to the next and for each subsequent stage under high light conditions required three to four months, and under low light conditions required four to five months. Higher light intensity therefore promotes the speedy recovery of plants.

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Resumen  El efecto de dos intensidades de luz (25 μmol m⁻²s⁻¹ y 50 μmol m⁻²s⁻¹) fue registrado en cuatro estados de desarrollo de embriones somáticos deCeratozamia mexicana cultivados en medio semisólido, 25°C. Los parámétras de crecimiento incluyeron peso fresco, peso seco, morfología, y actividad peroxidasa e invertasa. Bajo condiciones de baja iluminación, el peso fresco de los estados 1 y 2 fue mayor que en los estados 3 y 4. Además, hubo una alta frecuencia de hiperhidratación y poliembriogénesis en estados 1 y 2, mientras que los estados 3 y 4 no resultaron hiperhidratados ni ramificados. Los estados 2–4 fueron verdes. Bajo alta iluminación, los embriones tuvieron un menor peso fresco y menos hiperhidratación. En baja iluminación la actividad peroxidasa fue menor, aunque en los embriones en estado 1 en ambas condiciones de iluminación mostraron la más alta actividad. Los embriones en estado 1 requirieron tres o cuatro meses para desarrollarse hasta el estado 2 bajo condiciones de alta iluminación; y dos o tres meses en baja iluminación. La actividad invertasa en condiciones de baja iluminación fue minima en el estado 2. Todos los embriones tuvieron altos nivelés de glucosa. El desarrollo de los embriones de estado 2 al siguiente y a los subsecuentes, bajo alta iluminación, requirió tres o cuatro meses, y bajo condiciones de baja iluminación requirió cuatro o cinco meses. Una alta intensidad luminosa parece promover la velocidad de recuperación de plantas.

     

How does light and phosphorus fertilisation affect the growth and ectomycorrhizal community of two contrasting dipterocarp species?

Phosphorus concentrations in many south-east Asian tropical rain forest soils are very low. To determine the growth responses of seedlings of a light-demanding (Shorea leprosula) and a more shade-tolerant (Hopea nervosa) dipterocarp species to increasing P, we carried out a nursery fertilisation experiment. Responses of symbiotic ectomycorrhizal (EcM) fungi to the treatments were also determined. Seedlings were grown under high light (13 mol m⁻² d⁻¹) or moderate light (4 mol m⁻² d⁻¹) in shade-chambers and were fertilised with a solution containing 0, 1, 10 or 100 mg L⁻¹ P. The growth of Hopea and Shorea showed different responses to the light and P fertilisation treatments with Hopea having greater growth under moderate light conditions and Shorea having greater growth under high light conditions. Shorea responded to P fertilisation by increasing its foliar P concentrations and growth rates, whereas Hopea did not take up additional P and did not improve its growth rates. There was no effect of either light or P fertilisation on total EcM colonisation or EcM diversity, but around half of the EcM morphotypes observed were affected by one of these two abiotic perturbations, most notably for Riessiella sp. which increased with P fertilisation suggesting it may not be a mutualistic fungus. These results show how niche partitioning in both dipterocarp seedlings and EcM fungi can be divided along contrasting axes.     

Repetitive Somatic Embryogenesis of Ocotea catharinensis Mez. (Lauraceae): Effect of Somatic Embryo Developmental Stage and Dehydration

Repetitive embryogenesis of Ocotea catharinensis from globular/early cotyledonary somatic embryos was successfully supported by WPM supplemented with 22.7 g l⁻¹ sorbitol, 20 g l⁻¹ sucrose, 400 mg l⁻¹ glutamine and 2 g l⁻¹ Phytagel. The best medium to induce repetitive embryogenesis in cotyledonary somatic embryos was half strength WPM supplemented with 20 g l⁻¹ sucrose, 400 mg l⁻¹ glutamine, 1.5 g l⁻¹ activated charcoal and 2 g l⁻¹ Phytagel. The mature somatic embryos gradually air dehydrated showed repetitive embryogenesis after subculture on half strength B5 medium supplemented with 20 g l⁻ sucrose, 20 g l⁻¹ Phytagel, 1.5 g l⁻¹ activated charcoal, 115.6 μM gibberellic acid and 214.8 μM naphthaleneacetic acid. The early cotyledonary, cotyledonary and mature somatic embryos tolerated respectively 95, 86 and 54% fresh weight losses without losing their repetitive embryogenesis potential. Cotyledonary and mature somatic embryos gradually air dehydrated in sealed Petri dishes showed 40–41% repetitive embryogenesis respectively after 20 days and 12 weeks desiccation storage. Repetitive embryogenesis in cotyledonary somatic embryos was significantly stimulated by chemical dehydration with 0.5 M sorbitol and 56% repetitive embryogenesis was achieved even after exposure to 2 M sorbitol for 24 h. The cotyledonary somatic embryos when alginate-encapsulated showed 47% repetitive embryogenesis even after chemical dehydration in 1.5 M sorbitol for 4 days followed by 1 h air dehydration, but failed to survive to the same dehydration conditions without encapsulation. The optimized repetitive embryogenesis and desiccation protocols offer the possibility to use in vitro techniques for continuous reliable somatic embryo production and short term germplasm storage.     

Exogenous retinoic acid decreases in vivo and in vitro proliferative activity during the early migratory stage of neural crest cells

We have previously demonstrated that directional migration of neural crest cells (NCC) is associated with a high cell density, resulting from an active cell proliferation. It is also known that treatment with retinoic acid (RA) causes a dose-dependent inhibition of proliferation of some cell types, and that administration of RA during the early stages of embryonic development, induces cranio-facial abnormal patterns corresponding to NCC derivatives. In view of these findings, it was of interest to determine if exogenous RA is a potential modulator of the mitotic rate of NCC, and to explore the hypothesis of an inhibitory effect exerted by RA on the proliferative behaviour of NCC in vivo and in vitro. Homogenates of RA-treated chick embryos showed a low [³H]dT incorporation, indicating a generalized diminution of DNA synthesis. The labelling index (LI=number of labelled cells/total number of cells) revealed that NCC from RA-treated and control embryos had higher values of [³H]dT incorporation than neural tube cells (P < 0.0001). Autoradiographs of RA-treated chick embryos showed a significantly lower [³H]dT incorporation in NCC at the prosencephalic and mesencephalic levels, as well as in the neural tube cells at the prosencephalic, mesencephalic and rhombencephalic levels, than in control chick embryos (P < 0.0001). NCC cultures treated with 1 or 10 μm RA had a significantly lower LI than in cultures treated with 0.1 μm RA or control cultures (P < 0.04). In chick embryos, the mitotic index of NCC was 0.026 for RA-treated and 0.033 for controls, while the duration of the cell cycle was significantly longer in the NCC of RA-treated embryos (～ 40 h) than in controls (～ 25 h). The length of the cell cycle phases of NCC was similar in both experimental conditions, except for G₁ phase, which was significantly longer in the RA-treated group than in controls. These results show that RA blocks DNA synthesis and lengthens the proliferative behaviour of NCC both in early chick embryos and in vitro, effects that could modify the morphogenetic patterns of NCC distribution through a decreased cell population.     

EFFECTS OF UV RADIATION ON GROWTH AND PHLOROTANNINS IN FUCUS GARDNERI (PHAEOPHYCEAE) JUVENILES AND EMBRYOS1

Diminishing levels of atmospheric ozone are increasing UV stress on intertidal algae. Early developmental stages tend to be more susceptible to environmental stresses; however, little research has examined how these stages are protected from UV radiation (UVR). Many brown algae contain high levels of phlorotannins, which are thought to function in screening UVR. In this study, we tested the effects of ambient levels of UV‐B and UV‐A on growth and phlorotannin production in 1‐ to 2‐cm juvenile and microscopic postsettlement embryos of the intertidal alga Fucus gardneri Silva. Algae were grown in four light treatments: 1) ambient light; 2) under cellulose acetate, which lowered light quantity but did not affect light quality; 3) under Mylar^TM, which filtered UV‐B; and 4) under Plexiglas^TM, which blocked UV‐A and UV‐B. Over a 3‐week period, UV‐B inhibited and UV‐A enhanced the growth of F. gardneri embryos, whereas the growth of juveniles was not affected. Phlorotannin concentrations of both embryos and juveniles did not differ in any of the light treatments. Our results suggest that embryos of F. gardneri are susceptible to UV light but develop a tolerance to it as they mature. This tolerance may result from increases in phlorotannin concentrations that occur during maturation; however, phlorotannin production in embryonic or juvenile stages is either not induced by UV light or takes more than 3 weeks to occur.     

Lithium induces dorsal-type migration of mesodermal cells in the entire marginal zone of urodele amphibian embryos

Summary The effect of lithium (Li⁺) on gastrulation movements was investigated during the development of the urodele amphibianPleurodeles waltl. Attention was focused on mesodermal cell migration. Under conditions of Li⁺ treatment providing a maximal enhancement of dorsoanterior structures, it was found that the dorsoventral polarity of gastrulation was abolished. In particular, vital staining and scanning electron microscopy observations on embryo fractures showed that mesodermal cells migrated radially after Li⁺ treatment, which led to the formation of rounded embryos. Epiboly movements thus were accelerated. Nevertheless, contrasting with the precocious disappearance of the early-formed yolk plug, archenteron invagination was constantly retarded and commenced with a delay of several hours as compared to control gastrulae. Cell-lineage analysis of the progenies from ventral or dorsal equatorial blastomeres of 32-cell-stage embryos provided evidence that both dorsal and ventral mesoderm contributed to notochordal tissue after Li⁺ treatment. Dorsalization of the entire marginal zone was confirmed by the ability of the entire mesoderm rudiment to behave as a dorsal organiser after Li⁺ treatment. Comparison of the migratory behaviour of isolated animal hemispheres from Li⁺-treated or control embryos cultured on fibronectin-coated substrate indicated that all marginal cells acquired the autonomous capacity for migration of dorsal marginal cells under the action of lithium.     

Accumulation pattern and identification of seed storage proteins in zygotic embryos of Pinus strobus and in somatic embryos from different maturation treatments

Somatic embryogenesis (SE) of Pinus strobus L. has been greatly improved over the last few years with respect to both the initiation frequencies from a number of seed families and production of mature somatic embryos that readily convert to plants. However, there are no data on biochemical characterization of somatic embryos in relation to zygotic embryos of eastern white pine and on the optimal duration of the maturation stage. It is believed that somatic embryos closely resembling zygotic embryos not only morphologically but biochemically would display more vigorous growth. Hence, in this study the accumulation pattern of the most abundant seed storage proteins in zygotic and somatic embryos were characterized by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and identified by amino acid sequencing and tandem mass spectrometry (MS/MS). This showed that somatic embryos accumulated storage proteins in a similar manner to zygotic embryos and that the most abundant were the buffer‐insoluble 11S‐ globulins MW 59.6 kDa, which dissociated under reduced conditions to 38.2–40.0 and 22.5–23.5 kDa range polypeptides, and buffer‐soluble 7S vicilin‐like proteins MW 46.0–49.0 kDa, which did not separate under reduced conditions. Other relatively abundant soluble proteins were in the ranges of 25–27 and 27–29 kDa. The only group of proteins that showed different migration profiles in the presence of β‐mercaptoethanol (ME) were the low molecular mass proteins of 14.6–16.5 kDa. Somatic embryos that matured for 9 weeks on medium with 6% sucrose accumulated more storage proteins than those matured on medium with 3% sucrose and the extension of the maturation period to 12 weeks resulted in significant reduction of the storage proteins on both media. As expected, somatic embryos matured on medium with 6% sucrose had lower water potential (Ψ) than those from medium with 3% sucrose. Nonetheless, the somatic embryos matured under the best of tested conditions (6% sucrose for 9 weeks) had slightly higher water content; 1.35 ± 0.28 g H₂O g^?1 DM (mean ± sd ) than the mature non‐dried zygotic embryos; (1.16 ± 0.09 g H₂O g^?1 DM), and accumulated less storage proteins, whose amounts were either similar to (7S‐vicilins) or below (11S‐globulins) those found in the immature zygotic embryos collected 2 weeks prior to the usual cone collection. The implications of these results for further research and development of viable artificial seed is discussed.     

Ontogenetic Behavior and Migration of Volga River Russian sturgeon, Acipenser gueldenstaedtii, with a Note on Adaptive Significance of Body Color

We conducted laboratory experiments with Volga River Russian sturgeon, Acipenser gueldenstaedtii, to develop a conceptual model of early behavior. We daily observed fish from day-0 (embryos, first life interval after hatching) to day-29 feeding larvae for preference of bright habitat and cover, swimming distance above the bottom, up- and downstream movement, and diel activity. Hatchling embryos initiated a downstream migration, which suggests that predation risk of embryos at spawning sites is high. Migration peaked on days 0–5 and ceased on day 7 (8-day migration). Migrants preferred bright, open habitat and early migrants swam-up far above the bottom (maximum daily median, 140cm) in a vertical swim tube. Post-migrant embryos did not prefer bright illumination but continued to prefer white substrate, increased use of cover habitat, and swam on the bottom. Larvae initiated feeding on day 10 after 170.6 cumulative temperature degree-days. Larvae did not migrate, weakly preferred bright illumination, preferred white substrate and open habitat, and swam near the bottom (daily median 5–78cm). The lack of a strong preference by larvae for bright illumination suggests foraging relies more on olfaction than vision for locating prey. A short migration by embryos would disperse wild sturgeon from a spawning area, but larvae did not migrate, so a second later migration by juveniles disperses young sturgeon to the sea (2-step migration). Embryo and larva body color was light tan and tail color was black. The migration, behavior, and light body color of Russian sturgeon embryos was similar to species of Acipenser and Scaphirhynchus in North America and to Acipenser in Asia that migrate after hatching as embryos. The similarity in migration style and body color among species with diverse phylogenies likely reflects convergence for common adaptations across biogeographic regions.     

Direct somatic embryogenesis fromBegonia gracilis explants

Direct somatic embryogenesis ofBegonia gracilis was achieved from microcultured laminar segments and petioles on Murashige and Skoog medium with 0.5 mg 1^–1 kinetin and 2% coconut water. Somatic embryos were obtained with greater frequency from petiole explants than from leaf blade sections. Under red light (45 mol m^–2 s^–1), approximately 80% of the petiole explants successfully produced somatic embryos but only 30% of the leaf blade sections responded. However, somatic embryos were significantly more abundant on responding lamina explants (60–70 embryos/leaf section) than on petioles (40–50 embryos/petiole). These trends were similar for explants kept in the dark, but overall production was lower. Somatic embryos were produced more quickly (5 weeks) from petioles than from lamina explants (8 weeks). The somatic embryos germinated to produce plantlets and subsequently shoot cultures with the same appearance as the parental clone.Abbreviations BA benzyladenine - MS Murashige and Skoog (1962) - NAA naphthalene acetic acid - SE somatic embryo     

Migratory Responses of Benthic Diatoms to Light and Temperature Monitored by Chlorophyll Fluorescence

As an important adaptation for survival in the sediments of intertidal flats, benthic diatoms move up and down in response to a wide range of environmental stimuli. We investigated the vertical migration of two diatoms—Cylindrotheca closterium (Agradh) Kützing (B-25) and Nitzschia sp. (B-3)—under different combinations of light intensity and temperature conditions. An imaging pulse amplitude modulated (PAM) fluorometer was used to measure the minimum fluorescence (F ₀) in order to monitor variations in diatom biomass in surface sediments. Rapid light curves (RLCs) were applied to assess their photosynthetic activities. Both species had increased motility under higher temperatures, with the longer valved C. closterium being twice as fast as the shorter valved Nitzschia sp. The former was also influenced by exposure to light intensities of 100 or 250 μmol m⁻² s⁻¹, whereas the latter was not. Consequently, no light/temperature interaction effect was associated with the vertical migration of Nitzschia sp., perhaps because of its lower photosynthetic capacity and smaller cell size. Therefore, we conclude that motile benthic diatoms exhibit species-specific responses to light and temperature due to differences in their photosynthetic capability and morphological characteristics.     

Callus in Rosmarinus officinalis L. (Lamiaceae): A morphoanatomical,histochemical and volatile analysis

This study aims to evaluate the anatomy of calli and their chemico-morphological characteristics after in vitro culture. Callus was induced from leaf explants of Rosmarinus officinalis derived from in vitro grown plants on media containing 2,4-dichlorophenoxyacetic acid (2,4D) and thidiazuron (TDZ) supplementation. Both capitate and peltate glandular trichomes were recognized in R. officinalis callus by light and electron microscopy. Histochemical tests showed positive reactions to lipophilic and terpenoid compounds for capitate trichomes and parenchyma or meristematic cells. Volatiles were isolated by simultaneous distillation–extraction and analysed by gas chromatography. Specifically, the use of 2,4D was shown to have a positive effect on the capacity of R. officinalis friable callus to produce volatile monoterpene compounds relative to TDZ. The main volatiles produced by callus were α-pinene, β-pinene and camphor produced in media containing 0.5 and 1.0 mg L^? 1 of 2,4D. Significant quantities of α-pinene, β-pinene and camphor were produced with 0.5 mg L^? 1 of 2,4D. On the basis of this accumulated evidence, it can be concluded that R. officinalis callus produces volatiles in developing glandular trichomes and that cellular aggregates and in vitro culture can be used for the production of volatiles under in vitro controlled conditions from selected plant material.     

Multiple mechanisms mediate motor neuron migration in the zebrafish hindbrain

The transmembrane protein Van gogh‐like 2 (Vangl2) is a component of the noncanonical Wnt/Planar Cell Polarity (PCP) signaling pathway, and is required for tangential migration of facial branchiomotor neurons (FBMNs) from rhombomere 4 (r4) to r5‐r7 in the vertebrate hindbrain. Since vangl2 is expressed throughout the zebrafish hindbrain, it might also regulate motor neuron migration in other rhombomeres. We tested this hypothesis by examining whether migration of motor neurons out of r2 following ectopic hoxb1b expression was affected in vangl2^? (trilobite) mutants. Hoxb1b specifies r4 identity, and when ectopically expressed transforms r2 to an “r4‐like” compartment. Using time‐lapse imaging, we show that GFP‐expressing motor neurons in the r2/r3 region of a hoxb1b‐overexpressing wild‐type embryo migrate along the anterior‐posterior (AP) axis. Furthermore, these cells express prickle1b (pk1b), a Wnt/PCP gene that is specifically expressed in FBMNs and is essential for their migration. Importantly, GFP‐expressing motor neurons in the r2/r3 region of hoxb1b‐overexpressing trilobite mutants and pk1b morphants often migrate, even though FBMNs in r4 of the same embryos fail to migrate longitudinally (tangentially) into r6 and r7. These observations suggest that tangentially migrating motor neurons in the anterior hindbrain (r1‐r3) can use mechanisms that are independent of vangl2 and pk1b functions. Interestingly, analysis of tri; val double mutants also suggests a role for vangl2‐independent factors in neuronal migration, since the valentino mutation partially suppresses the trilobite mutant migration defect. Together, the hoxb1b and val experiments suggest that multiple mechanisms regulate motor neuron migration along the AP axis of the zebrafish hindbrain. © 2009 Wiley Periodicals, Inc. Develop Neurobiol, 2010     

The cell adhesion-associated protein Git2 regulates morphogenetic movements during zebrafish embryonic development

Signaling through cell adhesion complexes plays a critical role in coordinating cytoskeletal remodeling necessary for efficient cell migration. During embryonic development, normal morphogenesis depends on a series of concerted cell movements; but the roles of cell adhesion signaling during these movements are poorly understood. The transparent zebrafish embryo provides an excellent system to study cell migration during development. Here, we have identified zebrafish git2a and git2b, two new members of the GIT family of genes that encode ArfGAP proteins associated with cell adhesions. Loss-of-function studies revealed an essential role for Git2a in zebrafish cell movements during gastrulation. Time-lapse microscopy analysis demonstrated that antisense depletion of Git2a greatly reduced or arrested cell migration towards the vegetal pole of the embryo. These defects were rescued by expression of chicken GIT2, indicating a specific and conserved role for Git2 in controlling embryonic cell movements. Git2a knockdown embryos showed defects in cell morphology that were associated with reduced cell contractility. We show that Git2a is required for phosphorylation of myosin light chain (MLC), which regulates myosin II-mediated cell contractility. Consistent with this, embryos treated with Blebbistatin-a small molecule inhibitor for myosin II activity-exhibited cell movement defects similar to git2a knockdown embryos. These observations provide in vivo evidence of a physiologic role for Git2a in regulating cell morphogenesis and directed cell migration via myosin II activation during zebrafish embryonic development.