Germination ofMusa velutina seeds: Comparison ofin vivo andin vitro systems

Summary In the genusMusa, germination is extremely variable and relatively difficult. Even more difficulties are faced when producing hybrids. The seed yield of hybrids in breeding programs is usually low and often, to ensure the viability and survival of seeds, it is necessary to attempt to germinate a large excess of these seeds. In this context,in vitro embryo culture might be an invaluable tool for obtaining desirable hybrid plants in a short time. Seeds ofMusa velutina were sown in seed trays in a peat-based mixture. Thein vivo seed germination reached 78% but only after 9 mo. Because of this delayed and intermittent germination, embryos were excised from seeds and inoculated onto half-strength Murashige and Skoog (1962) medium, with or without supplementation with various concentrations of gibberellic acid. Light and dark conditions were also used to test their effect on embryo germination. After 2 wk, 82% of embryos germinated in the dark on medium containing 0.1 μM gibberellic acid. Addition of gibberellic acid increased the shoot length and root number over the gibberellic acid-free treatment. Similarly, dark conditions gave a significant increase over light conditions for all the parameters except root number where light or dark conditions did not make any difference. Thus, the present study highlights the importance of various components of thein vitro culture ofMusa embryos and the advantage over direct use of greenhouse-sown seeds both in terms of the time taken to germinate and the final percentage.     

Response by macrozoobenthos biomass to water level regulation in some Finnish lake littoral zones

The relationship of the macrozoobenthos biomass in the littoral area to the yearly fluctuation in water level and the characteristics of the area or lake are studied using data collected from sheltered bays in regulated and natural waters. Most of the lakes were clear and oligotrophic. The benthos biomass at all depths in the littoral decreased with increased water level fluctuation, provided that the transparency of the water was uniform.The macrozoobenthos biomass in the 0–3 m depth zone could be predicted fromlog macrozoobenthos biomass (mg ODW) m^-2=4.25-1.33 (log Biomass Index) in which the Biomass Index is calculated as% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaaeOqaiaabM% gacaqGVbGaaeyBaiaabggacaqGZbGaae4CaiaabccacaqGjbGaaeOB% aiaabsgacaqGLbGaaeiEaiaab2dacaqGGaGaaeiiaiaabccacaqGGa% GaaeiiaiaabccacaqGGaGaaeiiaiaabccacaqGGaGaaeiiaiaabcca% caqGGaGaaeiiaiaabccacaqGGaGaaeiiaiaabccacaqGGaGaaeiiai% aabccadaWcaaabaeqabaGaae4DaiaabggacaqG0bGaaeyzaiaabkha% caqGGaGaaeiBaiaabwgacaqG2bGaaeyzaiaabYgacaqGGaGaaeOzai% aabYgacaqG1bGaae4yaiaabshacaqG1bGaaeyyaiaabshacaqGPbGa% ae4Baiaab6gacaqGGaGaaeyAaiaab6gacaqGGaGaaeiDaiaabIgaca% qGLbGaaeiiaiaabchacaqGYbGaaeyzaiaabAhacaqGPbGaae4Baiaa% bwhacaqGZbGaaeiiaiaabMhacaqGLbGaaeyyaiaabkhaaeaacaqGOa% GaaeyBaiaabUdacaqGGaGaae4yaiaabggacaqGSbGaae4yaiaabwha% caqGSbGaaeyyaiaabshacaqGLbGaaeizaiaabccacaqGMbGaaeOCai% aab+gacaqGTbGaaeiiaiaab2gacaqGVbGaaeOBaiaabshacaqGObGa% aeiBaiaabMhacaqGGaGaaeyBaiaabwgacaqGHbGaaeOBaiaabccaca% qG2bGaaeyyaiaabYgacaqG1bGaaeyzaiaabohacaqGPaaaaeaacaqG% tbGaaeyzaiaabogacaqGJbGaaeiAaiaabMgacaqGGaGaaeizaiaabM% gacaqGZbGaae4AaiaabccacaqG2bGaaeyyaiaabYgacaqG1bGaaeyz% aiaabccacaqGPbGaaeOBaiaabccacaqG0bGaaeiAaiaabwgacaqGGa% Gaae4CaiaabggacaqGTbGaaeyzaiaabccacaqGVbGaaeiCaiaabwga% caqGUbGaaeiiaiaabEhacaqGHbGaaeiDaiaabwgacaqGYbGaaeiiai% aabohacaqGLbGaaeyyaiaabohacaqGVbGaaeOBaiaabccacaqGOaGa% aeyBaiaabMcaaaaccaGae8hiaaIaaKiEaiab-bcaGiaaigdacaaIWa% GaaGimaiaac6caaaa!CBD8!\[{\text{Biomass Index = }}\frac{\begin{gathered} {\text{water level fluctuation in the previous year}} \hfill \\ {\text{(m; calculated from monthly mean values)}} \hfill \\ \end{gathered} }{{{\text{Secchi disk value in the same open water season (m)}}}} \user1{x} 100.\]The whole illuminated littoral shifts due to water level fluctuation, which disturbs the zonation of the benthos. Such an increase or decrease in benthic biomass has been observed after one year of disturbance due to water level fluctuation. It need, however, a study based on the carefully planned and collected data, in which it can be taken account by a multivariate statistical analysis also the interactions between the important factors affected the littoral benthos.     

Cytological characterization of isolated sperm cells of perennial ryegrass (Lolium perenne L.)

Summary The cytoplasmic content and the distribution of intramembrane particles (IMPs) of the plasma membrane of isolated sperm cells of perennial ryegrass (Lolium perenne L.) have been characterized using flow cytometry, transmission electron microscopy, confocal scanning laser microscopy and freeze-fracture studies. The isolated haploid sperm cells contain a variety of cell organelles with the exception of microtubules. Proplastids and plastids with starch were observed, although only rarely. Vacuoles containing remnants of organelles and stacked lamellae of endoplasmic reticulum with cytoplasmic inclusions were observed frequently, indicating that autophagy takes place. The number of mitochondria varies from 11 to 26 with an average of 17. Generally, the nucleus has a lobed shape and displays various interphasic stages of chromatin condensation. The analysis of the number of mitochondria and the nuclear state did not show evidence of sperm cell dimorphism. The cytological variability observed, could be explained by differences in developmental stages already present in vivo at the moment of isolation. No correlation between the number of mitochondria and the nuclear cross-sectioned area and/or the condensation state of the chromatin could be found. The density of intramembrane particles of the plasma membrane on the exoplasmic fracture face is more than twice that on the protoplasmic fracture face. That is the opposite of what was found for sporophytic cells of perennial ryegrass. These results are discussed in relation to the potential use of these cells for biotechnology and developmental studies.     

Enhancement of extractable ethylene at light/dark transition in primary leaves of paclobutrazol-treated Phaseolus vulgaris seedlings

Paclobutrazol [(2RS,3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)pentan-3-ol], a triazole growth retardant, increased the 1-aminocyclopropane-1-carboxylic acid (ACC) level and resulted in reduced ethylene production, estimated as ethylene release in a closed system or by vacuum-extraction, in the primary leaves of Phaseolus vulgaris L. cv. Juliska seedlings exposed to light. At the light/dark transition, a definite enhancement of the endogenous ethylene level was observed by vacuum-extraction of primary leaves of treated plants and the ethylene deficiency of retardant-treated leaves ceased. The concentration of ACC after the light/dark transition followed the pattern for ethylene, and the increase in ACC content was paralleled by a decrease in malonyl-ACC.
It is concluded that the internal level of ethylene is not necessarily lower in the primary leaves of paclobutrazol-treated bean plants, but under special environmental conditions in vivo it may reach that of the control.     

The effects of continuous illumination on southern and northern Micrasterias strains

Different strains of Micrasterias (Chlorophyta, Conjugatophyceae); M. rotata (Grev.) Ralfs ex. Ralfs and M. denticulata Breb. ex. Ralfs var. angulosa (Hantzsch) W. & G. S. West from northern and southern Finland were treated with continuous illumination in order to study the cellular effects of the treatment and whether the tolerance to continuous light of the northern Finnish strains is related to the different daylenght conditions in northern and southern areas. During the growing season the Finnish strains normally live in long-day conditions or even in continuous light (between 60 and 70°N), and they also tolerated continuous illumination in the laboratory. Ultrastructural changes were found especially in the chloroplasts, where formation of calcium precipitates of different forms and sizes and also formation of plastoglobuli containing lipids appeared. However, even in 4-week treatments the ultrastructure of cells of these northern strains was not totally disrupted, contrary to what was found in southern M. torreyi , studied earlier. Southern and northern strains tolerated continuous illumination in different ways. They seem to differ from each other physiologically, and the differences are possibly located in their ionic metabolism and regulation. The injuries sustained during continuous illumination of Micrasterias may largely be caused by the accumulation of Ca²⁺ in cytoplasm and organelles, especially in the chloroplasts.     

旋毛虫肌幼虫ES抗原的基因克隆及高效表达

作者对编码旋毛虫肌幼虫ES抗原的部分结构基因进行了克隆、鉴定和表达。用RNA PCR技术直接从旋毛虫肌幼虫总RNA中反转录并扩增出0．7kh的靶DNA,酶切分析后将其克隆到融合表达载体pEx3lC中。SDS—PAGE电泳表明,含重组子的大肠杆菌能够表达出一分子量为37kDa的融合蛋白(P37),后者占菌体总蛋白的22%以上,并以包含体形式存在于菌体中。经对纯化后表达蛋白的ELlSA检测,证明它能被猪旋毛虫病阳性血清和抗旋毛虫单克隆抗体识别。研究结果揭示,重组蛋白P37对于研制旋毛虫病诊断抗原和免疫抗原具有潜在的应用价值。     

Dietary protein requirement for captive juvenile green turtles (Chelonia mydas)

Head-starting programs are extremely important for restoring the population of sea turtles in wild whereas husbandry conditions and feeding regimens of captive turtles are still limited. In the current study, the optimal dietary protein requirement for green turtle (Chelonia mydas) was investigated to support rearing in head-starting programs. Twenty-five-day-old turtles (44.5–46.2 g body weight, n = 45) were randomly distributed into 15 experimental plastic tanks, comprising three treatment replications of 3 turtles each. They were fed fishmeal-based feeds containing different levels of protein (30%, 35%, 40%, 45%, and 50%) for 8 weeks. At the end of feeding trial, growth performance (specific growth rate = 1.86% body weight/day) and feed utilization (protein efficiency ratio = 3.30 g gain/g protein) were highest in turtles fed with 40% protein in feed (p < .05). These nutritional responses were significantly supported by specific activities of fecal digestive enzymes, especially trypsin, chymotrypsin, amylase, and the amylase/trypsin ratio. Also, this dietary level improved the deposition of calcium and phosphorus in carapace, supporting a hard carapace and strong healthy bones. There were no negative effects in general health status of reared turtles, as indicated by hematological parameters. Based on a broken-line analysis between dietary protein levels and specific growth rate, the optimal protein level for green turtles was estimated as 40.6%. Findings from the current study support the use of artificial diets of specific protein levels to rear captive green turtle before release to natural habitats.     

Addressing high excitation conditions in time-resolved X-ray diffraction experiments and issues of biological relevance

One of the most important fundamental questions connecting chemistry to biology is how chemistry scales in complexity up to biological systems where there are innumerable possible pathways and competing processes. With the development of ultrabright electron and x-ray sources, it has been possible to literally light up atomic motions to directly observe the reduction in dimensionality in the barrier crossing region to a few key reaction modes. How do these chemical processes further couple to the surrounding protein or macromolecular assembly to drive biological functions? Optical methods to trigger photoactive biological processes are needed to probe this issue on the relevant timescales. However, the excitation conditions have been in the highly nonlinear regime, which questions the biological relevance of the observed structural dynamics.     

Influences of sea level changes and volcanic eruptions on Holocene vegetation in Tonga

Here, we investigate Mid- to Late-Holocene vegetation changes in low-lying coastal areas in Tonga and how changing sea levels and recurrent volcanic eruptions have influenced vegetation dynamics on four islands of the Tongan archipelago (South Pacific). To investigate past vegetation and environmental change at Ngofe Marsh (‘Uta Vava’u), we examined palynomorphs (pollen and spores), charcoal (fire), and sediment characteristics (volcanic activity) from a 6.7-m-long sediment core. Radiocarbon dating indicated the sediments were deposited over the last 7700 years. We integrated the Ngofe Marsh data with similar previously published data from Avai’o’vuna Swamp on Pangaimotu Island, Lotofoa Swamp on Foa Island, and Finemui Swamp on Ha’afeva Island. Plant taxa were categorized as littoral, mangrove, rainforest, successional/ disturbance, and wetland groups, and linear models were used to examine relationships between vegetation, relative sea level change, and volcanic eruptions (tephra). We found that relative sea level change has impacted vegetation on three of the four islands investigated. Volcanic eruptions were not identified as a driver of vegetation change. Rainforest decline does not appear to be driven by sea level changes or volcanic eruptions. From all sites analyzed, vegetation at Finemui Swamp was most sensitive to changes in relative sea level. While vegetation on low-lying Pacific islands is sensitive to changing sea levels, island characteristics, such as area and elevation, are also likely to be important factors that mediate specific island responses to drivers of change.     

Expression of wild-type GBSS transgenes in the off-spring of partially and fully complementedamylose-free transformants of potato

Theamylose-free (amf) potato mutant can easily be complemented through introduction of the wild-type gene coding for granule-bound starch synthase (GBSS). After iodine staining the starch of theamf mutant is red whereas that of the wild type and the complementedamf mutant is blue. The level of complementation of selected transformants and their sexual off-spring after backcrossing withamf was investigated using sporophytic tuber cells and gametophytic microspore cells. Two diploid and two tetraploid transformants with full complementation demonstrated the expected segregation patterns of 1:1 (one active insert) or 3:1 (two independently segregating active inserts) in the microspores and in the F1 offspring based on staining of tubers. All expected genotypes in the F1 generation were found, based on microspore segregation patterns of the individual F1 plants. Two transformants with partial complementation (mixed phenotypes) were investigated. One of them, B1, was tetraploid and duplex for the GBSS insert, which had originated through mitotic doubling of the transformed diploid cells. In the F1 generation three phenotypic classes were found:amf, fully complemented and partially complemented. The latter two classes exist independently of a simplex or duplex gene status. The second transformant with partial complementation, B10, appeared to have a complex molecular composition. One cluster of five transgenes caused the partial complementation. Fully and partially complemented phenotypic classes were found after crossing B10 with theamf mutant. Indications were found that the ploidy level of the tissue in which the genes were introduced and expressed played an important role. Firstly, partial complementation was found after transformation of the diploid and not of the tetraploidamf genotypes. Secondly, the level of complementation was higher in tissue with lower ploidy levels, as illustrated by the colour of the starch inin vitro tubers (2x–4x cells) versus field-grown tubers (16x–64x).