Efficient and stable regeneration from protoplasts of <Emphasis Type="Italic">Cyclamen coum</Emphasis> Miller via somatic embryogenesis

Embryogenic cultures of Cyclamen coum were established on solid media and in suspension, and their growth characteristics in response to different concentrations of plant growth regulators (PGRs) were evaluated. Embryogenic cultures exhibited a high regeneration capacity of 876 somatic embryos per gram fresh mass. Up to 4.24 × 10⁵ protoplasts per gram of fresh mass were isolated from somatic embryos and embryogenic suspension cultures. Protoplasts derived from both embryos and suspension cultures were successfully cultured in vitro and regenerated into plants via somatic embryogenesis. Phenotypic analyses and flow cytometric measurements revealed that some regenerated plants were tetraploid. About 20% of the protoplast-derived calluses used for regeneration were tetraploid, while tetraploidy was found in 0.9% of the plants regenerated from the embryogenic cultures.     

Multidimensional plasticity in the Glanville fritillary butterfly: larval performance is temperature,host and family specific

Variation in environmental conditions during development can lead to changes in life-history traits with long-lasting effects. Here, we study how variation in temperature and host plant (i.e. the consequences of potential maternal oviposition choices) affects a suite of life-history traits in pre-diapause larvae of the Glanville fritillary butterfly. We focus on offspring survival, larval growth rates and relative fat reserves, and pay specific attention to intraspecific variation in the responses (G × E × E). Globally, thermal performance and survival curves varied between diets of two host plants, suggesting that host modifies the temperature impact, or vice versa. Additionally, we show that the relative fat content has a host-dependent, discontinuous response to developmental temperature. This implies that a potential switch in resource allocation, from more investment in growth at lower temperatures to storage at higher temperatures, is dependent on the larval diet. Interestingly, a large proportion of the variance in larval performance is explained by differences among families, or interactions with this variable. Finally, we demonstrate that these family-specific responses to the host plant remain largely consistent across thermal environments. Together, the results of our study underscore the importance of paying attention to intraspecific trait variation in the field of evolutionary ecology.     

Avoidance of precipitation and carbohydrate breakdown in autoclaved plant tissue culture media

The extent of breakdown of fructose and glucose derived from sucrose in the medium of Murashige and Skoog (1962) during autoclaving was investigated by polarographic measurement. Although not present in the original MS medium but often used in place of FeSO₄ + Na₂-EDTA, FeNa-EDTA was found to be primarily responsible for catalyzing the breakdown of these monosaccharides. It would therefore be good practice to autoclave FeNa-EDTA separate from the carbohydrate constituents of the medium in order to reduce the formation of toxic substances derived from the latter's breakdown. Autoclaving FeNa-EDTA separately has the additional advantage of preventing precipitation of certain micronutrient elements. Further precipitation can be avoided by autoclaving FeNa-EDTA and KH₂PO₄ together, but separately, from other components of the medium. By eliminating precipitation and minimizing the breakdown of monosaccharides during autoclaving, it is possible to improve the quality of the medium without resorting to sterilization by filtering.Abbreviations HMF 5-(hydroxymethyl)-2-furaldehyde - FeNa-EDTA ferric monosodium ethylenediaminetetraacetic acid - MS Murashige and Skoog - NAA naphthylacetic acid     

The role of host promiscuity in the invasion process of a seaweed holobiont

Invasive species are co-introduced with microbiota from their native range and also interact with microbiota found in the novel environment to which they are introduced. Host flexibility toward microbiota, or host promiscuity, is an important trait underlying terrestrial plant invasions. To test whether host promiscuity may be important in macroalgal invasions, we experimentally simulated an invasion in a common garden setting, using the widespread invasive macroalga Agarophyton vermiculophyllum as a model invasive seaweed holobiont. After disturbing the microbiota of individuals from native and non-native populations with antibiotics, we monitored the microbial succession trajectories in the presence of a new source of microbes. Microbial communities were strongly impacted by the treatment and changed compositionally and in terms of diversity but recovered functionally by the end of the experiment in most respects. Beta-diversity in disturbed holobionts strongly decreased, indicating that different populations configure more similar –or more common– microbial communities when exposed to the same conditions. This decline in beta-diversity occurred not only more rapidly, but was also more pronounced in non-native populations, while individuals from native populations retained communities more similar to those observed in the field. This study demonstrates that microbial communities of non-native A. vermiculophyllum are more flexibly adjusted to the environment and suggests that an intraspecific increase in host promiscuity has promoted the invasion process of A. vermiculophyllum. This phenomenon may be important among invasive macroalgal holobionts in general.Subject terms: Symbiosis, Molecular ecology, Microbial ecology     

Characterization of a reverse gyrase from the extremely thermophilic hydrogen-oxidizing eubacterium Calderobacterium hydrogenophilum

Abstract Non-acid and acid glycolipids were isolated from the small intestine of a newborn calf and tested for the ability to bind Escherichia coli carrying K99 fimbriae. The bacteria did not bind to any of the non-acid glycolipids, whereas in the acid glycolipid fraction several gangliosides were detected which bind to K99 fimbriae. Gangliosides capable of binding K99 fimbriated E. coli were characterized as NeuGc-GM3, NeuGc-GM2, NeuGc-GD1a NeuAc-SPG and NeuAc-SPG. No binding was detected to NeuAc-GM3 and NeuGc-GM1.     

Host-Virus Interaction in Ribonucleic Acid Bacteriophage-Infected Escherichia coli: I. Location of “Late” MS2-Specific Ribonucleic Acid Synthesis

When actinomycin-treated, MS2-infected Escherichia coli are labeled during a brief period later than 16 min after infection, the newly synthesized MS2 ribonucleic acid (RNA) appears first in the 30,000 x g sediment, probably bound to fragments of bacterial membranes, since the radioactivity can be released from the sediment with deoxycholate or urea. With longer labeling times, radioactivity also appears in the 30,000 x g supernatant fluid. While on the membrane, the RNA is organized into particles with sedimentation coefficients of 40, 32, and 27S in the presence of low Mg(2+). In the presence of high Mg(+), MS2-specific RNA is found in polyribosomes. These data are interpreted to mean that MS2-specific RNA is synthesized and organized into larger structures on membrane. More than 8 min of labeling is required before radioactivity is found in the 81S virion which appears in the supernatant fluid.     

Comparison of methods for the analysis of therapeutic immunoglobulin G Fc-glycosylation profiles—Part 1: Separation-based methods

Immunoglobulin G (IgG) crystallizable fragment (Fc) glycosylation is crucial for antibody effector functions, such as antibody-dependent cell-mediated cytotoxicity, and for their pharmacokinetic and pharmacodynamics behavior. To monitor the Fc-glycosylation in bioprocess development, as well as product characterization and release analytics, reliable techniques for glycosylation analysis are needed. A wide range of analytical methods has found its way into these applications. In this study, a comprehensive comparison was performed of separation-based methods for Fc-glycosylation profiling of an IgG biopharmaceutical. A therapeutic antibody reference material was analyzed 6-fold on 2 different days, and the methods were compared for precision, accuracy, throughput and other features; special emphasis was placed on the detection of sialic acid-containing glycans. Seven, non-mass spectrometric methods were compared; the methods utilized liquid chromatography-based separation of fluorescent-labeled glycans, capillary electrophoresis-based separation of fluorescent-labeled glycans, or high-performance anion exchange chromatography with pulsed amperometric detection. Hydrophilic interaction liquid chromatography-ultra high performance liquid chromatography of 2-aminobenzamide (2-AB)-labeled glycans was used as a reference method. All of the methods showed excellent precision and accuracy; some differences were observed, particularly with regard to the detection and quantitation of minor glycan species, such as sialylated glycans.