Amino acids are general growth inhibitors of Nicotiana silvestris in tissue culture

The growth of Nicotiana silvestris in suspension culture is inhibited by all of the common protein amino acids at the millimolar level, except for L-glutamine. A defined experimental system for growth/inhibition studies has been established, and growth studies were carried out with cells that had been maintained in the exponential growth phase for at least 10 generations (EE cells). The following results were obtained after particularly detailed studies with aromatic amino acids. The onset of inhibition was preceded by a duration of normal growth rate which varied within a range of 12 to 48 h. The degree of inhibition was directly proportional to amino acid concentration and inversely related to the initial cell density of the inoculum. A slowed, but still exponential rate of growth persisted during an early phase of inhibition. Under sufficiently severe conditions, this was followed by progressive diminution of growth rate and eventual lysis. The most drastic inhibitory effects caused by aromatic amino acids were in the order: phenylalanine, tryptophan and tyrosine. When EE cells cultivated under conditions of growth inhibition were diluted into fresh medium, immediate resumption of growth at the uninhibited rate occurred and persisted. On the other hand, when growth-inhibited EE cells were diluted into medium containing the same concentration of amino acid used in the first round of growth, an initial burst of uninhibited growth lasting about 24 h was followed by a drastic, progressively declining growth rate which deteriorated to cell death and lysis. When cells in stationary phase were used as an inoculum, as is done in typical growth characterizations with suspension cultures, the sensitivity to inhibition during the subsequent exponential growth phase was several-fold greater than was the case with EE cells. Hypotheses that growth inhibition might be caused by ammonia toxicity, keto-acid toxicity, or by inhibition of nitrate utilization were ruled out. Observations that provide new insight are: (i)growth-inhibited cells undergo drastic plasmolysis, (ii) L-glutamine is an effective antagonist of amino-acid inhibitors, and (iii) growth-inhibited cells exhibit a transient restoration of normal growth rate upon dilution into fresh growth medium. These results implicate a linkage of amino acids with osmotic regulation and nitrogen metabolism.     

Strains of Colletotrichum coffeanum Noack Causing Coffee Berry Disease in Angola and Malawi with Characteristics Different to the Kenya Strain

Strains of Colletotrichum isolated from berries of Coffea arabica from Angola and Malawi were identified as C. coffeanum causing coffee berry disease (CBD). The isolates were highly aggressive to berries of the cultivar ‘Catimor’ and produced high numbers of acervuli and extremely large masses of pink conidia when cultivated in the dark. These strains are apparently different in aggressiveness and morphology to those described in the past from Kenya.     

A gustatory mutant of Drosophila defective in pyranose receptors

Summary Mutations in an X-linked gene, gust-A, block the responses of Drosophila melanogaster to a group of pyranose sugars. It is shown that the behavioural effects of this mutation are correlated with a loss of electrical responses in taste receptors. The mutation affects the chemoacceptors for pyranose sugars leaving the furanose acceptors intact.     

Studies of the mechanisms for the induction of in vivo tumor immunity. V. A comparison of the generation of the primary cell-mediated cytotoxic response using in vitro mixed-lymphocyte--tumor cell culture and an in vivo technique

In vivo immunization of C57BL/6 mice with FBL-3, a syngeneic Friend-virus-induced leukemia, can induce a specific, T-cell-mediated cytotoxic response, as measured by the ¹²⁵IUdR release assay. In vitro it was difficult to generate an analogous, primary cytotoxic response, using a normal spleen responder population. After modification of the splenic responders by adding normal peritoneal cells, this modified population then had the capacity to mount a primary cytotoxic response in the mixed-lymphocyte-tumor cell culture reaction to FBL-3. We have characterized the effector population as well as the helper (peritoneal) cells which were responsible for elevating the cytolytic response to FBL-3. The results indicate that there are at least two populations of cells which are essential for inducing a primary cell-mediated cytotoxic response. First, the effectors which are directly responsible for mediating the cytotoxic reactions and are derived from radiosensitive T cells and second, a helper cell population which is radioresistant and has the characteristics of macrophages.     

Effects of flask sealing and growth regulators on in vitro propagation of neem (Azadirachta indica A. Juss.)

In vitro propagation of neem (Azadirachta indica A. Juss.) may offer an efficient alternative to seed propagation of this species. For optimization of in vitro propagation, different basal salt formulations, growth regulators, and culture container sealants (polytetrafluoroethylene hydrophobic membranes [PTFE]) were evaluated. Nodal segments cultured on Murashige and Skoog (MS) medium showed the highest shoot formation per explant (1.67). Explants cultured in flasks containing MS medium with 0.5 mg L⁻¹ benzyladenine, 0.5 mg L⁻¹ kinetin, and 0.05 mg L⁻¹ naphthaleneacetic acid, and sealed with two PTFE membranes, produced the highest number of shoots (4.04). In contrast, explants cultured in flasks without membranes showed leaf chlorosis and senescence. For plant recovery, regenerants were acclimatized in a substrate of coconut fiber and eucalyptus bark (1:1) and showed 80% survival. Our results indicated that the use of flasks with vents was beneficial for in vitro propagation of this important plant.     

A novel cellular automata approach: seed input/output of the alien species Leucaena leucocephala in the soil and the effects of climate changes

The potential impact of the soil seed bank of a given species and subsequent population persistence will depend on the robustness of the seeds against depletion. Invasiveness is frequently linked to this behavior and successful control of invasive species depends on reducing the size of their seed banks. We propose a hybrid model that simulates the seed bank dynamics in association with their physical dormancy (PD) based on experimental data on seed bank depletion of a Leucaena leucocephala population during the dispersal phase. The hybrid model uses cellular automata (CA) and Monte Carlo techniques. The sensitivity analysis revealed that the model delivers reproducible and reliable results. Then, we discuss the ability of the hybrid model to predict the effect of seasonal climate change. A 2.0 °C increase in temperature was enough to cause a considerable impact in the seed depletion curve, and after 1 year, ~?75% of the seeds had already disappeared from the seed bank. The proposed hybrid CA model can be used to study soil-stored seed banks with physical dormancy, and could be applied to other Fabaceae species with similar seed traits. We believe that this computational model contributes to a better understanding of seed bank dynamics and will aid in the control of invasive species and for testing different scenarios to reduce or eliminate seed banks that have already been formed.

     

Role of AIP56 disulphide bond and its reduction by cytosolic redox systems for efficient intoxication

Apoptosis‐inducing protein of 56 kDa (AIP56) is a major virulence factor of Photobacterium damselae subsp. piscicida, a gram‐negative pathogen that infects warm water fish species worldwide and causes serious economic losses in aquacultures. AIP56 is a single‐chain AB toxin composed by two domains connected by an unstructured linker peptide flanked by two cysteine residues that form a disulphide bond. The A domain comprises a zinc‐metalloprotease moiety that cleaves the NF‐kB p65, and the B domain is involved in binding and internalisation of the toxin into susceptible cells. Previous experiments suggested that disruption of AIP56 disulphide bond partially compromised toxicity, but conclusive evidences supporting the importance of that bond in intoxication were lacking. Here, we show that although the disulphide bond of AIP56 is dispensable for receptor recognition, endocytosis, and membrane interaction, it needs to be intact for efficient translocation of the toxin into the cytosol. We also show that the host cell thioredoxin reductase‐thioredoxin system is involved in AIP56 intoxication by reducing the disulphide bond of the toxin at the cytosol. The present study contributes to a better understanding of the molecular mechanisms operating during AIP56 intoxication and reveals common features shared with other AB toxins.     

Ecosystem services provided by armadillos

Awareness of the natural ecological processes provided by organisms that benefit human well‐being has significantly progressed towards the goal of making conservation a mainstream value. Identifying different services and the species that provide them is a vital first step for the management and maintenance of these so‐called ecosystem services. Herein, we specifically address the armadillos, which play key functional roles in terrestrial ecosystems, including as ecosystem engineers, predators, and vectors of invertebrates and nutrients, although these roles have often been overlooked. Armadillos can control pests, disperse seeds, and be effective sentinels of potential disease outbreaks or bioindicators of environmental contaminants. They also supply important material (meat, medicines) and non‐material (learning, inspiration) contributions all over the Americas. We identify key gaps in the understanding of ecosystem services provided by armadillos and areas for future research required to clarify their functional role in terrestrial ecosystems and the services they supply. Such information will produce powerful arguments for armadillo conservation.