Metschnikowia andauensis as a new biocontrol agent of fruit postharvest diseases

Potential antagonists were isolated from the epiphytic flora associated with oranges and pome fruit. A total of 1465 microorganisms were tested in a preliminary screening against blue and green moulds on pome and citrus fruit, respectively. Among them, approximately 3% reduced incidence and severity by more than 50% and 4 microorganisms fulfilled the selection criteria of reduction in severity and incidence by 75%. The most effective was a yeast identified as Metschnikowia andauensis, strain NCYC 3728 (PBC-2), isolated from the surface of ‘Bravo de Esmolfe’ apple fruit cultivated in North Portugal. The biocontrol activity of M. andauensis PBC-2 was dependent on its applied concentration. At 5 × 10⁶ cfu/mL incidence (% of infected wounds) and severity (lesion diameter) were reduced by 62 and 70%, respectively and at 1 × 10⁷ cfu/mL, the greatest reduction was achieved, 90% of incidence and 95% of severity. The broad spectrum of action of M. andauensis PBC-2 was evaluated with effective control being achieved against Rhizopus stolonifer, Penicillium expansum and Botritys cinerea, on ‘Rocha’ pears and on different apple cultivars and against Penicillium digitatum and Penicillium italicum on mandarins and oranges. In semi-commercial trials in cold storage, the reduction of blue mould was 90%. Rapid colonization of fresh apple fruit wounds was observed during the first 24 h of cold storage, followed by a significant population increase during the first 15 days of storage and then the population remained stable until the end of storage.     

Enantioselective organocatalysis using SOMO activation

The asymmetric alpha-addition of relatively nonpolar hydrocarbon substrates, such as allyl and aryl groups, to aldehydes and ketones remains a largely unsolved problem in organic synthesis, despite the wide potential utility of direct routes to such products. We reasoned that well-established chiral amine catalysis, which activates aldehydes toward electrophile addition by enamine formation, could be expanded to this important reaction class by applying a single-electron oxidant to create a transient radical species from the enamine. We demonstrated the concept of singly occupied molecular orbital (SOMO) activation with a highly selective alpha-allylation of aldehydes, and we here present preliminary results for enantioselective heteroarylations and cyclization/halogenation cascades.     

Control of crystal size and lattice formation by starmaker in otolith biomineralization

The stone-like otoliths from the ears of teleost fishes are involved in balance and hearing and consist of calcium carbonate crystallites embedded in a protein framework. We report that a previously unknown gene, starmaker, is required in zebrafish for otolith morphogenesis. Reduction of starmaker activity by injection of modified antisense oligonucleotides causes a change in the crystal lattice structure and thus a change in otolith morphology. The expression pattern of starmaker, along with the presence of the protein on the growing otolith, suggest that the expression levels of starmaker control the shape of the otoliths.     

Beef authentication and retrospective dietary verification using stable isotope ratio analysis of bovine muscle and tail hair

Stable isotope ratio analysis (SIRA) was used as an analytical tool to verify the preslaughter diet of beef cattle. Muscle and tail hair samples were collected from animals fed either pasture (P), a barley-based concentrate (C), silage followed by pasture (SiP), or silage followed by pasture with concentrate (SiPC) for 1 year (n = 25 animals per treatment). The (13)C/(12)C, (15)N/(14)N, (2)H/(1)H, and (34)S/(32)S isotope ratios in muscle clearly reflected those of the diets consumed by the animals. By applying a stepwise canonical discriminant analysis, a good discrimination of bovine meat according to dietary regimen was obtained. On the basis of the classification success rate, the (13)C/(12)C and (34)S/(32)S ratios in muscle were the best indicators for authentication of beef from animals consuming the different diets. Analysis of (13)C/(12)C and (15)N/(14)N in tail hair sections provided an archival record of changes to the diet of the cattle for periods of over 1 year preslaughter.     

Identification of the molecular make-up of the Potato virus Y strain PVY(Z): genetic typing of PVY(Z)-NTN

Potato virus Y (PVY) strains were originally defined by interactions with different resistance genes in standard potato cultivars. Five distinct strain groups are defined that cause local or systemic hypersensitive responses (HRs) in genetic background with a corresponding N gene: PVY(O), PVY(N), PVY(C), PVY(Z), and PVY(E). The nucleotide sequences of multiple isolates of PVY(O) and PVY(N) differ from each other by ≈8% along their genomes. Additionally, complete genome sequences of multiple recombinant isolates are composed of segments of parental PVY(O) and PVY(N) sequences. Here, we report that recombinant isolate PVY-L26 induces an HR in potato 'Maris Bard' carrying the putative Nz gene, and is not recognized by two other resistance genes, Nc and Ny(tbr). These genetic responses in potato, combined with the inability of PVY-L26 to induce vein necrosis in tobacco, clearly define it as an isolate from the PVY(Z) strain group and provide the first information on genome structure and sequence of PVY(Z). The genome of PVY-L26 displays typical features of European NTN-type isolates with three recombinant junctions (PVY(EU-NTN)), and the PVY-L26 is named PVY(Z)-NTN. Three typical PVY(NTN) isolates and two PVY(N) isolates, all inducing vein necrosis in tobacco, were compared with PVY-L26. One PVY(NTN) isolate elicited HR reactions in Maris Bard, similar to PVY-L26, while two induced a severe systemic HR-like reaction quite different from the quasi-symptomless reaction induced by two PVY(N) isolates. 'Yukon Gold' potato from North America produced HR against several PVY(NTN) isolates, including PVY-L26, but only late and limited systemic necrosis against one PVY(N) isolate. Consequently, according to symptoms in potato indicators, both PVY(Z) and PVY(NTN) isolates appeared biologically very close and clearly distinct from PVY(O) and PVY(N) strain groups.     

ROS and NO production in compatible and incompatible tomato-<Emphasis Type="Italic">Meloidogyne incognita</Emphasis> interactions

Nitric oxide (NO) has been shown to be an essential regulatory molecule in plant response to pathogen infection in synergy with reactive oxygen species (ROS). At the present, nothing is known about the role of NO in disease resistance to nematode infection. We used a resistant tomato cultivar with different sensitivity to avirulent and virulent populations of the root-knot nematode Meloidogyne incognita to investigate the key components involved in oxidative and nitrosative metabolism. We analyzed the superoxide radical production, hydrogen peroxide content, and nitric oxide synthase (NOS)-like and nitrate reductase activities, as potential sources of NO. A rapid NO accumulation and ROS production were found at 12 h after infection in compatible and incompatible tomato-nematode interactions, whereas the amount of NO and ROS gave different results 24 and 48 h after infection amongst compatible and incompatible interactions. NOS-like arginine-dependent enzyme rather than nitrate reductase was the main source of NO production, and NOS-like activity increased substantially in the incompatible interaction. We can envisage a functional overlap of both NO and ROS in tomato defence response to nematode invasion, NO and H₂O₂ cooperating in triggering hypersensitive cell death. Therefore, NO and ROS are key molecules which may help to orchestrate events following nematode challenge, and which may influence the host cellular metabolism.     

Vertical advection of extracellular DNA by water capillarity in soil columns

The fate of extracellular DNA in the environment concerns both the fate of transgenes from genetically modified organisms and the evolution of active bacteria capable of incorporating this DNA into their genomes. This study addressed the possibility that DNA, like other organic molecules, could move vertically in the capillary fringe of groundwater aquifers. The targeted gene fragment used here was the 35S-nptII sequence, which was below detection levels in controls. Initial microcosm studies detected the DNA target molecule by PCR during the entire experiment. The vertical advection of water and DNA were monitored for a period of 3 days in soil columns. DNA was added as a water solution at the bottom of the unsaturated soil column, and then DNA-free water was added at the bottom after 12 and 24 h. After the addition of the DNA solution, capillary water rose 4 cm within the soil column and the target DNA was detected up to that height. After 60 min, the entire soil column (10 cm) was wetted and the target sequence was detected up to a height of 7.5 cm. After the second wetting (12 h later), the target sequence was detected up to the top of the soil column (10 cm). However, after the third wetting (24 h later), the marker sequence was only found at heights from 0.5 to 4 cm. Results clearly show the vertical movement of DNA due the capillary rise and suggest the possibility of DNA degradation within the soil column.     

Heavy Metals in Water Percolating Through Soil Fertilized with Biodegradable Waste Materials

The influence of manure and composts on the leaching of heavy metals from soil was evaluated in a model lysimeter experiment under controlled conditions. Soil samples were collected from experimental fields, from 0- to 90-cm layers retaining the layout of the soil profile layers, after the second crop rotation cycle with the following plant species: potatoes, spring barley, winter rapeseed, and winter wheat. During the field experiment, 20 t DM/ha of manure, municipal sewage sludge composted with straw (SSCS), composted sewage sludge (SSC), dried granular sewage sludge (DGSS), “Dano” compost made from non-segregated municipal waste (CMMW), and compost made from municipal green waste (CUGW) was applied, i.e., 10 t DM/ha per crop rotation cycle. The concentrations (μg/dm³) of heavy metals in the leachate were as follows: Cd (3.6–11.5)?<?Mn (4.8–15.4)?<?Cu (13.4–35.5)?<?Zn (27.5–48.0)?<?Cr (36.7–96.5)?<?Ni (24.4–165.8)?<?Pb (113.8–187.7). Soil fertilization with organic waste materials did not contaminate the percolating water with manganese or zinc, whereas the concentrations of the other metals increased to the levels characteristic of unsatisfactory water quality and poor water quality classes. The copper and nickel content of percolating water depended on the concentration of those metals introduced into the soil with organic waste materials. The concentrations of Cd in the leachate increased, whereas the concentrations of Cu and Ni decreased with increasing organic C content of organic fertilizers. The widening of the C/N ratio contributed to Mn leaching. The concentrations of Pb, Cr, and Mn in the percolating water were positively correlated with the organic C content of soil.