Near infrared spectra indicate specific mutant endosperm genes and reveal a new mechanism for substituting starch with (1→3,1→4)-β-glucan in barley

Near Infrared Reflectance spectroscopy was tested as a screening method to characterise high lysine mutants from a barley collection by classification through Principal Component Analysis (PCA). Mean spectra of the samples within each cluster identified gene-specific patterns in the 2270–2360 nm region. The characteristic spectral signatures representing the lys5 locus (Risø mutants 13 and 29) were found to be associated with large changes in percentage of starch and (1→3,1→4)-β-glucan. These alleles compensated for a low level of starch (down to 30%) by a high level of (1→3,1→4)-β-glucan (up to 15–20%), thus, maintaining a constant production of polysaccharides at 50–55%, within the range of normal barley.The spectral tool was tested by an independent data set with six mutants with unknown polysaccharide composition. Spectral data from four of these were classified within the high (1→3,1→4)-β-glucan BG lys5 cluster in a PCA. Their high (1→3,1→4)-β-glucan and low starch content was verified. It is concluded that genetic diversity such as from gene regulated polysaccharide and storage protein pathways in the endosperm tissue can be discovered directly from the phenotype by chemometric classification of a spectral library, representing the digitised phenome from a barley gene bank.     

A scenario tree model for the Canadian Notifiable Avian Influenza Surveillance System and its application to estimation of probability of freedom and sample size determination

In 2008, Canada designed and implemented the Canadian Notifiable Avian Influenza Surveillance System (CanNAISS) with six surveillance activities in a phased-in approach. CanNAISS was a surveillance system because it had more than one surveillance activity or component in 2008: passive surveillance; pre-slaughter surveillance; and voluntary enhanced notifiable avian influenza surveillance. Our objectives were to give a short overview of two active surveillance components in CanNAISS; describe the CanNAISS scenario tree model and its application to estimation of probability of populations being free of NAI virus infection and sample size determination. Our data from the pre-slaughter surveillance component included diagnostic test results from 6296 serum samples representing 601 commercial chicken and turkey farms collected from 25 August 2008 to 29 January 2009. In addition, we included data from a sub-population of farms with high biosecurity standards: 36,164 samples from 55 farms sampled repeatedly over the 24 months study period from January 2007 to December 2008. All submissions were negative for Notifiable Avian Influenza (NAI) virus infection. We developed the CanNAISS scenario tree model, so that it will estimate the surveillance component sensitivity and the probability of a population being free of NAI at the 0.01 farm-level and 0.3 within-farm-level prevalences. We propose that a general model, such as the CanNAISS scenario tree model, may have a broader application than more detailed models that require disease specific input parameters, such as relative risk estimates.     

Starved bacteria retain their size but lose culturability - Lessons from a 5000 years old undisturbed A-horizon

The vast majority of soil bacteria are unable to form visible colonies on agar media. One hypothesis is that unculturable soil bacteria are dwarf cells that may either be small starved forms derived from larger species or represent inherently small species. We test the hypotheses that cells of extremely starved soil bacterial communities are smaller and less culturable than cells of bacterial communities from a richer soil, and that culturability is related to cell size by comparing an extremely starved community from a 5200-year-old A-horizon buried under a burial mound with a community from a modern agricultural A-horizon.We serially filtered cell suspensions through filters with successively smaller pore sizes (0.8 μm, 0.6 μm and 0.4 μm) and assessed total cell number and culturability, i.e. the ability to form colonies on two types of agar media, in each size fraction. Cell size distributions were assessed in unfiltered suspensions. Average cell size was only moderately reduced in the starved community, where culturability was low for all size classes. In contrast, culturability was much higher in the modern community, where culturability decreased dramatically with decreasing cell sizes.     

Societal Costs of Late Blight in Potato and Prospects of Durable Resistance Through Cisgenic Modification

In the European Union almost 6 Mha of potatoes are grown representing a value of close to €6,000,000,000. Late blight caused by Phytophthora infestans causes annual losses (costs of control and damage) estimated at more than €1,000,000,000. Chemical control is under pressure as late blight becomes increasingly aggressive and there is societal resistance against the use of environmentally unfriendly chemicals. Breeding programmes have not been able to markedly increase the level of resistance of current potato varieties. New scientific approaches may yield genetically modified marker-free potato varieties (either trans- and/or cisgenic, the latter signifying the use of indigenous resistance genes) as improved variants of currently used varieties showing far greater levels of resistance. There are strong scientific investments needed to develop such improved varieties but these varieties will have great economic and environmental impact. Here we present an approach, based on (cisgenic) resistance genes that will enhance the impact. It consists of five themes: the detection of R-genes in the wild potato gene pool and their function related to the various aspects in the infection route and reproduction of the late blight causing pathogen; cloning of natural R-genes and transforming cassettes of single or multiple (cisgenic) R-genes into existing varieties with proven adaptation to improve their value for consumers; selection of true to the wild type and resistant genotypes with similar qualities as the original variety; spatial and temporal resistance management research of late blight of the cisgenic genetically modified (GM) varieties that contain different cassettes of R-genes to avoid breaking of resistance and reduce build-up of epidemics; communication and interaction with all relevant stakeholders in society and transparency in what research is doing. One of the main challenges is to explain the different nature and possible biological improvement and legislative repercussions of cisgenic GM-crops in comparison with transgenic GM-crops. It is important to realize that the present EU Directive 2001/18/EC on GM crops does not make a difference between trans- and cisgenes. These rules were developed when only transgenic GM plants were around. We present a case arguing for an updating and refinement of these rules in order to place cisgenic GM-crops in another class of GM-plants as has been done in the past with (induced) mutation breeding and the use of protoplast fusion between crossable species.     

Development of bio-assays and screening for resistance to beet armyworm (Spodoptera Exigua Hübner) in Allium cepa L. and its wild relatives

The beet armyworm (Spodoptera exigua Hübner)is the most important pest in tropical Alliumcultivations. All shallot (Allium cepa L. group Aggregatum) cultivars are susceptible to this pest. Therefore accessions from three wild Alliumspecies, namely A. galanthum Kar. et Kir., A. fistulosum L. and A. royleiStearn, next to A. cepa L. were used to screen for resistance. First of all, a reliable bio-assay had to be developed. To this end transparent plastic cages with in total 5 plants of one accession per cage were placed on per lite in a heated greenhouse. Five 3-day old larvae were inoculated on each plant. Eight days after inoculation the number of surviving larvae per cage and the mean fresh weightper larva was determined. The lowest larval survival (36%) was found on A. roylei. This was not, however, significantly different from other Allium accessions. Significant differences were found in the fresh weight per larva fed on different Allium accessions. The larvae survived on A. roylei had a very low fresh weight (10.3 mg per larva), while those on an accession of A. fistulosum had the highest fresh weight (45.1 mg per larva). The larval fresh weight on A. roylei was lower than all the other accessions except from the tropical shallot cultivar Bawang Bali. To check whether or not a toxic compound was involved in the resistance present in A. roylei, tenaccessions from four Allium species were screened. Five 3-day old larvae were inoculated on regularly replaced leaf material of each accession ofAllium species. No significant differences were found in mean fresh weight per larva and mean survival of larvae among different accessions. There were also no significant differences in pupal weight and developmental time. All larvae became pupae 10 days after inoculation. The data indicate that there is no toxic compound present in A. roylei. These results are underlined by the observation in the greenhouse bio-assay that A. roylei plants were equally damaged by the beet armyworm compared to otherAllium species. The results obtained so far therefore suggest that introduction of resistance to S. exigua via the exploitation of variation for resistance to the beet armyworm in A. roylei is unclear and that genetic engineering using Cry sequences could provide a way forward. This revised version was published online in July 2006 with corrections to the Cover Date.     

Organic Halogens in Landfill Leachates

Using a group parameter, totalorganic halogens (TOX), high TOX concentrationswere found in leachates and leachate contaminatedgroundwaters at two Danish mixed sanitary andhazardous waste sites. With commonly used screeningprocedures for organic contaminants, the individualhalogenated organic compounds behind the high TOXcould not be found. Conversely, part of the TOX couldbe attributed to halogenated humic substances that cannot be analysed with conventional organic contaminantanalyses. Even though TOX can still be considered anindicator of leachate contamination in the aquifers,attempts to identify the halogenated contaminantsbehind the TOX using screening methods such as GC-MSwill be unsuccessful in most cases.     

Screening for New Cosmeceuticals from Brown Algae Fucus vesiculosus with Antioxidant and Photo-Protecting Properties

Phlorotannins play a role in biological functions to protect the cells against UV and oxidative damage in brown algae. We hypothesized that these compounds can function as photo-protectors and antioxidants in skin care formulations. Two types of extracts (water (FV-WE) and 67% v/v ethanol (FV-EE)) from Fucus vesiculosus were obtained with a phlorotannin content between 7−14% in dry extract. Exposure to sun light during growth was included as a factor on the phlorotannin content but did not influence the phlorotannin content. However, green colored F. vesiculosus had lower total phenolic content (TPC) (FV-WE = 6.9 g GAE 100 g⁻¹ dw, FV-EE = 7.8 g GAE 100 g⁻¹ dw) compared to those with a yellow/brownish color (FV-WE = 10.4–13.7 g GAE 100 g⁻¹ dw, FV-EE = 11.2–14.0 g GAE 100 g⁻¹ dw). UVA and UVB photo protective capabilities of the extracts through different biological effective protection factors (BEPFs) were evaluated using in vitro methods; the Mansur method for sun protection factor (SPF) and calculation of effective solar absorption radiation (%ESAR) to determine SPF and UVA protection factor (UVA-PF) of the extract and in seaweed enriched lotion. The SPF was negligible, when evaluating FV-WE in lotion (10 and 20% w/w). Moreover, %ESAR of the FV-WE showed SPF and some UVA-PF, but not enough to give sufficient SPF in lotions (10% w/w). It was concluded that the concentration of UV protecting compounds in the extracts was too low to and that further fractionation and purification of phlorotannins is needed to increase the SPF.     

Surgical treatment of a large congenital cavernous haemangioma on the thorax of a foal

A 3‐day‐old male foal was presented with a fluctuant 25 × 15 cm mass on the thorax. The mass had increased in size since birth. The mass did not respond to conservative treatment consisting of aspiration of serohaemorrhagic contents and compression bandages, and it was therefore surgically removed when the foal was approximately 2½ weeks. A histopathological diagnosis of cavernous haemangioma was made. Healing progressed without complications despite a large surgical wound left to heal by second intention. Recurrence was not observed during the 1.5‐year follow‐up period.