Effects of method of straw disposal and depth of cultivation on populations of Fusarium spp. in soil and on brown foot rot in continuous winter wheat

Soil samples (0–10 cm depth) taken from plots of continuous winter wheat at intervals between January 1993 and July 1996 were dilution-plated on PCNB agar to identify and enumerate colony-forming units (cfus) of Fusarium spp. Cfus of the main wheat pathogen, F. culmorum, were more numerous in soil from plots that were shallow-cultivated to 10 cm after each harvest than in those that were ploughed. They were also usually more numerous in plots in which straw had been chopped and incorporated (by shallow cultivation or ploughing) than in those in which straw had been burnt. Increasing amounts of straw tended to increase the number of cfus. Deeper sampling (15–25 cm) showed that ploughing resulted in a more even, but diluted, distribution of cfus down the soil profile. Populations of some commonly found species that are not pathogenic on wheat {F. oxysporum, F. equiseti, F. flocciferum and F. merismoides) were sometimes also affected by straw disposal or cultivation treatments. Alternative methods of estimating population densities (baiting soil with wheat kernels or wheat seedlings, or plating-out straw fragments) showed generally similar effects of treatments on populations of F. culmorum; the relative merits of the methods are considered. Large differences in population densities of all species occurred within and between seasons. The smallest populations were found in January 1995 but, in the case of F. culmorum, they increased to relatively large numbers in July 1995. The summer of 1995 was warm and dry and especially favourable for development of fusarium foot rot. Severe disease occurred only in this year and was less frequent after ploughing than after tine-cultivation but more frequent after straw burning than after straw incorporation. The implications of these observations for understanding the role of propagules of F. culmorum in soil as inoculum for foot rot are discussed.     

A new cost-effective, selective and differential medium for the isolation of Cronobacter spp

The objective of the present study was to develop a new selective, differential and cost-effective medium (Kim and Rhee — KR-medium) for the isolation of Cronobacter spp. In this new medium, which contained salicin as a differential agent, Cronobacter spp. generated typical colonies with characteristic violet-colored centers surrounded by a transparent to opalescent border, and the growth of other microorganisms (40 strains) was inhibited or produced visually distinguishable colonies. Using healthy and heat- and desiccation-injured cells, the quantity of nutrients was adjusted to determine the optimal recovery rate, selectivity, differentiation and cost-effectiveness. Peptone and salicin concentrations were established as 10 and 8 g/L, respectively. The KR medium was then validated using salicin fermenting organisms, including Cronobacter spp. (52 strains), Enterobacter cloacae (50 strains) and Klebsiella pneumonia (10 strains) isolated from clinical and food specimens. All strains of Cronobacter spp. produced typical colonies and other salicin fermenting organisms were easily distinguishable from Cronobacter spp. with the exception of 2 E. cloacae strains. The verification of KR medium was carried out in powdered infant formula artificially inoculated with healthy, heat-injured, and desiccation-injured Cronobacter spp. and the expected typical colonies were appeared. The KR medium had a high specificity (98%) and sensitivity (100%), with no false-negative results. Moreover, we show that the cost of the KR medium is much lower than that of other selective and differential media. The use of the KR medium for the selective isolation of Cronobacter spp. in laboratories and food industry settings may therefore lessen the financial burden of Cronobacter spp. detection.     

Strain-dependent proliferation in response to human gastric mucin and adhesion properties of Helicobacter pylori are not affected by co-isolated Lactobacillus sp

Background: Helicobacter pylori colonize the mucus layer that covers the gastric epithelium and can cause gastritis, ulcers, and gastric cancer. Recently, Lactobacillus sp. have also been found to reside in this niche permanently. This study compares adhesive properties and proliferation of co‐isolated lactobacilli and H. pylori in the presence of mucins and investigates possibilities for lactobacilli‐mediated inhibition of H. pylori. Materials and methods: Binding and proliferation of four H. pylori and four Lactobacillus strains, simultaneously isolated after residing in the stomachs of four patients for >4 years, to human gastric mucins were investigated using microtiter‐based methods. Results: The H. pylori strains co‐isolated with lactobacilli exhibited the same mucin binding properties as demonstrated for H. pylori strains previously. In contrast, no binding to mucins was detected with the Lactobacillus strains. Proliferation of mucin‐binding H. pylori strains was stimulated by the presence of mucins, whereas proliferation of non‐binding H. pylori and Lactobacillus strains was unaffected. Associative cultures of co‐isolated H. pylori and Lactobacillus strains showed no inhibition of H. pylori proliferation because of the presence of whole bacteria or supernatant of lactobacilli. Conclusions: The presence of lactobacilli in the stomach did not select for different mucin binding properties of H. pylori, and Lactobacillus sp. did neither compete for binding sites nor inhibit the growth of co‐isolated H. pylori. The effects of human gastric mucins on H. pylori proliferation vary between strains, and the host–bacteria interaction in the mucus niche thus depends on both the H. pylori strain and the microenvironment provided by the host mucins.     

Dung Beetle and Terrestrial Mammal Diversity in Forests, Indigenous Agroforestry Systems and Plantain Monocultures in Talamanca, Costa Rica

In order to explore the importance of indigenous agroforestry systems for biodiversity conservation, we compared the abundance, species richness and diversity of dung beetles and terrestrial mammals across a gradient of different land use types from agricultural monocultures (plantains) to agroforestry systems (cocoa and banana) and forests in the BriBri and Cabécar indigenous reserves in Talamanca, Costa Rica. A total of 132,460 dung beetles of 52 species and 913 tracks of 27 terrestrial mammal species were registered. Dung beetle species richness and diversity were greatest in the forests, intermediate in the agroforestry systems and lowest in the plantain monocultures, while dung beetle abundance was greatest in the plantain monocultures. The number of mammal tracks per plot was significantly higher in forests than in plantain monocultures, whereas mammal species richness was higher in forests than in either cocoa agroforestry systems or plantain monocultures. Species composition of both terrestrial mammals and dung beetles also varied across the different land use types. Our study indicates that indigenous cocoa and banana agroforestry systems maintain an intermediate level of biodiversity (which is less than that of the original forest but significantly greater than that of plantain monocultures) and provide suitable habitat for a number of forest-dependent species. Although the agroforestry systems appear to serve as favorable habitats for many terrestrial mammal species, their potential positive contribution to mammal conservation is being offset by heavy hunting pressure in the reserves. As in other agricultural landscapes, the conservation of biodiversity in Talamanca will depend not only on maintaining the existing forest patches and reducing the conversion of traditional agroforestry systems to monocultures, but also on reducing hunting pressure.     

Stomatal control of transpiration from a developing sugarcane canopy

Abstract. Stomatal conductance of single leaves and transpiration from an entire sugarcane (Saccharum spp. hybrid) canopy were measured simultaneously using independent techniques. Stomatal and environmental controls of transpiration were assessed at three stages of canopy development, corresponding to leaf area indices (L) of 2.2, 3.6 and 5.6. Leaf and canopy boundary layers impeded transport of transpired water vapour away from the canopy, causing humidity around the leaves to find its own value through local equilibration rather than a value determined by the humidity of the bulk air mass above the canopy. This tended to uncouple transpiration from direct stomatal control, so that transpiration predicted from measurement of stomatal conductance and leaf-to-air vapour pressure differences was increasingly overestimated as the reference point for ambient vapour pressure measurement was moved farther from the leaf and into the bulk air. The partitioning of control between net radiation and stomata was expressed as a dimensionless decoupling coefficent ranging from zero to 1.0. When the stomatal aperture was near its maximum this coefficient was approximately 0.9, indicating that small reductions in stomatal aperture would have had little effect on canopy transpiration. Maximum rates of transpiration were, however, limited by large adjustments in maximum stomatal conductance during canopy development. The product of maximum stomatal conductance and L. a potential total canopy conductance in the absence of boundary layer effects, remained constant as L increased. Similarly, maximum canopy conductance, derived from independent micrometeorological measurements, also remained constant over this period. Calculations indicated that combined leaf and canopy boundary layer conductance decreased with increasing L such that the ratio of boundary layer conductance to maximum stomatal conductance remained nearly constant at approximately 0.5. These observations indicated that stomata adjusted to maintain both transpiration and the degree of stomatal control of transpiration constant as canopy development proceeded.     

Effects of biocontrol agents on lipid and protein composition of Indian mustard seeds from plants infected with Alternaria species

Field experiments were carried out with Indian mustard (Brassica juncea L. Cv RLM 1359) to investigate the influence of biocontrol agents on seeds from plants infected with Alternaria blight. The biocontrol agents viz, Trichoderma harzianum, Pseudomonas fluorescens and Bacillus subtilis were applied as seed treatment/seed treatment coupled with spray on 30 and 60 days after sowing of seeds in experimental fields. The plants treated with different biocontrol agents were more developed than non-treated plants throughout the experiment. Biochemical analysis revealed that application of biocontrol agents resulted in increase in lipid and protein content in seeds from treated plants. The proportion of various lipidic fractions i.e. phospholipids, glycolipids and sterol content in seeds increased with a corresponding decrease in total glycerides. The proportion of 18:3, 20:1 and 22:1 fatty acids increased while that of 18:1 and 18:2 fatty acids decreased in seeds with application of biocontrol agents. There were both qualitative and quantitative differences in the banding patterns of albumin and globulin proteins after application of biocontrol agents. The data suggested that biochemical alterations in the host induced by treatment with biocontrol agents could be associated with defence mechanisms and enhanced growth of the plant.     

Cyclization of All-E- and 2Z-Geranylfarnesols by a Bacterial Triterpene Synthase: Insight into Sesterterpene Biosynthesis in Aleuritopteris Ferns

Aleuritopteris ferns produce triterpenes and sesterterpenes with tricyclic cheilanthane and tetracyclic 18-episcalarane skeletons. The structural and mechanistic similarities between both classes of fern terpene suggest that their biosynthetic enzymes may be closely related. We investigate here whether a triterpene synthase is capable of recognizing geranylfarnesols as a substrate, and is able to convert them to cyclic sesterterpenes. We found that a bacterial triterpene synthase converted all-E-geranylfarnesol (1b) into three scalarane sesterterpenes with 18αH stereochemistry (5, 7 and 8), as well as mono- and tricyclic sesterterpenes (6 and 9). In addition, 2Z-geranylfarnesol (4) was converted into an 18-episcalarane derivative (10), whose skeleton can be found in sesterterpenes isolated from Aleuritopteris ferns. These results provide insight into sesterterpene biosynthesis in Aleuritopteris ferns.     

Mate guarding in relation to seasonal changes in the energy reserves of two freshwater amphipods (Gammarus fossarum and G. pulex)

1. We assessed sex‐specific seasonal changes in major energy storage compounds (triglycerides, glycogen) in Gammarus fossarum and Gammarus pulex collected from the field, with respect to their reproductive activity. 2. The dynamics of stored energy followed a seasonal pattern in both species and sexes. Moreover, over a 4‐year period, these changes were independent of the year in which they were investigated. Stored energy reached a peak in late winter, but was depleted in late summer and early autumn, coinciding with the reproductive periods. 3. Triglyceride (annual mean ± SD) accounted for 79.7 ± 11.9% of the total stored energy and was responsible for the seasonal pattern. In contrast, glycogen contributed a lesser percentage (20.3 ± 11.9%). Over the study period, the amount of stored energy ranged between 0.39 and 4.08 kJ g^?1 dry mass (triglyceride: 0.19–3.69 kJ g^?1 dry mass; glycogen: 0.14–0.80 kJ g^?1 dry mass). 4. In both species, the energy reserves of males were drastically depleted shortly before the cessation of precopulatory mate guarding in the field, thus offering a bioenergetic explanation for the reproductive period in these two widespread species.     

Expressed sequence tag-derived microsatellite markers of perennial ryegrass (Lolium perenne L.)

An expressed sequence tag (EST) library of the key grassland species perennial ryegrass (Lolium perenne L.) has been exploited as a resource for microsatellite marker development. Out of 955 simple sequence repeat (SSR) containing ESTs, 744 were used for primer design. Primer amplification was tested in eight genotypes of L. perenne and L. multiflorum representing (grand-) parents of four mapping populations and resulted in 464 successfully amplified EST-SSRs. Three hundred and six primer pairs successfully amplified products in the mapping population VrnA derived from two of the eight genotypes included in the original screening and revealed SSR polymorphisms for 143 ESTs. Here, we report on 464 EST-derived SSR primer sequences of perennial ryegrass established in laboratory assays, providing a dedicated tool for marker assisted breeding and comparative mapping within and among forage and turf grasses. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.