The long way down--are carbon and oxygen isotope signals in the tree ring uncoupled from canopy physiological processes?

The carbon (δ(13)C) and oxygen (δ(18)O) stable isotope composition is widely used to obtain information on the linkages between environmental drivers and tree physiology over various time scales. The tree-ring archive can especially be exploited to reconstruct inter- and intra-annual variation of both climate and physiology. There is, however, a lack of information on the processes potentially affecting δ(13)C and δ(18)O on their way from assimilation in the leaf to the tree ring. As a consequence, the aim of this study was to trace the isotope signals in European beech (Fagus sylvatica L.) from leaf water (δ(18)O) and leaf assimilates (δ(13)C and δ(18)O) to tree-ring wood via phloem-transported compounds over a whole growing season. Phloem and leaf samples for δ(13)C and δ(18)O analyses as well as soil water, xylem water, leaf water and atmospheric water vapour samples for δ(18)O analysis were taken approximately every 2 weeks during the growing season of 2007. The δ(13)C and δ(18)O samples from the tree rings were dated intra-annually by monitoring the tree growth with dendrometers. δ(18)O in the phloem organic matter and tree-ring whole wood was not positively related to leaf water evaporative enrichment and δ(18)O of canopy organic matter pools. This finding implies a partial uncoupling of the tree-ring oxygen isotopic signal from canopy physiology. At the same time, internal carbon storage and remobilization physiology most likely prevented δ(13)C in tree-ring whole wood from being closely related to intra-annual variation in environmental drivers. Taking into account the post-photosynthetic isotope fractionation processes resulting in alterations of δ(13)C and δ(18)O not only in the tree ring but also in phloem carbohydrates, as well as the intra-annual timing of changes in the tree internal physiology, might help to better understand the meaning of the tree-ring isotope signal not only intra- but also inter-annually.     

Effects of Foliar Fertilization on Incidence of Berry Shrivel and Bunch Stem Necrosis in Vitis vinifera L. CVS ‘Pinot Blanc’ and ‘Zweigelt’

Berry shrivel and bunch stem necrosis are the economically most important physiological disorders of grapevine. The nutritional status of the plant is considered as one of the main elicitors for the occurrence of these disorders. For foliar fertilization trials, two vineyards that had previously been affected by both diseases were selected in Southwest Germany, one with the variety ‘Zweigelt’ and one with Pinot Blanc. By the use of potassium, magnesium, and calcium foliar fertilizers, a rapid increase of different nutrients in the plants should be caused and the effects on the diseases observed. In 2010, no significant differences in the fertilization treatments were found for both varieties. In ‘Pinot Blanc’, significantly more berries were affected by berry shrivel in 2011 in plots treated with calcium fertilizer compared to those treated with potassium and magnesium. Plots with magnesium fertilizer showed significantly less berries affected by berry shrivel than the untreated control.     

Fungal colonization patterns in necrotic rootstocks and stem bases of dieback‐affected Fraxinus excelsior L.

European ash (Fraxinus excelsior) trees currently face the major threat of ash dieback caused by an invasive fungus, Hymenoscyphus fraxineus. Collar rots in F. excelsior have been increasingly associated with infections by this pathogen. However, the aetiology of the collar rots is still unclear and remains heavily debated. In contrast to most studies of this kind, entire rootstocks of four diseased ash trees were dug out to examine necrotic tissues in these rootstocks and stem bases in detail and to sample necrotic wood for fungal isolation. With the aid of morphological and molecular identification techniques, five to twelve fungal taxa were detected per tree. Members of the Nectriaceae family and Botryosphaeria stevensii, the causal agent of stem and branch cankers on many tree species, were frequently isolated from outer xylem. In contrast, H. fraxineus was the dominating species in interior wood layers. Microsatellite genotyping of 77 H. fraxineus isolates helped to identify up to six different genotypes per tree. The role of H. fraxineus and other isolated fungi in the aetiology of ash collar rots are discussed.     

Antioxidant,ethylene and membrane leakage responses to powdery mildew infection of near-isogenic barley lines with various types of resistance

Leaves of powdery mildew-susceptible barley (Hordeum vulgare cv. Ingrid) and related near-isogenic lines bearing various resistance genes (Mla12, Mlg or mlo5) were inoculated with Blumeria graminis f. sp. hordei race A6. Fungal attack induced several-fold increases in ethylene emission and electrolyte leakage in leaves of susceptible Ingrid beginning 3 days after inoculation. Activities of peroxidase, superoxide dismutase, glutathione S-transferase, ascorbate peroxidase and glutathione reductase enzymes were induced markedly in susceptible leaves 5–7 days after inoculation. Similar, but less pronounced pathogen-induced changes were detected in inoculated leaves of Mla-type resistant plants that show hypersensitive cell death upon inoculation, and, to an even lesser extent, in the Mlg and mlo lines, where no visible symptoms accompanied the incompatible interaction. Glutathione content increased only in susceptible barley 7 days after inoculation. Catalase activity, total ascorbate content and redox state were not influenced by inoculation in any of the genotypes. The activity of dehydroascorbate reductase was significantly reduced 3–5 days after inoculation in the susceptible parental plants and after 5 days in Mla and Mlg lines, while it was stable in the mlo barley. Slightly elevated levels of H₂O₂ were observed in the inoculated resistant plants. In contrast, H₂O₂ content decreased in the susceptible line 7 days after pathogen attack. These data indicate that high levels of antioxidants are involved in the compatible interaction of susceptible barley and powdery mildew by protecting the pathogen from oxidative damage.     

The effects of patulin from Penicillium vulpinum on seedling growth,root tip ultrastructure and glutathione content of maize

A culture of Penicillium vulpinum CM1 isolated from a soil sample cultivated with maize was examined for the production of the mycotoxins cyclopiazonic acid, griseofulvin, patulin (PAT), and roquefortine C. The fungal strain was positive for PAT- and roquefortine C–producing ability, while it showed negative producing ability for cyclopiazonic acid, and griseofulvin. Both PAT and roquefortine C toxins were tested for their inhibitory effect on the germination of maize seeds. Roquefortine C showed no phytotoxicity up to a concentration of 100 μg ml^?1. However, a notable reduction in the average values of fresh weight and length of roots and shoots was observed following application of PAT and a culture filtrate of P. vulpinum to maize seedlings. The phytotoxic effect was found to be dose-dependent. To analyse the cellular effects of PAT, maize root tips were treated with PAT at different concentrations. The higher dose of 25 μg ml^?1 exhibited morphological changes in the nature of cytoplasm and cytoplasmic organelles as was shown by transmission electron microscopy. Furthermore, an increase in the vacuolation with localized tonoplast dissolution was observed. At this concentration, there was evidence of a disruption of lipid metabolism. On the metabolic level, the phytotoxicity of PAT and the culture filtrate of P. vulpinum led to alterations in the total reduced glutathione (GSH) concentration in maize seedlings. The GSH concentrations examined in roots and shoots after PAT, fungal filtrate or fungal crude extract treatments were enhanced compared with control treatments. Findings further showed higher total GSH levels in shoots than in roots following treatments either with PAT or the fungal crude extract. The results are discussed in the context of known herbicide and metal effects on GSH synthesis and transport mechanisms.     

Well-defined multispecies probiotic and enzyme combination outperforms traditional fermented probiotic applications in an intensive Pacific white shrimp,Litopenaeus vannamei (Boone, 1931), culture system

A series of treatments were designed to evaluate the efficacy of feeding commercial multispecies probiotics feeding with enzymes and fermentation process on the growth parameters and culture environment of Pacific white shrimp, Litopenaeus vannamei (Boone, 1931), in an intensive culture system. Commercial multispecies probiotics and enzymes (PEs) were continuously applied in three different doses, namely (i) 0.2, (ii) 0.4, and (iii) 0.6 mg L⁻¹ and designated as 0.2, 0.4, and 0.6 PE during the first 30 days of intensive culture of Pacific white shrimp, L. vannamei (Boone, 1931). The probiotics were continually applied every alternate day, while the enzymes were added every sixth day throughout the trial period. The PE dose for all treated tanks was increased by 0.2 ppm after 30 days of culture and another 0.2 ppm after day 60. Meanwhile, fermentation technique, which has become the common method applied in Indonesia in the control treatment, was added on the same day with the PE group with increasing dosage, following the same trend with the 0.6 PE group. Results showed that the group receiving 0.6 PE showed higher final biomass, higher mean weight, and protein retention efficiency as well as a lower feed conversion ratio compared with the control treatment. Abundance of Vibrio spp. remained below 10³ cfu mL⁻¹ throughout the trial. Water quality indicators TAN, NO₂ N and NO₃ N peaked in weeks 3–5 and then declined after that until the end of the culture period in all tanks. This decline was significantly faster in PE-treated tanks. Multispecies PEs have potential applications in controlling Vibrio spp., maintaining proper water quality condition, and enhancing the growth of shrimp in intensive culture system.     

Association mapping and marker development of the candidate genes (1 → 3),(1 → 4)-β-d-Glucan-4-glucanohydrolase and (1 → 4)-β-Xylan-endohydrolase 1 for malting quality in barley

Cell wall degradation is a crucial process within the malting process of barley. Therefore, the haplotype diversity of genes for two cell wall degrading enzymes, (1 → 3),(1 → 4)-β-d-Glucan-4-glucanohydrolase and (1 → 4)-β-Xylan-endohydrolase 1, was investigated and associations to malting quality parameters were performed. The (1 → 3),(1 → 4)-β-d-Glucan-4-glucanohydrolase gene glb2 had two major haplotypes defined by three SNPs and one INDEL, which explained 8.9 and 9.5% of the total variation of malt extract content and viscosity in the spring barley gene pool, respectively. The most significant associations of (1 → 4)-β-Xylan-endohydrolase 1 gene X-1 were found for diastatic power, saccharification VZ45 and soluble nitrogen with 18, 12 and 8% of the total variation explained by SNP3 in the spring barleys. High-throughput markers were developed for both genes which can be used for marker assisted selection.     

Detection of Mycobacterium avium ssp. paratuberculosis in ileocaecal lymph nodes collected from elderly slaughter cows using a semi-nested IS900 polymerase chain reaction

The aim of this study was to investigate the occurrence of subclinical Mycobacterium avium spp. paratuberculosis (MAP) infections at slaughter by testing ileocaecal lymph nodes with a semi-nested IS900 PCR. Tissue samples were available within the framework of a parallel study investigating BSE-susceptibility factors in members of BSE-cohorts in the German Federal State of Lower Saxony. Ileocaecal lymph nodes were collected over a 2-year sampling period from 99 slaughter cattle of a mean age of 6.5 years (5.5-7.5 years). A recently developed IS900 semi-nested polymerase chain reaction (snPCR) assay offering a sensitivity of 1 genome equivalent was used for the detection of MAP-DNA. Based on this snPCR, 17 out of the 99 samples gave positive results, indicating a MAP occurrence of 17.17% in the random sample. All PCR products were sequenced for screening of polymorphisms. Nucleotide homologies of 98.5-100% were found with respect to the MAP K10 reference sequence IS900 (GenBank: AE16958). PCR analysis of ileocaecal lymph nodes collected from slaughter cattle proved to be a suitable technique to determine MAP occurrence in the local cattle population.