Detection of phloridzin in strawberries (Fragaria x ananassa Duch.) by HPLC-PDA-MS/MS and NMR spectroscopy

The phenolic profile of strawberry fruits (Fragaria x ananassa Duch., Rosaceae) was investigated by high-performance liquid chromatography with photodiode array detection. A peak displaying retention time and UV spectral data identical to those of phloridzin (phloretin 2'-O-beta-d-glucoside), a dihydrochalcone glucoside so far considered characteristic of apples, was monitored. For further characterization, crude extracts of strawberries were purified on polyamide, and the target compound was isolated by preparative and analytical HPLC. Structure elucidation was performed on the basis of APCI- and ESI-MS in the negative ion mode as well as by 1D and 2D NMR spectroscopy using authentic phloridzin for comparison. The d-configuration of the sugar moiety was established by HPLC analysis of the corresponding acyclic 1-deoxy-1-(N-acetyl-alpha-methylbenzylamino)alditol acetate. Apart from its chemotaxonomic relevance, this first report on the occurrence of phloridzin in strawberries is of particular interest for the authenticity control of strawberry products such as juices, jams, and fruit preparations since phloridzin has so far been used for the detection of fraudulent admixtures.     

RADIX: rhizoslide platform allowing high throughput digital image analysis of root system expansion

Background

Phenotyping of genotype-by-environment interactions in the root-zone is of major importance for crop improvement as the spatial distribution of a plant’s root system is crucial for a plant to access water and nutrient resources of the soil. However, so far it is unclear to what extent genetic variations in root system responses to spatially varying soil resources can be utilized for breeding applications. Among others, one limiting factor is the absence of phenotyping platforms allowing the analysis of such interactions.

Results

We developed a system that is able to (a) monitor root and shoot growth synchronously, (b) investigate their dynamic responses and (c) analyse the effect of heterogeneous N distribution to parts of the root system in a split-nutrient setup with a throughput (200 individual maize plants at once) sufficient for mapping of quantitative trait loci or for screens of multiple environmental factors. In a test trial, 24 maize genotypes were grown under split nitrogen conditions and the response of shoot and root growth was investigated. An almost double elongation rate of crown and lateral roots was observed under high N for all genotypes. The intensity of genotype-specific responses varied strongly. For example, elongation of crown roots differed almost two times between the fastest and slowest growing genotype. A stronger selective root placement in the high-N compartment was related to an increased shoot development indicating that early vigour might be related to a more intense foraging behaviour.

Conclusion

To our knowledge, RADIX is the only system currently existing which allows studying the differential response of crown roots to split-nutrient application to quantify foraging behaviour in genome mapping or selection experiments. In doing so, changes in root and shoot development and the connection to plant performance can be investigated.

     

In silico prediction of Gallibacterium anatis pan-immunogens

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0080-0) contains supplementary material, which is available to authorized users.     

Yield decrease in sugar beet caused by reduced tillage and direct drilling

In a long-term series of on-farm tillage trials (10 loessial sites in southern and eastern Germany; annual mouldboard ploughing 0.25–0.3 m deep, mulching with a rigid-tine cultivator 0.1–0.15 m deep, direct drilling with no tillage except seedbed preparation for sugar beet solely) sugar beet yield was significantly decreased by direct drilling compared to ploughing. This study was conducted to (i) show that the lower plant density caused by mulching and direct drilling contributes to yield decrease but explains effects just partially, and (ii) determine the relation between soil structural properties and sugar beet yield. In 2003–2005 plant density experiments (53,000, 65,000 and 82,000 plants ha^?1) were introduced to tillage plots on five selected environments. Yield and soil structural properties of four layers representing 0–0.43 m soil depth were determined.White sugar yield (WSY) significantly declined with direct drilling compared to ploughing treatment, whereas mulching treatment diminished WSY less pronounced. Moreover, decreasing plant density significantly lowered WSY. No interactions between tillage and plant density occurred, revealing that both factors additively affected WSY.Decreasing tillage depth increased penetration resistance (PR) and dry bulk density (DBD), and diminished air filled pore volume (AFPV) in the topsoil down to 0.27 m depth. Several soil structural parameters were closely correlated with each other as well as WSY. Variation of single parameters explained up to 60% of WSY variance attributed to tillage. Combining DBD from 0.03 to 0.07 m depth, average PR from 0.03 to 0.27 m and AFPV from 0.03 to 0.18 m soil depth explained 77% of the tillage effect. Nevertheless, multi-collinearity of soil physical parameters allowed no clear conclusions on the cause-and-effect mechanisms.Conclusively, lowered plant density and soil structure degradation due to reduced tillage may independently decrease sugar beet yield. When grown on loessial soils this crop requires mechanical loosening down to 0.15–0.20 m depth to produce high yields.     

Pathogens of bark beetles (Coleoptera: Curculionidae) in Bulgarian forests

The occurrence and prevalence of bark beetle pathogens in forest stands in Bulgaria were investigated in 944 specimens belonging to 21 bark beetle species. Protozoa, microsporidia, fungi and nematodes occurred in 19 of all investigated species. The infections were found in the gut (nematodes, gregarines, microsporidia), gonads (microsporidia) and hemolymph (nematodes) of the infected insects. Protozoan species (Gregarina typographi, Gregarina spp.) were detected in eight bark beetle species. Morphometric data about G. typographi and Gregarina spp. are presented. The prevalence of the gregarines varied between 1.4% and 64.2%. Microsporidia of the genera Nosema and Chytridiopsis were revealed in three bark beetle species. The prevalence of microsporidia ranged between 1.5% and 11.8%. This is the first report of a microsporidium in Taphrorychus villifrons and of gregarines in T. villifrons, Pityogenes bistridentatus, P. conjunctus, and Orthotomicus erosus. The fungus Beauveria bassiana was found in 3.4% of Hylurgops palliatus specimens. Nematodes (in gut and haemolymph) were revealed in 19 bark beetle species and their prevalence varied between 10% and 98.5%.     

Tillage system affects fertilizer-induced nitrous oxide emissions

Since the development of effective N₂O mitigation options is a key challenge for future agricultural practice, we studied the interactive effect of tillage systems on fertilizer-derived N₂O emissions and the abundance of microbial communities involved in N₂O production and reduction. Soil samples from 0–10 cm and 10–20 cm depth of reduced tillage and ploughed plots were incubated with dairy slurry (SL) and manure compost (MC) in comparison with calcium ammonium nitrate (CAN) and an unfertilized control (ZERO) for 42 days. N₂O and CO₂ fluxes, ammonium, nitrate, dissolved organic C, and functional gene abundances (16S rRNA gene, nirK, nirS, nosZ, bacterial and archaeal amoA) were regularly monitored. Averaged across all soil samples, N₂O emissions decreased in the order CAN and SL (CAN?=?748.8?±?206.3, SL?=?489.4?±?107.2 μg kg^?1) followed by MC (284.2?±?67.3 μg kg^?1) and ZERO (29.1?±?5.9 μg kg^?1). Highest cumulative N₂O emissions were found in 10–20 cm of the reduced tilled soil in CAN and SL. N₂O fluxes were assigned to ammonium as source in CAN and SL and correlated positively to bacterial amoA abundances. Additionally, nosZ abundances correlated negatively to N₂O fluxes in the organic fertilizer treatments. Soils showed a gradient in soil organic C, 16S rRNA, nirK, and nosZ with greater amounts in the 0–10 than 10–20 cm layer. Abundances of bacterial and archaeal amoA were higher in reduced tilled soil compared to ploughed soils. The study highlights that tillage system induced biophysicochemical stratification impacts net N₂O emissions within the soil profile according to N and C species added during fertilization.     

The Climate-Species-Matrix to select tree species for urban habitats considering climate change

Two hundred and fifty urban woody species are used in Central European parks and gardens. Based on reliable publications we attempt, for the first time, to extensively classify and assess them with regards to their usability after predicted climate changes. A new Climate-Species-Matrix has been developed for this purpose. In a two-dimensional assessment, four degrees of drought resistance and winter robustness are the decisive criteria. The woody species (divided into trees of more than 10 m in potential height and trees/shrubs of up to 10 m in height) were placed into 16 categories by decreasing tolerance, ranging from 1-1 (very suitable) to 4-4 (very limited usability). For many of these species, as well as many others that are not mentioned in this study, not all questions could be answered. Nonetheless, this categorization provides a sound basis for decisions in planning the use of woody species in cities. This planning process must also include additional criteria, dependent on individual requirements such as, soil parameters, shade resistance, aesthetics, etc. This paper is therefore meant to be a basis for discussion and supplementation for further research.     

Concept and feasibility study for the integrated evaluation of environmental monitoring data in forests

In the 1970s unexpected forest damages, called “new type of forest damage” or “forest decline”, were observed in Germany and other European countries. The Federal Republic of Germany and the German Federal States implemented a forest monitoring system in the early 1980s, in order to monitor and assess the forest condition. Due to the growing public awareness of possible adverse effects of air pollution on forests, in 1985 the ICP Forests was launched under the convention on long-range transboundary air pollution (CLRTAP) of the United Nations Economic Commission for Europe (UN-ECE). The German experience in forest monitoring was a base for the implementation of the European monitoring system. In 2001 the interdisciplinary case study “concept and feasibility study for the integrated evaluation of environmental monitoring data in forests”, funded by the German Federal Ministry of Education and Research, concentrated on in-depths evaluations of the German data of forest monitoring. The objectives of the study were: (a) a reliable assessment of the vitality and functioning of forest ecosystems, (b) the identification and quantification of factors influencing forest vitality, and (c) the clarification of cause-effect-relationships leading to leaf/needle loss. For these purposes additional data from external sources were acquired: climate and deposition, for selected level I plots tree growth data, as well as data on groundwater quality. The results show that in particular time series analysis (crown condition, tree growth, and tree ring analysis), in combination with climate and deposition are valuable and informative, as well as integrated evaluation of soil, tree nutrition and crown condition data. Methods to combine information from the extensive and the intensive monitoring, and to transfer process information to the large scale should be elaborated in future.

植保无人机低空低量施药雾滴沉积飘移分布立体测试方法

随着植保无人机在中国的广泛使用,植保无人机的沉积分布均匀性与雾滴飘移流失也引起各方面的重视。目前,针对植保无人机施药雾滴沉积飘移的测试方法较少,且着重于从沉积或飘移中某一方面分析植保无人机雾滴沉积飘移规律,未对作业中全方位的雾滴的沉积飘失规律进行系统测试。该文基于国际标准ISO22866和ISO24253建了1套针对低空低量植保无人机的立体测试方法,分别在地面布置沉积和飘移收集器,在空中架设立体沉积和空中飘移收集器,结合航拍影像所获取的植保无人机准确作业参数,对4个型号植保无人机分别搭载德国Lechler公司的IDK120-015和TR80-0067喷头进行了测试,系统分析了无人机周边的总沉积以验证方法准确性,计算了总地面沉降以表征可利用部分和空中耗散以评估环境风险。结果表明,各植保无人机地面沉积率在53.6%~76.6%,地面飘移率最高17.4%,空中飘移率可高达14.7%;该测试系统可收集62.4%~101.7%无人机喷洒出的雾滴。测试的4种植保无人机在搭载IDK喷头后均明显降低了雾滴飘移,但也同时降低地面沉积率;各植保无人机在搭载2种喷头时沉积规律不同,不同植保无人机设计需要选择不同喷头。该测试方法能够有效的收集并分析植保无人机在作业区域的雾滴立体分布状态,可为植保无人机综合评估提供新的参考依据。