Bovine foetal sex determination—Different DNA extraction and amplification approaches for efficient livestock production

Foetal sex determination using polymerase chain reaction (PCR) in mammals is based on the amplification of gender‐specific foetal DNA sequences circulating in maternal blood. The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal DNA and, consequently, its very small amounts in maternal bloodstream. Circulating cell‐free foetal DNA (ccffDNA) encompasses short nucleotide fragments (300–600 bp) in maternal circulation. The aim of this study was to assess this non‐invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various DNA isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non‐pregnant heifer was the controls. Extraction of foetal DNA was accomplished by three different methods: using tubes with silicone membranes, a single‐tube extraction without silicone membranes and phenol–chloroform extraction. Following each extraction method, foetal DNA was amplified using PCR and real‐time PCR with both bAML and TSPY primers in a separate reaction. Positive results were obtained only after amplification of foetal DNA extracted with a single‐tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the PCR test was 90% in Group I, but the technique failed to detect male foetuses in Group II. The real‐time PCR test sensitivity in Group I was 90% and in Group II 91.6%.     

基于TM影像的黄陵县土地利用变化动态监测

以1987年和2002年两期TM影像为信息源,应用计算机自动识别分类与GIS空间分析相结合的方法,提取了黄陵县不同时期的土地利用现状信息及其动态变化。结果表明:研究区森林覆盖率高,林地占区域土地总面积的70%以上;耕地面积较小,占地比例不到7%。1987年至2002年,耕地和草地面积明显减少,园地增加突出,林地、居民点及工矿用地略有增长,其中面积变化最大的是耕地,减少111.13 km2;变化率最大的是园地,达到329.44%。     

Establishment of a comprehensive indicator system for the assessment of biodiversity and ecosystem services

Context

Common indicators are needed to monitor biodiversity loss and the implications for the sustainable provision of ecosystem services (ES). A plethora of different sets of indicators may hinder the identification of major endpoints for large-scale assessments of biodiversity and ecosystem services (BES).

Objectives

We aim to describe the main challenges of indicators for BES assessment and provide suggestions for establishing a comprehensive indicator system.

Methods

An extensive literature review was conducted in this study. We review the main challenges of indicators for BES assessment and propose corresponding improvements from our perspectives of theory and practical applications.

Results

The main theoretical challenges of existing indicators include inconsistent definitions and classifications of ES, misunderstanding of linkages among biodiversity, ecosystem structure, functions and services, and practical problems relate to such issues as indicator representativeness, data availability, and uncertainty. Our suggested improvements include clarifying the main terms, concepts and classification of indicators, establishing a robust conceptual framework with clear interactions among different components and indicators, selecting indicators based on ecosystem properties, streaming existing data into one platform, and strengthening validation of proxies. The steps for constructing a comprehensive indicator system for BES assessment are summarized.

Conclusions

Clear definitions of key terms are indispensable to classify indicators and construct a conceptual framework. Improved understanding of the relations among indicators of biodiversity, ecosystem functions, and ES across multiple scales can guide the development of the indicator system. The integrated indicator system is an important tool for BES assessment to support decision making for sustainable development.

     

The joint action of fungicides in mixtures: comparison of two methods for synergy calculation

Two methods for evaluating the joint action of fungicides in mixtures were analysed. In order to obtain a relatively rapid answer on the type of interaction between fungicides in a mixture (additivity, synergism or antagonism), one can apply the Abbott formula to data on fungus survival. Tests with this method will not be accurate at high effective dose values. A more accurate determination of the joint action of fungicides can be made by the Wadley method, applied to data on effective doses. This involves more experimental work than the Abbott method. Optimization of mixing ratios of fungicides required a set of experimental data on effective doses with several mixing ratios.     

Evaluation of the temporal distribution of <Emphasis Type="Italic">Fusarium graminearum</Emphasis> airborne inoculum above the wheat canopy and its relationship with Fusarium head blight and DON concentration

With the aim of unravelling the role of airborne Fusarium graminearum inoculum in the epidemic of Fusarium head blight (FHB) caused by this species in wheat spikes, a network of Burkard air samplers was set up in five wheat fields distributed in Belgium from 2011 to 2013. Each year from April to July, the daily amounts of F. graminearum inoculum above the wheat canopy were quantified using a newly developed TaqMan qPCR assay. The pattern of spore trapping observed was drastically different per year and per location with a frequency of detection between 9 and 66% and a mean daily concentration between 0.8 and 10.2 conidia-equivalent/m³. In one location, air was sampled for a whole year. Inoculum was frequently detected from the wheat stem elongation stage until the end of the harvesting period, but high inoculum levels were also observed during the fall. Using a window-pane analysis, different periods of time around wheat flowering (varying in length and starting date) were investigated for their importance in the relation between airborne inoculum and FHB parameters (FHB severity, frequency of F. graminearum infection and DON). For almost all the combinations of variables, strong and significant correlations were found for multiple window lengths and starting times. Inoculum quantities trapped around flowering were highly correlated with F. graminearum infection (up to R?=?0.84) and DON (up to R?=?0.9). Frequencies of detection were also well correlated with both of these parameters. DON concentrations at harvest could even be significantly associated with the F. graminearum inoculum trapped during periods finishing before the beginning of the anthesis (R?=?0.77). Overall, these results highlight the key role of the airborne inoculum in F. graminearum epidemics and underline the importance of monitoring it for the development of disease forecasting tools.     

Wax matters: absence of very‐long‐chain aldehydes from the leaf cuticular wax of the glossy11 mutant of maize compromises the prepenetration processes of Blumeria graminis

Conidial germination and differentiation, the so‐called prepenetration processes, of the barley powdery mildew fungus (Blumeria graminis f.sp. hordei) are triggered in vitro by very‐long‐chain aldehydes, minor constituents of barley leaf wax. However, until now it has not been demonstrated that these cuticle‐derived molecules also play a significant role in the initiation and promotion of the fungal prepenetration processes in vivo, on the surface of a living plant leaf. In the maize (Zea mays) wax mutant glossy11, which is completely devoid of cuticular very‐long‐chain aldehydes, germination and appressorial differentiation of B. graminis were strongly impeded. Spraying the mutant leaf surface with aldehyde‐containing wild‐type wax or pure n‐hexacosanal (C₂₆‐aldehyde) fully restored fungal prepenetration, whereas maize wild‐type leaf surfaces coated with n‐docosanoic acid exhibited reduced conidial germination rates of 23%, and only 5% of the conidia differentiated infection structures. In vitro studies were performed to further corroborate the extensive prevention of fungal germination and differentiation in response to artificial surfaces coated with aldehyde‐deficient maize wax. Because of its phenotype affecting the B. graminis prepenetration processes, the glossy11 mutation of maize may become a valuable molecular target and genetic tool that could provide a means of developing basal powdery mildew resistance in the globally important crops wheat and barley.     

Evaluation of the durability of the <Emphasis Type="Italic">Barley yellow dwarf virus</Emphasis>-resistant <Emphasis Type="Italic">Zhong ZH</Emphasis> and <Emphasis Type="Italic">TC14</Emphasis> wheat lines

Serial passage experiments (SPE) of a Barley yellow dwarf virus-PAV (BYDV-PAV) isolate were performed on Zhong ZH and TC14 wheat lines to evaluate the durability of their resistance to BYDV. At different passage numbers (from the 2nd to the 114th), biological properties of the produced isolates were recorded either by monitoring infection percentages and virus titers of the first 3 weeks of viral infection or by measuring their impact on yield components. Statistical analyses using the area under pathogen progress curves and the area under concentration progress curves demonstrated that these two resistant lines induce, after only a few passages, a selection of variant(s) with significantly modified infection abilities. Isolates resulting from SPE performed on these lines induced important decreases of yield components. These results indicate that the use of Zhong ZH and TC14 lines in BYDV-resistant breeding programmes should be approached with caution.     

Variability in response to toosendanin in a laboratory colony of<Emphasis Type="Italic">Trichoplusia ni</Emphasis>

Laboratory rearing of insects for scientific research ensures a constant supply of insects. However, laboratory colonies may diverge genetically and/or phenotypically from wild populations due to selection pressures in their artificial environment. This study examined how variation in a colony ofTrichoplusia ni influenced reproducibility over time of bioassays of toosendanin (a limonoid extracted from the bark ofMelia azedarach). In four of five experiments insect growth was reduced significantly (P=0.05) by the presence of toosendanin in the diet. The fifth experiment showed the same trend, but the effect was not significant. Mean larval mass of both control and treated insects also varied greatly among experiments. The considerable variability observed in this population highlights the importance of replicating experiments in time, and not just space, to confirm reproducibility of treatment effects. http://www.phytoparasitica.org posting Jan. 23, 2007.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002     

Impact of environmental factors on infestation of tea leaves byHelopeltis theivora, and associated changes in flavonoid flavor components and enzyme activities

Attack of tea plant (Camellia sinensis (L.) O. Kuntze) leaves by the tea mosquito bugHelopeltis theivora (Hemiptera: Miridae) was positively correlated to temperature and rainfall, and partially to humidity, as determined in 12 varieties during the period 2000–2002. The insect attack was severe during the months of May–September, which had high temperature and rainfall, and led to severe loss of biomass due to curling and drying up of the leaves. The biochemical response of these 12 varieties of tea to attack by the insect was determined with special reference to oxidative enzymes and flavonoid flavor components. Insect attack led to an increase in the activities of the oxidative enzymes peroxidase, ascorbate peroxidase and polyphenol oxidase. Activities of phenyl alanine ammonia lyase generally decreased as a result of insect attack, which was significant in the United Planters Association of South India (UPASI) varieties. A significant decrease in polyphenols was also obtained in UPASI varieties. HPLC analysis of catechins revealed a decrease in some of the catechins in the infected leaves. Analysis of theaflavins from infusion of healthy andHelopeltis-infested tea leaves revealed no changes. http://www.phytoparasitica.org posting Dec. 19, 2004.