The need of an epidemio-surveillance network to prevent Huanglongbing arrival in the South of the Mediterranean Basin

HuangLongBing (HLB) originated in the 1900's in Asia where it is transmitted by the psyllid Diaphorina citri. In South Africa another form of HLB, transmitted by Trioza erytreae was described in the 1960's. The "African" and "Asian" forms may occasionally occur in a same area (Peninsula Arabia), where both species of the psyllids vectors are present, and each psyllid can carry both forms of the bacteria associated with HLB - Ca. L. asiaticus and Ca.L africanus. In the 2000's the arrival of HLB on the American continent re-boosted research on HLB. In Africa, after South Africa, HLB has caused severe damages in East Africa, mainly Kenya and Tanzania. Recently Ca. L asiaticus was identified in Ethiopia (1).On the west side, in Cameroon, syndrome of HLB was described and T. eythreae was present. This psyllid also invaded Madeira and the Canary Islands ten years ago. The Mediterranean basin produces around 18 million tons of citrus yearly. Because of the importance of movements, trade, tourism, pilgrims, Citrus production is in danger. The threat could come from the East (Saudi Arabia, Iran, Yemen) the South (Ethiopia, Somalia) and the West (Cameroon, Canarias). Cirad in collaboration with Embrapa -Brazil, Cameroon and partners in Morocco, Tunisia, Egypt and Turkey will start actions for a prevention strategy. A training curse on symptoms, psyllids identification and molecular diagnosis is scheduled on early 2011 and will be the foundation of an epidemio-surveillance network in the South Mediterranean basin. (Texte intégral

Spatial imaging of Zn and other elements in Huanglongbing-affected grapefruit by synchrotron-based micro X-ray fluorescence investigation

Huanglongbing (HLB) is a highly destructive, fast-spreading disease of citrus, causing substantial economic losses to the citrus industry worldwide. Nutrient levels and their cellular distribution patterns in stems and leaves of grapefruit were analysed after graft-inoculation with lemon scions containing 'Candidatus Liberibacter asiaticus' (Las), the heat-tolerant Asian type of the HLB bacterium. After 12 months, affected plants showed typical HLB symptoms and significantly reduced Zn concentrations in leaves. Micro-XRF imaging of Zn and other nutrients showed that preferential localization of Zn to phloem tissues was observed in the stems and leaves collected from healthy grapefruit plants, but was absent from HLB-affected samples. Quantitative analysis by using standard references revealed that Zn concentration in the phloem of veins in healthy leaves was more than 10 times higher than that in HLB-affected leaves. No significant variation was observed in the distribution patterns of other elements such as Ca in stems and leaves of grapefruit plants with or without graft-inoculation of infected lemon scions. These results suggest that reduced phloem transport of Zn is an important factor contributing to HLB-induced Zn deficiency in grapefruit. Our report provides the first in situ, cellular level visualization of elemental variations within the tissues of HLB-affected citrus. © 2014 © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology

Gene regulatory networks elucidating huanglongbing disease mechanisms.

Next-generation sequencing was exploited to gain deeper insight into the response to infection by Candidatus liberibacter asiaticus (CaLas), especially the immune disregulation and metabolic dysfunction caused by source-sink disruption. Previous fruit transcriptome data were compared with additional RNA-Seq data in three tissues: immature fruit, and young and mature leaves. Four categories of orchard trees were studied: symptomatic, asymptomatic, apparently healthy, and healthy. Principal component analysis found distinct expression patterns between immature and mature fruits and leaf samples for all four categories of trees. A predicted protein - protein interaction network identified HLB-regulated genes for sugar transporters playing key roles in the overall plant responses. Gene set and pathway enrichment analyses highlight the role of sucrose and starch metabolism in disease symptom development in all tissues. HLB-regulated genes (glucose-phosphate-transporter, invertase, starch-related genes) would likely determine the source-sink relationship disruption. In infected leaves, transcriptomic changes were observed for light reactions genes (downregulation), sucrose metabolism (upregulation), and starch biosynthesis (upregulation). In parallel, symptomatic fruits over-expressed genes involved in photosynthesis, sucrose and raffinose metabolism, and downregulated starch biosynthesis. We visualized gene networks between tissues inducing a source-sink shift. CaLas alters the hormone crosstalk, resulting in weak and ineffective tissue-specific plant immune responses necessary for bacterial clearance. Accordingly, expression of WRKYs (including WRKY70) was higher in fruits than in leaves. Systemic acquired responses were inadequately activated in young leaves, generally considered the sites where most new infections occur

Gas Biosensor Arrays Based on Single-Stranded DNA-Functionalized Single-Walled Carbon Nanotubes for the Detection of Volatile Organic Compound Biomarkers Released by Huanglongbing Disease-Infected Citrus Trees.

Volatile organic compounds (VOCs) released by plants are closely associated with plant metabolism and can serve as biomarkers for disease diagnosis. Huanglongbing (HLB), also known as citrus greening or yellow shoot disease, is a lethal threat to the multi-billion-dollar citrus industry. Early detection of HLB is vital for removal of susceptible citrus trees and containment of the disease. Gas sensors are applied to monitor the air quality or toxic gases owing to their low-cost fabrication, smooth operation, and possible miniaturization. Here, we report on the development, characterization, and application of electrical biosensor arrays based on single-walled carbon nanotubes (SWNTs) decorated with single-stranded DNA (ssDNA) for the detection of four VOCs-ethylhexanol, linalool, tetradecene, and phenylacetaldehyde-that serve as secondary biomarkers for detection of infected citrus trees during the asymptomatic stage. SWNTs were noncovalently functionalized with ssDNA using π-π interaction between the nucleotide and sidewall of SWNTs. The resulting ssDNA-SWNT hybrid structure and device properties were investigated using Raman spectroscopy, ultraviolet (UV) spectroscopy, and electrical measurements. To monitor changes in the four VOCs, gas biosensor arrays consisting of bare SWNTs before and after being decorated with different ssDNA were employed to determine the different concentrations of the four VOCs. The data was processed using principal component analysis (PCA) and neural net fitting (NNF)

Natural parasitism of the Citrus Leafminer (Lepidoptera: Gracillariidae) over eight years in seven citrus regions of São Paulo, Brazil

The citrus leafminer (CLM) Phyllocnists citrella Stainton (Lepidoptera: Gracillariidae) was frst recorded in Brazil in 1996. In 1998, the parasitoid Ageniaspis citricola Logvinovskaya (Hymenoptera: Encyrtdae) was introduced and established in many regions of the country. In this study, 130 onehour-samplings of sweet orange leaves (Citrus sinensis [L.] Osbeck) hostng CLM pupal chambers were carried out to estmate the CLM parasitsm rate (%) by its parasitoids in 7 regions of São Paulo State between 2000 and 2008. The sample sizes varied from 10 to 275 leaves (mean = 65). The most abundant parasitoid was the encyrtd A. citricola (found in 91.8% of the samplings). The highest level of CLM parasitsm by A. citricola was recorded in the southern region (Botucatu), 70.2 ± 6.6 (mean ± SEM), and the lowest level was recorded in the northern region (Barretos), 12.8 ± 5.7%. CLM parasitsm by A. citricola and by natve parasitoids (Galeopsomyia fausta LaSalle, Cirrospilus spp. and Elasmus sp.) did not differ between seasons. The 6-fold increase in the use of insectcides in citrus groves, afer 2004 when the Huanglongbing (HLB) disease was found in São Paulo State, did not reduce the level of CLM parasitsm. The level of parasitsm was 50.8 ± 4.2% before the advent of HLB (2000–2004) and 56.0 ± 4.4% thereafer (2005–2008), indicatng adaptaton of A. citricola in a disturbed agroecosystem.A minadora das folhas dos citros (MFC), Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae), foi encontrada pela primeira vez no Brasil em 1996. Em 1998, o parasitoide Ageniaspis citricola Logvinovskaya (Hymenoptera: Encyrtidae) foi introduzido e se estabeleceu em várias regiões do país. Nesse estudo, foram feitas130 amostragens, de uma hora, de folhas de laranjeiras doces [Citrus sinensis (L.) Osbeck] com câmaras pupais da MFC, para se estimar o parasitismo da MFC em 7 regiões do estado de São Paulo, entre 2000 e 2008. O tamanho das amostras variou de 10 a 275 folhas (média = 65). O parasitoide mais abundante foi o encirtídeo A. citricola (encontrado em 91.8% das amostragens). O maior parasitismo da MFC por A. citricola foi observado na região sul do estado (Botucatu), 70,2 ± 6,6 (média ± EPM), e o menor parasitismo na região norte (Barretos), 12,8 ± 5,7%. O parasitismo da MFC por A. citricola e seus parasitoides nativos (Galeopsomyia fausta LaSalle, Cirrospilus spp. and Elasmus sp.) não diferiram entre as estações do ano. O aumento de seis vezes no uso de inseticidas nos pomares de citros, após 2004, quando o Huanglongbing (HLB) foi encontrado no estado de São Paulo, não reduziu o nível de parasitismo da MFC. O nível médio de parasitismo foi de 50,8 ± 4,2%, antes do HLB (2000-2004), e 56,0 ± 4,4%, após o HLB (2005-2008), indicando a adaptação de A. citricola a um agroecossistema perturbado.info:eu-repo/semantics/publishedVersio

Optimization of Oil-in-Water Emulsion Stability: Experimental Design, Multiple Light Scattering, and Acoustic Attenuation Spectroscopy

To find an optimal formulation of oil-in-water (O/W) emulsions (o = 0.05), the effect of emulsifier nature and concentration, agitation speed, emulsifying time, storage temperature and their mutual interactions on the properties and behavior of these dispersions is evaluated by means of an experimental design (Nemrodw software). Long-term emulsion stability is monitored by multiple light scattering (Turbiscan ags) and acoustic attenuation spectroscopy (Ultrasizer). After matching surfactant HLB and oil required HLB, a model giving the Sauter diameter as a function of emulsifier concentration, agitation speed and emulsification time is proposed. The highest stability of C12E4-stabilized O/W emulsions is observed with 1% emulsifier

Hamiltonians for the Quantum Hall Effect on Spaces with Non-Constant Metrics

The problem of studying the quantum Hall effect on manifolds with nonconstant metric is addressed. The Hamiltonian on a space with hyperbolic metric is determined, and the spectrum and eigenfunctions are calculated in closed form. The hyperbolic disk is also considered and some other applications of this approach are discussed as well.Comment: 16 page

Quantification of sirolimus by liquid chromatography-tandem mass spectrometry using on-line solid-phase extraction

Quantification of the new immunosuppressant sirolimus (syn. rapamycin; Rapamune((R))) in whole blood by chromatography is essential for its clinical use since no immunoassay is available although monitoring is mandatory. Here we report on a rapid and convenient liquid chromatography (LC)-tandem mass spectrometry method and describe our practical experience with its routine use. Whole blood samples were hemolyzed and deproteinized using an equal volume (150 mul) of a mixture of methanol/zinc sulfate solution containing the internal standard desmethoxy-rapamycin. After centrifugation, the clear supernatants were submitted to an on-line solid-phase extraction procedure using the polymeric Waters Oasis HLB(R) material, with elution of the extracts onto the analytical column in the back-flush mode by column switching. For analytical chromatography a RP-C18 column was used with 90/10 methanol/2 mM ammonium acetate as the mobile phase. A 1:10 split was used for the transfer to the mass spectrometer, a Micromass Quattro LC-tandem mass spectrometry system equipped with a Z-spray((R)) source and used in the positive electrospray ionization mode. The following transitions were recorded: sirolimus, 931>864 m/z, and desmethoxy-rapamycin (I.S.), 901>834 m/z. The analytical running time was 5 min, including on-line extraction. The method has a linear calibration curve (r>0.99; range 1.6-50 mug/l) and is rugged and precise with monthly CVs <7% at a sirolimus concentration of 13.1 mug/l in routine use; the instrumentation proved to be reliable and required minimal maintenance