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Reina M.,CSIC - Institute of Natural Products and Agrobiology | Ruiz-Mesia W.,CSIC - Institute of Natural Products and Agrobiology | Ruiz-Mesia W.,University of the Amazon | Lopez-Rodriguez M.,University of La Laguna | And 3 more authors.
Journal of Natural Products | Year: 2012

Ten indole alkaloids were isolated from Geissospermum reticulatum, seven (1-7) from the leaves and three (8-10) from the bark. Seven were aspidospermatan-type alkaloids (1-3, 5-9), including four (5-8) with a 1-oxa-3-cyclopentene group in their molecule, which we named geissospermidine subtype. Compounds 1-3, 5-8, and 10 had not been reported previously as natural products, while 4 and 9 were the known alkaloids O-demethylaspidospermine and flavopereirine. Their structures were determined by spectroscopic techniques including 1D and 2D NMR experiments (COSY, NOESY, HSQC, HMBC). Additionally, X-ray crystallographic analyses of 1, 2, and 6 were performed. Antiparasitic activities of the ethanolic and alkaloidal extracts and of the pure alkaloids were tested against Trypanosoma cruzi and Leishmania infantum. In general, the extracts exhibited selective action and were more active against Leishmania than against Trypanosoma. Alkaloid 4 was also very active against L. infantum. © 2012 The American Chemical Society and American Society of Pharmacognosy.

Rodriguez Martin J.A.,Instituto Nacional Of Investigacion Y Tecnologia Agraria Y Alimentaria Inia | Carbonell G.,Instituto Nacional Of Investigacion Y Tecnologia Agraria Y Alimentaria Inia | Nanos N.,Technical University of Madrid | Gutierrez C.,Institute Ciencias Agrarias ICA
Archives of Environmental Contamination and Toxicology | Year: 2013

This study spatially analysed the relation between mercury (Hg) content in soil and Hg in rock fragment for the purpose of assessing natural soil Hg contribution compared with Hg from human inputs. We present the Hg content of 318 soil and rock fragment samples from 11 islands distributed into two Spanish archipelagos (the volcanic Canary Islands [Canaries] and the Mediterranean Balearic [Balearic] islands). Assumedly both are located far enough away from continental Hg sources to be able to minimise the effects of diffuse pollution. Physical and chemical soil properties were also specified for the samples. Hg contents were significantly greater in the Balearic limestone soils (61 μg kg-1) than in the volcanic soils of the Canaries (33 μg kg -1). Hg levels were also greater in topsoil than in rocky fragments, especially on the Balearics. The soil-to-rock ratios varied between 1 and 30. Interestingly, the highest topsoil-to-rock Hg ratio (>16 ×) was found in the vicinity of a coal-fired power plant in Majorca, whereas no similar areas in the Canary archipelago were identified. © 2012 Springer Science+Business Media New York.

Campos-Herrera R.,University of Florida | Campos-Herrera R.,Institute Ciencias Agrarias ICA | El-Borai F.E.,University of Florida | El-Borai F.E.,Zagazig University | Duncan L.W.,University of Florida
Molecular Ecology Resources | Year: 2012

Quantitative real-time PCR (qPCR) is a powerful tool to study species of cryptic organisms in complex food webs. This technique was recently developed to detect and quantify several species of entomopathogenic nematodes (EPNs), which are widely used for biological control of insects, and some natural enemies of EPNs such as nematophagous fungi and the phoretic bacteria Paenibacillus sp. and Paenibacillus nematophilus. A drawback to the use of primers and TaqMan probes designed for Paenibacillus sp. is that the qPCR also amplified Paenibacillus thiaminolyticus and Paenibacillus popilliae, two closely related species that are not phoretically associated with EPNs. Here, we report that the detection of Paenibacillus sp. DNA in nematode samples was two orders of magnitude greater (P<0.001) when the bacterium was added to soil together with its EPN species-specific host Steinernema diaprepesi than when it was added concomitantly with other EPNs or with species of bacterial-feeding nematodes. Just 6% of samples detected trace amounts of P. thiaminolyticus and P. popilliae exposed to the same experimental conditions. Thus, although the molecular assay detects Paenibacillus spp. DNA in nonphoretic associations, the levels are essentially background compared to the detection of Paenibacillus sp. in association with its nematode host. © 2012 Blackwell Publishing Ltd.

Campos-Herrera R.,University of Florida | Campos-Herrera R.,Institute Ciencias Agrarias ICA | El-Borai F.E.,University of Florida | El-Borai F.E.,Zagazig University | Duncan L.W.,University of Florida
Journal of Invertebrate Pathology | Year: 2012

Entomopathogenic nematodes (EPNs) are promising biological control agents of soil-dwelling insect pests of many crops. These nematodes are ubiquitous in both natural and agricultural areas. Their efficacy against arthropods is affected directly and indirectly by food webs and edaphic conditions. It has long been suggested that a greater understanding of EPN ecology is needed to achieve consistent biological control by these nematodes and the development of molecular tools is helping to overcome obstacles to the study of cryptic organisms and complex interactions. Here we extend the repertoire of molecular tools to characterize soil food webs by describing primers/probe set to quantify certain free-living, bactivorous nematodes (FLBNs) that interact with EPNs in soil. Three FLBN isolates were recovered from soil baited with insect larvae. Morphological and molecular characterization confirmed their identities as Acrobeloides maximum (RT-1-R15C and RT-2-R25A) and Rhabditis rainai (PT-R14B). Laboratory experiments demonstrated the ability of these FLBNs to interfere with the development of Steinernema diaprepesi, Steinernema riobrave and Heterorhabditis indica parasitizing the weevil Diaprepes abbreviatus (P< 0.001), perhaps due to resource competition. A molecular probe was developed for the strongest competitor, A. maximum. We selected the highly conserved SSU rDNA sequence to design the primers/probe, because these sequences are more abundantly available for free-living nematodes than ITS sequences that can likely provide better taxonomic resolution. Our molecular probe can identify organisms that share ≥98% similarity at this locus. The use of this molecular probe to characterize soil communities from samples of nematode DNA collected within a citrus orchard revealed positive correlations (P< 0.01) between Acrobeloides-group nematodes and total numbers of EPNs (S. diaprepesi, H. indica and Heterorhabditis zealandica) as well as a complex of nematophagous fungi comprising Catenaria sp. and Monachrosporium gephyropagum that are natural enemies of EPNs. These relationships can be broadly interpreted as supporting Linford's hypothesis, i.e., decomposition of organic matter (here, insect cadavers) greatly increases bactivorous nematodes and their natural enemies. © 2012 Elsevier Inc.

Bonani J.P.,University of Sao Paulo | Fereres A.,Institute Ciencias Agrarias ICA | Garzo E.,Institute Ciencias Agrarias ICA | Miranda M.P.,Fundecitrus | And 2 more authors.
Entomologia Experimentalis et Applicata | Year: 2010

Detailed information on probing behavior of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is critical for understanding the transmission process of phloem-limited bacteria (Candidatus Liberibacter spp.) associated with citrus 'huanglongbing' by this vector. In this study, we investigated stylet penetration activities of D. citri on seedlings of Citrus sinensis (L.) Osbeck cv. Pêra (Rutaceae) by using the electrical penetration graph (EPG-DC system) technique. EPG waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration into plant tissues. The main waveforms were correlated with histological observations of salivary sheath termini in plant tissues, to determine the putative location of stylet tips. The behavioral activities were also inferred based on waveform similarities in relation to other Sternorrhyncha, particularly aphids and whiteflies. In addition, we correlated the occurrence of specific waveforms with the acquisition of the phloem-limited bacterium Ca. Liberibacter asiaticus by D. citri. The occurrence of a G-like xylem sap ingestion waveform in starved and unstarved psyllids was also compared. By analyzing 8-h EPGs of adult females, five waveforms were described: (C) salivary sheath secretion and other stylet pathway activities; (D) first contact with phloem (distinct from other waveforms reported for Sternorrhyncha); (E1) putative salivation in phloem sieve tubes; (E2) phloem sap ingestion; and (G) probably xylem sap ingestion. Diaphorina citri initiates a probe with stylet pathway through epidermis and parenchyma (C). Interestingly, no potential drops were observed during the stylet pathway phase, as are usually recorded in aphids and other Sternorrhyncha. Once in C, D. citri shows a higher propensity to return to non-probing than to start a phloem or xylem phase. Several probes are usually observed before the phloem phase; waveform D is observed upon phloem contact, always immediately followed by E1. After E1, D. citri either returns to pathway activity (C) or starts phloem sap ingestion, which was the longest activity observed. © 2009 The Netherlands Entomological Society.

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