Characterisation and bioactivity of oosporein produced by endophytic fungus Cochliobolus kusanoi isolated from Nerium oleander L.

Bioactive compounds comprising secondary metabolites produced by endophytic fungi have wide applications in pharmacology and agriculture. Isolation, characterisation and evaluation of biological activities of secondary metabolites were carried out from Cochliobolus kusanoi an endophytic fungus of Nerium oleander L. The fungus was identified based on 18S rDNA sequence analysis. There are no reports available on the compounds of C. kusanoi hence, antimicrobial metabolite produced by this fungus was extracted and purified by fractionation using hexane, diethyl ether, dichloromethane, ethyl acetate and methanol. Out of all the solvent fractions, the methanol fraction exhibited better antimicrobial activity which was further purified and characterised as oosporein. Oosporein from C. kusanoi exhibited broad spectrum in vitro antimicrobial, antioxidant and cytotoxic activities. The characterisation and antioxidant activity of oosporein from C. kusanoi are reported for the first time.

Análise multielementar em folhas de Nerium oleander L. usando fluorescência de raios X por dispersão em energia

Desde o início do século XX, a poluição do ar nos grandes centros piorou em consequência processo de industrialização e urbanização, juntamente com o rápido crescimento populacional e do transporte motorizado. Algumas espécies de plantas absorvem os poluentes atmosféricos pelas suas folhas e então, fixa-os em sua matriz, tornando-se assim um biomonitor de poluição nessa área. Assim, a análise foliar dessas espécies de vegetal pode ser usado como monitoramento ambiental. Uma das plantas que tem a habilidade de reter certos elementos químicos do ambiente e pode ser usada como biomonitor é a Nerium oleander L.. Neste estudo utilizou-se folhas de Nerium oleander L. para avaliar os níveis de poluição ambiental em uma sub-região da Região Metropolitana do Rio de Janeiro através da Fluorescência de Raios X (EDXRF). O sistema de EDXRF foi desenvolvido no próprio laboratório e consiste de um sistema portátil de XRF formado por um mini tubo raio X de baixa potência (anodo de Ag e operação em 20 kV/50 μA) e um detector de SiPIN. As amostras de Nerium oleander L. foram coletadas de plantas adultas. As amostras foram coletadas durante as quatros estações do ano (verão, outono, inverno e primavera). Todas as folhas foram coletadas a uma distância superior de 1,5 m em relação ao solo. As amostras foram acondicionadas em sacos plásticos e depois da chegada ao laboratório foram colocados sob refrigeração a 5 C. No laboratório, as amostras foram limpas com um pincel com cerdas macias para retirar a poeira. Depois disso, as amostras foram colocadas na estufa a 60 C por 48 h. Em seguida, as amostras foram pulverizadas (44 μm). Depois desse processo, alíquotas de 500 mg de massa foram prensadas a uma pressão de 2.32×108 por cerca de 15 minutos, afim de se obter pastilhas finas com diâmetro de 2,54 cm e densidade superficial de 100 mg/cm2. Foi possível detectar a concentração de 13 elementos: S, Cl, K, Ca, Mn, Fe, Cu, Zn, Br, Rb, Sr, Ba e Pb. A partir da concentração de cada elemento foram obtidos os mapas de distribuição elementar da área de estudo para cada estação. A análise da correlação de Pearson mostrou que existe uma correlação significativa entre os elementos Fe, Zn, Ba e Pb, entre os elementos Ca e Sr e entre os elementos Cl, K, Rb. A análise do PCA (Análise por Componentes Principais) mostrou que existem dois fatores principais da emissão de poluição ambiental: emissão por ressuspensão do solo (Cl, K, Ca, Mn, Rb e Sr) e emissões veiculares e industriais (Fe, Zn, Ba e Pb). O estudo da poluição ambiental através da técnica de EDXRF utilizando folhas de Nerium oleander L. como biomonitor se mostrou uma técnica de baixo custo e eficiência substancial na determinação da concentração elementar dos poluentes atmosféricos.

Fructose-1, 6-diphosphate (FDP) as a novel antidote for yellow oleander-induced cardiac toxicity: a randomized controlled double blind study

BACKGROUND: Cardiac toxicity due to ingestion of oleander plant seeds in Sri Lanka and some other South Asian countries is very common. At present symptomatic oleander seed poisoning carries a mortality of 10% in Sri Lanka and treatment of yellow oleander poisoning is limited to gastric decontamination and atropine administration. The only proven effective antidote is digoxin antibodies but these are not available for routine use because of the high cost. The main objective of this study is to investigate the effectiveness of a new and inexpensive antidote for patients with life threatening arrhythmias due oleander poisoning. METHOD/DESIGN: We set up a randomised double blind clinical trial to assess the effectiveness of Fructose 1, 6 diphosphate (FDP) in acute yellow oleander poisoning patients admitted to the adult medical wards of a tertiary hospital in Sri Lanka. Patients will be initially resuscitated following the national guidelines and eligible patients will be randomised to receive either FDP or an equal amount of normal saline. The primary outcome measure for this study is the sustained reversion to sinus rhythm with a heart rate greater than 50/min within 2 hours of completion of FDP/placebo bolus. Secondary outcomes include death, reversal of hyperkalaemia on the 6, 12, 18 and 24 hour samples and maintenance of sinus rhythm on the holter monitor. Analysis will be on intention-to-treat. DISCUSSION: This trial will provide information on the effectiveness of FDP in yellow oleander poisoning. If FDP is effective in cardiac glycoside toxicity, it would provide substantial benefit to the patients in rural Asia. The drug is inexpensive and thus could be made available at primary care hospitals if proven to be effective. TRIAL REGISTRATION: Current Controlled trial ISRCTN71018309.

Época de coleta e ácido indolbutírico no enraizamento de estacas de espirradeira (Nerium oleander L.)

A espirradeira (Nerium oleander L.) é uma importante espécie arbórea ornamental, muito utilizada no meio urbano. É propagada por estacas, porém a porcentagem de enraizamento é baixa e não há estudos sobre fatores que influenciam nesse processo. Este trabalho teve, portanto, o objetivo de estudar o efeito da época de coleta e do ácido indolbutírico (AIB) no enraizamento de estacas de duas variedades de espirradeira (Nerium oleander L.), determinadas pela coloração das flores (rosa e branca). O experimento foi instalado na UNESP, Campus de Jaboticabal/SP, no verão e no inverno. O delineamento experimental foi em blocos casualizados em esquema fatorial 2 x 2 x 4 (duas variedades combinadas com duas estações do ano e quatro concentrações de AIB - 0, 1.000, 2.000 e 4.000 mg kg-1). As avaliações foram realizadas 60 dias após a estaquia, sendo estas as variáveis: porcentagem de enraizamento, número médio, comprimento e massa de matéria seca de raízes. Concluiu-se que o enraizamento de ambas as variedades de espirradeira (rosa e branca) foi superior no verão. A variedade de flores rosas apresentou maior número e comprimento médio de raízes no verão, porém as maiores porcentagens de enraizamento e massa de matéria seca de raízes foram encontradas no inverno. O ácido indolbutírico foi efetivo para aumentar a porcentagem de enraizamento nas concentrações testadas de 1.000 e 2.000 mg kg-1; maior número, comprimento e massa de matéria seca de raízes foram obtidos na concentração de 2.000 mg kg-1.

Influence of hydric stress in the production of toxic principles in Nerium oleander L.

Many plants utilized in the urban center shows substances considered toxic whose production could be influenced by some factors, like hydric stress, including ornamental Nerium oleander L., widely used in gardens in various parts of the world, which presents production of cardioative glucosides, considered toxic. This study had the objective to evaluate the effect of field capacity in the biomass and the level of cardioative glucosides in seedlings of Nerium oleander. The experiment was carried at UNIDERP, in Campo Grande City, Mato Grosso do Sul State, Brazil, at the University for Development of State and Pantanal Region, using the experimental delineation in randomized blocks. There were 4 treatments (25%; 50%; 75% and 100% of the field capacity), 5 replications and 4 plants by parcel, totalling 80 plants. The evaluations were realized 60 days after the seedlings were planted. The quantitative analysis of the cardioative glucosides was realized by gravimetric test, after selective extraction of the glucosides. Were conclude that increase of the quantity of water in the soil raised the biomass production until 75% of the field capacity and increased the level of cardioative glucosides, showing that water management is very important and should be provided only the necessary to development of the plant.

Intoxicação experimental de cães com folhas verdes de Nerium oleander e uso da frutose 1,6 difosfato e da glicose como tratamentos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Host recognition by a polyphagous lepidopteran (Helicoverpa armigera): Primary host plants, host produced volatiles and neurosensory stimulation

An important question in the host-finding behaviour of a polyphagous insect is whether the insect recognizes a suite or template of chemicals that are common to many plants? To answer this question, headspace volatiles of a subset of commonly used host plants (pigeon pea, tobacco, cotton and bean) and nonhost plants (lantana and oleander) of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) are screened by gas chromatography (GC) linked to a mated female H. armigera electroantennograph (EAG). In the present study, pigeon pea is postulated to be a primary host plant of the insect, for comparison of the EAG responses across the test plants. EAG responses for pigeon pea volatiles are also compared between females of different physiological status (virgin and mated females) and the sexes. Eight electrophysiologically active compounds in pigeon pea headspace are identified in relatively high concentrations using GC linked to mass spectrometry (GC-MS). These comprised three green leaf volatiles [(2E)-hexenal, (3Z)-hexenylacetate and (3Z)-hexenyl-2-methylbutyrate] and five monoterpenes (α-pinene, β-myrcene, limonene, E-β-ocimene and linalool). Other tested host plants have a smaller subset of these electrophysiologically active compounds and even the nonhost plants contain some of these compounds, all at relatively lower concentrations than pigeon pea. The physiological status or sex of the moths has no effect on the responses for these identified compounds. The present study demonstrates how some host plants can be primary targets for moths that are searching for hosts whereas the other host plants are incidental or secondary targets.

Evaluating the efficacy of the EZ-Ject herbicide system in Queensland, Australia

The EZ-Ject herbicide system was evaluated as a stem injection method for controlling woody weeds in a range of situations where traditional chemical application methods have limited scope. The equipment was trialled on three Queensland weed species; pond apple (Annona glabra), velvety tree pear (Opuntia tomentosa) and yellow oleander (Cascabela thevetia); at five different cartridge densities (0, 1, 2, 3 and 4) and with two herbicides (glyphosate and imazapyr). Cartridges filled with imazapyr were significantly more effective at controlling the three woody weed species than those filled with glyphosate. Injecting plants with three imazapyr cartridges resulted in plant kills ranging from 93 to 100%, compared with glyphosate kills of 17 to 100%. Pond apple was the most susceptible species, requiring one imazapyr cartridge or two glyphosate cartridges to kill 97 and 92% of the treated plants. Plant mortality increased as the number of cartridges injected increased. Mortality did not differ significantly for treatments receiving three and four imazapyr cartridges, as these cartridge densities met the criterion of injecting one cartridge per 10-cm basal circumference, a criterion recommended by the manufacturers for treating large plants (>6.35 cm in diameter at breast height). The cost of treating a weed infestation of 1500 plants ha–1 with three cartridges per tree is $1070 ha–1, with labour costs accounting for 16% of the total. The high chemical costs would preclude this technique from broad-scale use, but the method could have application for treating woody weeds in sensitive, high conservation areas.

Stem injection: a control technique often overlooked for exotic woody weeds

In the rangelands of northern Australia, basal bark, cut stump, hand applied residual herbicides and foliar spraying have traditionally been the main herbicide techniques for control of individual exotic woody weeds growing within scattered to medium density infestations. In this paper we report on the preliminary results of stem injection as an alternate technique for the control of yellow oleander ( Cascabela thevetia (L.) Lippold), a woody weed that is difficult to kill. A randomised complete block experiment comprising 12 herbicide treatments (including a control) and three replicates was undertaken. Two rates of triclopyr + picloram, hexazinone, glyphosate, 2,4- D + picloram and metsufuron methyl and one rate of imazapyr were tested. At 15 months after application, triclopyr + picloram, glyphosate, 2,4-D + picloram and imazapyr all recorded high mortality (>90%) for at least one application rate. These results suggest that stem injection warrants further investigation as a control technique for other exotic woody weeds growing in rangelands.

Germinación de semillas de especies adaptadas a un medio extremófilo: Río Tinto

Se presentan los porcentajes de germinación de 12 especies representativas de la vegetación de ribera y matorral del ecosistema del río Tinto en diferentes condiciones de pH (5,4; 3,2; 1,8 y agua del río Tinto). Se observan distintas respuestas de germinación en las especies estudiadas, encontrando los mayores porcentajes de germinación en Nerium oleander y Salicornia ramosissima. Dada la respuesta positiva de estas dos especies, se discute su utilidad para la restauración de los territorios afectados en los alrededores del río Tinto. Para el resto de especies, se observan bajos porcentajes de germinación y en la mayoría de las especies analizadas el pH no influye en la germinación. Hay muchos factores que limitan la germinación de las semillas tales como la latencia que requiere de un pre-tratamiento (de frío o de calor), variación en las condiciones naturales que son difíciles de recrear en el laboratorio o la baja calidad de las semillas.

Strain variability in the DNA immigration control region (ICR) of Xylella fastidiosa

The genome of the bacterium Xylella fastidiosa contains four ORFs (XF2721, XF2725, XF2739 and XF0295) related to the restriction modification type I system, ordinarily named R-M. This system belongs to the DNA immigration control region (ICR). Each CIRF is related to different operon structures, which are homologues among themselves and with subunit Hsd R from the endonuclease coding genes. In addition, these ORFs are highly homologous to genes in Pseudomonas aeruginosa, Methylococcus capsulatus str. Bath, Legionella pneumophila, Helicobacter pylori, Xanthomonas oryzae pv. Oryzae and Silicibacter pomeroyi, as well as to genes from X. fastidiosa strains that infect grapevine, almond and oleander plants. This study was carried out on R-M ORFs from forty-three X. fastidiosa strains isolated from citrus, coffee, grapevine, periwinkle, almond and plum trees, in order to assess the genetic diversity of these loci through PCR-RFLP. PCR-RFLP analysis of the four ORFs related to the R-M system from these strains enabled the detection of haplotypes for these loci. When the haplotypes were defined, wide genetic diversity and a large range of similar strains originating from different hosts were observed. This analysis also provided information indicating differences in population genetic structures, which led to detection of different levels of gene transfer among the groups of strains. (c) 2005 Elsevier SAS. All rights reserved.

Aspectos biológicos, morfológicos e comportamentais de Aspidiotus nerii Bouché, 1833 (Hemiptera: Diaspididae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modeling the age-specific per capita growth and reproduction of Rhizobius lophanthae (Col: Coccinellidae)

A per capita model for the growth, development and reproduction of the coccinellid predator Rhizobius lophanthae (Blaisd) feeding on the oleander scale (Aspidiotus nerii Bouche (Homoptera: Diaspididae)) was developed. A thermal threshold for development of 9.4 degrees C was found. Under conditions of unlimited food, the relationship of mass at time t+1 to that at t (in days at 25 degrees C) suggests an 8.7 percent growth rate per mg larvae per day at 25 degrees C. An adult female beetle produces approximately 20 eggs per day while consuming an average of 8.5 scales/day. This is approximately 2.16 eggs per scale consumed above the maintenance level of 1.88 scales per day. More precisely, this compensation point is 0.12 mg of prey/mg of predator/day at 25 degrees C and the egestion rate is 1 - beta = 0.63.

Comparative analyses of the complete genome sequences of Pierce's disease and citrus variegated chlorosis strains of Xylella fastidiosa

Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X.fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grape-growing region of California. Comparative analyses with a previously sequenced X.fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X.fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X.fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.

Atividade inseticida e Modos de Ação de Extratos Vegetais Sobre Mosca Branca (Bemisia tabaci)

Insecticide potential and efficaciousness of vegetal extracts concentrations from six botanic families in three different modes were analyzed on the nymphs of the silverleaf whitefly Bemisia tabaci (Genn. 1889) on a tomato plantation Lycopersicon esculentum Mill. Nymph breeding was kept in cherry tomato shrubs. Test solutions were obtained from successive extractions with ethanol and evaporation in a rotary evaporator. Tests consisted of 10 solutions in four concentrations (500; 1,000; 1,500 and 3,000 μg/mL) and control treatment (distilled water + 1% DMSO) under three different modes of activity, namely, contact, translaminar and systemic. Experimental design was totally randomized with four repetitions and eleven treatments. Means were compared by Scott-Knott test at 5% probability and lethal concentrations (CL50) were calculated by POLO. In the case of contact activity mode, all extracts had efficiency above 50% with the highest concentration (3,000 μg/mL) in which the highest efficiency for all extracts tested was verified. There was a need for dose increase in translaminar activity mode for similar results, whereas in the case of systemic activity the best performance was obtained with extracts of Nerium oleander, Derris amazonica and Ipomoea carnea. Extracts of the last two caused a higher percentage when compared with that of other extracts in all activity modes lower than CL50.