Chromosomal organization of the 18S and 5S rRNAs and histone H3 genes in Scarabaeinae coleopterans: insights into the evolutionary dynamics of multigene families and heterochromatin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chromosomal organization and evolutionary history of Mariner transposable elements in Scarabaeinae coleopterans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

若尔盖湿地3种两栖类的种间竞争及畜牧业对其食性的影响

从空间、时间、食物3个方面研究了若尔盖湿地3种两栖类的种间竞争，运用生态位理论探讨了3种两栖类利用环境资源的能力以及它们之间的共存模式，研究了3种两栖类年周期食性，并考察了畜牧业对3种两栖类食性及食物竞争格局的影响；此外，还通过实验室研究对2种两栖类幼体的种间竞争策略进行了考察。主要结果如下： 1、两栖类的空间资源利用状况：在3种两栖类成体生态位宽度的比较中，岷山蟾蜍（Bufo minshanicus）成体在牧场性质(0.41)、牛粪数量(0.42)、植被盖度(0.20)、地表温度(0.50)、地表湿度(0.51) 5个维度上的生态位宽度最窄；而倭蛙（Nanorana pleskei）成体在牛粪数量(0.81)、植被高度(0.63)、植被盖度(0.47)、小水体距离(0.68) 4个维度上的生态位宽度最宽。在3种两栖类亚成体生态位宽度的比较中，岷山蟾蜍亚成体在牧场性质(0.66)、牛粪数量(0.58)、植被高度(0.64)、小水体距离(0.51)、地表湿度(0.79) 5个维度的上生态位宽度最宽；倭蛙亚成体在牧场性质(0.39)、牛粪数量(0.30)、地表温度(0.18)、地表湿度(0.33) 4个维度上的生态位宽度最窄。高原林蛙（Rana kukunoris）在地表温度（成体：0.62；亚成体：0.56）、地表湿度（成体：0.84；亚成体：0.60）两个维度上具有较大的生态位宽度值，而在小水体距离维度上（成体：0.27；亚成体：0.14）的生态位宽度值则很小。比较3种无尾两栖类在不同生长阶段（成体、亚成体）的生态位宽度，发现高原林蛙和倭蛙的亚成体对栖息环境的要求更高。3种两栖类空间资源利用的相似程度很高，高原林蛙与倭蛙之间的生态重叠度(0.87)较之它与岷山蟾蜍(0.81)的生态位重叠度更大。 2、两栖类的日活动节律：高原林蛙成体、亚成体、岷山蟾蜍亚成体活动的最低气温为0℃、2℃、8℃；岷山蟾蜍和高原林蛙亚成体出现的数量与气温成极显著的正相关（r=0.797, p<0.001；r=0.794, p<0.001），高原林蛙成体出现的数量与气温有一定相关性（r=0.456, p<0.05）；晴天时两栖类的活动性明显高于阴天（p<0.001）；多云转晴天气，高原林蛙和岷山蟾蜍亚成体出现两次日活动高峰，分别为中午12:30左右和下午15:30～16:30之间；多云天气，高原林蛙和岷山蟾蜍亚成体出现两次日活动高峰，分别为9:30～10:30之间和15:30～16:30之间。 3、两栖类的食物资源利用状况：春、秋两季，高原林蛙最主要的食物是蜉金龟科(Aphodiidae)昆虫，相对重要性指数(IRI)最高（春季：35.28%，秋季：28.57%），其次为昆虫的幼虫，以及双翅目的毛蚊科(Bibionidae)、蝇科(Muscidae)、丽蝇科(Calliphoridae)昆虫，秋季，蝗虫是高原林蛙食物组成中的重要部分；岷山蟾蜍最主要的食物是蚂蚁(IRI，春季：85.54%，秋季：49.70%)，其次为蜉金龟科、象甲科(Curculionidae)、步甲科(Carabidae)、粪金龟科(Geotrupidae) 等鞘翅目昆虫；倭蛙春季的最主要食物也是蜉金龟科昆虫(IRI，春季：13.41%)，其次为蚂蚁、毛蚊科昆虫、昆虫的幼虫以及狼蛛科(Lycosidae)。3种两栖类中，倭蛙的食性生态位宽度相对较宽（0.43），而岷山蟾蜍（0.09）和高原林蛙（0.22）的生态位宽度较窄，与春季相比，两栖类在秋季的食谱更宽。以利用食物种类为标准，春季高原林蛙与倭蛙的生态位重叠度(0.40)比它与岷山蟾蜍的生态位重叠度（0.33）更大。 4、畜牧业对两栖类食性及食物竞争格局的影响：以藏牦牛粪为食物或寄居场所的昆虫，如蜉金龟科、粪金龟科、毛蚊科、蝇科、丽蝇科昆虫和某些昆虫幼虫，是3种两栖类食物谱中最主要的组成部分，蜉金龟科昆虫在高原林蛙食谱中的比例更高，高原林蛙可能从畜牧业发展中获得更多的好处，使之在食物竞争方面处于优势地位。与无放牧样地相比，在有放牧样地的中，两栖类食谱中的蜉金龟科昆虫数量更多(有放牧：31.94%；无放牧：21.32%)、出现频率更高(有放牧：76.38%；无放牧：44%)。然而在不同样地上（有放牧/无放牧），两栖类的食物组成无显著性差异(P=0.188)，两栖类的数量(P=0.075)、肥满度(P=0.537)均没有显著差别。 5、两栖类幼体的竞争策略：实验室条件下，通过活动性水平，变态时的体重、增长率和完成变态所需时间考察自然条件下常同水塘分布的中华蟾蜍（Bufo gargarizans）和高原林蛙蝌蚪的竞争策略。结果表明：中华蟾蜍蝌蚪在不同食物资源条件下，所选择的生存策略可能不同，即食物资源充足时，增加活动性获取更多食物，食物资源有限时，降低活动性且提前完成变态；与中华蟾蜍蝌蚪相比，在食物资源有限时高原林蛙蝌蚪获取食物能力可能更强。 This paper presented the study of competition of three amphibians (Rana kukunoris, Nanorana pleskei, Bufo minshanicus) based on spatial, temporal and dietary scales in Zoige wetland. We measured coexistence patterns of three amphibians and analyzed their ability of exploiting resource. Effects of grazing on the diet composition and diet competition of amphibians were analyzed by their diet composition during spring and autumn. Furthermore, we examined the competitive ability of larval common frogs (Rana kukunoris)and common toads(Bufo gargarizans) in a laboratory experiment, and analyzed their competitive strategies respectively. The results were as follows: 1 .The status of using spatial resource Niche breadths of B. minshanicus adults on 5 dimensional axes including character of pasture(0.41), number of yaks dung(0.42), vegetation coverage(0.20), temperature (0.50)and humidity(0.51) of ground surface were narrower than adults of R. kukunoris and N. pleskei. Niche breadths of B. minshanicus subadults were broader than R.kukunoris subadults and N.pleskei subadults on 5 dimensional axes including character of pasture (0.66), number of yaks dung (0.58), vegetation height (0.64), distance to small waterbodies (0.51), humidity of ground surface (0.79). Niche breadths of N. pleskei subadults were the narrowest in three anurans subadults on 4 dimensional axes including character of pasture (0.39), number of yaks dung (0.30), temperature (0.18) and humidity (0.33) of ground surface, niche breadths of N. pleskei adults were the broadest in three anurans adults on 4 dimensional axes including number of yaks dung (0.81), vegetation height (0.63) and coverage(0.47), distance to small waterbodies(0.68).Comparatively, niche breadths of R. kukunoris were broader on the two microclimate factors including temperature(adults：0.62；subadults：0.56) and humidity (adults：0.84；subadults：0.60)of ground surface, but was narrow on distance to small waterbodies(adults：0.27；subadults：0.14). Strategies for using habitat resource of adults and subadults of the three species anuran were different. Generally, subadults of R. kukunoris and N. pleskei needs better habitat condition. It was quite similar that three anurans exploited spatial resource, Niche overlap between R. kukunoris and N. pleskei (0.87) was greater than that between R. kukunoris and B.minshanicus(0.81). 2.Daily activity rhythm R. kukunoris audlts were active when air temperatures were as low as 0℃, R. kukunoris subadults were active at 2℃, B.minshanicus subaudlts were active at 8℃. Positive correlation was found between activities of amphibians and air temperature, Subadults of R.kukunoris, (r=0.797, p<0.001), Subadults,of,B.minshanicus, (r=0.794, p<0.001), andbadults,of,R.kukunoris(r=0.456, p<0.05).Amphibians were more active during sunny days than cloudy days. In cloudy turning into sunny, R. kukunoris and B.minshanicus subadults had two active peak: at noon about 12:30 and 15:30～16:30 pm; in cloudy, R. kukunoris and B.minshanicus subadult had two active peak too : 9:30～10:30am,15:30～16:30pm. 3.Diet analysis Aphodiidae was the most commonly consumed food item by R. kukunoris based on index of relative importance (IRI) during spring (35.28%) and autumn (28.57%) in Zogie wetland. Besides Aphodiidae, larval insect, dipterans such as Bibionidae, Muscidae, Calliphoridae also were important food item for R. kukunoris, in autumn, locust was one of important food item for R. kukunoris. The most important food item for B.minshanicus during spring (IRI:85.54%) and autumn (IRI:49.70%) was ants, following, was coleopterans, such as Aphodiidae, dung beetle. Aphodiidae (IRI:13.41%) were the most important consumed food item by N. pleskei during spring too, following, was ants and Bibionidae. Dietary breadth of N. pleskei (0.43) were greater than R. kukunoris (0.22) and B. minshanicus (0.09). As a whole, Dietary breadth of amphibians during aurumn were greater than spring. Based on prey item, dietary overlap between R. kukunoris and N. pleskei (0.40) was greater than that between R. kukunoris and B.minshanicus (0.33) during spring. 4.Effects of grazing on the diet composition and diet competition of amphibians Amphibians are an important part of the pasture ecosystems as prey and predator. In Zogie wetland, major diet of amphibians was closely associated with dung of yaks, for example, Aphodiidae, Bibionidae, Muscidae, dung beetle. Dung of yaks was major diet and habitat of these insects. Proportion of Aphodiidae was higher in diet composition of R. kukunoris than N. pleskei and B.minshanicus, with development of pasturage, R. kukunoris may have a diet competitive advantage over N. pleskei and B.minshanicus. Number of Aphodiidae in diet composition of amphibians was higher in samples with grazing (31.94%) than in those without grazing (21.32%). Occurrence Frequency of Aphodiidae in diet composition of amphibians was higher in samples with grazing (76.38%) than in those without grazing (44%). However, There was not significantly different on diet composition (P=0.188), and number (P=0.075) and the relative fatness (P=0.537) of amphibians between grazing samples and without grazing. 5.Competitive strategies of amphibian larvae I examined the competitive ability of larval toads (Bufo gargarizans) and frogs (Rana kukunoris) which co-occur in the nature pond by activity level, the growth rate and mass at metamorphosis and larval period in a laboratory experiment. The results suggest: In laborary, B.gargarizans adapted himself to different food level by changing activity. At high food level, B. gargarizans increased activity to gain more diet. At low food level, B. gargarizans decreased activity and achieved early metamorphosis. When food resource was limit, R. kukunoris could gain more food than B. gargarizans.

The effect of farming system and fertilisers on pests and natural enemies: A synthesis of current research

Organic agriculture is becoming widespread due to increased consumer demand and regulatory and political support. Organic agriculture can increase arthropod diversity but the response of pests and their natural enemies is variable. Fertiliser is an important component of agricultural systems and its effects on pests and natural enemies will influence agroecosystems. In this study, meta-analysis and vote-counting techniques were used to compare farming system (organic and conventional) and fertiliser effects on arthropod pests and their natural enemies. The meta-analyses indicated that pests generally benefitted from organic techniques, this is particularly evident when experiments were carried out on a smaller scale. Pest responses to organic and conventional fertiliser types were divergent, plant composts benefitted pest arthropods while the opposite was true for manures, this has implications for pest management. Most natural enemy groups responded positively to organic farming although this was not true for Coleopterans. Experimental scale had a prominent impact on natural enemy responses with farm scale studies showing particularly positive effects of organic agriculture on natural enemies. This suggests that it is large scale features of organic agriculture such as landscape heterogeneity that are beneficial to natural enemies. Natural enemy responses to organic fertilisers were positive indicating that field scale management practices including fertiliser can also be important in pest management.

Identificação de genes cry de Bacillus thuringiensis no controle de Sphenophorus levis, o bicudo da cana-de-açúcar

A bactéria B. thuringiensis caracteriza-se pela produção de proteínas tóxicas a representantes de diversas ordens de insetos, as quais são codificadas por genes cry. Este trabalho foi realizado com objetivo de selecionar isolados de B. thuringiensis, por meio da caracterização morfológica e molecular, identificando as diferentes subclasses dos genes cry3 e cry35 e determinar a patogenicidade contra Sphenophorus levis, uma das mais importantes pragas da cultura da cana-de-açúcar. Foram utilizados 1163 isolados de B. thuringiensis e com a observação em microscópio com contraste de fases foram confirmadas como pertencentes à espécie de B. thuringiensis. O material genético foi purificado pela matriz de troca iônica Instagene Matrix e submetido a PCR com iniciadores gerais cry3 e cry35 identificando-se 30 isolados contendo genes com potencial para o controle de coleópteros, os quais juntamente com as linhagens-padrão de B. thuringiensis var. tenebrionis, B. thuringiensis var. morrissone e B. thuringiensis var. tolworthi foram utilizados para a realização do bioensaio. Através de análise discriminante alocaram-se os isolados em quatro grupos quanto à toxicidade de B. thuringiensis. Os grupos ficaram assim definidos: um grupo que promovem até 10% de mortalidade contendo as testemunhas e duas linhagens;um grupo que causou 39% de mortalidade contendo três linhagens padrão e dez isolados; um grupo com 52% de mortalidade contendo treze isolados e um grupo com 70% de mortalidade contendo cinco isolados, os quais devem ser considerados promissores no controle biológico de S. levis.

Caracterização e seleção de isolados de Bacillus thuringiensis efetivos contra Sitophilus oryzae L., 1763

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização molecular e seleção de isolados de Bacillus thuringiensis com potencial inseticida para Sphenophorus levis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ecologia de dois grupos de sagüis-brancos, mico argentatus (Linnaeus 1771) em um fragmento florestal natural, Santarém-Pará

Este trabalho apresenta os resultados de um estudo de longo-prazo sobre ecologia e comportamento de dois grupos silvestres de Mico argentatus em um fragmento florestal natural, isolado por uma matriz de vegetação de savana, na região de Alter do Chão, Amazônia central. Os grupos, denominados Cm1 e Cm2 foram habituados e monitorados durante 12 meses em 2000 e quatro meses em 2001 (Cm1) e sete meses em 2000 e três meses em 2001 (Cm2). Análises de variação temporal envolveram três períodos sazonais distintos (Chuvosa-Início. Chuvosa-Final e Seca) em 2000, e a comparação longitudinal dos meses julho e setembro em 2000-2001. Análises das relações entre as variáveis ecológicas (abundância de principais recursos: frutos e insetos) e os padrões ecológicos e comportamentais entre os grupos foram estabelecidos. Os grupos apresentaram composições e tamanhos diferentes, que variaram entre 4-8 e 8-11 membros, em Cm1 e Cm2, respectivamente. Os dados quantitativos foram coletados utilizando-se a amostragem de varredura instantânea com amostras de um minuto de duração a cada intervalo de cinco minutos, durante todo o período de atividade diária do grupo. Os resultados mostraram que ambos os grupos gastam mais da metade do tempo em forrageio e alimentação. A dieta foi constituída de frutos, néctar, gomas e presa animal (invertebrados, principalmente ortópteros, formicídeos e coleópteros, além de lagartos e anfíbios). O tamanho total da área dos grupos Cm1 e Cm2 em 2000 foram respectivamente, 11,5 ha e 14,6 ha. As comparações sazonais mostraram uma similaridade entre grupos na alocação sistemática de mais tempo ao comportamento de forrageio e alimentação na medida em que os recursos não gomíferos se tornavam mais escassos. Já a análise longitudinal, além de apresentar uma redução de 30% na precipitação em 2001, mostrou uma mudança considerável na composição e distribuição espacial dos dois grupos. Assim, diferentes aspectos da comparação entre grupos pareceram refletir a influência destes diferentes fatores, com padrões opostos nos dois grupos. O aspecto geral mais importante parece ter sido o registro de padrões bem diferentes, além de similaridades importantes em grupos que ocupam quase o mesmo espaço, e a variabilidade das condições do hábitat entre as estações e entre os anos. As estratégias comportamentais identificadas pareceram ser reflexo, tanto da variabilidade e flexibilidade comportamental já conhecidas para os calitriquídeos, como de condições ecológicas impostas pela variabilidade na disponibilidade dos recursos num fragmento reduzido.

Diet Seasonality and Feeding Preferences of Brachycephalus pitanga (Anura: Brachycephalidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Recombinant expression, localization and in vitro inhibition of midgut cysteine peptidase (Sl-CathL) from sugarcane weevil, Sphenophorus levis

A cDNA coding for a digestive cathepsin L, denominated Sl-CathL, was isolated from a cDNA library of Sphenophorus levis larvae, representing the most abundant EST (10.49%) responsible for proteolysis in the midgut. The open reading frame of 972 bp encodes a preproenzyme similar to midgut cathepsin L-like enzymes in other coleopterans. Recombinant Sl-CathL was expressed in Pichia pastoris, with molecular mass of about 42 kDa. The recombinant protein was catalytically activated at low pH and the mature enzyme of 39 kDa displayed thermal instability and maximal activity at 37 degrees C and pH 6.0. Immunocytochemical analysis revealed Sl-CathL production in the midgut epithelium and secretion from vesicles containing the enzyme into the gut lumen, confirming an important role for this enzyme in the digestion of the insect larvae. The expression profile identified by RT-PCR through the biological cycle indicates that Sl-CathL is mainly produced in larval stages, with peak expression in 30-day-old larvae. At this stage, the enzyme is 1250-fold more expressed than in the pupal fase, in which the lowest expression level is detected. This enzyme is also produced in the adult stage, albeit in lesser abundance, assuming the presence of a different array of enzymes in the digestive system of adults. Tissue-specific analysis revealed that Sl-CathL mRNA synthesis occurs fundamentally in the larval midgut, thereby confirming its function as a digestive enzyme, as detected in immunolocalization assays. The catalytic efficiency of the purified recombinant enzyme was calculated using different substrates (Z-Leu-Arg-AMC, Z-Arg-Arg-AMC and Z-Phe-Arg-AMC) and rSl-CathL exhibited hydrolysis preference for Z-Leu-Arg-AMC (k(cat)/K-m = 37.53 mM S-1), which is similar to other insect cathepsin L-like enzymes. rSl-CathL activity inhibition assays were performed using four recombinant sugarcane cystatins. rSl-CathL was strongly inhibited by recombinant cystatin CaneCPI-4 (K-i = 0.196 nM), indicating that this protease is a potential target for pest control. (C) 2011 Elsevier Ltd. All rights reserved.

Evaluation of the pathogenicity of multiple isolates of Beauveria bassiana (Hypocreales: Clavicipitaceae) on Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) for the assessment of a solid formulation under simulated field conditions

A solid state formulation of Beauveria bassiana (Balsamo) Vuillemin has been developed for biological control of the Red Palm Weevil (RPW), Rhynchophorus ferrugineus (Olivier, 1790). Two kinds of bioassays (dry conidia and dipping) using 10 isolates from several coleopterans in Mediterranean environments, identified 2 RPW derived isolates (193 and 203) as most pathogenic to RPW larvae and adults (zero survival within first 4–5 d for dry conidia, and 14 and 23 d for dipping bioassays). Isolate 203 (5.1 × 108 ± 1.9 × 108 conidia g-1) was formulated with fragmented date seed into solid granules and tested in palms infested with RPW under semi-field conditions in Feb, Apr/May and Jun of both 2007 and 2008. Beauveria bassiana significantly reduced RPW adult survival with respect to controls in May 2007 and in the Apr/Jun 2008 experiments. Total RPW adult mortality was achieved within 30 days for all B. bassiana treatments, and was associated with increasing numbers of insects with signs of mycosis in 2008 experiments. Beauveria bassiana formulation reduced RPW multiplication in artificially infested palms compared to controls, and a positive correlation between numbers of larvae and time post-infestation was recorded. The suppression of RPW adult populations by B. bassiana persisted for at least 3 months under semi-field conditions. The Beauveria bassiana solid formulation, which induces great adult mortality and persistence in the field, could be applied as a preventive as well as a curative treatment for the integrated management of RPW.

Arthropods associated with the crown of Mauritia flexuosa (Arecaceae) palm trees in three different environments from Brazilian Cerrado

Artrópodos associados à copa de árvores, principalmente palmeiras, são pouco conhecidos no Cerrado. Para descrever a estrutura da comunidade de artrópodos à copa de Mauritia flexuosa (Arecaceae) foram amostradas 150 palmeiras em seis veredas” do Distrito Federal, de áreas silvestres, rurais e periurbanas na estação chuvosa. Os artrópodos presentes nos ninhos abandonados de aves, refúgios de mamíferos, folhas e matéria orgânica foram coletados manualmente, fixados em etanol 70% e separados em ordem, família, morfoespécie e guildas alimentares. As características das palmeiras medidas foram altura da estipe, diâmetro da copa, número de folhas e de ninhos de aves nas palmeiras. Foram coletados 3.862 indivíduos, pertencentes a 15 ordens, 45 famílias e 135 morfoespécies. As ordens mais abundantes foram Coleoptera (28,6%), Blattodea (21,8%), Collembola (11,4%) e Hemiptera (10,2%). As famílias Blaberidae, Tenebrionidae, Entomobryidae, Reduviidae, Oniscidae, Staphylinidae, Carabidae e Formicidae representaram 82,1% de todos os indivíduos coletados. A maioria das morfoespécies foi pouco abundante, 71 (52,6%) apresentaram uma abundância média igual ou menor que 1 indivíduo/palmeira. Coleoptera compreendeu o maior número de morfoespécies (43,7%) seguida de Araneae (20,0%). A análise das guildas alimentares mostrou prevalência de predadores e hematófagos (36,0%). A riqueza e a abundância de artrópodos foram menores no ambiente periurbano. O número de ninhos de aves apresentou correlação positiva com abundância e riqueza, o que não ocorreu com as medidas das palmeiras. A importância de M. flexuosa para a manutenção da artropodofauna nas “veredas” no bioma Cerrado é discutida. _______________________________________________________________________________________ ABSTRACT