Permanent Genetic Resources added to Molecular Ecology Resources Database 1 May 2009-31 July 2009

This article documents the addition of 512 microsatellite marker loci and nine pairs of Single Nucleotide Polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Alcippe morrisonia morrisonia, Bashania fangiana, Bashania fargesii, Chaetodon vagabundus, Colletes floralis, Coluber constrictor flaviventris, Coptotermes gestroi, Crotophaga major, Cyprinella lutrensis, Danaus plexippus, Fagus grandifolia, Falco tinnunculus, Fletcherimyia fletcheri, Hydrilla verticillata, Laterallus jamaicensis coturniculus, Leavenworthia alabamica, Marmosops incanus, Miichthys miiuy, Nasua nasua, Noturus exilis, Odontesthes bonariensis, Quadrula fragosa, Pinctada maxima, Pseudaletia separata, Pseudoperonospora cubensis, Podocarpus elatus, Portunus trituberculatus, Rhagoletis cerasi, Rhinella schneideri, Sarracenia alata, Skeletonema marinoi, Sminthurus viridis, Syngnathus abaster, Uroteuthis (Photololigo) chinensis, Verticillium dahliae, Wasmannia auropunctata, and Zygochlamys patagonica. These loci were cross-tested on the following species: Chaetodon baronessa, Falco columbarius, Falco eleonorae, Falco naumanni, Falco peregrinus, Falco subbuteo, Didelphis aurita, Gracilinanus microtarsus, Marmosops paulensis, Monodelphis Americana, Odontesthes hatcheri, Podocarpus grayi, Podocarpus lawrencei, Podocarpus smithii, Portunus pelagicus, Syngnathus acus, Syngnathus typhle,Uroteuthis (Photololigo) edulis, Uroteuthis (Photololigo) duvauceli and Verticillium albo-atrum. This article also documents the addition of nine sequencing primer pairs and sixteen allele specific primers or probes for Oncorhynchus mykiss and Oncorhynchus tshawytscha; these primers and assays were cross-tested in both species.

Legume type and temperature effects on the toxicity of insecticide to the genus Callosobruchus (Coleoptera: Bruchidae)

Three bruchid pest species, Callosobruchus maculatus, Callosobruchus chinensis and Callosobruchus rhodesianus, were studied for their response to insecticide toxicity taking into account the separate and interactive effects of temperature and pre-adult food. The food types used were cowpea (Vigna unguiculata) and mungbean (Vigna radiata). Callosobruchus maculatus was the most tolerant to malathion and the least affected by temperature change while C. rhodesianus was the least tolerant. Over a 4 C range (23, 25, 27 C), there was generally a significant impact of temperature on the tolerance of the three species to the insecticide. The food type on which the insects developed influenced considerably the degree of insecticide tolerance. Callosobruchus maculatus and C. chinensis populations reared onmungbean had higher tolerance to malathion than their counterparts reared on cowpea, but the opposite was observed in C. rhodesianus populations. The food influence in this study suggested an ancestral cause or fitness cost depending on the species. The interaction of food-by-temperature had no significant effect on malathion toxicity to this genus. Correlation analysis showed C. chinensis to be relatively less sensitive to insecticide concentration over the range studied compared with the other two species.

Variation in the sensitivity of Callosobruchus (Coleoptera: Bruchidae) acetylcholinesterase to the organophosphate insecticide malaoxon: effect of species, geographical strain and food type

BACKGROUND: Bruchid beetles, Callosobruchus species, are serious pests of economically important grain legumes; their activity in stores is often controlled by use of synthetic insecticides. Esterases are known to be involved in insecticide resistance in insects. However, there is dearth of information on esterase activity in the genus Callosobruchus. In this study we investigated the effect of species, geographical strain and food type on the variation of acetylcholinesterase (AChE) activity and its inhibition by malaoxon (malathion metabolite) using an in vitro spectrophotometric method. RESULT: AChE activity varied significantly among species and strains and also among legume type used for rearing them. Generally irrespective of species, strain or food type, the higher the AChE activity of a population, the higher its inhibition by malaoxon. C. chinensis had the highest AChE activity of the species studied and in the presence of malaoxon it had the lowest remaining AChE activity, while C. rhodesianus retained the highest activity. CONCLUSION: A firsthand knowledge of AChE activity in regional Callosobruchus in line with the prevailing food types should be of utmost importance to grain legume breeders, researchers on plant materials for bruchid control and pesticide manufacturer/applicators for a robust integrated management of these bruchids.

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 May 2009-31 July 2009

This article documents the addition of 512 microsatellite marker loci and nine pairs of Single Nucleotide Polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Alcippe morrisonia morrisonia, Bashania fangiana, Bashania fargesii, Chaetodon vagabundus, Colletes floralis, Coluber constrictor flaviventris, Coptotermes gestroi, Crotophaga major, Cyprinella lutrensis, Danaus plexippus, Fagus grandifolia, Falco tinnunculus, Fletcherimyia fletcheri, Hydrilla verticillata, Laterallus jamaicensis coturniculus, Leavenworthia alabamica, Marmosops incanus, Miichthys miiuy, Nasua nasua, Noturus exilis, Odontesthes bonariensis, Quadrula fragosa, Pinctada maxima, Pseudaletia separata, Pseudoperonospora cubensis, Podocarpus elatus, Portunus trituberculatus, Rhagoletis cerasi, Rhinella schneideri, Sarracenia alata, Skeletonema marinoi, Sminthurus viridis, Syngnathus abaster, Uroteuthis (Photololigo) chinensis, Verticillium dahliae, Wasmannia auropunctata, and Zygochlamys patagonica. These loci were cross-tested on the following species: Chaetodon baronessa, Falco columbarius, Falco eleonorae, Falco naumanni, Falco peregrinus, Falco subbuteo, Didelphis aurita, Gracilinanus microtarsus, Marmosops paulensis, Monodelphis Americana, Odontesthes hatcheri, Podocarpus grayi, Podocarpus lawrencei, Podocarpus smithii, Portunus pelagicus, Syngnathus acus, Syngnathus typhle,Uroteuthis (Photololigo) edulis, Uroteuthis (Photololigo) duvauceli and Verticillium albo-atrum. This article also documents the addition of nine sequencing primer pairs and sixteen allele specific primers or probes for Oncorhynchus mykiss and Oncorhynchus tshawytscha; these primers and assays were cross-tested in both species.

MICROPROPAGAÇÃO do KIWI CV. HAYWARD

O presente trabalho teve como objetivo desenvolver um protocolo para a obtenção de mudas de kiwi (Actinidia chinensis Planch), cv. Hayward, por meio do cultivo in vitro de cotilédones. Utilizou-se o meio de MURASHIGE & SKOOG (1962) -- MS, suplementado com dois tipos de auxina (AIA e AIB) e uma citocinina (BAP). Foram verificados os efeitos de três doses de auxinas (0,125; 0,250 e 0,375 mg.L-1), combinadas com três doses de citocinina (0,5; 1,0 e 1,5 mg.L-1) na capacidade morfogênica dos explantes. Procedeu-se o estudo histológico dos órgãos das plântulas obtidas in vitro, e verificou-se, também, a capacidade de aclimatação das mudas ex vitro. A menor dose de AIB (0,125 mg.L-1), independentemente das doses de BAP, foi a mais eficaz na morfogênese dos explantes. Não foram verificadas alterações histológicas e anatômicas das plântulas obtidas in vitro. Aos três meses após o cultivo ex vitro dos explantes, verificou-se a sobrevivência de 88% das plantas transplantadas em condições de campo.

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 May 2009-31 July 2009

This article documents the addition of 512 microsatellite marker loci and nine pairs of Single Nucleotide Polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Alcippe morrisonia morrisonia, Bashania fangiana, Bashania fargesii, Chaetodon vagabundus, Colletes floralis, Coluber constrictor flaviventris, Coptotermes gestroi, Crotophaga major, Cyprinella lutrensis, Danaus plexippus, Fagus grandifolia, Falco tinnunculus, Fletcherimyia fletcheri, Hydrilla verticillata, Laterallus jamaicensis coturniculus, Leavenworthia alabamica, Marmosops incanus, Miichthys miiuy, Nasua nasua, Noturus exilis, Odontesthes bonariensis, Quadrula fragosa, Pinctada maxima, Pseudaletia separata, Pseudoperonospora cubensis, Podocarpus elatus, Portunus trituberculatus, Rhagoletis cerasi, Rhinella schneideri, Sarracenia alata, Skeletonema marinoi, Sminthurus viridis, Syngnathus abaster, Uroteuthis (Photololigo) chinensis, Verticillium dahliae, Wasmannia auropunctata, and Zygochlamys patagonica. These loci were cross-tested on the following species: Chaetodon baronessa, Falco columbarius, Falco eleonorae, Falco naumanni, Falco peregrinus, Falco subbuteo, Didelphis aurita, Gracilinanus microtarsus, Marmosops paulensis, Monodelphis Americana, Odontesthes hatcheri, Podocarpus grayi, Podocarpus lawrencei, Podocarpus smithii, Portunus pelagicus, Syngnathus acus, Syngnathus typhle,Uroteuthis (Photololigo) edulis, Uroteuthis (Photololigo) duvauceli and Verticillium albo-atrum. This article also documents the addition of nine sequencing primer pairs and sixteen allele specific primers or probes for Oncorhynchus mykiss and Oncorhynchus tshawytscha; these primers and assays were cross-tested in both species.

Potencial alelopático de extrato etanólico de Anadenanthera macrocarpa e Astronium graveolens

The objective of this study was to evaluate the allelophatic potential ethanol extract of leaves of Anadenanthera macrocarpa (Benth.) Brenan (angico) and Astronium graveolens Jacq (aroeira) on the biomonitor germination of Brassica chinensis (Malaysia's cabbage) and Lactuca sativa (lettuce). For the test of allelopathy was the bioassay of germination of B. chinensis and L. sativa using solutions of ethanol extracts in different concentrations (5, 10 and 20 mg / mL) with continuous light for 11 days. The experimental and control groups were divided into the Petri dish containing 50 seeds of B. chinensis and L. sativa each. Were performed six replicates for each concentration of extract of leaves of A. macrocarpa and A. graveolens and the negative control (water). The rolling action of the treatments was performed by quantitative observation and monitoring of germination every six hours. The results show that the ethanol extract of A. macrocarpa and A. graveolens presents allelopathic potential, regardless of the concentration used, but with answers dose-dependent, and thus able to interfere directly in germination, average speed and time of germination biomonitor of B. chinensis and L. sativa.

Efeito de auxinas e boro no enraizamento de estacas caulinares de kiwi retiradas em diferentes épocas

This work was carried out to study the effects of some synthetical auxins and boron on the rooting of stem cuttings of kiwi (Actinidia deliciosa Pl. cv. Tomuri). Cuttings of semi-woody stems with two knots and leaves divided in two, with approximately 10 cm of length were utilized. The base of the cuttings received the following treatments: 1) water only; 2) NAA 300 ppm; 3) IBA 300 ppm; 4) NAA 300 ppm + B; 5) IBA 300 ppm + B; 6) NAA 0,5%-talc and 7) IBA 0,5%-talc. After these treatments, the stems were placed in suitable rooting dishes, with pure vermiculite in misty nebulization chamber for 120 days. Evaluations were made based on the following observations: percentage of rooted stem cuttings; reductor sugar and total sugar analyses and tryptophan analyses. The results showed that the autumm season is the best for rooting for kiwi stem cuttings. The exogenous application of 0.5% of IBA talc on the bases of the cuttings showed positive results.

Effect of different plot coverings on emergence and early development of seven palm species (arecaceae)

The Arecaceae family comprises plants with economical importance in many Brazilian regions, for agricultural exploration or for landscaping. In great portion, species of this family present low germination velocity and percentage. This work meant to evaluate the germination and early development of seven palm species (Archontophoenix alexandrae H. Wendl. et Drude, Copernicia prunifera (Miller) H.E. Moore, Latania commersonii Gmel., Livistona chinensis R. Br., Syagrus campos-portoana Bondar, Syagrus coronata (Mart.) Beccari, Syagrus picrophylla Barb. Rod.), submitted to three kinds of seed bed plot coverings. Three 10 x 2 m seedbeds were built and filled with a mixture of sand, soil and chicken manure (1:3:0.5 proportion), where two lines were sown with each specie. On top of each seedbed, plastic covering and fifty percent screen were set allowing one third of the seedbed to full sunlight exposure. Seedbeds were irrigated by dripping system. All species had the same germination rate, regardless of the covering, by the end of the experiment (146 days after sowing), eventhough, A. alexandrae under plastic covering conditions, L. commersonii at full sunlight exposure and Syagrus campos-portoana under fifty percent shade, had reached that percentage around 51 days after sowing. The remaining species reached the greatest germination percentage earlier with some of the coverings, rather than at full sunlight exposure. For the studied conditions, covering type had no effect in leaf length and width. For leaf number, there was interaction between species x covering type for Livistona chinensis and Copernicia prunifera.

Aceria litchii (Keifer) em lichia: ocorrência sazonal, danos provocados e identificação de possíveis agentes de controle biológico

Pós-graduação em Agronomia - FEIS

Variação espaço temporal da comunidade zooplanctônica em viveiros de cultivo de camarão branco, Litopenaeus vannamei (Boone, 1931), no município Curuçá, Pará-Brasil

Ao longo das últimas décadas, a carcinicultura vem apresentando um grande crescimento em diversas partes do mundo, com o Brasil seguindo esta tendência mundial (FAO, 2004). Nesta atividade três espécies de camarão têm se destacado como as mais cultivadas, sendo elas Penaeus monodon (Fabricius, 1798), Fenneropenaeus chinensis (Osbeck, 1765) e Litopenaeus vannamei (Boone, 1931), responsáveis por cerca de 80% da produção mundial (FAO, 2004). No Brasil L. vannamei é a espécie mais cultivada, com a produção brasileira correspondendo a 5% da produção mundial (FAO, 2004). L. vannamei é uma espécie marinha originária do Oceano Pacífico, distribuída do México ao Peru. Por ser eurihalino, este camarão pode se adaptar às mais diversas condições de cultivo, desde águas salgadas até de menores salinidades (BRAY et al., 1993; PONCE-PALAFOX et al., 1997), característica que tem aumentado o interesse dos produtores. Embora seja exótica no Brasil, L. vannamei, mostra maior resistência à variação de temperatura e salinidade do que outros camarões peneídeos nativos (BRITO et al., 2000). O alimento do camarão e as estratégias de seu fornecimento têm merecido uma atenção especial do setor, gerando novas técnicas ou seu aperfeiçoamento. A ração nos sistemas de cultivo intensivo e semi-intensivo, por exemplo, é responsável por 50-60% dos custos totais de produção, demonstrando a importância de novas estratégias para minimizar sue uso. O aumento da biomassa do plâncton (alimento natural), e conseqüentemente, da cadeia alimentar, reduz os custos com a alimentação suplementar, influenciando diretamente os custos finais de produção (AVAULT, 2003). Segundo Nunes (1995), o incremento da produtividade natural é tão importante quanto o uso de uma ração nutricionalmente completa e bem balanceada. Logo após a introdução nos viveiros de cultivo, a base da alimentação de L. vannamei é composta, em parte, pelo alimento natural disponível (NUNES et al. 1997; MARTINEZ-CORDOVA et al. 1997; ROTHLISBERG, 1998) complementada com ração comercial. Martinez-Cordova et al. (2002) mostraram que as concentrações de clorofila ‘a’ diminuem cerca de 50% do início ao fim do cultivo, provavelmente devido a pastagem pelo zooplâncton e por alguns invertebrados bentônicos. Além da importância do zooplâncton como alimento para as pós-larvas de camarão nos viveiros de engorda, o uso destes organismos (principalmente copépodes) como alimento vivo na aqüicultura marinha vem recebendo grande atenção nos últimos anos (DELBARE et al. 1996). Tal fato ocorre por serem ricos em fosfolipídios, ácidos graxos essenciais altamente insaturados e antioxidantes naturais, sendo nutricionalmente superiores aos rotíferos e aos náuplios de artemia, comumente usados na larvicultura marinha (SARGENT et al. 1997, STOTTRUP e NOSKER, 1997) promovendo o sucesso as larviculturas de camarão (PAYNE et al. 1998; SCHIPP et al. 1999; PAYNE e RIPPINGALE, 2000). Desta forma, estudos sobre o cultivo intensivo de camarões marinhos que enfoquem a composição da comunidade planctônica, as variáveis bióticas e abióticas no sistema, e a característica dos efluentes gerados, são de grande importância. Assim, os resultados obtidos podem incrementar a produtividade aquática no cultivo, alem de fornecer subsídios para pesquisas posteriores de avaliação e mitigação dos impactos ambientais causados por esta atividade.

Avaliação de progênies de longan

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Jasmonates and abscisic acid influence fruit ripening and plant water use: practical, physiological and morphological aspects

The aim of the present thesis was to better understand the physiological role of the phytohormones jasmonates (JAs) and abscisic acid (ABA) during fruit ripening in prospect of a possible field application of JAs and ABA to improve fruit yield and quality. In particular, the effects of exogenous application of these substances at different fruit developmental stages and under different experimental conditions were evaluated. Some aspects of the water relations upon ABA treatment were also analysed. Three fruit species, peach (Prunus persica L. Batsch), golden (Actinidia chinensis) and green kiwifruit (Actinidia deliciosa), and several of their cvs, were used for the trials. Different experimental models were adopted: fruits in planta, detached fruit, detached branches with fruit, girdled branches and micropropagated plants. The work was structured into four sets of experiments as follows: (i) Pre-harvest methyl jasmonate (MJ) application was performed at S3/S4 transition under field conditions in Redhaven peach; ethylene production, ripening index, fruit quality and shelf-life were assessed showing that MJ-treated fruit were firmer and thus less ripe than controls as confirmed by the Index of Absorbance Difference (IAD), but exhibited a shorter shelf-life due to an increase in ethylene production. Moreover, the time course of the expression of ethylene-, auxin- and other ripening-related genes was determined. Ripening-related ACO1 and ACS1 transcript accumulation was inhibited though transiently by MJ, and gene expression of the ethylene receptor ETR2 and of the ethylene-related transcription factor ERF2 was also altered. The time course of the expression of several auxin-related genes was strongly affected by MJ suggesting an increase in auxin biosynthesis, altered auxin conjugation and release as well as perception and transport; the need for a correct ethylene/auxin balance during ripening was confirmed. (ii) Pre- and post-harvest ABA applications were carried out under field conditions in Flaminia and O’Henry peach and Stark Red Gold nectarine fruit; ethylene production, ripening index, fruit quality and shelf-life were assessed. Results show that pre-harvest ABA applications increase fruit size and skin color intensity. Also post-harvest ABA treatments alter ripening-related parameters; in particular, while ethylene production is impaired in ABA-treated fruit soluble solids concentration (SSC) is enhanced. Following field ABA applications stem water potential was modified since ABA-treated peach trees retain more water. (iii) Pre- and post-harvest ABA and PDJ treatments were carried out in both kiwifruit species under field conditions at different fruit developmental stages and in post-harvest. Ripening index, fruit quality, plant transpiration, photosynthesis and stomatal conductance were assessed. Pre-harvest treatments enhance SSC in the two cvs and flesh color development in golden kiwifruit. Post-harvest applications of either ABA or ABA plus PDJ lead to increased SSC. In addition, ABA reduces gas exchanges in A. deliciosa. (iv) Spray, drench and dipping ABA treatments were performed in micropropagated peach plants and in peach and nectarine detached branches; plant water use and transpiration, biomass production and fruit dehydration were determined. In both plants and branches ABA significantly reduces water use and fruit dehydration. No negative effects on biomass production were detected. The present information, mainly arising from plant growth regulator application in a field environment, where plants have to cope with multiple biotic and abiotic stresses, may implement the perspectives for the use of these substances in the control of fruit ripening.

Detection and molecular characterization of viruses infecting Actinidia spp.

Kiwifruit (genus Actinidia) is an important horticultural crop grown in the temperate regions. The four world’s largest producers are China, Italy, New Zealand and Chile. More than 50 species are recognized in the genus but the principal species in cultivation are A. deliciosa and A. chinensis. In Italy, as well as in many other countries, the kiwifruit crop has been considered to be relatively disease free and then no certification system for this species has been developed to regulate importation of propagation plant material in the European Union. During the last years a number of fungal and bacterial diseases have been recorded such as Botrytis cinerea and Pseudomonas syringae pv. actinidiae. Since 2003, several viruses and virus-like diseases have been identified and more recent studies demonstrated that Actinidia spp can be infected by a wide range of viral agents. In collaboration with the University of Auckland we have been detected thirteen different viral species on kiwifruit plants. During the three years of my PhD I worked on the characterization of Cucumber mosaic virus (CMV) and Pelargonium zonate spot virus (PZSV). The determination of causal agents has been based on host range, symptom expression in the test plant species and morphological properties of the virus particles using transmission electron microscopy (TEM) and using specific oligonucleotide primers in reverse transcription-polymerase chain reaction (RT-PCR). Both viruses induced several symptoms on kiwifruit plants. Moreover with new technologies such as high-throughput sequencing we detected additional viruses, a new member of the family Closteroviridae and a new member of the family Totiviridae. Taking together all results of my studies it is clear that, in order to minimize the risk of serious viral disease in kiwifruit, it is vital to use virus-free propagation material in order to prevent the spread of these viruses.

Gain of chromosome 8q is associated with metastases and poor survival of patients with clear cell renal cell carcinoma

The aim of this study was to evaluate the prevalence of chromosome 8q gain in clear cell renal cell carcinoma (CCRCC) and to correlate the findings with tumor phenotype and disease-specific survival (DSS).