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The objective of this work was to evaluate the development of Davidiella sp. and its asexual form, Cladosporium sp., under different environmental conditions in the rubber tree (Hevea brasiliensis). Rubber tree leaves were inoculated with a spore suspension and kept in a humid chamber under different temperatures and wetness periods. The behavior of the fungi was evaluated using a scanning electron microscope (SEM) and an ultraviolet light microscope (UV). In the images obtained in SEM, four hours after inoculation of the fungus, it was possible to verify the germination and penetration of conidia at temperatures of 10 to 20 °C. The formation of conidiophores was verified from six hours after inoculation, indicating that it is in the reproductive period. In the sexual phase, in SEM, from four hours after inoculation, it was possible to verify the formation of small protuberances at temperatures between 10 and 20 °C. These black dots evolve into circular, protruding black spots, like the symptoms of black crust, with apparent spore formation on them. The data obtained from the UV analyses corroborate those from SEM, showing that the fungus has good development in its two phases between temperatures of 20 and 25 °C and that the period of wetness on the leaf can contribute to the initial development of the pathogen.

The fungus Austropuccinia psidii infects young tissues of Eucalyptus plants until they are two years old in the nursery and field, causing Myrtaceae rust. The characteristics making older eucalypt leaves resistant to A. psidii and the reason for the low levels of this pathogen in older plants need evaluations. The aim of this study was to evaluate the morphological differences between Eucalyptus grandis leaves of different growth stages and two plant ages to propose a visual phenological scale to classify E. grandis leaves according to their maturation stages and to evaluate the time of leaf maturation for young and adult plants. A scale, based on a morphological differentiation for E. grandis leaves, was made. The color, shape and size distinguished the leaves of the first five leaf pairs. Anatomical analysis showed a higher percentage of reinforced tissue, such as sclerenchyma-like tissue and collenchyma, greater leaf blade thickness, absence of lower palisade parenchyma in the mature leaves and a higher number of cavities with essential oils than in younger ones. Changes in anatomical characteristics that could reduce the susceptibility of older E. grandis leaves to A. psidii coincide with the time of developing leaf resistance. Reduced infection of this pathogen in older plants appears to be associated with a more rapid maturation of their leaf tissues.

Anthracnose, caused by fungi of the genus Colletotrichum, is present in the main areas where rubber trees (Hevea brasiliensis) are planted. Thus, considering that biological agents can be an alternative for disease control, the present study aimed to carry out initial studies to investigate the response of rubber tree seedlings inoculated with Colletotrichum and treated with saprobes fungi from the semiarid region of Northeast Brazil (Curvularia eragrostidis, Memnoniella levispora, Myrothecium roridum and Phialomyces macrosporus). Seedlings of the rubber tree clone RRIM600 were sprayed with biocontrol agents as preventive and curative treatments seven days before and after C. tamarilloi inoculation, respectively. Assessments included plant response to disease expression based on the percentage of symptomatic area on treated leaves, percentage of graft death, and percentage of apical death in seedlings 30 days after inoculation with C. tamarilloi. In addition, the enzymes peroxidase and phenylalanine ammonia lyase (PAL) had their activity quantified by their association with plant resistance to pathogens. The fungus C. eragrostidis had the best result in controlling anthracnose when applied as a preventive treatment, showing 10% less disease than the untreated plant. The same was observed for the fungus P. macrosporus when used in the curative form. These fungi also reduced the graft death. In these cases, PAL activity was higher and may be linked to the induction of resistance against the pathogen. The peroxidase activity was not expressive for treatments with saprobic fungi in the periods studied. Therefore, among the tested fungi, C. eragrostidis and P. macrosporus are promising for the control of anthracnose, deserving further studies.

The ascomycete fungus Geosmithia morbida (GM) and the walnut twig beetle (WTB), Pityophthorus juglandis, constitute a recently discovered fungal–insect complex responsible for the harmful Thousand Cankers Disease (TCD) in walnut trees. Key aspects of the epidemiology and ecology of the fungus and its insect vector were investigated in a disease outbreak found in central Italy, currently the southernmost introduction area of TCD on the Old Continent. Walnut trees with symptoms of crown dieback and epicormic branching with beetle galleries were sampled for pathogen isolation, while funnel traps were used to catch beetles and obtain adult flight curves. Growth-temperature relationships were determined for the fungus; optimum growth temperature for GM was 25°C, but it thrived up to 37°C. The phloem-boring bark beetle showed a longer flight period than in northern Italy. The beetle haplotype identified in Tuscany (H1) differed from haplotypes H2 and H21 found in northern Italy, suggesting possible multiple anthropogenic introductions. The substantial high temperature tolerance shown by the two organisms suggests they will not be constrained by the high temperatures of warmer areas. As a consequence, the impact of TCD in southern Europe, where the native Juglans regia is widely cultivated for its nuts and wood, would potentially be disastrous to local economies. In fact, although J. regia is not considered as susceptible as J. nigra, it can nevertheless be attacked in conditions of high bark beetle population density, a harbinger of high fungus’ propagule pressure.

Anthracnose, caused by fungi of the genus Colletotrichum, is present in the major rubber tree crop areas in Brazil, especially in São Paulo, Mato Grosso do Sul, Paraná, Minas Gerais, Espírito Santo, and northern states. This disease can affect different tissues of the rubber tree, leading to production losses. Thus, a better understanding of the pathosystem Colletotrichum x rubber tree can provide evidence to subsequent epidemiological research and phytosanitary management studies of this disease in the field. The present study aimed to investigate C.tamarilloi colonization and reproduction steps in resistant clones (IAC 502, IAC 507, RRIM 937) and in one susceptible clone (RRIM 600) of the rubber tree, verifying the influence of temperature up to 48 h after inoculation of the fungus, under 24 h wetness. Samples were analyzed under a light, a UV and a scanning electron microscope. Data indicated that the fungus had a delay in its development in resistant clones and, although colonization was expressive 48 h after inoculation, the new spore formation rate in the analyzed samples was lower in resistant clones. For RRIM 600, rapid colonization and intensive sporulation could be observed.

Lindgren funnel traps were used to monitor Pityophthorus juglandis occurrence. Traps were placed directly on walnut trees, with the top tied to one of the lower branches (about 2m high). An 8-funnel model was used instead of a 4-funnel trap, with the specific pheromone bait positioned between the fourth and the fifth funnel. Traps were customized with a 5mm metal mesh which was placed inside the bottom funnel so that debris (mainly foliage) and larger non-target insects would not end up inside the collecting jar. Geosmithia morbida was isolated from beetle adults, larvae and necrotic woody tissue around beetle galleries. Contaminant-free colonies were subcultured in purity and identified by: a) colony phenotyping [morphology, texture and pigmentation; margin type (regular/irregular; lobed/non-lobed); mycelium compactness; surface bumpiness; growth/temperature relationships]; b) micromorphology: type, morphology and ontogeny of conidiophores, metulae and phialides; conidiogenesis; shape, dimension and pigmentation of conidia; c) DNA fingerprinting. • Our protocol was customized to prevent traps from swinging in the wind and to optimize beetle catches by transversely fixing the bottom of funnel traps to the tree trunk with wooden shafts for stability. • To enhance fungus isolation in purity, a semi-selective Potato Dextrose Agar (PDA) medium, enriched with the antibiotics Ampicillin (Policillin-N) and Rifampicin (Rifamycin), was devised to prevent contamination by Gram-positive and Gram-negative bacteria and by mycobacteria.

The aim of the present study was to evaluate structural and biochemical aspects related to the interaction of resistant (RRIM 937, IAC 502 and 507) and susceptible (RRIM 600) rubber tree clones with C. tamarillo. For such analysis, ultrathin sections of the leaf limb were embedded in historesin and differently stained to verify structural alterations and presence of starch grains, arginine, lipids, tannins and lignins. The total proteins and activity of the enzymes peroxidase and (PAL) were quantified. Stomatal density was also analyzed under a scanning electron microscope. Data indicated alterations in the cell content of resistant clones inoculated with the pathogen, as well as greater lignin and lipid accumulation in these samples. For tannins, there was no difference between inoculated and non-inoculated clones. Arginine was found at greater quantities in IAC 502 and 507. Starch grains were not detected in any of the analyzed samples. Protein level and stomatal density were lower in resistant clones. Peroxidase activity was more expressive in resistant clones. PAL activity, there was no significant difference between clones. The lignin and lipids, total protein, peroxidase activity and stomatal density may be related to the resistance of rubber tree clones to anthracnose.

El objetivo de este trabajo fue monitorear su infestación en plantío de seringueira y posteriormente realizar la distribución espacial de la población en las raíces de los clones GT 1 y RRIM 600, , sin identificación del porta-injerto, en dos épocas de 2013. El experimento se ubicó en el municipio de Itiquira, MT, en las coordenadas geográficas de 17o17'40,29 "y 17o26'17,73" de latitud sur y 54o44'28 , 6 "y 54o44'43,55" de longitud oeste. En mayo se recogieron 264 muestras y en noviembre se recogieron 227 muestras en el área total de 6.646 hectáreas plantadas con 22 clones, utilizados para la interpolación por el método del krigado. Para los dos clones evaluados fueron 55 puntos en mayo (clone GT 1 = 29 y RRIM 600 = 26 muestreos) y 52 muestras en noviembre (clone GT 1 = 27 y RRIM 600 = 25 muestreos), en área total de 2.409 hectáreas. Con un aparato GNSS se identificó cada punto muestral, recogiendo raíces de la caucho a 0,10 m de profundidad. Para el análisis de las raíces se utilizó metodología de Coolen y D'Herde (1972). Las infestaciones en mayo, en los dos clones, fueron mayores que en noviembre. El Índice de Dependencia Espacial (IDE) de mayo y noviembre de 2013 fue moderado.

This study investigated the genetic variability in progenies of Eucalyptus dunnii Maiden for resistance against rust (Puccinia psidii). Field experiments were installed in two regions with different soil-climatic conditions. Open-pollinated progenies were established in a randomized complete block design. Sixty and 48 progenies were evaluated under field conditions at two sites, respectively, with six replications and eight trees per plot. In another experiment in a controlled environment, 53 progenies were evaluated in randomized blocks with six replications and nine plants per plot. The following traits were evaluated: plant height, severity of pest attack and the most susceptible stage to the leaf disease. The genetic variability for rust resistance in the E. dunnii population under study was high, with a genetic coefficient of variation of 36.07%; 7% of the evaluated progenies were rust-resistant. It indicates a high potential for selection and breeding of the species.

Eucalyptus plantation area has been increasing in Brazil, with 29% of the total plantation area being located in Minas Gerais state, which currently is being utilized primarily for charcoal production. However, diseases often increase the production costs of Eucalyptus. The objective of this study was to evaluate the effect of the fungus Ceratocystis fimbriata Ellis & Halsted on Eucalyptus wood for charcoal production. The basic density, volume, extractives, lignin, and holocellulose content of the wood were determined, as well as the gravimetric yield, volatile matter, fixed carbon, ash, and gross calorific values of charcoal. The introduction of the fungus C. fimbriata to Eucalyptus decreased the wood production and holocellulose content, but it also increased the wood’s lignin and extractives content. The chemical changes in the wood did not affect the charcoal produced. Volume of wood losses due to C. fimbriata can result in a loss of up to 3478.43 US$/ha.

http://dx.doi.org/10.5902/198050989296The leaf spot (Mycosphaerella leaf disease = MLD) caused by Teratosphaeria nubilosa has caused damage in eucalypt plantations in southern and southeastern Brazil. The need to assess the disease in the field to evaluate of this damage, efficiency control, evaluation of germplasm induces to the necessity of having a visual scale for evaluation of disease. The objective was to develop a diagrammatic scale for young leaves and one for adult leaves of Eucalyptus globules for MLD. To do so, the leaves collected in the field were scanned for image analysis. The damaged area, the healthy leaf area and the external area of the same scale RGB (Red, Green, Blue) were determined. Subsequently, it was determinate the levels of severity depending on the sample distribution with seven levels for young leaves and six for adult leaves. For the visual acuity test and validate the scale, the leaves were evaluated for severity, with and without scale. With this proposed scales, the assessors showed good accuracy both for young and adult leaves with R2=0,98 and R2=0,80, respectively. The importance of the development of diagrammatic scales for assessing MLD in eucalyptus must to the fact that allows quantification of the symptoms accurately and precisely.

The leaf spot (Mycosphaerella leaf disease = MLD) caused by Teratosphaeria nubilosa has caused damage in eucalypt plantations in southern and southeastern Brazil. The need to assess the disease in the field to evaluate of this damage, efficiency control, evaluation of germplasm induces to the necessity of having a visual scale for evaluation of disease. The objective was to develop a diagrammatic scale for young leaves and one for adult leaves of Eucalyptus globules for MLD. To do so, the leaves collected in the field were scanned for image analysis. The damaged area, the healthy leaf area and the external area of the same scale RGB (Red, Green, Blue) were determined. Subsequently, it was determinate the levels of severity depending on the sample distribution with seven levels for young leaves and six for adult leaves. For the visual acuity test and validate the scale, the leaves were evaluated for severity, with and without scale. With this proposed scales, the assessors showed good accuracy both for young and adult leaves with R2=0,98 and R2=0,80, respectively. The importance of the development of diagrammatic scales for assessing MLD in eucalyptus must to the fact that allows quantification of the symptoms accurately and precisely.

O objetivo deste estudo foi buscar in silico os ESTs do Banco de dados do Genoma de Eucalyptus (FORESTs, 2001) correlacionados com genes de alongamento celular envolvidos com crescimento de forma geral. Utilizando bibliotecas de cDNA, nós identificamos agrupamentos de ESTs semelhantes às proteínas controlando alongamento celular registradas no National Center of Biotechnologies Information – NCBI (2002). A busca mostrou similaridade para os seguintes agrupamentos de ESTs: o agrupamento EGEZLV2207D07.g com a enzima esterol delta redutase, EGEQRT4200A01.g com a proteína LKB biossintética de brassinosteróide, EGJMLV2220C06.g com EMBL-transportador mitocondrial half ABC, EGACST6260F07.g e EGMCFB1086B12.g com esterol 22 alfa hidroxilase (CYP90) (P450) e EGEQSL1051D09.g com EGEZSL5201E08.g para aproteína copine. The objective of study was in silico mining of ESTs from Eucalyptus ESTs database (FORESTs, 2001) correlated with cell elongation genes of growth character. Using cDNA libraries, we identified similar EST clusters to the proteins involved on control of cell elongation have been registered on National Center of Biotechnologies Information – NCBI (2002). The data mining has shown similarities for the following ESTs clusters: EGEZLV2207D07.g with delta sterol reductase, EGEQRT4200A01.g with LKB brassinosteroid biosynthetic protein, EGJMLV2220C06.g with EMBL-transporter of mitochondrial half ABC, EGACST6260F07.g and EGMCFB1086B12.g with steroid 22-α hydroxylase, and EGEQSL1051D09.g with EGEZSL5201E08.g for copine protein.

No melhoramento genético de espécies florestais, uma população base ouindivíduos superiores pré-selecionados tem importância fundamental para a manutenção do programa. Indivíduos de melhores procedências e de ampla base genética propiciam a obtenção de ganhos de forma contínua. O objetivo deste trabalho foi avaliar a diversidade genética em duas populações-núcleo de Eucalyptus grandis. Foram avaliados 39 indivíduos, sendo 19 pertencentes à população 1 e 20, à população 2, utilizando-se 14 primers microssatélite. Os fragmentos foram identificados e analisados a partir dos programas GeneScan e Genotyper, utilizando-se um sequenciador automático ABI Prism 3100. Onúmero de alelos encontrados para cada primer variou de cinco a 15 para a população 1 e, de 8 a 18 para a população 2. A heterozigosidade estimada foi maior na população 2, 0,869, contra 0,843 na população 1. A média da distância genética entre os indivíduos da população 1 foi 0,6220 e na população 2 foi 0,6112. Com a caracterização molecular dos indivíduos destas populações foi construído um banco de dados que permitirá, a partir dos parâmetrosde genética de populações, monitorar esses programas de melhoramento em diferentes ciclos de seleção.In genetic breeding of forest species, a base population or pre-selected higher individuals have a fundamental importance to program maintenance due to their better origins and large genetic basis, which continuously propitiates gains. The aim of this study was to verify the variability level in two Eucalyptus grandis nucleus populations. Thus, 39 individuals were evaluated – 19 in population 1, and 20 in population 2. Fourteen microsatellite primers were measured, identified and analyzed using GeneScan and Genotyper software through an ABI Prism 3100 automatic sequencer. The number of alleles in each primer varied between 5 and 15 in population 1, and from 8 to 18 in population 2. Heterozygosity was higher in population 2 – 0.869, versus 0.843 in population 1. Mean genetic distance amongindividuals was 0.6220 in population 1 and 0.6112 in population 2. After individual molecular characterization, a database was compiled to allow the control of these improvement programs in different selection cycles based on population genetic parameters.

Na agricultura moderna, diversas tecnologias auxiliam no aumento daprodutividade, sendo o herbicida uma delas, mas existem consequências atreladas ao seu uso, como os diversos efeitos sobre organismos não alvos. Neste trabalho, objetivou-se verificar esses efeitos sobre agentes fitopatogênicos, assim como avaliar o efeito do herbicida glyphosate sobre diversas doenças, em plantas de soja transgênicas.Verificou-se forte ação fungicida com o uso do herbicida glyphosate, assim como os outros avaliados “in vitro”, sobre os fungos testados, e os mesmos resultados foram observados nas plantas em condição de campo.In modern agriculture, several technologies have helped increase productivity, and herbicide is one of them. However, there are consequences linked to its use, such as the various effects on non-target organisms. The purpose of this work was to verify these effects on phytopathogenic agents, as well as assess the effect of glyphosate on diseases in transgenic soybean. There was a strong fungicide action using glyphosate herbicide as well as with the others evaluated in vitro regarding fungi tested. The same results were observed in plants in field conditions.

Trata-se de um estudo sobre “datamining”envolvendo genes ligados ao crescimento decontrole não hormonal, utilizando o banco dedados de ESTs de eucalipto, efetuado atravésdo Projeto Genoma do Eucalipto (FORESTs)comparados ao nível de aminoácidos. Foramidentificados os clusters de ESTsEGBGFB1211D01.g, EGEZRT6201E10.g,EGCCFB1220G07.g, EGRFCL1206E01.g,EGEQST2006A06.g, EGRFCL1206E01.g,EGEQRT3001H05.b e EGBFRT3106G11.g,similares às proteínas de celulose sintase e suassubunidades controlando o crescimento emArabidopsis thaliana, Gossipium hirsutum,Populus tremuloides, Zea mays e Nicotiana alata,registradas no National Center of BiotechnologiesInformation - NCBI, informação valiosa parafuturos programas de melhoramento genético dogênero Eucalyptus.This is a study about data mining ofexpressed sequence tags (ESTs) involved withcellulose synthase growth effect genes resultedfrom the Eucalyptus ESTs Genome Project(FORESTs) compared at aminoacids level. Using asequencing of derived from cDNAs librariesinduced and not induced by bacteria, wereidentified EST clusters EGBGFB1211D01.g,EGEZRT6201E10.g, EGCCFB1220G07.g,EGRFCL1206E01.g, EGEQST2006A06.g,EGRFCL1206E01.g, EGEQRT3001H05.b, andEGBFRT3106G11.g, similar to cellulose synthaseproteins controlling growth effect in Arabidopsisthaliana, Gossipium hirsutum, Populustremuloides, Zea mays, and Nicotiana alata,registered on National Center of BiotechnologiesInformation - NCBI. These mining results areimportant to improve Eucalyptus breeding programs.

RESUMO: Entre janeiro e fevereiro de 2013 ocorreu uma epidemia de mal das folhas em regiões consideradas como de "escape" à doença, causando preocupações e indagações nos produtores de borracha e ao setor heveícola como um todo. Uma possível adaptação do fungo a essas zonas foi levantada e a consistência do controle da doença pela utilização do binômio plantas com troca uniforme das folhas + período seco, típicos de regiões de "escape", foi questionada. O objetivo deste trabalho foi trazer informações importantes aos heveicultores sobre a interação clima-patógeno-hospedeiro e comparar variáveis de normais climatológicas (média histórica) e dados climáticos deste período de ocorrência da epidemia (janeiro e fevereiro de 2013). Concluiu-se que não foi o patógeno que se adaptou às plantas de seringueira em clima mais seco, nas regiões de "escape", mas que o clima mais úmido ocorreu nessas regiões mais secas, nesse período.

The present study investigated myrtle rust and gall wasp from non-commercial eucalypt taxa with potential to hybridization. We selected 38 genotypes from 18 species of Corymbia and Eucalyptus (Symphyomyrtus) genus. Two rust inoculations were evaluated using inoculums from rose apple and eucalypt, and natural gall occurrence in a high pest infestation condition. Corymbia was more susceptible to rust than Eucalyptus genus, and it was observed severity variation due to the sources of rust inoculum. Species that belong to Exsertaria and Transversaria (Latoangulatae) sections were more susceptible to gall wasp, and Corymbia species showed no development of gall.

Os objetivos deste trabalho foram mensurar a área da nervura secundária e quantificar o número de tricomas da superfície foliar de cultivares de feijoeiro suscetível (Carioca), moderadamente resistente (Pérola) e resistente (IAPAR-81) à antracnose, e observar as relações entre tais características com o estágio pré-infeccional. Para mensurar a área da nervura secundária, fragmentos foliares foram amostrados, fixados em FAA 50, conservados em álcool 70%, infiltrados em resina glicol-metacrilato, cortados transversalmente e corados com azul de toluidina. Os tricomas foram quantificados em impressões epidérmicas em microscópio de projeção. Na análise pré-infeccional, o material foi fixado em glutaraldeído 2,5% e processado para o estudo ao microscópio eletrônico de varredura. A cultivar resistente apresentou menor área de nervura secundária e maior pilosidade que a cultivar moderadamente resistente e a suscetível. Na cultivar resistente, por causa da menor área de nervura e maior pilosidade, o patógeno permaneceu sobre a nervura e envolvido com o tricoma. Na cultivar suscetível, com maior área de nervura e menor pilosidade, foram observadas necrose e estruturas fúngicas na superfície foliar. Há relação entre estas características foliares e a resistência à antracnose durante o estágio pré-infeccional do patógeno.The objectives of this work were to measure the secondary vein area and to quantify the trichomes of leaf surface in bean cultivar susceptible (Carioca), medium resistant (Pérola) and resistant (IAPAR-81) to anthracnose, and to observe relationships between leaf blade characteristics with the preinfectional stages. In order to measure the secondary vein area, sample leaf pieces were fixed in FAA 50 and conserved in 70% alcohol. These leaf samples were infiltrated in glycol metacrylate resin, cut transversally and stained with toluidine blue. The trichomes were quantified in epidermical impressions in projection microscopy. The material for preinfection analysis was fixed in 2.5% glutaraldehyde and processed to study the scanning electron microscopy. The resistant cultivar showed minor secondary vein area and high pilosity than the cultivar medium resistant and susceptible. In the resistant cultivar, due to minor vein area and high pilosity, the pathogen stayed on the vein and involved on the trichomes. In susceptible cultivar, with high vein and minor pilosity, necrosis and fungal structure on the leaf surface were observed. The results show a relation between the minor vein and high leaf trichomes quantity with the resistance to the anthracnose in pathogen preinfection stages.

Boron (B) is a low mobility plant micronutrient whose molecular mechanisms of absorption and translocation are still controversial. Many factors are involved in tolerance to Boron excess or deficiency. Recently, the first protein linked to boron transport in biological systems, BOR1, was characterized in Arabidopsis thaliana. This protein is involved in boron xylem loading and is similar to bicarbonate transporters found in animals. There are indications that BOR1 is a member of a conserved protein family in plants. In this work, FORESTS database was used to identify sequences similar to this protein family, looking for a probable BOR1 homolog in eucalypt. We found five consensus sequences similar to BOR1; three of them were then used in multiple alignment analysis. Based on amino acid similarity and in silico expression patterns, a consensus sequence was identified as a candidate BOR1 homolog, helping deeper experimental assays that could identify the function of this protein family in Eucalyptus.

This work was aimed at locating Eucalyptus ESTs corresponding to the PROTOX or PPO enzyme (Protoporphyrinogen IX oxidase, E.C. 1.3.3.4) directly related to resistance to herbicides that promote oxidative stress, changing the functionality of this enzyme. PROTOX, which is the site of action of diphenyl-ether (oxyfluorfen, lactofen, fomesafen), oxadiazole (oxadiazon and oxadiargyl), and aryl triazolinone (sulfentrazone and carfentrazone) herbicides, acts on the synthesis route of porphyrins which is associated with the production of chlorophyll a, catalases, and peroxidases. One cluster and one single read were located, with e-values better than e-70, associated to PROTOX. The alignment results between amino acid sequences indicated that this enzyme is adequately represented in the ESTs database of the FORESTs project.

Herbicides inhibit enzymatic systems of plants. Acetolactate synthase (ALS, EC = 4.1.3.18) and 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, EC 2.5.1.19) are key enzymes for herbicide action. Hundreds of compounds inhibit ALS. This enzyme is highly variable, enabling the selective control of weeds in a number of crops. Glyphosate, the only commercial herbicide inhibiting EPSPS is widely used for non-selective control of weeds in many crops. Recently, transgenic crops resistant to glyphosate were developed and have been used by farmers. The aim of this study was the data mining of eucalypt expressed sequence tags (ESTs) in the FORESTs Genome Project database (https://forests.esalq.usp.br) related to these enzymes. Representative amino acid sequences from the NCBI database associated with ALS and EPSPS were blasted with ESTs from the FORESTs database using the tBLASTx option of the blast tool. The best blasting reads and clusters from FORESTs, represented as nucleotide sequences, were blasted back with the NCBI database to evaluate the level of similarity with available sequences from different species. One and seven clusters were identified as showing high similarity with EPSPS and ALS sequences from the literature, respectively. The alignment of EPSPS sequences allowed the identification of conserved regions that can be used to design specific primers for additional sequencings.

This work was aimed at locating Eucalyptus ESTs corresponding to the GS enzyme (Glutamine Synthetase, EC = 6.3.1.2) and to the D1 protein, which are directly related to resistance to herbicides that promote oxidative stress. Glutamine Synthetase corresponds to the site of action of the herbicide glufosinate. Herbicides that belong to groups such as ureas, uracils, triazines and triazinones act on the D1-Qb complex (receptor of electrons from the Photosystem II) by inactivating it. The clusters EGEQRT3302E01.g, EGEQRT3001F12.b; EGEZLV1203B04.g; EGBGFB1211H06.g and EGEZLV1205F09.g enclosed complete sequences (with 356 amino acids) of the Glutamine Synthetase enzyme. The cluster EGEQSL1054G06.g is a consensus of four reads and enclosed a complete sequence of D1 Protein (with 353 amino acids). The comparison of the sequences of Protein D1 from different species showed that the substitutions of serine (S) by glycine (G) or serine (S) by threonine (T) at the position 264 could produce plants resistant to herbicides that act on electron flow on Photosystem II. The sequence of amino acids corresponding to the cluster EGEQSL1054G06.g had a serine in position 264 indicating sensitivity of the Eucalyptus plants to herbicides that act on this site.

O objetivo deste trabalho foi selecionar clones de seringueira (Hevea spp.) promissores para a região do planalto do Estado de São Paulo. De uma população de 70 clones, de polinização controlada, integrantes do experimento de avaliação em pequena escala, foram avaliados 16 da série IAC 300. O experimento foi instalado na Estação Experimental de Pindorama, no espaçamento de 7 x 3 m, em delineamento de blocos casualizados, com três repetições, tendo os clones RRIM 600 e GT 1 como testemunhas. Quanto à produção de borracha seca, destacaram-se oito e dez clones superiores ao RRIM 600 e GT 1, respectivamente. Todos os clones avaliados apresentaram-se vigorosos em relação ao perímetro do caule na abertura do painel. Sete deles mostraram alta resistência; sete, resistência moderada, e dois, suscetíveis à antracnose do painel. Sugere-se sua avaliação em experimentos de grande escala, para avaliar, além da produção, os demais caracteres secundários, em diferentes ambientes, para futuras recomendações em larga escala para o Estado de São Paulo.

O objetivo deste trabalho foi avaliar a adaptabilidade e expressão fenotípica de caracteres superiores de dez clones amazônicos de seringueira (Hevea spp.) no planalto do Estado de São Paulo em um período de 10 anos, obedecendo ao delineamento de blocos ao acaso com três repetições e parcelas lineares de seis plantas. O clone IAN 3156 foi o mais produtivo, com média de 65,57 g de borracha seca/árvore/sangria, no período de quatro anos, seguido pelo clone RO 45 com 52,29 g de borracha seca/árvore/sangria, enquanto o clone-testemunha, RRIM 600, produziu 41,04 g/árvore/sangria. Todos os clones apresentaram crescimento vigoroso. O perímetro do caule na abertura do painel variou de 37,01 cm (IAN 3193) a 49,41 cm (IAN 4493). A porcentagem de plantas aptas à sangria variou de 30,0% (IAN 3703) a 93,75% (IAN 6323). Exceto os clones IAN 3156 e IAN 4493 com 7,00 mm e 6,32 mm, respectivamente, todos os outros clones apresentaram espessura de casca virgem inferior ao clone RRIM 600, que apresentou 6,18 mm. O clone IAN 3193 apresentou maior incidência de quebra do ponteiro pelo vento. Todos os clones estudados apresentaram baixa incidência de secamento de painel. O bom desempenho dos clones IAN 3156, RO 45, Fx 3899 e IAN 4493 permite que possam ser experimentados em larga escala, envolvendo diferentes ambientes no Estado de São Paulo.

É apresentado o desempenho de 19 novos clones de seringueira [Hevea brasiliensis (Willd. exAdr. de Juss.) Muell. Arg.], resultantes de hibridações conduzidas no Instituto Agronômico e avaliados em experimento de pequena escala, tendo o clone RRIM 600 como testemunha. O experimento em campo obedeceu ao delineamento em blocos ao acaso, com três repetições. Com relação à produção de borracha seca, o clone IAC 40 apresentou a maior média (62,22 g/árvore/sangria) nos três anos de avaliação, seguido pelo IAC 301 (57,67 g/árvore/sangria) e pelo IAC 300 (50,61 g/árvore/sangria), com produções 154%, 138% e 123% superiores em relação ao RRIM 600 (41,04 g/árvore/sangria). Todos os clones selecionados foram vigorosos, com perímetro do caule na abertura do painel variando de 37,81 cm (IAC 317) a 50,90 cm (IAC 315). A porcentagem de plantas aptas a sangria variou de 20,0% (IAC 317) a 100% (IAC 315). Todos os clones apresentaram baixas incidências de quebra pelo vento e de secamento do painel. Não foi detectada nenhuma doença foliar em caráter epidêmico. Dos clones estudados, 15 apresentaram alta resistência à antracnose do painel, e foram superiores ao RRIM 600; os outros cinco apresentaram resistência moderada semelhante ao RRIM 600.The present paper shows the performance of 19 rubber tree [Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell. Arg.] originated from Hevea breeding programme conducted at the Instituto Agronômico de Campinas (IAC) and evaluated in a small scale trial in the northwestern of São Paulo State, Brazil. The old popular clone RRIM 600 of Malaysian origin was used as control. The trial was laid out in a randomized block design with three replications. Concerned to yielding the clone IAC 40 recorded highest yield (62.22 g/tree/tap) over three years of tapping which was followed by IAC 301 (57.67 g/tree/tap) and IAC 300 (50.61 g/tree/tap) yielding 154%, 138% and 123% superior to the control clone and RRIM 600 whereas recorded 41.04 tree/tap. All selected clones were of vigorous growing, with the girth at opening ranging from 37.81 cm for IAC 317 to 50.90 cm for IAC 315 and tappability ranging from 20.0% for IAC 317 to 100% for IAC 315. All the clones experienced low to very low incidence of wind damage. All selected clones showed low to very low incidence of dry trees. Fifteen clones showed high resistance and five presented moderate resistance to anthrachnose panel canker disease similarly to RRIM 600.

Este trabalho objetivou apresentar resultados da primeira seleção de clones de seringueira (Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell.Arg.) da série IAC 300, e amazônicos das séries IAN e Fx, em experimento de pequena escala, visando produção, crescimento e resistência ao mal-das-folhas. A produção e o vigor de 20 clones foram avaliados por dois e nove anos, respectivamente, em PariqüeraAçú, no Vale do Ribeira, SP. Os clones IAC 301, IAC 304, IAC 306 e IAC 319 produziram satisfatoriamente nos dois primeiros anos de sangria. Os clones amazônicos IAN 6323, Fx 3864 e IAN 2903, com produções de 1.078 kg, 945 kg e 900 kg/ha/ano, respectivamente, foram superiores à testemunha IAN 873 (878 kg/ha/ano). Os clones selecionados apresentaram crescimento vigoroso, com extremos de perímetro do caule, na abertura do painel, de 37,40 cm (IAN 4493) a 53,75 cm (IAN 6323), e percentual de plantas aptas à sangria de 7,0% (IAN 4493) a 100% (IAN 6323 e IAC 302), exceto os clones Fx 3899 e IAN 3044. O IAC 315, com 7,37 mm, mostrou maior espessura de casca virgem que o IAN 873 (6,44 mm). Os clones IAC 320, IAC 306 e IAC 315 foram os mais resistentes ao Microcyclus ulei.

Rubber tree [Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell-Arg] anthracnose panel canker disease resistance, caused by Colletotrichum gloeosporioides (Penz.) Sacc., and growth and yield characters were assessed at three years old in the nursery, in 18 half-sib progenies. There were highly significant (P < 0.01) genetic differences among progenies for most characters. The genetic component of variance accounted for 3.37, 6.07, 12.74, 12.13, 19.82 and 3.58% of the phenotypic variance for anthracnose-infected area, anthracnose lesion stripe length, yield, girth, virgin bark thickness and total number of latex vessel rings, respectively. Narrow-sense heritability estimates for the above characters were 13.45, 24.30, 50.97, 48.52, 79.30 and 14.30%, respectively. No significant genetic correlations were found among anthracnose panel canker resistance, yield and growth characters. The frequency distribution of anthracnose-infected area and anthracnose lesion stripe length suggests polygenic control. Selecting the best three progenies out of 18 progenies would result in a genetic gain of 12.87 and 24.24% for infected area and lesion stripe length, respectively. The best five selected individuals out of 50 individuals within each progeny would result in a genetic gain of 9.47 and 24.62%, with a total gain of 22.34 and 48.87% for these two characters, respectively.Estudos genéticos da relação da resistência da antracnose do painel, caracteres da produção e vigor da seringueira [Hevea brasiliensis (Willd. ex. Adr. de Juss.) Muell-Arg] foram conduzidos em 18 progenies de meios-irmãos na Estação Experimental de Pindorama, Estado de São Paulo. Os resultados obtidos mostraram diferenças genéticas significativas entre progenies para a maioria dos caracteres estudados. Os componentes genéticos da variância contribuíram com 3,37% da variância fenotípica para área infectada, 6,07% para comprimento de estrias, ambos causados pela anthracnose, 12,74% para produção, 12,13% para perímetro do caule, 19,82% para espessura de casca virgem e 3,58% para número total de vasos laticíferos. As herdabilidades no sentido restrito para os caracteres acima foram: 13,45%, 24,30%, 50,97%, 48,52%, 79,30% e 14,30%, respectivamente. Correlações significativas não foram encontradas entre antracnose do painel e caracteres da produção e perímetro do caule. A distribuição de freqüência de antracnose sugere que o caráter é poligênico. A seleção das três melhores progênies resultaria em um ganho genético para antracnose de 12,87% e 24,24% para área infectada e comprimento das estrias, respectivamente. A seleção dos cinco melhores indivíduos dentre os 50 indivíduos de cada progênie, para ambos os caracteres, resultaria em ganho genético de 9,47% e 24,62% com ganho total de 22,34% e 48,87%, respectivamente.

No melhoramento genético de espécies florestais, uma população base ou indivíduos superiores pré-selecionados tem importância fundamental para a manutenção do programa. Indivíduos de melhores procedências e de ampla base genética propiciam a obtenção de ganhos de forma contínua. O objetivo deste trabalho foi avaliar a diversidade genética em duas populações-núcleo de Eucalyptus grandis. Foram avaliados 39 indivíduos, sendo 19 pertencentes à população 1 e 20, à população 2, utilizando-se 14 primers microssatélite. Os fragmentos foram identificados e analisados a partir dos programas GeneScan e Genotyper, utilizando-se um sequenciador automático ABI Prism 3100. O número de alelos encontrados para cada primer variou de cinco a 15 para a população 1 e, de 8 a 18 para a população 2. A heterozigosidade estimada foi maior na população 2, 0,869, contra 0,843 na população 1. A média da distância genética entre os indivíduos da população 1 foi 0,6220 e na população 2 foi 0,6112. Com a caracterização molecular dos indivíduos destas populações foi construído um banco de dados que permitirá, a partir dos parâmetros de genética de populações, monitorar esses programas de melhoramento em diferentes ciclos de seleção.

Na agricultura moderna, diversas tecnologias auxiliam no aumento da produtividade, sendo o herbicida uma delas, mas existem consequências atreladas ao seu uso, como os diversos efeitos sobre organismos não alvos. Neste trabalho, objetivou-se verificar esses efeitos sobre agentes fitopatogênicos, assim como avaliar o efeito do herbicida glyphosate sobre diversas doenças, em plantas de soja transgênicas. Verificou-se forte ação fungicida com o uso do herbicida glyphosate, assim como os outros avaliados “in vitro”, sobre os fungos testados, e os mesmos resultados foram observados nas plantas em condição de campo.

O controle biológico de uma doença fúngica consiste no emprego de um organismo (predador, parasita ou patógeno) que afete o outro fungo que esteja causando danos econômicos. Dentre os possíveis agentes de biocontrole, um fungo vem se destacando como promissor, sendo conhecido como Clonostachys rosea, forma assexuada de Bionectria ochroleuca. Para isto há a necessidade da produção in vitro deste fungo, para poder ser usado como antagonista de fungos fitopatogênicos. Efetuou-se ensaios objetivando verificar meio de cultura que melhor favorece-se a produção de conídios pelo fungo. O trabalho foi realizado no Setor de Defesa Fitossanitária, do Departamento de Produção Vegetal, FCA - UNESP, campus Botucatu. Foi utilizado o isolado CCR64 (EMBRAPA-CNPMA).Os meios de culturas testados foram: BDA; Aveia-agar; Mazeina-agar; Arroz-agar; V8-5%; V8-10%; V8-20%; TJ-5%; TJ-10; TJ-20%. A esporulação do fungo, nos diferentes meios de culturas, foi avaliada no 8Ëš dia de incubação. Os dados foram analisados utilizando método de comparação entre médias, através do teste Tukey a 5% de probabilidade, sendo os dados transformados para (X+1)0,5. Todos os meios de culturas testados se mostraram aptos para a produção de conídios, verificou-se que o melhor meio para produção de conídios de Bionectrai ochroleuca é o TJ-5%, seguido do TJ-20%, com uma esporulação média de 3,5 x106 conídios/mL.

A mamoneira é uma espécie tropical e está sujeita a diversas doenças, que causam grandes prejuízos, dentre essas, o mofo cinzento (Botryotinia ricini) é uma das mais importantes. A esporulação deste fungo foi avaliada nos meios de culturas BDA; Aveia-agar; Mazeina-agar; Arroz-agar; Folhas de mamona triturada-ágar (FM); FM-CaCO3; suco V8 a 5% (V8-5%); V8-10%; V8-20%; suco de tomate a 5% (TJ-5%); TJ-10%; TJ-20%. A esporulação do fungo, nos diferentes meios de culturas, foi avaliada no 8Ëš dia de incubação. Os dados foram analisados utilizando método de comparação entre médias, através do teste Tukey a 5% de probabilidade, sendo os dados transformados para (X+1)0,5. Todos os meios de culturas testados se mostraram aptos para a produção de conídios, mas obteve-se os melhores resultados no meio de cultura V8-20%, com 5,7 x 106 conídios/mL, ficando em segundo lugar o meio BDA, com uma esporulação média de 3,5 x106 conídios/mL.

Objetivou-se neste trabalho determinar a relação entre a avaliação da incidência da clorose variegada dos citros (CVC) e de Xylella fastidiosa por meio da sintomatologia e serologia (DIBA – Dot immunobinding assay), para estudos de focos. As avaliações foram realizadas em pomar comercial de laranja Pera, Citrus sinensis (L.) Osbeck, enxertada sobre limoeiro Cravo (Citrus limonia Osbeck) (Rutaceae). A área experimental constitui-se de duas parcelas, com 15 linhas e 10 plantas/linha, totalizando-se 300 plantas. Foram feitos quatro levantamentos em um período de nove meses. Todas as plantas do talhão foram avaliadas pela sintomatologia, procedendo-se à coleta de 20 folhas por planta nos quatro quadrantes para análise serológica. Pela sintomatologia, o número de focos foi menor do que o obtido por DIBA, apenas na primeira avaliação, e foi 3,1, 4,1 e 4,5 vezes maior na segunda, terceira e quarta avaliações, respectivamente. O tamanho dos focos definidos por DIBA foi maior do que aqueles definidos por sintomas. Assim, 92% dos focos avaliados por sintomas e 74% dos avaliados por DIBA foram menores do que 10 plantas. Os valores médios do índice de compactação de foco (ICF) foram 0,86 por sintomas e 0,85 por DIBA. O valor médio do índice de forma de foco (IFF) foi 1,17 na avaliação por sintomas e 0,94 por DIBA. A maior eficiência de detecção da incidência pelo DIBA em relação à sintomatologia é comprovada pelo menor número de focos e de focos unitários, assim como pelo maior número de plantas por foco, ressaltando-se a importância da utilização de métodos mais sensíveis para o estudo da epidemiologia da CVC

O objetivo do presente trabalho é apresentar as espécies de Uredinales sobre membros de Blechnaceae, Thelypteridaceae, Schizaeaceae, Myrtaceae, Oxalidaceae, Rhamnaceae, Rubiaceae, Sapindaceae, Smilacaceae e Vitaceae da Reserva Florestal da Cidade Universitária "Armando de Salles Oliveira" São Paulo, SP, Brasil. As espécies descritas e ilustradas bem como seus respectivos hospedeiros são: Desmella anemiae Syd. & P. Syd. sobre Blechnum occidentale L. (Blechnaceae) e Thelypteris dentata (Forssk.) E. St. John (Thelypteridaceae), Endophyllum circumscriptum (Schwein.) Whetzel & Olive sobre Cissus selloana Planch. (Vitaceae), Hemileia vastatrix Berk. & Broome sobre Coffea arabica L. (Rubiaceae), Phakopsora colubrinae Viégas sobre Hovenia dulcis Thunb. (Rhamnaceae), Puccinia arechavaletae Speg. sobre Serjania caracasana (Jacq.) Willd. e Serjania communis Cambess. (Sapindaceae), Puccinia lygodii Arthur sobre Lygodium volubile Sw. (Schizaeaceae), Puccinia oxalidis Dietel & Ellis sobre Oxalis latifolia Kunth (Oxalidaceae), Puccinia psidii G. Winter sobre Psidium guajava L. e Syzygium jambos (L.) Alston (Myrtaceae) e Sphenospora smilacina Syd. sobre Smilax quinquenervia Vell. (Smilacaceae).