Estado nutricional e avaliação de caracteres secundários da produção em dois clones jovens de seringueira em São José do Rio Preto, SP

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

Efeito Pesticida de Ésteres de Sacarose Sobre Calacarus heveae Feres (Acari: Eriophyidae) em Dois Clones de Seringueira

Sucrose esters are biodegradable, non pollutant and safe for health; moreover, they have shown great potential in the pest control. We tested a crude mix of sucrose esters on the survivorship and oviposition of Calacarus heveae Feres (Acari: Eriophyidae) females, an important rubber tree pest mite. The females were collected from rubber leaflets of the GT 1 and RRIM 600 clones. The mites were kept on stock arenas placed in rearing chamber at 28 + 0,1°C, 80 + 10% of relative humidity and 12h of light phase about two weeks before the assays. After this period, the mites were transferred to assay arenas made with leaflets from the same clone of stock arenas. The survivorship of females sprayed with sucrose esters in the concentrations of 1, 2, 3 and 4 g/L and their oviposition effect with 0.5 and 1 g/L were analyzed. The mites sprayed with 4 g/L had about 80% of mortality, while those with 1 g/L around 60%. All mites that died in the treatments with sucrose ester became dark with wrinkled tegument and decreased their body volume, suggesting dehydration. We also observed the efficacy of 1 g/L concentration to decrease the female oviposition in about 50%, in the third day after spraying. No differences were observed in the mortality and oviposition between females kept on both clones.

Clones de Eucalyptus sp. submetidos a estresse hídrico em dois tipos de solo

Pós-graduação em Agronomia (Irrigação e Drenagem) - FCA

Desenvolvimento inicial de clones IAC de seringueira em São José do Rio Preto, SP

In this study the development of four clones (IAC 35; 40; 300 e 301) of rubber tree [Hevea brasiliensis (Willd. ex A. Juss.) Müell. Arg.] selected by Instituto Agronômico de Campinas were evaluated during the first 24 months of cultivation. The experiment was laid out in randomized block design with three replications and four plants for parcel following the 1.5mx1.0m spacing at São José do Rio Preto, SP. The clone RRIM 600 was used as check. The variables analyzed were the trunk girth, the number of matured whorl, the photosynthesis rate and the concentrations of chlorophyll a, b, total and carotenoids. At the end of the experiment the values of trunk girth at 0.5m above the budgrafting union varied from 5.85 cm (IAC 301) to 10.53 cm (IAC 300) and the mean number of matured whorl varied from 2.58 (IAC 301) to 3.91 (RRIM 600). During the dry season, when the plants were 22 month old, the mean value of photosynthesis rate of IAC 40 and IAC 301 (12mol m-2 s-1) were lower compared to the other clones (15mol m-2 s-1). The chlorophyll a, b and total carotenoids concentrations were equal or superior to the check, never lower. Considering the variables analyzed, excluding the IAC 301, the other clones show performance comparable to RRIM 600.

Teores de rotenona em Clones de Timbó (Derris Spp. Fabaceae) de diferentes regiões da Amazônia e os seus efeitos na emergência de Imagos em Musca Domestica L

The rotenone contents analysis in clones of three timbó species permited the classification of these plants based on their efficiency to control Musca domestica larvae. The results showed significant correlations between the rotenone contents in the Derris urucu and D. nicou clones with the capacity to control the larvae. The plants with high rotenone contents were more effective. The rotenone contents, the effect of Derris species clones on the flics and the localization of the plants origin, showed that among the plants could have happened a populational isolation in the Amazonia during the pleistocene epoch. In Derris sp., that presented lower rotenone contents and was ineffective to control of the larvae, these differences were not observed.

Differential response of clones of eucalypt to glyphosate

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

Performance of different rubber tree clones against South American leaf blight (Microcyclus ulei)

This study was aimed to evaluate the behaviour of eighteen clones of Hevea brasiliensis (rubber) against South American leaf blight and tostudy progress of the disease. The experiment was conducted in Cachoeiro de Itapemirim-ES, where 18 clones were evaluated. The experi-mental design was a randomized block with four replicates; each experimental unit consisted of three plants. Evaluations were performed at15-day intervals on 30 leaflets per tree. Disease incidence was quantified and infection classified according to the stages of development andtype of damage. Leaf blight occurred during the entire experimental period; however, disease intensity varied with the resistance level of theclones and the time of year. Clones FX 3864, RRIM 725, RRIM 711, IAC 300 and IAN 873 exhibited the highest resistance to leaf blight.

Composição físico-química e cor de clones de mandioquinha-salsa

This study aimed to characterize the physical and chemical composition of ten items of arracacha grown in the municipality of São Manuel for the 2009 harvest. In the roots of the clones BGH (4560, 5741, 5744, 5746, 5747, 6414, 6513, 6525 and 7609) and the cultivar Amarela de Senador Amaral the characteristics evaluated were: color (L *, a * and b *) and moisture, ash, crude fiber, raw grease, protein, reducing sugars, total sugars and starch. After obtaining the data, an analysis was performed for the variance of test F and comparisons between the means made by the Tukey test at 5% probability. There was no significant difference to the results of luminosity (L *) while BGH 6414 and BGH 5744 showed the highest values for chroma and * BGH 5741, BGH 6414, BGH 7609, 'Amarela de Senador Amaral' BGH 5747 presented the highest chroma values for b *. Clones BGH 7609 and BGH 6414 showed significantly higher levels of dry matter and with the potential yield of agro-industrial processes it would be best suited in the form of frying. The materials that showed significantly larger amounts of ash were BGH 6525, BGH 5747, 'Amarela de Senador Amaral ", BGH 4560, BGH 5746, BGH 6513. Regarding the contents of fatty matter BGH 6525, BGH 5741 and BGH 5744 showed the highest levels. The results of BGH 7609 showed crude fiber significantly higher than the other materials tested, it can be used in diets composed of fibers. BGH 4560 and cultivar had the highest crude protein. BGH 5741 showed the lowest reducing sugar content among the clones, but not significantly different from results found for the cultivar. All clones showed total sugar levels were higher in the cultivar, which may have more flavor. BGH 5741, BGH 5746, BGH 6525 and BGH 6513 showed significantly higher starch content than the cultivar Amarela de Senador Amaral. From these results we conclude that the clones have similar color characteristics, and are potentially a nutritionally adequate substitute for the cultivar.

Effects of abiotic factors on growth and chemical defenses in cultivated clones of Laurencia dendroidea J. Agardh (Ceramiales, Rhodophyta)

Laurencia dendroidea shows high inter- and intrapopulation variability in the amount of the sesquiterpene elatol, caused by genetic variation as well as environmental factors. To test the independent effect of physical and nutritional conditions, the growth and the levels of elatol in L. dendroidea clones were evaluated under different conditions of temperature, salinity, irradiance, and culture medium in the laboratory. Growth of L. dendroidea was clearly affected by all these factors, but elatol levels were influenced only by temperature and salinity. Better conditions for growth did not produce a similar effect on elatol production in L. dendroidea, contradicting the carbon/nutrient balance and growth/differentiation balance models. On the contrary, severe conditions of temperature and salinity promoted a decrease in elatol levels, as predicted by the environmental stress model. Our results using clones indicated that abiotic factors clearly take part in fostering chemical variations observed in natural populations, in addition to genetic factors, and can promote differential susceptibility of plant specimens to natural enemies.

Yield and chemical properties of rubber of Hevea clones according to phenological stages

The objective of this work was to evaluate the yield performance and macronutrient content of rubber extracted from four Hevea brasiliensis clones, under different tapping systems and plant phenological stages. The experiment was carried out in the 2010 and 2011 crop seasons, in a split-plot randomized complete block design, with four replicates. The main treatments - GT 1, PB 235, IAN 873, and RRIM 600 clones - were allocated in the plots, and the secondary treatments, which were the tapping systems 1/2S d/2, 1/2S d/4 ET 2.5%, and 1/2S d/7 ET 2.5%, were allocated in the subplots. The analyzed variables were natural rubber yield and macronutrient contents. Samples of natural rubber were obtained in the leaf development, mature leaf, and leaf senescence phenological stages. Rubber yield and its macronutrient contents are more influenced by tapping practice than by genetic material in the restrictive phenological stages of foliage.

FIG CLONES SELECTION OF cv. ROXO DE VALINHOS OBTAINED BY IRRADIATED BUDS

The fig tree (Ficus carica L.) is a fruit tree of great world importance and, therefore, the genetic improvement becomes an important field of research for the crop improvement, being necessary to gather information on this species, mainly regarding its genetic variability so that appropriate propagation projects and management are made. However, the fig, in Brazil, is all produced from only one cultivar, Roxo de Valinhos, which produces seedless fruit, making impossible the conventional breeding. So, the fig breeding through induced mutagenic becomes a very important research line, greatly contributing to the fig culture development. The objective of this study was to select fig plants formed by cuttings treated with gamma ray. The plants used were obtained from buds of the cv. Roxo de Valinhos. The cuttings were irradiated with gamma rays in an irradiator Gamma Cell at 10 cm from the tip of the cutting, at doses of 30 Gy with dose rate of 238 Gy/h. The experiment consisted of 450 treatments, where each formed plant was a treatment. The treatments were numbered sequentially from 1 to 450 and spaced 2.5 x 1.5 m. It was evaluated the vegetative and the fruits characteristics, and the incidence of major crop pests and diseases. The analysis data showed that there is genetic variability among treatments and that the plants under numbers 1, 5, 20, 79, 164, 189, 194, 201, 221, 214, 258, 301, 322, 392, 433 and 440 are probably genetic mutants that should be tested as commercial orchards.

Chemical composition and enzymatic digestibility of sugarcane clones selected for varied lignin content

Abstract Background The recalcitrance of lignocellulosic materials is a major limitation for their conversion into fermentable sugars. Lignin depletion in new cultivars or transgenic plants has been identified as a way to diminish this recalcitrance. In this study, we assessed the success of a sugarcane breeding program in selecting sugarcane plants with low lignin content, and report the chemical composition and agronomic characteristics of eleven experimental hybrids and two reference samples. The enzymatic digestion of untreated and chemically delignified samples was evaluated to advance the performance of the sugarcane residue (bagasse) in cellulosic-ethanol production processes. Results The ranges for the percentages of glucan, hemicellulose, lignin, and extractive (based on oven-dry biomass) of the experimental hybrids and reference samples were 38% to 43%, 25% to 32%, 17% to 24%, and 1.6% to 7.5%, respectively. The samples with the smallest amounts of lignin did not produce the largest amounts of total polysaccharides. Instead, a variable increase in the mass of a number of components, including extractives, seemed to compensate for the reduction in lignin content. Hydroxycinnamic acids accounted for a significant part of the aromatic compounds in the samples, with p-coumaric acid predominating, whereas ferulic acid was present only in low amounts. Hydroxycinnamic acids with ester linkage to the hemicelluloses varied from 2.3% to 3.6%. The percentage of total hydroxycinnamic acids (including the fraction linked to lignin through ether linkages) varied from 5.0% to 9.2%, and correlated to some extent with the lignin content. These clones released up to 31% of glucose after 72 hours of digestion with commercial cellulases, whereas chemically delignified samples led to cellulose conversion values of more than 80%. However, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during the enzymatic treatment. Conclusion Some of the experimental sugarcane hybrids did have the combined characteristics of high biomass and high sucrose production with low lignin content. Conversion of glucan to glucose by commercial cellulases was increased in the samples with low lignin content. Chemical delignification further increased the cellulose conversion to values of more than 80%. Thus, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during the enzymatic treatment.

Seleção de clones de figueira cv. roxo de Valinhos formados por gemas irradiadas

A figueira (Ficus carica L.), pertencente à família das Moráceas, constitui-se numa das mais importantes frutíferas cultivadas, elevando o Brasil à condição de décimo maior produtor e exportador de figos do mundo. Porém, a ficicultura apresenta alguns problemas fitossanitários, além de, no Brasil, estar toda implantada com uma única cultivar, a Roxo de Valinhos, que produz frutos sem sementes, inviabilizando o melhoramento convencional. Nesse sentido, o melhoramento genético, com o uso de mutagênicos, passa a ser uma linha de pesquisa altamente importante, podendo contribuir enormemente para o desenvolvimento da cultura. Diante disto, o objetivo do presente trabalho foi selecionar mutantes em plantas de figueira formadas por estacas irradiadas com raios gama, a fim de aumentar sua variabilidade genética com relação ao desenvolvimento vegetativo e reprodutivo. Utilizaram-se plantas formadas por estacas originadas de gemas da cultivar Roxo de Valinhos irradiadas com raios gama, no irradiador tipo Gamma a 0,10 m do ápice, na dose de Gy com taxa de dose de 238 Gy/h. O experimento constou de 450 tratamentos, sendo cada planta formada considerada um tratamento, numerando-as sequencialmente de 1 a 450 e cultivadas em espaçamento de 2,5 x 1,5 m.. As avaliações foram realizadas a partir das características tanto das folhas quanto dos frutos, bem como da incidência das principais pragas e doenças da cultura nestas plantas. Da análise dos dados, conclui-se que há variabilidade genética entre os tratamentos e que algumas plantas são prováveis mutantes, mostrando-se assim com potencial para posteriores estudos, devendo ser testadas em plantios comerciais.

In vitro generation and characterization of acute myeloid leukemia-reactive CD8 + cytotoxic T-lymphocyte clones from healthy donors

Donor-derived CD8+ cytotoxic T lymphocytes (CTLs) eliminating host leukemic cells mediate curative graft-versus-leukemia (GVL) reactions after allogeneic hematopoietic stem cell transplantation (HSCT). The leukemia-reactive CTLs recognize hematopoiesis-restricted or broadly expressed minor histocompatibility and leukemia-associated peptide antigens that are presented by human leukocyte antigen (HLA) class I molecules on recipient cells. The development of allogeneic CTL therapy in acute myeloid leukemia (AML) is hampered by the poor efficiency of current techniques for generating leukemia-reactive CTLs from unprimed healthy donors in vitro. In this work, a novel allogeneic mini-mixed lymphocyte/leukemia culture (mini-MLLC) approach was established by stimulating CD8+ T cells isolated from peripheral blood of healthy donors at comparably low numbers (i.e. 10e4/well) with HLA class I-matched primary AML blasts in 96-well microtiter plates. Before culture, CD8+ T cells were immunomagnetically separated into CD62L(high)+ and CD62L(low)+/neg subsets enriched for naive/central memory and effector memory cells, respectively. The application of 96-well microtiter plates aimed at creating multiple different responder-stimulator cell compositions in order to provide for the growth of leukemia-reactive CTLs optimized culture conditions by chance. The culture medium was supplemented with interleukin (IL)-7, IL-12, and IL-15. On day 14, IL-12 was replaced by IL-2. In eight different related and unrelated donor/AML pairs with complete HLA class I match, numerous CTL populations were isolated that specifically lysed myeloid leukemias in association with various HLA-A, -B, or -C alleles. These CTLs recognized neither lymphoblastoid B cell lines of donor and patient origin nor primary B cell leukemias expressing the corresponding HLA restriction element. CTLs expressed T cell receptors of single V-beta chain families, indicating their clonality. The vast majority of CTL clones were obtained from mini-MLLCs initiated with CD8+ CD62L(high)+ cells. Using antigen-specific stimulation, multiple CTL populations were amplified to 10e8-10e10 cells within six to eight weeks. The capability of mini-MLLC derived AML-reactive CTL clones to inhibit the engraftment of human primary AML blasts was investigated in the immunodeficient nonobese diabetic/severe combined immune deficient IL-2 receptor common γ-chain deficient (NOD/SCID IL2Rγnull) mouse model. The leukemic engraftment in NOD/SCID IL2Rγnull was specifically prevented if inoculated AML blasts had been pre-incubated in vitro with AML-reactive CTLs, but not with anti-melanoma control CTLs. These results demonstrate that myeloid leukemia-specific CTL clones capable of preventing AML engraftment in mice can be rapidly isolated from CD8+ CD62L(high)+ T cells of healthy donors in vitro. The efficient generation and expansion of these CTLs by the newly established mini-MLLC approach opens the door for several potential applications. First, CTLs can be used within T cell-driven antigen identification strategies to extend the panel of molecularly defined AML antigens that are recognizable by T cells of healthy donors. Second, because these CTLs can be isolated from the stem cell donor by mini-MLLC prior to transplantation, they could be infused into AML patients as a part of the stem cell allograft, or early after transplantation when the leukemia burden is low. The capability of these T cells to expand and function in vivo might require the simultaneous administration of AML-reactive CD4+ T cells generated by a similar in vitro strategy or, less complex, the co-transfer of CD8-depleted donor lymphocytes. To prepare clinical testing, the mini-MLLC approach should now be translated into a protocol that is compatible with good manufacturing practice guidelines.

In vitro characterization of human AML-reactive CTL clones generated from the naive subset of healthy donors and adoptive transfer into leukemia-engrafted NSG mice

Acute myeloid leukemia (AML) is a very aggressive cancer of the hematopoietic system. Chemotherapy and immunotherapeutical approaches including hematopoietic stem cell transplantation (HSCT) and donor lymphocyte infusion (DLI) are the only curative options available. The beneficial graft-versus-leukemia (GVL) effect of cellular immunotherapy is mostly mediated by donor-derived CD8+ T lymphocytes that recognize minor histocompatibility antigens (mHags) and leukemia-associated antigens (LAAs) presented on the surface of AML blasts (Falkenburg et al. 2008; Kolb 2008). A main complication is graft-versus-host disease (GVHD) that can be induced when cytotoxic T lymphocytes (CTLs) recognize broadly expressed antigens. To reduce the risk of GVHD, specific allogeneic T-cell therapy inducing selective GVL responses could be an option (Barrett & Le Blanc 2010; Parmar et al. 2011; Smits et al. 2011). This requires efficient in vitro strategies to generate AML-reactive T cells with an early differentiation phenotype as well as vigorous effector functions and humanized mouse models to analyze the anti-leukemic potential of adoptively transferred T cells in vivo. In this study, AML-reactive CTL clones and oligoclonal T-cell lines could be reliably generated from the naive subset of healthy HLA-class I-identical donors by stimulation with primary AML blasts in mini-mixed-lymphocyte / leukemia cultures (MLLCs) in eight different patient / donor pairs. These CTLs were promising candidates for cellular immunotherapy because of their relatively early differentiation phenotype and strong proliferative and lytic capabilities. The addition of the common γ-chain cytokine IL-21 to the stimulation protocol enabled more precursors to develop into potent leukemia-reactive CTLs, presumably by its beneficial effects on cell survival and antigen-specific proliferation during the first weeks of cultures. It also strengthened the early-stage phenotype. Three long-term cultured CTLs exemplarily transferred into leukemia-engrafted immunodeficient NSG mice mediated a significant reduction of the leukemic burden after a single transfusion. These results demonstrate that CTL clones with reactivity to patient-derived AML blasts can be isolated from the naive compartment of healthy donors and show potent anti-leukemic effects in vivo. The herein described allo-MLLC approach with in vitro “programmed” naive CTL precursors independent of a HSCT setting is a valuable alternative to the conventional method of isolating in vivo primed donor CTLs out of patients after transplantation (Kloosterboer et al. 2004; Warren et al. 2010). This would make leukemia-reactive CTLs already available at the time point of HSCT, when residual leukemia disease is minimal and the chances for complete leukemia eradication are high. Furthermore, leukemia-reactive CTLs effectively expanded by this in vitro protocol can be used as screening populations to identify novel candidate LAAs and mHags for antigen-specific immunotherapy.