Emissão de óxido nitroso em sistemas de lavoura contínua e integrada com pecuária no Cerrado e sua relação com frações da matéria orgânica do solo

Tese (doutorado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, Programa de Pós-Graduação em Agronomia, 2017.A integração lavoura-pecuária (ILP) é um sistema que tem sido cada vez mais implementado em várias regiões no Brasil. O acúmulo de matéria orgânica do solo (MOS) e a mitigação de emissão de gases de efeito estufa (GEE) são benefícios dos sistemas integrados. O óxido nitroso (N2O) é um dos principais gases do efeito estufa e sua emissão para a atmosfera é influenciada por diversos fatores, entre eles a disponibilidade de carbono (C) orgânico no solo. Para compreender como o C interfere na emissão desse gás em um sistema integrado, o presente estudo foi desenvolvido com o objetivo de avaliar a emissão de N2O do solo em sistema de integração lavoura-pecuária após 24 anos de duração de experimento e relacionar essas emissões com as frações da MOS em condições tropicais no Cerrado brasileiro. A pesquisa foi desenvolvida na Embrapa Cerrados, em Planaltina, DF, em um experimento instalado em 1991, cujo delineamento estatístico foi em blocos casualizados, com duas repetições. Os agrossistemas estudados foram: integração lavoura-pecuária sob plantio direto (ILP-PD), lavoura contínua sob plantio convencional (LC-PC), lavoura contínua sob plantio direto (LC-PD), e o Cerrado nativo como área de referência. No Capítulo I foram monitoradas as emissões de N2O nos diferentes agroecossistemas em um ano de avaliação, onde houve sucessão de culturas nas áreas agrícolas nesse período (sorgo consorciado com braquiária Piatã– soja – milho), e foram relacionadas com as co-variáveis edafoclimáticas (Nmineral – nitrato e amônio; espaço poroso saturado por água - EPSA). Conclui-se nesse capítulo que os maiores fluxos de N2O foram observados após a fertilização e os eventos de precipitação. Também foi possível verificar um efeito combinado do manejo do solo com a rotação de culturas nas emissões de N2O. Nos sistemas com lavoura contínua, as emissões foram maiores do que no sistema integrado durante o ciclo do sorgo e safrinha, em virtude da decomposição dos resíduos da cultura da soja, de baixa relação C/N e rica em N, principalmente dos nódulos com rizóbios. No Capítulo II, considerando que a MOS e a agregação do solo interferem na emissão de N2O, objetivou-se caracterizar as frações da MOS nos diferentes agroecossistemas. As emissões de N2O foram monitoradas por um ano e meio (~509 dias). Foram determinados os teores totais de carbono orgânico (COT) e nitrogênio (NT) e as frações da MOS: carbono oxidado por permanganato de potássio (CL), carbono da biomassa microbiana (CBM), carbono orgânico particulado (COP), carbono orgânico associado aos minerais (COAM), substâncias húmicas e suas relações (humina – HUM, ácido fúlvico – AF e ácido húmico – AH), carbono inerte (CI) e carbono e nitrogênio associado aos macro e microagregados (C-MACRO, C-MICRO, N-MACRO e N-MICRO). Os resultados desse capítulo mostraram que a emissão acumulada de N2O foi maior no LC-PC (4,56 kg ha-1 ), intermediária no LC-PD (3,73 kg ha-1 ) e no ILP-PD (1,75 kg ha-1 ), e menor no Cerrado (0,63 kg ha-1 ) em um ano e meio de monitoramento. A análise de componentes principais realizada entre o N2O acumulado no período avaliado e as frações de carbono demonstraram maior associação entre a menor emissão, que foi verificada no Cerrado, com as frações CBM, COAM e HUM, essas duas últimas representando as frações estáveis da MOS. A avaliação do acúmulo de frações de C nos diferentes agroecossistemas demonstra que apesar do Cerrado nativo apresentar maior população microbiana e altos teores de C total, há predominância das frações estáveis, que são menos disponíveis para a microbiota. Isso ajuda a explicar as menores taxas de mineralização, e consequentemente, a menor produção de N2O. Entre os sistemas agrícolas, as menores emissões de N2O foram verificadas no ILP-PD, o sistema que mais se aproximou da área de referência (Cerrado nativo). No geral, os maiores teores de C nas frações estáveis e em agregados também foram verificados no ILP-PD, confirmando seu potencial como sistema mitigador de emissão de N2O e acumulador de carbono no solo.The integrated crop-livestock (CLS) is a system which is increasingly being implemented in various regions of Brazil. The build-up of soil organic matter (SOM) and the mitigation of greenhouse gases (GHG) are benefits of the integrated systems. The nitrous oxide (N2O) is one of the major gases responsible for the greenhouse effect and its emission into the atmosphere is influenced by a number of factors, among which is the availability of organic carbon (C) in the soil. To better comprehend how C interferes in the emission of this gas in an integrated system, the present study was developed with the objective of evaluating N2O soil emission in a crop-livestock integrated system after a 24-year experiment and relate these emissions to SOM fractions in tropical conditions in the Brazilian Cerrado. The research was developed at Embrapa Cerrados, in Planaltina, DF, in an experiment installed in 1991, with statistical outline in randomized blocks, with two repetitions. The studied areas consisted of a system of continuous cropping under no tillage (CC-NT), a system of continuous cropping with annual heavy disc harrow (CC-CT), an integrated crop-livestock system under no tillage (CLSNT) and an adjacent area of native Cerrado as reference. In Chapter I, N2O emissions in the different agroecosystems were monitored in a one-year evaluation, with culture sequences in agricultural areas during this period (sorghum intercropped with Brachiaria brizantha cv. Piatã - soy – maize), and were related to the soil and climate co-variables (N-mineral – nitrate and ammonium; water filled pore space - WFPS). This chapter concludes that the larger N2O fluxes were observed after fertilization and precipitation events. It was also possible to verify a combined effect of soil management and crop rotation in N2O emissions. In continuous crop systems, the emissions were greater than that of the integrated system during the sorghum and interim harvest cycle, due to the decomposition of residues of soy crops, which presents a low C/N ratio and is rich in N in rhizobia. In Chapter II, considering that SOM and soil aggregation interfere in N2O emissions, the objective was to characterize SOM fractions in the different crop systems. N2O emissions were monitored for a year and a half (~509 days). The total organic carbon (TOC) and total nitrogen (TN) and SOM fractions were determined: permanganate oxidizable carbon (COxP), microbial biomass carbon (CMIC), particulate organic carbon (POC), mineral-associated organic carbon (MOC), humic fractions (humic acid – HA, fulvic acid – FA, humin – HUM), inert carbon (IC), Carbon and nitrogen levels in macro and microaggregates (C - MACRO, C - MICRO, N-MACRO and N-MICRO). The results of this chapter showed that the cumulative N2O emission was greater in the CC-CT (4.56 kg ha-1 ), intermediary in the CC-NT (3.73 kg ha-1 ) and the CLS-NT (175 kg ha-1 ), and lesser in the Cerrado (0.63 kg ha-1 ) during the year-and-a-half monitoring. The analysis of main components between the cumulative emission of N2O in the evaluated period and the carbon fractions showed greater association between the lower emissions, which was verified in the Cerrado, and the CMIC, MOC and HUM fractions, the last two representing SOM stable fractions. The evaluation of C fractions accumulation in the different crop systems demonstrates that although the native Cerrado presents a larger microbial population and high levels of total C, there is a predominance of stable fractions, which are less available to the microbiota. This helps explain the lower mineralization rates and, consequently, the lower N2O production. Among the crop systems, the N2O emissions were verified in the CLS-NT, system which more closely resembled the reference area (native Cerrado). In general, the greatest levels of C in stable fractions and in aggregates were also verified in the CLS-NT, confirming its potential as system which mitigates N2O emissions and propitiates carbon accumulation in the soil

CRISPR/Cas9-based generation of knockdown mice by intronic insertion of artificial microRNA using longer single-stranded DNA.

Knockdown mouse models, where gene dosages can be modulated, provide valuable insights into gene function. Typically, such models are generated by embryonic stem (ES) cell-based targeted insertion, or pronuclear injection, of the knockdown expression cassette. However, these methods are associated with laborious and time-consuming steps, such as the generation of large constructs with elements needed for expression of a functional RNAi-cassette, ES-cell handling, or screening for mice with the desired knockdown effect. Here, we demonstrate that reliable knockdown models can be generated by targeted insertion of artificial microRNA (amiRNA) sequences into a specific locus in the genome [such as intronic regions of endogenous eukaryotic translation elongation factor 2 (eEF-2) gene] using the Clustered Regularly Interspaced Short Palindromic Repeats/Crispr associated 9 (CRISPR/Cas9) system. We used in vitro synthesized single-stranded DNAs (about 0.5-kb long) that code for amiRNA sequences as repair templates in CRISPR/Cas9 mutagenesis. Using this approach we demonstrate that amiRNA cassettes against exogenous (eGFP) or endogenous [orthodenticle homeobox 2 (Otx2)] genes can be efficiently targeted to a predetermined locus in the genome and result in knockdown of gene expression. We also provide a strategy to establish conditional knockdown models with this method

Métodos para determinação do carbono orgânico em solos do Cerrado

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, Programa de Pós-Graduação em Agronomia, 2013.O bioma Cerrado tem se destacado mundialmente pelos índices de produtividade de grãos elevados. No entanto, a transformação das áreas de vegetação nativa dessa região para sistemas de exploração agropecuária tem sido acompanhada por perdas do estoque original de matéria orgânica do solo. Com a ampliação de programas governamentais de incentivo ao uso de sistemas conservacionistas de baixa emissão de carbono, surge a necessidade de se acompanhar as variações de carbono no solo, aumentando assim a necessidade de padronização dos diferentes métodos existentes. Diversos métodos tem sido empregados na determinação dos teores de carbono nos solos do Cerrado, não sendo raras discrepâncias entre os resultados obtidos para uma mesma amostra de solo. Características como a textura e mineralogia dos solos podem interferir no resultado obtido. Desta forma, é preciso aprofundar o espectro de análises envolvendo diferentes procedimentos laboratoriais e essas características de solos. O objetivo desse trabalho foi avaliar diferentes métodos analíticos empregados na determinação do carbono orgânico numa ampla diversidade de solos do Cerrado, buscando determinar aqueles que apresentem maior precisão e exatidão, bem como visa buscar possíveis correlações existentes entre os protocolos atualmente em uso e suas relações com a mineralogia e a textura. Foram analisadas amostras representativas do bioma Cerrado em áreas nativas e sob uso agropecuária, coletadas na camada de 0-20 cm. Os teores de carbono orgânico foram determinados pelos métodos por combustão úmida (Walkley & Black, Colorímetro, Mebius) e por combustão seca (Gravimétrico e Análise elementar -CHNS/O). Espectros na região do infravermelho próximo foram também obtidos com a perspectiva de obtenção de um modelo de predição do carbono. O método Mebius foi o que mais se aproximou do método de referência (análise elementar) apresentando coeficiente de regressão de 87%, demonstrando ser o método mais preciso em relação aos demais. A mineralogia do solo interferiu na determinação de carbono, principalmente quando utilizado o método Gravimétrico. Os modelos de predição de carbono obtidos apresentaram elevada relação com os métodos mais utilizados em laboratórios de rotina (Walkley & Black e Colorimétrico). ______________________________________________________________________________ ABSTRACTThe Cerrado biome is widely known for its high productivity index. However, the rapid transformation of native vegetation for agriculture in this region has been accompanied by the loss of the original stocks of soil organic matter. With the expansion of government programs to encourage the use of conservation tillage systems for low carbon comes the need to monitor changes in soil carbon, thereby increasing the necessity to standardize carbon values determined by different analysis methods. Several techniques have been used for the determination of carbon in soils of the Cerrado biome and the analytical procedure was influenced by different factors such as the soil texture and mineralogy. This study aimed to evaluate different analytical procedures employed in determining organic carbon in a wide diversity of Cerrado soils, to determine those with greater precision and accuracy, as well as aims to look for possible correlations between the protocols currently in use and their relationships with mineralogy and texture. Representative samples of the Cerrado biome in native areas under agricultural use were analyzed, they were collected at a depth of 0-20cm. The organic carbon was determined by wet combustion methods - Walkley & Black, Colorimeter, Mebius, and dry combustion - Gravimetric and elemental analysis (CHNS / O). These samples were also collected spectra by near infrared spectroscopy (NIRS) and through reference methods have been proposed prediction models carbon using the PLS. The Mebius was the one closest to the standard method (elemental analysis) showing a correlation of 87%. The colorimetric method is more practical and has advantages such as the use of smaller amounts of reagents; it may be an option to replace the Walkley & Black. The mineralogy of the soil interferes with the determination of carbon, particularly when using the gravimetric method. Regarding carbon prediction models, methods were used as reference elemental analysis and methods used in routine laboratories (Walkley and Black & Colorimetric)

On-off selectivity and asymmetry in apparent contrast: An adaptation study

Using textures composed of sparse bright/dark elements that can activate either on or off sensors selectively, Sato, Motoyoshi, and Sato (2012) reported simultaneous contrast-contrast effects tuned for contrast polarity. As with contrast-contrast effects, prolonged viewing of high-contrast stimuli reduces the perceived contrast of a subsequently presented stimulus. The present study examined whether contrast aftereffects are also selective for luminance polarity using texture patterns composed of sparse bright/dark elements. Results revealed that contrast aftereffects are selective for luminance polarity (polarity selectivity) but that adaptation aftereffects occur asymmetrically depending on the polarity of the adapter (polarity asymmetry). Polarity selectivity and asymmetry in adaptation aftereffects are reduced but not completely diminished if adapter and test stimuli are presented to separate eyes (dichoptically). Our results support the idea that suprathreshold contrast perception and its adaptational shifts are jointly determined by responses between monocular and binocular units

Allosteric Regulation of HIV-1 Reverse Transcriptase by ATP for Nucleotide Selection

Background: Human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) is a DNA polymerase that converts viral RNA genomes into proviral DNAs. How HIV-1 RT regulates nucleotide selectivity is a central issue for genetics and the nucleoside analog RT inhibitor (NRTI) resistance of HIV-1. Methodology/Principal Findings: Here we show that an ATP molecule at physiological concentrations acts as an allosteric regulator of HIV-1 RT to decrease the Km value of the substrate, decrease the kcat value, and increase the Ki value of NRTIs for RT. Computer-assisted structural analyses and mutagenesis studies suggested the positions of the ATP molecule and NRTIresistance mutations during a catalytic reaction, which immediately predict possible influences on nucleotide insertion into the catalytic site, the DNA polymerization, and the excision reaction. Conclusions/Significance: These data imply that the ATP molecule and NRTI mutations can modulate nucleotide selectivity by altering the fidelity of the geometric selection of nucleotides and the probability of an excision reaction

Peroxisome proliferator-activated receptor δ as a molecular target to regulate lung cancer cell growth

AbstractIt has been assumed that prostaglandin (PG)I2 signaling contributes to the negative growth control of lung cancer cells; however, the mechanism remains unresolved. PGI2 functions through a cell surface G protein-coupled receptor (prostaglandin I2-binding receptor, IP) and also exerts an effect by interacting with a nuclear hormone receptor, peroxisome proliferator-activated receptor δ (PPARδ). We found that PPARδ was a key molecule of PGI2 signaling to give negative growth control of lung cancer cells (A549), using carbarprostacyclin, a PGI2 agonist for IP and PPARδ, and L-165041, a PPARδ agonist. Furthermore, PPARδ-induced cell growth control was reinforced by the inhibition of cyclooxygenase. These results suggest that PPARδ activation under the suppression of PG synthesis is important to regulate lung cancer cell growth

Verbal Noun Complements and Complex Predicates in Japanese

Focusing on the complex predicate formation in Japanese, particularly on the complex predicates involving the verbal noun, the light verb su(nl) and an affixal predicate, this paper presents new data and additional arguments for Saito and Hoshi\u27s (1998, 2000) proposal on theta-role assignment by a predicate in the incorporated position, and it also proposes some significant modification to their proposal. The paper argues that the quasi-light verb construction is shown to involve the same process as the light verb construction without assuming the theta-absorption/merger in the former. It also discusses instances of the light verb su(nt) , where it functions not as the head of VP but rather as morphological "glue" between a verbal noun and a V-taking affixal predicate. The proposed analysis makes correct predications about constructions involving further complex predicates and those involving unaccusative verbal nouns

視覚情報処理における輝度極性の意義

学位の種別: 課程博士審査委員会委員 : （主査）東京大学教授 佐藤 隆夫, 東京大学教授 横澤 一彦, 東京大学教授 今水 寛, 東京大学准教授 村上 郁也, 東京大学准教授 本吉 勇University of Tokyo(東京大学