Metabolizable Protein: 1. Predicting Equations to Estimate Microbial Crude Protein Synthesis in Small Ruminants

Microbial crude protein (MCP) produced in rumen could be estimated by a variety of protocols of experimental sampling and analysis. However, a model to estimate this value is necessary when protein requirements are calculated for small ruminants. This model could be useful to calculate rumen degradable protein (RDP) requirements from metabolizable protein (MP). Then, our objective was to investigate if there is a difference in MCP efficiency between sheep and goats, and to fit equations to predict ruminal MCP production from dietary energy intake. The database consisted of 19 studies with goats (n = 176) and sheep (n = 316), and the variables MCP synthesis (g/day), total digestible nutrients (TDN), and organic matter (OM) intakes (g/day), and OM digestibility (g/kg DM) were registered for both species. The database was used for two different purposes, where 70% of the values were sorted to fit equations, and 30% for validation. A meta-analytical procedure was carried out using the MIXED procedure of SAS, specie was considered as the fixed dummy effect, and the intercept and slope nested in the study were considered random effects. No effect of specie was observed for the estimation of MCP from TDN, digestible Organic Matter (dOM), or metabolizable energy (ME) intakes (P > 0.05), considering an equation with or without an intercept. Therefore, single models including both species at the same fitting were validated. The following equations MCP (g/day) = 12.7311 + 59.2956 × TDN intake (AIC = 3,004.6); MCP (g/day) = 15.7764 + 62.2612 × dOM intake (AIC = 2,755.1); and MCP (g/day) = 12.7311 + 15.3000 × ME intake (AIC = 3,007.3) presented lower values for the mean square error of prediction (MSEP) and its decomposition, and similar values for the concordance correlation coefficient (CCC) and for the residual mean square error (RMSE) when compared with equations fitted without an intercept. The intercept and slope pooled test was significant for equations without an intercept (P < 0.05), indicating that observed and predicted data differed. In contrast, predicted and observed data for complete equations were similar (P > 0.05)

Integrated analysis of plant hormones role during witches¿ broom progression in cocoa

Orientadores: Gonçalo Amarante Guimarães Pereira, Maria Carolina Scatolin do RioTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A Vassoura-de-bruxa, causada pelo fungo hemibiotrofico Moniliophthora perniciosa, e uma das principais doenças que acometem o cacaueiro (Theobroma cacao L.). Durante a fase biotrófica, o patôgeno coloniza o espaço intercelular e estimula a formação de ramos hipertróficos denominados vassoura verde. No estagio avançado da doença, o fungo passa a colonizar também o interior das células e observa-se a morte do tecido originando a vassoura seca. A semelhança destes sintomas aos observados em outras fitopatologias sugere um desequilíbrio hormonal em resposta a infecção. Neste contexto, este trabalho apresenta um estudo integrado dos genes relacionados a diferentes hormônios vegetais e uma caracterização dos genes relacionados às citocininas. A possível participação dos hormônios no estabelecimento e na progressão da Vassoura-de-bruxa do cacaueiro e discutida. Dados de RNA-seq foram gerados durante as diferentes fases da interação. Em seguida, genes diferencialmente expressos foram classificados de acordo com a terminologia do Gene Ontology. Além disso, a capacidade de biossíntese de citocininas pelo fungo M. perniciosa foi avaliada através do teste de complementação utilizando a levedura TM128 sln1?. Em paralelo, foi realizada uma caracterização detalhada dos genes que constituem o mecanismo de percepção de citocininas em cacau, sistema denominado two-component. Genes relacionados às respostas hormonais e de defesa foram os mais representados entre o grupo de diferencialmente expressos. Durante a fase assintomática, e observada a indução de genes relacionados a auxinas e etileno. Por outro lado, no estagio avançado, foram identificados genes relacionados à degradação de citocininas e as respostas de defesa mediadas por acido salicílico e jasmonato/etileno. Paralelamente, foi identificado no M. perniciosa um gene para a síntese de citocininas, entretanto, a funcionalidade deste não foi confirmada. Em T. cacau, a arquitetura do sistema two-component e similar a encontrada em Arabidopsis sp., porem, nenhuma alteração significativa na expressão dos genes que constituem este sistema foi identificada. Por essa razão, não foi possível estabelecer uma relação clara entre as citocininas e os sintomas da Vassoura-de-bruxa. Contudo, o padrão de expressão de genes relacionados à homeostase de auxinas, sugere uma regulação negativa das respostas de defesa contra patogenos biotroficos durante fase inicial da doençaAbstract: The Witches' Broom disease, caused by the hemibiotrophic fungus Moniliophthora perniciosa, is the major disease affecting cacao (Theobroma cacao L.). During its biotrophic phase, the pathogen colonizes the intercellular space and stimulates the formation of hypertrophic branches called green broom. In the advanced stage of the disease, the fungus starts also to colonize the interior of the cells and it is noted the death of tissue originating the dry broom. The similarity of these symptoms to those observed in other plant diseases suggests a hormonal imbalance in response to the infection. In this context, this work presents an integrated study of plant hormone-related genes and a characterization of cytokinin-related genes. The possible participation of these genes in the establishment and progression of Witches' Broom in cacao is discussed. RNA-seq data regarding different phases of the interaction were generated. After this, differentially expressed genes were classified according to Gene Ontology terminology. Furthermore, the ability of cytokinin biosynthesis by M. perniciosa was assessed by a complementation test using the yeast TM128 sln1?. In parallel, we performed a detailed characterization of genes that constitute the mechanism of perception of cytokinins in cacao, the two-component system. For these, the gene expression values have been accessed through the previously generated RNA-seq data. Defense and hormone-related genes were the most represented among the differentially expressed genes. During asymptomatic phase, we observe the induction of auxin and ethylene-related genes. During the symptomatic stage, genes related to the degradation of cytokinins and defense responses mediated by salicylic acid and jasmonate/ethylene was identified. Simultaneously, a putative gene for the synthesis of cytokinins was identified in M. perniciosa genome; however, the participation of this gene in cytokinin synthesis was not confirmed. In T. cocoa, the architecture of two-component system is similar to that one found in Arabidopsis sp.; however, no significant change in the expression of genes that constitute this system was identified. For this reason, it was not possible to establish a clear relationship between cytokinins and the symptoms of the Witches' Broom disease. Moreover, the expression pattern of homeostasis auxin-related genes suggests a negative regulation of defense responses against biotrophic pathogens during the initial stage of the diseaseDoutoradoGenetica de MicroorganismosDoutor em Genetica e Biologia Molecula

G-quadruplex Formation Enhances Splicing Efficiency Of Pax9 Intron 1.

G-quadruplexes are secondary structures present in DNA and RNA molecules, which are formed by stacking of G-quartets (i.e., interaction of four guanines (G-tracts) bounded by Hoogsteen hydrogen bonding). Human PAX9 intron 1 has a putative G-quadruplex-forming region located near exon 1, which is present in all known sequenced placental mammals. Using circular dichroism (CD) analysis and CD melting, we showed that these sequences are able to form highly stable quadruplex structures. Due to the proximity of the quadruplex structure to exon-intron boundary, we used a validated double-reporter splicing assay and qPCR to analyze its role on splicing efficiency. The human quadruplex was shown to have a key role on splicing efficiency of PAX9 intron 1, as a mutation that abolished quadruplex formation decreased dramatically the splicing efficiency of human PAX9 intron 1. The less stable, rat quadruplex had a less efficient splicing when compared to human sequences. Additionally, the treatment with 360A, a strong ligand that stabilizes quadruplex structures, further increased splicing efficiency of human PAX9 intron 1. Altogether, these results provide evidences that G-quadruplex structures are involved in splicing efficiency of PAX9 intron 1.13437-4

Adubação verde: uma alternativa sustentável para o Brasil

A adubação verde constitui uma das principais práticas que sustentam tanto as culturas como o solo e o meio ambiente. É uma técnica utilizada há milhares de anos por inúmeros povos. Promove inúmeros benefícios para as culturas, aumentando sua produtividade, garantindo renda e benefícios para os produtores que se acham excluídos do mercado competitivo. Além destes benefícios há de citar sua influência positiva sobre as propriedades químicas, físicas e biológicas dos solos e sobre o meio ambiente, já que com essa técnica há a dispensa do uso de adubos sintéticos. A presente revisão trás uma abordagem sistêmica sobre o que é, e quais são os benefícios da adubação verde, bem como trás algumas características de algumas leguminosas.The green manure is practices that sustain both crops as soil and the environment. It is a technique used for thousands of years by many people. Promotes many benefits to the crops, increasing their productivity by ensuring, income and benefits for producers who feel excluded from the competitive market. In addition to these benefits must quote their positive influence on the chemical properties, physical and biological soil and the environment, as with this technique there is a waiver of the use of synthetic fertilizers. This systemic review back an approach on what is and what are the benefits of green manure, as well as behind some characteristics of some legumes

Degradação ruminal da silagem de capim-elefante aditivado com cana-de-açúcar e farelo de cacau Ruminal degradation of elephant grass sillage added with sugarcane and cocoa meal

Objetivou-se avaliar a degradação ruminal da matéria seca (MS), da proteína bruta (PB) e da fibra em detergente neutro (FDN) das silagens de capim-elefante; capim-elefante ensilado com 30% de cana-de-açúcar; 15% de farelo de cacau; e 45% de cana mais 15% de farelo de cacau. Amostras de cada silagem foram incubadas no rúmen de três novilhos mestiços holandês x zebu durante os períodos de 0, 12, 24, 48, 72 e 96 horas. O delineamento experimental utilizado foi o de parcelas subdivididas e os coeficientes não lineares a, b e c foram estimados por meio de procedimentos iterativos de Gauss-Newton. Para o capim-elefante ensilado com farelo de cacau, foram observados maiores valores da fração a da MS e da PB, próximos a 60%, além de maiores taxas da degradação da fração b e da degradabilidade efetiva (DE) da PB para as taxas de passagem 2, 5 e 8% h-1. As silagens que apresentaram as maiores frações não-degradáveis i da FDN foram as silagens de capim-elefante sem aditivo e com 30% de cana, enquanto a silagem aditivada com 45% de cana e 15% de farelo de cacau destacou-se pela maior degradação potencial da MS, da PB e da FDN, em todos os períodos estudados, e maiores DE da MS e da PB para as taxas de passagem 2, 5 e 8% h-1. A inclusão associada da cana-de-açúcar com o farelo de cacau, no momento da ensilagem do capim-elefante, apresenta-se como alternativa para aumentar a degradabilidade da forragem em estádio de crescimento avançado.The objective was to evaluate the dry matter (DM), crude protein (CP) and neutral detergent fiber (NDF) ruminal degradation of silages of elephant grass; elephant grass ensiled with 30% of sugarcane; 15% of cocoa meal; and 45% of sugarcane plus 15% of cocoa meal. Samples of each silage were incubated in the rumen of three Holstein x Zebu crossbred steers during the periods of 0, 12, 24, 48, 72 and 96 hours. The experimental design used was the one of subdivided parcels and the non linear coefficients a, b and c were estimated through Gauss-Newton interactive procedures. For the elephant grass ensiled with cocoa meal, larger values of DM and CP fraction a were observed, next to 60%, besides the greatest b fraction degradation rates and effective degradability (ED) of CP to passage rates of 2, 5 and 8% h-1. The silages that showed the largest non-degradable i fractions of NDF were the elephant grass silages without additive and with 30% of sugarcane, while the silage addicted with 45% of sugarcane and 15% of cocoa meal showed up by the largest DM, CP and NDF potential degradation in all of the studied periods, and larger DM and CP ED for passage rates of 2, 5 and 8% h-1. The associated inclusion of sugarcane with cocoa meal at the moment of the elephant grass ensilage comes as an alternative to increase the degradability of forage in advanced growth stage