A rapid in vitro protocol for propagation of Piper aduncum and Piper hispidinervum, two species from Amazon region with multipurpose uses

The morphogenic potential of nodal explants of Piper aduncum and Piper hispidinervum (Piperaceae) was investigated and a protocol for rapid micropropagation is described. An experiment based on the saline formulation of Murashige and Skoog (MS) and Wood Plant medium (WPM) combined with different N6-benzylaminopurine (BAP) and 1-naphthalene acetic acid (NAA) concentrations was evaluated. After determining the optimal concentration of growth regulators, the multiplication rates for the species, which were distributed in five subcultures, was assessed, and the number of plantlets produced in this period was recorded. After assessing the plants from all five subcultures, plantlets with well-developed root and shoot systems were transferred to pots containing substrate for acclimatization. The culture of nodal segments of P. aduncum and P. hispidinervum on hormone-free medium was shown to be a suitable method for micropropagation due to the high multiplication rate and good plant development. The use of BAP or BAP + NAA resulted to formation of vitrified multiple shoots and callus formation at the base of the microcuttings. Even at concentrations lower than 1 mg L-1, the use of BAP resulted in vitrified multiple shoot and callus formation, without significantly improving the multiplication rates. For both species, the first subculture resulted in the greatest number of axillary buds, and mainly for P. hispidinervum, the MS medium was the most appropriate for in vitro multiplication of microcuttings. The species showed 100% root formation, and acclimatization of plants from the fifth subculture in a greenhouse resulted in 100% survival.Key words: Spiked pepper, long pepper, micropropagation, dillapiol, safrole, morphogenesis,  organogenesis

Estudo da identificação dos problemas rotineiros e cálculo do nível de eficiência nos processos industriais da Cooperativa Mista de Tomé-Açu (CAMTA)/ Study of the identification of routine problems and calculation of the efficiency level in industrial processes of the Tomé-Açu Mixed Cooperative (CAMTA)

Um meio eficaz de se atingir o objetivo e garantir resultados positivos dentro de uma empresa é administrando a rotina de trabalho. O gerenciamento de rotina define as tarefas diárias juntamente com as obrigações de cada colaborador, para que todo processo seja executado da melhor forma. Para a gestão de indústria de uma empresa, o mesmo também ocorre, é possível utilizar ferramentas para melhor administrar o tempo e as rotinas das equipes, entre elas, uma supervisão eficiente e melhores produtos adquiridos de fornecedores externos. O objetivo desse estudo é identificar os problemas do dia a dia da Cooperativa Agrícola Mista de Tomé-Açu (CAMTA), e, ainda, apresentar possíveis soluções para tais disfunções. Na metodologia, de caráter descritivo, fez-se o uso da pesquisa bibliográfica e de campo, as conclusões dos resultados foram dadas pelas análises destes dados e visitas na Cooperativa. Frente a isto foram diagnosticados problemas que se repetem com maior frequência, sendo, na fábrica de produção de polpas, o grande número de faltas de funcionários que, possivelmente, ocorre devido à tolerância da direção; já na fábrica de extração de óleo, a oscilação de energia se destaca por está diretamente ligada a todos os outros problemas

Transformação genética de laranjas doces com o gene D4E1 dirigido pelo promotor AtPP2

The objective of this work was to produce transgenic 'Pêra' and 'Valência' sweet orange plants using the D4E1 gene driven by the Arabidopsis thaliana phloem protein (AtPP2) promoter and to quantify transgene expression in different transformation events. Genetic transformation experiments were carried out with epicotyl segments co‑cultivated with Agrobacterium tumefaciens. Six plants from 'Pêra' sweet orange and seven plants from 'Valência' sweet orange were confirmed as different transgenic events by means of the polymerase chain reaction (PCR) and the Southern blot techniques. Transgene expression was quantified using real‑time quantitative PCR. D4E1 gene expression levels vary from 5 up to 50 times among different transformation events.O objetivo deste trabalho foi obter plantas transgênicas de laranja 'Pêra' e 'Valência' por meio do gene D4E1 dirigido pelo promotor Arabidopsis thaliana phloem protein (AtPP2) e quantificar a expressão dos transgenes nos diferentes eventos de transformação. Os experimentos de transformação genética foram realizados com segmentos de epicótilo cocultivados com Agrobacterium tumefaciens. Seis plantas de laranja 'Pêra' e sete plantas de laranja 'Valência' foram confirmadas como distintos eventos transgênicos por meio das técnicas de reação em cadeia da polimerase (PCR) e Southern blot. A expressão dos transgenes foi quantificada por PCR quantitativo em tempo real. Os níveis de expressão do gene D4E1 variam de 5 a 50 vezes entre os diferentes eventos de transformação

Sweet orange genetic transformation with a hairpin type construction gene fragment of V-ATPase-A of Diaphorina citri Kuwayama

O Brasil é o maior produtor de laranja doce no mundo. Entretanto, a cultura enfrenta grandes problemas, devido ao ataque de pragas e doenças, o que reduz a produtividade da cultura. Entre estas doenças, destaca-se o huanglongbing (HLB), doença associada a três espécies da bactéria Candidatus Liberibacter. No Brasil, o psilídeo Diaphorina citri é o inseto transmissor do HLB. A falta de cultivares de laranja doce resistentes ao HLB torna a transformação genética de plantas uma medida em potencial para o controle desta doença. Plantas transgênicas de laranja doce expressando um RNA de dupla fita (dsRNA) de um gene essencial à sobrevivência de D. citri podem resultar no controle do inseto por meio do mecanismo de RNA de interferência. Tal mecanismo resulta na degradação do RNAm homólogo ao dsRNA. Este trabalho teve por objetivo produzir plantas transgênicas de laranja doce, expressando um fragmento do gene DcV-ATPase-A de D. citri, em uma construção gênica tipo hairpin. Dessa forma, o silenciamento gênico por RNA de interferência seria ativado no psilídeo quando este for submetido à alimentação nas plantas transgênicas. O trabalho foi iniciado com a elaboração da construção gênica contendo uma sequência repetida e invertida do gene DcV-ATPase-A de D. citri, para formação de um hairpin. Segmentos de epicótilo, provenientes de sementes germinadas in vitro das laranjas \'Hamlin\', \'Pêra\' e \'Valência\' (Citrus sinensis L. Osbeck) foram utilizados como fontes de explantes para a transformação genética via Agrobacterium tumefaciens. Paralelamente aos experimentos de transformação genética, insetos adultos de D. citri foram submetidos a experimentos de alimentação artificial, contendo RNA de dupla fita (dsRNA) ou pequenos RNA interferentes (siRNA) da DcV-ATPase-A. Ao final do período de alimentação, foram avaliadas o número de insetos vivos e a expressão relativa do RNAm da DcV-ATPase-A. Através de PCR e Southern blot, a transgenia foi confirmada em 26 e 39 plantas de laranja \'Hamlin\' e \'Valência\', respectivamente. A transgenia não foi confirmada nas plantas regeneradas de laranja \'Pêra\'. O número de inserções no transgene variou de 1 a 4 cópias. A produção dos siRNAs foi confirmada em 10 plantas de laranja \'Valência\', através de siRNA blot. O uso de dietas artificiais contendo dsRNA ou siRNA do gene DcV-ATPase-A não resultou em diferenças significativas no número de insetos vivos, ou no nível de expressão relativa do RNAm do gene DcV-ATPase-A em insetos adultos de D. citri.Brazil is the largest sweet orange producer in the world. However, this crop faces major problems due to the attack of pests and diseases that reduce its productivity. Among the diseases, stands out the huanglongbing (HLB), disease associated with three different species of the Candidatus Liberibacter bacteria. In Brazil, the psyllid Diaphorina citri is the insect vector of HLB. The absence of resistant sweet orange cultivars to HLB makes the genetic transformation of plants a potential methodology to control this disease. Sweet orange transgenic plants engineered to express double strand RNA (dsRNA) of an essential gene for D. citri survival could result in insect control by RNA interference. This mechanism results in degradation of homologous RNAm to dsRNA. The aim of this work was to produce transgenic sweet orange plants expressing a fragment of D. citri DcV-ATPase-A gene, in a hairpin construction aiming gene silencing by RNA interference in D. citri, when fed on sweet orange transgenic plants. The work started developing a gene construct containing an inverted and repeated sequence of DcV-ATPase-A gene, to form a hairpin. Epicotyl segments collected from in vitro germinated seedlings of \'Hamlin\', \'Pêra\' and \'Valência\' sweet oranges (Citrus sinensis L. Osbeck) were used as explants for the genetic transformation experiments via Agrobacterium tumefaciens. At the same time, adults of D. citri underwent artificial diet experiments containing dsRNA or siRNA of DcV-ATPase-A. At the end of feeding time, the survival of insects and the relative expression of DcV-ATPase-A RNAm were evaluated. Through PCR and Southern blot analysis, 26 \'Hamlin\' and 39 \'Valência\' sweet orange transgenic lines were confirmed. None of the regenerated \'Pêra\' plants were transgenic. The transgenic plants had 1 to 4 T-DNA insertions. The siRNA products were observed in 10 \'Valência\' plants, through siRNA blot. The artificial diets containing dsRNA or siRNA of DcV-ATPase-A resulted in no differences in the number of live insects and in the relative expression of DcV-ATPase-A RNAm in adult insects

Propagação in vitro de sacaca (Croton cajucara Benth.): entendimentos sobre a dificuldade no desenvolvimento de protocolos de micropropagação da espécie

http://dx.doi.org/10.5007/2175-7925.2015v28n4p41Sacaca is a medicinal plant from the Amazonian biome and it has been regarded as a substitute for rosewood (Aniba roseaodora) to produce linalool. This paper aimed to evaluate in vitro vegetative propagation of sacaca, including the establishment of propagules from the field, decontamination protocols, and determination of multiplication rates, besides describing limiting aspects for the culture during in vitro experiments. We used 1.0 cm microcuttings with an axillary bud, collected from adult plants in the field. Disinfestation treatments were tested in the establishment, and there is an evaluation of the collecting month influence on the contamination rates. After disinfestation, microcuttings were placed in test tubes containing MS medium, added with BAP (0, 1, 2 and 3 mg L-1) and GA3 (0 and 0.5 mg L-1). In vitro establishment of sacaca with 41.9% of sprouted microcuttings was obtained. The contamination rate reached 58.1% (65.4% caused by fungi and 34.6% by bacteria), with greater occurrence when propagules were collected between October and January, the rainiest months in the Amazon region. The increased BAP and GA3 concentrations in the culture medium provided significant improvements in the material multiplication rates. In spite of the results obtained, the species shows peculiarities and limitations to in vitro cultivation that were identified and described in this paper.http://dx.doi.org/10.5007/2175-7925.2015v28n4p41A sacaca é uma planta medicinal do bioma Amazônico e tem sido considerada como substituta do pau-rosa (Aniba roseodora) na produção de linalol. Este trabalho objetivou avaliar a propagação vegetativa in vitro de sacaca, incluindo o estabelecimento de propágulos do campo, protocolos de descontaminação e a determinação de taxas de multiplicação, além de descrever aspectos limitantes da cultura durante os trabalhos in vitro. Foram utilizadas microestacas de 1,0 cm, com uma gema axilar, coletadas de plantas adultas do campo. Tratamentos de desinfestação foram testados no estabelecimento, além de se avaliar a influência do mês do ano da coleta sobre as taxas de contaminação. Após desinfestadas, as microestacas foram inoculadas em tubos de ensaio com meio MS, suplementado com BAP (0, 1, 2 e 3 mg L-1) e AG3 (0 e 0,5 mg L-1). Foi obtido o estabelecimento in vitro da sacaca com 41,9% das microestacas brotadas. A taxa de contaminação alcançou 58,1% (65,4% de origem fúngica e 34,6% de origem bacteriana), com maior ocorrência quando os propágulos foram coletados entre os meses de outubro e janeiro, os mais chuvosos da região amazônica. O aumento nas concentrações de BAP e AG3 no meio de cultura melhorou as taxas de multiplicação do material. Apesar dos resultados obtidos, a espécie apresenta uma série de peculiaridades e limitações ao cultivo in vitro que foram identificadas e relatadas neste trabalho.

Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter

The objective of this work was to produce transgenic 'Pêra' and 'Valência' sweet orange plants using the D4E1 gene driven by the Arabidopsis thaliana phloem protein (AtPP2) promoter and to quantify transgene expression in different transformation events. Genetic transformation experiments were carried out with epicotyl segments co‑cultivated with Agrobacterium tumefaciens. Six plants from 'Pêra' sweet orange and seven plants from 'Valência' sweet orange were confirmed as different transgenic events by means of the polymerase chain reaction (PCR) and the Southern blot techniques. Transgene expression was quantified using real‑time quantitative PCR. D4E1 gene expression levels vary from 5 up to 50 times among different transformation events

Training Mode Comparisons on Cardiorespiratory, Body Composition and Metabolic Profile Adaptations in Reproductive Age Women: A Systemic Review and Meta-Analysis

Purpose: This study aimed to compare the effects of high-intensity interval training (HIT), sprint interval training (SIT) and moderate-intensity continuous training (MICT) on cardiorespiratory fitness (CRF), weight (kg), body fat mass (%), plasma glucose (fasting) and lipid levels in reproductive-age women. Method: The search was conducted in Pubmed, Cochrane Library, Virtual Health Library and Scielo. The meta-analyses were conducted using Review Manager software for random-effects models. The results were presented as standardized mean differences and 95%CI, which were calculated to determine the effect size of HIT/SIT and MICT interventions. Results: Eleven articles meet the inclusion criteria. The analyses demonstrated that all exercise modes improved body composition and metabolic profile, but nevertheless, MICT was significantly better at improving CRF (mL·min−1·kg−1) compared with HIT (2.45 mL·min−1·kg−1 (95% CI: 1.15 to 3.75 mL·min−1·kg−1); p < 0.05; I2 = 0%) and with SIT (0.98 mL·min−1·kg−1 (95% CI: −0.98 to 2.93 mL·min−1·kg−1); p = 0.33; I2 = 53%). Conclusion: Both HIT and SIT have the potential to be used as a training modality in reproductive-age women, with similar effects to MICT on body composition/metabolic markers but inferior effects on CRF, suggesting that HIT/SIT may be considered a “time-efficient component″ of weight management programs. However, the variability in the secondary outcome measures, coupled with the small sample sizes in studies, limits this finding

Training Mode Comparisons on Cardiorespiratory, Body Composition and Metabolic Profile Adaptations in Reproductive Age Women: A Systemic Review and Meta-Analysis

Purpose: This study aimed to compare the effects of high-intensity interval training (HIT), sprint interval training (SIT) and moderate-intensity continuous training (MICT) on cardiorespiratory fitness (CRF), weight (kg), body fat mass (%), plasma glucose (fasting) and lipid levels in reproductive-age women. Method: The search was conducted in Pubmed, Cochrane Library, Virtual Health Library and Scielo. The meta-analyses were conducted using Review Manager software for random-effects models. The results were presented as standardized mean differences and 95%CI, which were calculated to determine the effect size of HIT/SIT and MICT interventions. Results: Eleven articles meet the inclusion criteria. The analyses demonstrated that all exercise modes improved body composition and metabolic profile, but nevertheless, MICT was significantly better at improving CRF (mL·min−1·kg−1) compared with HIT (2.45 mL·min−1·kg−1 (95% CI: 1.15 to 3.75 mL·min−1·kg−1); p −1·kg−1 (95% CI: −0.98 to 2.93 mL·min−1·kg−1); p = 0.33; I2 = 53%). Conclusion: Both HIT and SIT have the potential to be used as a training modality in reproductive-age women, with similar effects to MICT on body composition/metabolic markers but inferior effects on CRF, suggesting that HIT/SIT may be considered a “time-efficient component″ of weight management programs. However, the variability in the secondary outcome measures, coupled with the small sample sizes in studies, limits this finding

Identification of genes from the general phenylpropanoid and monolignol-specific metabolism in two sugarcane lignin-contrasting genotypes

The phenylpropanoid pathway is an important route of secondary metabolism involved in the synthesis of different phenolic compounds such as phenylpropenes, anthocyanins, stilbenoids, flavonoids, and monolignols. The flux toward monolignol biosynthesis through the phenylpropanoid pathway is controlled by specific genes from at least ten families. Lignin polymer is one of the major components of the plant cell wall and is mainly responsible for recalcitrance to saccharification in ethanol production from lignocellulosic biomass. Here, we identified and characterized sugarcane candidate genes from the general phenylpropanoid and monolignol-specific metabolism through a search of the sugarcane EST databases, phylogenetic analysis, a search for conserved amino acid residues important for enzymatic function, and analysis of expression patterns during culm development in two lignin-contrasting genotypes. Of these genes, 15 were cloned and, when available, their loci were identified using the recently released sugarcane genomes from Saccharum hybrid R570 and Saccharum spontaneum cultivars. Our analysis points out that ShPAL1, ShPAL2, ShC4H4, Sh4CL1, ShHCT1, ShC3H1, ShC3H2, ShCCoAOMT1, ShCOMT1, ShF5H1, ShCCR1, ShCAD2, and ShCAD7 are strong candidates to be bona fide lignin biosynthesis genes. Together, the results provide information about the candidate genes involved in monolignol biosynthesis in sugarcane and may provide useful information for further molecular genetic studies in sugarcane