Micropropagation studies were carried out using the seeds for establishing an in vitro culture of Vriesea gigantea and Vriesea philippocoburgii. Germination rate of V. gigantea was higher than of V. philippocoburgii. Plantlets of V. philippocoburgii gave rise to many adventitious shoots when cultivated in Knudson basal medium. In contrast, for V. gigantea, a higher salts-concentration was needed, so that the number of shoots was increased by Murashige and Skoog medium. Addition of activated charcoal and naphthaleneacetic acid in regeneration medium allowed the growth of shoots and the formation of roots, confirming the success of in vitro culture. The differences in expression of the genotypes reinforce the need of more research in order to set up the conditions that could offer the best response of the specific tissues.
-Somatic embryogenesis is an efficient method for the production of target cells for soybean genetic transformation. However, this method still offers low percentages of plant regeneration, and perhaps is related to the maturation process and high morphological abnormalities of the matured embryos. This study aimed to identify a maturation medium that could contribute to the outcome of more efficient plant regeneration results. Embryogenic clusters, derived from cotyledons of immature seeds of the soybean cultivars Bragg and IAS5, were used as starting material for embryos development. Different maturation media were tested by using 6% maltose, 3% sucrose or 6% sucrose, combined with or without 25 g L -1 of the osmotic regulator polyethylene glycol (PEG-8000). The histodifferentiated embryos were quantified and classified in morphological types. Percentages of converted embryos were analyzed. Cultivar Bragg resulted in higher matured embryo quantities, but lower percentages were obtained for the conversion in comparison to cultivar IAS5. While the addition of PEG did not affect the number of embryos converted into plants, 6% sucrose enhanced the conversion percent significantly.Index terms: Glycine max, PEG-8000, sucrose, osmotic regulation. Fontes de carbono e polietilenoglicol na conversão de embriões somáticos de sojaResumo -A embriogênese somática é um eficiente método na produção de células-alvo para a transformação genética de soja. Entretanto, este método ainda apresenta baixas porcentagens de regeneração de plantas, o que pode estar relacionado ao processo de maturação e às altas anormalidades morfológicas dos embriões maturos. Este estudo visou a identificar um meio de maturação que pudesse contribuir com resultados mais eficientes de regeneração de plantas. Foram utilizados conjuntos embriogênicos derivados de cotilédones imaturos de sementes de soja das cultivares Bragg e IAS5 para o desenvolvimento de embriões. Testaram-se diferentes meios de maturação, com a utilização de 6% de maltose, 3% de sacarose ou 6% de sacarose, combinadas ou não com 25 g L -1 do regulador osmótico polietilenoglicol (PEG-8000). Os embriões histodiferenciados foram quantificados e classificados em tipos morfológicos. A porcentagem de embriões convertidos em plantas foi analisada. A cultivar Bragg apresentou maior quantidade de embriões maturos, mas foram obtidas menores porcentagens de conversão em comparação à cultivar IAS5. A adição de PEG não afetou o número de embriões convertidos em plantas. Entretanto, a concentração de 6% de sacarose aumentou significativamente a porcentagem de conversão.Termos para indexação: Glycine max, PEG-8000, sacarose, regulação osmótica.
The genotoxicity of untreated and treated sewage from two municipal wastewater treatment plants (WTP BN and WTP SJN) in the municipality of Porto Alegre, in the southern Brazilian state of Rio Grande do Sul, was evaluated over a one-year period using the Tradescantia pallida var. purpurea (Trad-MCN) bioassay. Inflorescences of T. pallida var. purpurea were exposed to sewage samples in February (summer), April (autumn), July (winter) and October (spring) 2009, and the micronuclei (MCN) frequencies were estimated in each period. The high genotoxicity of untreated sewage from WTP BN in February and April was not observed in treated sewage, indicating the efficiency of treatment at this WTP. However, untreated and treated sewage samples from WTP SJN had high MCN frequencies, except in October, when rainfall may have been responsible for reducing these frequencies at both WTPs. Physicochemical analyses of sewage from both WTPs indicated elevated concentrations of organic matter that were higher at WTP SJN than at WTP BN. Chromium was detected in untreated and treated sewage from WTP SJN, but not in treated sewage from WTP BN. Lead was found in all untreated sewage samples from WTP SJN, but only in the summer and autumn at WTP BN. These results indicate that the short-term Trad-MCN genotoxicity assay may be useful for regular monitoring of municipal WTPs.
In an attempt to establish an alternative plant regeneration system for soybean [Glycine max (L.) Merrill] cultivars used in Brazilian breeding programs, ten genotypes were tested for their embryogenic potential. Cotyledons were removed as explants from immature seeds harvested from field-grown plants. After 45 days on induction medium, the number of responding cotyledons and the number of somatic embryos per immature cotyledon were evaluated. The percentage of explants that produced somatic embryos varied from 1 to 70% among cultivars. The average number of somatic embryos produced per cotyledon pair ranged from 0.01 to 10.3 with a mean of 3.4. Suspension cultures were initiated with three Agrobacterium tumefaciens susceptible cultivars. Suspensions were successfully developed from Bragg and IAS5 cultivars. The packed cell volume, in one-month growth, increased 8.1 fold for Bragg and 3.5 fold for IAS5 and the fresh weight increased 6.6 and 2.8 fold, respectively. The cultivars differed for the analysed parameters. All tissue from each cultivar was transferred to the maturation medium and subsequently to the germination medium. The germination frequency was 45.7 and 54.9% for Bragg and IAS5, respectively. Plants were gradually exposed to ambient humidity over one week and then planted in soil. All plants yielded seeds in the greenhouse.
Cattleya intermedia is an Atlantic Forest species endemic to Brazil that is classed as vulnerable on the list of threatened species. In this study, C. intermedia plantlets were micropropagated in an asymbiotic culture and the influence of different concentrations of sucrose (15, 30, 45 and 60 g L -1 , plus a zero sucrose medium) and macronutrient salts (complete Murashige and Skoog (MS) medium and half MS medium (with half-strength macronutrients)) on survival and development of the plantlets was evaluated. In all treatments 100% plantlet survival was achieved. The integrated analysis of height of aerial part, number of leaves per plantlet, fresh mass, number of roots per plantlet and length of the longest root showed that the plantlets exhibited greatest development at the half-strength macronutrient concentrations with 45 or 60 g L -1 of sucrose, as well as at the complete macronutrient concentration with 60 g L -1 of sucrose. Plantlets acclimatized and reintroduced to an environment in which the species occurs naturally exhibited 98.6% survival. The results obtained in this study allowed the establishment of optimal conditions for asymbiotic micropropagation, which is a requisite for future studies focused on conservation of C. intermedia.
-(Megaspore germination and initial development of Regnellidium diphyllum Lindman (Pteridophyta, Marsileaceae) sporophytes in the presence of cadmium). Regnellidium diphyllum has its distribution restricted to Southern Brazil and adjoining localities in Uruguay and Argentina. Currently it is on the list of threatened species of Rio Grande do Sul. The conversion of wetlands into agricultural areas or soil contamination by the introduction of waste products and fertilizers may compromise the establishment and survival of this species. Among the pollutants are heavy metals, such as cadmium (Cd). Megaspores were germinated in liquid culture medium, with concentrations 0 (control), 0.39; 0.78; 1.56; 3.12; 6.25; 12.5; 25; 50 and 100 mg L . In apomictical sporophytes, the growth of primary root and leaf was significantly reduced and no secondary leaf was formed at Cd concentrations of 12.5 and higher than this. The results indicated that R. diphyllum is tolerant to the presence of Cd up to considerably higher concentrations (0.78 mg L -1 ) than that normally found in unpolluted aquatic ecosystems (0.01 mg L -1 ), although the sensitivity to higher concentrations might endanger the establishment and permanence of this species in habitats exposed to contamination with this metal.Key words -ecophysiology, heavy metals, pollution, pteridophytes, reproduction RESUMO -(Germinação de megásporos e desenvolvimento inicial de esporófitos de Regnellidium diphyllum Lindman (Pteridophyta, Marsileaceae) na presença de cádmio). Regnellidium diphyllum tem sua distribuição restrita ao Sul do Brasil e localidades vizinhas no Uruguai e na Argentina. Atualmente, ela faz parte da lista de espécies ameaçadas do Rio Grande do Sul. A conversão de áreas úmidas em espaços para agricultura ou a contaminação do solo pela introdução de resíduos tóxicos e fertilizantes podem comprometer o estabelecimento e a sobrevivência da espécie. Entre os poluentes, estão metais pesados, como o cádmio (Cd). Megásporos foram germinados em meio de cultura líquido, com concentrações de 0 (controle), 0,39; 0,78; 1,56; 3,12; 6,25; 12,5; 25; 50 e 100 mg L . Em esporófitos apomíticos, houve redução significativa do crescimento da raiz e da folha primárias e ausência de formação de folha secundária a partir de 12,5 mg L -1 de Cd. Os resultados indicaram que R. diphyllum é tolerante à presença de Cd em concentrações consideravelmente mais altas (0,78 mg L -1 ) do que aquelas normalmente encontradas em ecossistemas aquáticos não poluídos (0,01 mg L -1 ), embora a sensibilidade a concentrações mais altas possa ameaçar o estabelecimento e a permanência dessa espécie em habitats expostos à contaminação com esse metal.Palavras-chave -ecofisiologia, metais pesados, poluição, pteridófitas, reprodução
The deterioration of environmental quality in the Sinos River basin is directly associated with the impacts of intense industrialization and urbanization. An integrated environmental assessment (IEA) was conducted in July and September of 2012, in areas along the sources of the EstânciaVelha/Portão, Pampa and Schmidt streams using physical, chemical and biological methods. The water in the three sampling sites was not proper for human consumption, presented a low toxic contamination index (TCI) and mesotrophic characteristics. One site was included in Class 4, and two, in Class 3, according to current legislation. The rapid assessment protocol (RAP) indicated a natural environmental condition for habitat diversity and environmental impact in the three sites. The Tradescantia pallida (Rose) D.R. Hunt var. purpurea Boom biomarker showed water genotoxicity in two of the sites. The integrated diagnosis of water quality in these streams is fundamentally important to ensure the sustainable management of water resources and their multiple uses, as well to estimate their contribution to pollution in this river basin.Keywords: genotoxicity, water quality indices, source, rapid assessment protocol. Avaliação Ambiental Integrada de arroios da bacia do Rio dos Sinos no Estado do Rio Grande do Sul, Brasil ResumoA deterioração da qualidade ambiental da bacia do Rio dos Sinos está diretamente associada a impactos oriundos da intensa industrialização e urbanização. A avaliação ambiental integrada (AAI) foi realizada nos meses de julho e setembro de 2012, em áreas junto às nascentes dos arroios Estância Velha/Portão, Pampa e Schmidt por meio de metodologias que utilizam parâmetros físicos, químicos e biológicos. Os três sítios amostrais apresentaram águas impróprias para o consumo humano, baixo índice de contaminação por tóxicos (ICT) e características mesotróficas. Um sítio foi enquadrado na Classe 4 e dois na Classe 3, segundo a legislação vigente. Os resultados da aplicação do protocolo de avaliação rápida (PAR) indicaram uma situação ambiental natural, quanto à diversidade de habitats e aos impactos ambientais, nos três sítios amostrados. O biondicador Tradescantia pallida (Rose) D.R. Hunt var. purpurea Boom evidenciou genotoxicidade da água em dois sítios amostrados. O diagnóstico integrado da qualidade das águas destes arroios é de fundamental importância para assegurar o gerenciamento sustentado dos recursos hídricos e seus múltiplos usos, além de estimar sua contribuição à poluição desta bacia hidrográfica.Palavras-chave: genotoxicidade, índices de qualidade da água, nascente, protocolo de avaliação rápida.
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