ABSTRACT
Anthurium magnificum Linden es una importante planta ornamental perteneciente a la familia Araceae. Esta especie es muy demandada como planta de maceta para la decoración de interiores y jardines, así como follaje de corte. Los métodos de propagación tradicionales en esta especie presentan ciertas desventajas por lo que no permiten satisfacer la creciente demanda. Estas limitantes convierten al cultivo de tejidos vegetales en el método más eficiente para propagar plantas de Anthurium magnificum a un ritmo más rápido que los métodos de propagación convencionales. En el cultivo de tejidos vegetales el nitrato de plata adicionado a los medios de cultivo actúa como un inhibidor de la acción del etileno y desempeña un papel crucial en la regulación de procesos fisiológicos esenciales de las plantas. El presente trabajo tuvo como objetivo determinar el efecto de diferentes concentraciones de nitrato de plata en el crecimiento in vitro de brotes de Anthurium magnificum. Para ello, se cultivaron brotes in vitro de Anthurium magnificum en medios de cultivo enriquecidos con diferentes concentraciones de nitrato de plata. Los resultados mostraron que los mayores valores en las variables altura de la planta, número de raíces y longitud de las raíces se obtuvieron en un medio de cultivo con 1,0 mg L-1 de nitrato de plata. En la variable área foliar de la planta los mayores valores se presentaron en los medios de cultivo que contenían nitrato de plata independientemente de la concentración empleada.
Anthurium magnificum Linden is an important ornamental plant belonging to the Araceae family. This species is in great demand as a pot plant for interior and garden decoration, as well as cut foliage. The traditional propagation methods in this species have certain disadvantages, which is why they cannot satisfy the growing demand. These limitations make plant tissue culture the most efficient method to propagate Anthurium magnificum plants at a faster rate than conventional propagation methods. In the culture of plant tissues, silver nitrate added to the culture media acts as an inhibitor of the action of ethylene and plays a crucial role in the regulation of essential physiological processes in plants. The objective of this work was to determine the effect of different concentrations of silver nitrate on the in vitro growth of Anthurium magnificum shoots. For this, in vitro shoots of Anthurium magnificum were cultivated in culture media enriched with different concentrations of silver nitrate. The results showed that the highest values in the variables plant height, number of roots and length of the roots were obtained in a culture medium with 1.0 mg L-1 of silver nitrate. In the variable leaf area of the plant, the highest values were presented in the culture media that contained silver nitrate, regardless of the concentration used.
ABSTRACT
Abstract Objective Our study aimed to elucidate the effect of PAI-1 (Plasminogen Activator Inhibitor-1) and t-PA (Tissue-type Plasminogen Activator) in tissue remodeling in nasal polyps patients. Methods Samples were streamed as early Nasal Polyps (eNP, n = 10) and inferior tissue from the same patient, mature Nasal Polyps (mNP, n = 14), and Control group (n = 15), respectively. Immunohistochemistry and immunofluorescence were applied to detect localization. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Western blot were used to measure different levels among three groups. The mNP tissue was cultured in vitro and treated with TGF-β1 (Transforming Growth Factor-beta 1) activator, TGF-β1 inhibitor (SB431542), and PAI-1 inhibitor (TM5275); then Western blot, qRT-PCR, and ELISA were used to assess changes. Results The immunohistochemistry and immunofluorescence showed that PAI-1 expression decreased in eNP and mNP, mainly in epithelium and glands. The transcriptional expression and protein level of TGF-β1/t-PA/PAI-1/Collagen1 were lower in eNP than IT while mNP group demonstrated lower mRNA expression and protein level of TGF-β1/t-PA/PAI-1/Collagen1 than Control group. In mNP tissue culture in vitro, TGF-β1 activator elevated t-PA, PAI-1, and Collagen1 with higher release of PAI-1 and Collagen1 in supernatant, whereas SB431542 suppressed above reactions; TM5275 lowered transcriptional and protein level of Collagen1 in supernatant. Conclusion Early Nasal polyps' formation in middle meatus mucous is related with fibrillation system PAI-1/t-PA and tissue remodeling; moreover, nasal polyps' development is regulated by TGF-β1-mediated PAI-1 reduction. Level of evidence 3b.
ABSTRACT
Chinese medicinal resources are the cornerstone of the sustainable development of traditional Chinese medicine industry. However, due to the fecundity of species, over-exploitation, and limitations of artificial cultivation, some medicinal plants are depleted and even endangered. Tissue culture, a breakthrough technology in the breeding of traditional Chinese medicinal materials, is not limited by time and space, and can allow the production on an annual basis, which plays an important role in the protection of Chinese medicinal resources. The present study reviewed the applications of tissue culture of medicinal plants in the field of Chinese medicinal resources, including rapid propagation of medicinal plant seedlings, breeding of novel high-yield and high-quality cultivars, construction of a genetic transformation system, and production of secondary metabolites. Meanwhile, the current challenges and suggestions for the future development of this field were also proposed.
Subject(s)
Sustainable Development , Plants, Medicinal/genetics , Plant Breeding , Medicine, Chinese Traditional , TechnologyABSTRACT
Abstract Objective To evaluate the reproductive and histological characteristics of fresh cultured ovarian tissue from transgender male patients. Methods An in vitro pilot study in which samples were collected during sex reassignment surgery for transgender male patients. The ovarian cortex was cut into fragments of 2 mm, 3mm, and 4 mm, and placed in a 96-well plate suitable for cultivation at days 0, 2, 4, 6, and 8, when the histology was analyzed. Results Stromal hyperplasia was observed in all samples, and it was not associated with the obtainment of primordial or primary follicles. Peripheral reduction in cell count was also a recurrent finding. Primordial and primary follicles were identified with a heterogeneous pattern in fragments from the same patient and from different patients, and follicles in more advanced stages of development (secondary and antral) were not found. There was an association between the diameter of the ovarian fragments and the identification of primary follicles (p=0.036). The number of days in culture was associated with histological signs of tissue damaging in the fragments (p=0.002). The total number of follicles identified in the samples with 2mm in diameter was significantly lower than in those that measured 4mm in diameter (p=0.031). Conclusion A diameter of 4mm is suitable for ovarian tissue culture with the benefit of ease of handling. Even after prolonged exposure to testosterone, the ovarian fragments presented primordial and primary follicles, maintaining viability throughout the days they were exposed to the culture. Freezing the ovarian cortex of transgender patients who will undergo surgery for gender reassignment would be an interesting option, in the future, for the preservation of fertility.
Resumo Objetivo Avaliar as características reprodutivas e histológicas de tecido ovariano cultivado a fresco de pacientes transexuais masculinos. Métodos Estudo experimental in vitro e piloto, no qual amostras foram coletadas durante a cirurgia de redesignação de sexo para pacientes transexuais masculinos. O córtex ovariano foi cortado em fragmentos de 2mm, 3mm, e 4mm, e colocado em placa de 96 poços própria para cultivo nos dias 0, 2, 4, 6 e 8, quando a histologia foi analisada. Resultados Hiperplasia estromal foi observada em todas as amostras, e não esteve associada à obtenção de folículos primordiais ou primários. A redução periférica no número de células também foi um achado recorrente. Folículos primordiais e primários foramidentificados com padrão heterogêneo emfragmentos domesmo paciente e em fragmentos de pacientes diferentes, não sendo encontrados folículos em estágios mais avançados de desenvolvimento (secundários e antrais). Houve associação entre o diâmetro dos fragmentos ovarianos e a identificação dos folículos primários (p=0,036). O número de dias de cultura esteve associado a sinais histológicos de lesão tecidual nos fragmentos (p=0,002). O número total de folículos identificados nas amostras de 2mm de diâmetro foi significativamente menor do que nas de 4mm de diâmetro (p=0,031). Conclusão O diâmetro de 4mm parece ser mais adequado para a cultura de tecido ovariano com a vantagem de fácil manejo. Mesmo após exposição prolongada à testosterona, os fragmentos ovarianos apresentavam folículos primordiais e primários, e manteve a viabilidade ao longo dos dias de exposição à cultura. No futuro, o congelamento da cortical do ovário de pacientes transgêneros que se submeterão à cirurgia de redesignação sexual poderia ser uma opção interessante para a preservação da fertilidade.
Subject(s)
Humans , Male , Ovary , Tissue Culture Techniques , Sex Reassignment Surgery , Fertility Preservation , Ovarian ReserveABSTRACT
The objective of this work was to carry out the in vitro establishment of Echynochloa polystachya aiming at obtaining a micropropagation protocol for works involving the selection of superior genotypes and the cultivation of the species. E. polystachya stems were collected in the municipality of Manaus-AM. Explants were inoculated in test tubes containing Murashige and Skoog (MS) medium. Thirty days after in vitro establishment, the rate of sprouting and contamination were evaluated. Experiments were also carried out to assess the effects of sucrose and 6-benzylaminopurine (BAP) concentrations on the tillering rate of explants. It was found that during the successive subcultures there was a decrease in internodes and the consequent loss of vigor. There were responses in the multiplication rate at concentrations starting from 45 g L-1 sucrose. In addition, BAP and sucrose interfered the development and in vitro multiplication. Sucrose in conjunction with BAP was harmful and shortened internodes. The physiological state of the explants for the species under study was intrinsically linked to the concentrations of sucrose used for the culture medium and the concentrations of BAP. However, the sucrose and BAP concentrations suggested for in vitro cultivation of E. Polystachya must be adjusted during successive subcultures. Absence of contamination in the in vitro establishment occurred at concentrations 15, 30 and 60 g L-1 sucrose. The combination of 1.5 mg L-1 BAP and 30 g L-1 sucrose promoted greater induction of sprouts. In addition, the in vitro rooting of E. polystachya was 45%.
Subject(s)
In Vitro Techniques , Brachiaria , Tissue Culture TechniquesABSTRACT
Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.
O lúpulo é uma nova cultura no Brasil. A cultura de tecidos pode ser uma técnica importante para a propagação rápida do lúpulo. Este artigo tem como objetivo caracterizar respostas de diferentes genótipos sob diferentes reguladores de crescimento através da inter-relação de variáveis de resposta importantes para o crescimento in vitro. Três genótipos foram cultivados em seis meios de cultura com diferentes combinações de reguladores de crescimento, BAP (6-benzilaminopurina), AIA (ácido 3-indolacético) e GA3 (ácido giberélico). As médias foram comparadas por contrastes ortogonais e a inter-relação das variáveis de resposta foi realizada por análise de trilha. Os genótipos americanos apresentaram desenvolvimento radicular favorável sob a combinação BAP + AIA, enquanto o uso do AIA melhorou o desenvolvimento da parte aérea. A origem dos genótipos foi importante para definir o melhor protocolo para o cultivo in vitro. O coeficiente de trilha mostrou que a variável número de brotos tem um efeito direto mais forte no número de segmentos nodais. Adicionalmente, em experimentos com cultura de tecidos, o uso de uma covariável e distribuição de erro adequada aumentou significativamente a precisão experimental.
Subject(s)
Plant Growth Regulators , Brazil , Plant Shoots/genetics , Culture Media , GenotypeABSTRACT
Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.
Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.
Subject(s)
Germination , Clusiaceae , Plant Extracts/toxicity , Plant Leaves , Seedlings , AllelopathyABSTRACT
ABSTRACT: Couroupita guianensis Aubl. is an Amazonian forest species with important medicinal and ornamental value. This study evaluated the effect of different culture media and light spectra on the in vitro germination and development of the zygotic embryos of C. guianensis. The culture media, MS and WPM, were evaluated without the addition of plant growth regulators and were associated with four LED light spectra: white (CW), 70% red + 30% blue (R2B), 100% red (R), and 100% blue (B). One hundred percent of the seeds successfully underwent in vitro germination, and the culture media did not interfere with embryo development. In addition to this, the different light spectra induced in vitro morphogenesis and R2B treatment significantly promoted the production of secondary roots. This effect may aid in the rooting and acclimatization of seedlings of this species.
RESUMO: Couropita guianensis Aubl. é uma espécie florestal de origem amazônica, de importância no uso medicinal e ornamental. O objetivo do presente estudo foi avaliar o efeito de diferentes meios de cultivo e espectros de luz na germinação e desenvolvimento in vitro de embriões zigóticos de Couropita guianensis. Os meios de cultura avaliados foram o MS e o WPM, sem adição de reguladores de crescimento, associados a quatro espectros de luz de LED branca (CW), 70% vermelha + 30% azul (R2B), 100% vermelha (R) e 100% azul (B). Ocorreu 100% de germinação in vitro e os meios de cultivo não interferiram no desenvolvimento dos embriões. Os diferentes espectros de luz induziram a morfogênese in vitro e o tratamento R2B promoveu a produção de raízes secundárias em número significativo. Este efeito poderá auxiliar no processo de enraizamento e aclimatização de mudas dessa espécie.
ABSTRACT
Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.
Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.
Subject(s)
Lettuce/drug effects , Anthraquinones , Clusiaceae/chemistry , Clusiaceae/toxicity , Terpenes , In Vitro TechniquesABSTRACT
Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.
O lúpulo é uma nova cultura no Brasil. A cultura de tecidos pode ser uma técnica importante para a propagação rápida do lúpulo. Este artigo tem como objetivo caracterizar respostas de diferentes genótipos sob diferentes reguladores de crescimento através da inter-relação de variáveis de resposta importantes para o crescimento in vitro. Três genótipos foram cultivados em seis meios de cultura com diferentes combinações de reguladores de crescimento, BAP (6-benzilaminopurina), AIA (ácido 3-indolacético) e GA3 (ácido giberélico). As médias foram comparadas por contrastes ortogonais e a inter-relação das variáveis de resposta foi realizada por análise de trilha. Os genótipos americanos apresentaram desenvolvimento radicular favorável sob a combinação BAP + AIA, enquanto o uso do AIA melhorou o desenvolvimento da parte aérea. A origem dos genótipos foi importante para definir o melhor protocolo para o cultivo in vitro. O coeficiente de trilha mostrou que a variável número de brotos tem um efeito direto mais forte no número de segmentos nodais. Adicionalmente, em experimentos com cultura de tecidos, o uso de uma covariável e distribuição de erro adequada aumentou significativamente a precisão experimental.
Subject(s)
Humulus/growth & development , Humulus/genetics , Genetic Enhancement/methods , Plant Growth Regulators/analysis , In Vitro TechniquesABSTRACT
Abstract Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.
Resumo Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.
ABSTRACT
Abstract Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.
Resumo O lúpulo é uma nova cultura no Brasil. A cultura de tecidos pode ser uma técnica importante para a propagação rápida do lúpulo. Este artigo tem como objetivo caracterizar respostas de diferentes genótipos sob diferentes reguladores de crescimento através da inter-relação de variáveis de resposta importantes para o crescimento in vitro. Três genótipos foram cultivados em seis meios de cultura com diferentes combinações de reguladores de crescimento, BAP (6-benzilaminopurina), AIA (ácido 3-indolacético) e GA3 (ácido giberélico). As médias foram comparadas por contrastes ortogonais e a inter-relação das variáveis de resposta foi realizada por análise de trilha. Os genótipos americanos apresentaram desenvolvimento radicular favorável sob a combinação BAP + AIA, enquanto o uso do AIA melhorou o desenvolvimento da parte aérea. A origem dos genótipos foi importante para definir o melhor protocolo para o cultivo in vitro. O coeficiente de trilha mostrou que a variável número de brotos tem um efeito direto mais forte no número de segmentos nodais. Adicionalmente, em experimentos com cultura de tecidos, o uso de uma covariável e distribuição de erro adequada aumentou significativamente a precisão experimental.
ABSTRACT
Safflower(Carthamus tinctorius), a valuable traditional Chinese medicinal plant, has attracted much attention in recent years. This study established a stable tissue culture system of safflower and analyzed the chromatogram of its secondary metabolites, providing high-quality experimental materials for further research on natural products in safflower. The calluses were established from the safflower seeds germinated in a sterile environment, and then they were differentiated into the aseptic seedlings, or cultured to obtain suspension cells in liquid medium. The ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS), Progenesis QI, and principal component analysis(PCA) were used to detect and analyze the secondary metabolites in the suspension cells before and after induction with different elicitors(methyl jasmonate, silver nitrate, salicylic acid and yeast extract). A total of 23 secondary metabolites including flavonoids, phenylpropanoids, alkaloids, fatty acids and aromatic glycosides were detected in safflower suspension cells. In response to the four elicitors, 11 compounds showed increased or decreased relative content. The results indicate that different elicitors have various effects on the accumulation of secondary metabolites in safflower suspension cells, and yeast extract shows more obvious positive induction. Therefore, different elicitors may play a role in the expression of related genes in the biosynthetic pathway of specific secondary metabolites. The results facilitate the discovery of targeted elicitors and the large-scale production of valuable secondary metabolites in the future.
Subject(s)
Carthamus tinctorius , Chromatography, High Pressure Liquid , Chromatography, Liquid , Flavonoids , Glycosides , Mass SpectrometryABSTRACT
The biotechnological interest in genus Physalis has increased in the last decades, however, there are still few micropropagation studies of this genus. The objective of this study was to evaluate P. angulata photoautotrophic and photomixotrophic micropropagation with gas exchange under seven light spectra and five concentrations of sucrose. Lighting were yellow, blue, white, red, green, red + blue LEDs and natural light filtered by mesh. Sucrose concentrations were 0, 7.5, 15, 22.5 and 30 g.L-1. Phytotechnical, anatomical features and photopigment contents were evaluated through stem and root segment length, leaf number, leaf area, chlorophyll a and b contents, carotenoids, adaxial epidermis, palisadic and spongy parenchyma and abaxial epidermis. The data were compared by Scott-Knott's mean test and principal components analysis using the R software. Comparing the variables within lighting types, it was observed that only the screen treatment, screen-filtered natural illumination, obtained assessment in all variables. Comparing the levels of sucrose, it was observed that the treatment 15 g.L-1 sucrose obtained the highest number of averages with maximum evaluation. It was concluded that the natural light filtered by screen with 50% of shading allowed the photoautotrophic micropropagation of P. angulata. Better development results were observed in photomixotrophic micropropagation with 15 g.L-1 of sucrose.
O interesse biotecnológico em Physalis aumentou nas últimas décadas, porém, ainda existem poucos trabalhos de micropropagação desse gênero. Objetivou-se avaliar sua micropropagação fotoautotrófica e fotomixotrófica com troca gasosa sob sete tipos de iluminação e cinco concentrações de sacarose. Foram utilizados LEDs amarelo, azul, branco, vermelho, verde, vermelho + azul e luz natural filtrada por malha. As concentrações de sacarose foram 0, 7,5, 15, 22,5 e 30 g.L-1. Características fitotécnicas, anatômicas e teor de fotopigmentos foram avaliados através de comprimento de segmento de caule e raíz, número de folhas, área foliar, teores de clorofilas a e b, carotenoides, epiderme adaxial, parênquimas paliçádico e esponjoso e epiderme abaxial. Os dados foram comparados por teste de média Scott-Knott e análise de componentes principais utilizando-se o software R. Comparando-se as variáveis dentro de tipos de iluminação, observou-se que apenas o tratamento screen, iluminação natural filtrada por malha, obteve avaliação máxima em todas as variáveis. Comparando-se os níveis de sucrose, observou-se que o tratamento 15 g.L-1 sacarose obteve o maior número de médias com avaliação máxima. Concluiu-se que a luz natural filtrada por tela com 50% de sombreamento permitiu a micropropagação fotoautotrófica de P. angulata. Observou-se melhores resultados de desenvolvimento na micropropagação fotomixotrófica com 15 g.L-1 de sacarose.
Subject(s)
Botany , Physalis , Tissue Culture TechniquesABSTRACT
RESUMEN Prosopis pallida, conocido como algarrobo, es una especie emblemática de los bosques secos del norte del Perú. Es de gran importancia económica por su uso en la producción de leña y carbón, así como en la producción de algarrobina proveniente de sus frutos. Actualmente las actividades humanas han deforestado grandes poblaciones de algarrobo en el bosque seco, por lo que es muy importante una propagación masiva para planes de reforestación a gran escala en esos ecosistemas, con la finalidad de conservar la especie y también las características genéticas de individuos élite. El objetivo de la investigación fue establecer un protocolo para la propagación in vitro de algarrobo. Previo a la siembra in vitro, se evaluaron tres tratamientos pregerminativos para poder acelerar la germinación de las semillas con la finalidad de obtener mayor material de propagación. Luego se evaluó el efecto del medio de plantas leñosas con la adición de cuatro concentraciones (0; 0,5; 1,0 y 1,5 mg/L) de citoquininas (BAP y ZEA) sobre la propagación in vitro de Prosopis pallida, habiendo realizado tres ensayos debido a la poca efectividad de brotación de las yemas apicales, resultando mejor la ausencia de citoquininas en yemas apicales con sus dos cotiledones, usando tapas de algodón en los tubos de ensayo. En esta etapa se evaluó el número de nudos, altura de plántula y número de brotes. Para el enraizamiento se ensayó con tres concentraciones (0; 0,5 y 1,0 mg/L) de auxinas (NAA, IBA y IAA), y se evaluó el porcentaje de enraizamiento, longitud de la raíz y número de raíces; obteniéndose mejores resultados con 0,5 mg/L IBA. En la fase de aclimatación se evaluó el porcentaje de aclimatación en dos tipos de sustratos, obteniéndose mejores resultados con sustrato comercial de turba y vermiculita.
ABSTRACT Prosopis pallida, known as algarrobo, is an emblematic species of the dry forests of northern Peru. It is of great economic importance for its use in the production of firewood and coal, as well as in the production of carob from its fruits. Currently human activities have deforested large populations of algarrobo in the dry forest, so it is very important a massive propagation for large-scale reforestation plans in these ecosystems, in order to conserve the species and also the genetic characteristics of elite individuals. The objective of the research was to establish a protocol for the in vitro propagation of algarrobo. Before in vitro propagation, It was evaluated the effect of three seeds treatments to accelerate seeds germination in order to get more propagation material. Then, the effect of woody plants medium with the addition four concentrations (0, 0.5, 1.0 and 1.5 mg /L) of cytokinins (BAP and ZEA) on the in vitro propagation of Prosopis was evaluated and it has performed three trials for the ineffective budding of apical buds, resulting in a better absence of cytokinins in apical buds with their two cotyledons, and using cotton lids in the test tubes. In this stage the number of nodes, seedling height and number of shoots was evaluated. For rooting, it was tested with three concentrations (0, 0.5 and 1.0 mg /L) of auxins (NAA, IBA and IAA), and the percentage of rooting, root length and amount of roots was evaluated, obtaining better results with 0.5 mg / L IBA. In the acclimation phase, the acclimation percentage was evaluated using two substrates, and the best results were commercial substrate of peat and vermiculite.
ABSTRACT
Physalis alkekengi is an ornamental plant that can also be used as a medicinal plant due to its anti-inflammatory, bactericidal, antitumor and fungicidal properties. Polyploidization can be an important tool in the genetic improvement of this species. The objective this work was to obtain tetraploids in vitro and to evaluate the phytotechnical traits of P. alkekengi. For this, nodal segments of P. alkekengi var. Franchettii were inoculated into petri dishes containing 100 ml of MS medium supplemented with colchicine at concentrations 0; 0.04; 0.08; 0.12; and 0.16% and kept in the dark for 24 and 48h. After the respective treatment periods with colchicine the segments were inoculated into test tubes. The tetraploids were identified by flow cytometry and classical cytogenetics. In vitro seedlings were measured: root length, nodal segment length, leaflet number and total leaf area. In the acclimatization phase, the area of the second leaf and total leaf, petiole radius, stem length, fruit weight with calyx, without calyx, fruit diameter, number of seeds and brix of the pulp were evaluated. Chlorophyll a, chlorophyll b, total chlorophyll, total carotenoid, total chlorophyll / total carotenoid ratio and chlorophyll a / b ratio were also estimated. The treatment that most produced tetraploid seedlings was with 0.08% colchicine per 24h. No significant difference was observed in 7 (seven) variables, these being all variables of photopigments, stem diameter (steam) and brix. In general, diploid (2x) plants were better in 9 (nine) while tetraploid seedlings were better in 6 (six) of the phytotechnical variables. It was concluded that the MS medium supplemented with 0.08% colchicine for 24 h allowed P. alkekengi tetraploides to be obtained with better phytotechnical qualities.
Physalis alkekengi é uma planta ornamental que também pode ser usada como planta medicinal devido às suas propriedades anti-inflamatórias, bactericidas, antitumorais e fungicidas. A poliploidização pode ser uma ferramenta importante para o melhoramento genético dessa espécie. O objetivo deste trabalho foi obter tetraplóides in vitro e avaliar as características fitotécnicas de P. alkekengi. Para isso, segmentos nodais de P. alkekengi var. Franchettii foram inoculados em placas de Petri contendo 100 ml de meio MS suplementado com colchicina nas concentrações 0; 0,04; 0,08; 0,12; e 0,16% e mantido no escuro por 24 e 48h. Após os respectivos períodos de tratamento com colchicina, os segmentos foram inoculados em tubos de ensaio. Os tetraplóides foram identificados por citometria de fluxo e citogenética clássica. As plântulas in vitro foram medidas: comprimento da raiz, comprimento do segmento nodal, número de folhetos e área foliar total. Na fase de aclimatação foram avaliadas a área da segunda folha e área foliar total, raio do pecíolo, comprimento do caule, peso do fruto com cálice, sem cálice, diâmetro do fruto, número de sementes e brix da polpa. Também foram estimadas clorofila a, clorofila b, clorofila total, carotenóides totais, razão clorofila total / carotenóide total e razão clorofila a / b. O tratamento que mais produziu mudas tetraplóides foi com colchicina a 0,08% por 24 horas. Não foi observada diferença significativa em 7 (sete) variáveis, sendo todas variáveis de fotopigmentos, diâmetro do caule (vapor) e brix. Em geral, as plantas diplóides (2x) foram melhores em 9 (nove) variáveis fitotécnicas, enquanto as mudas tetraplóides foram melhores em 6 (seis). Concluiu-se que o meio MS suplementado com colchicina a 0,08% por 24 h permitiu obter tetraploides de P. alkekengi com melhores qualidades fitotécnicas.
Subject(s)
Aneugens , Physalis , Tissue Culture TechniquesABSTRACT
BACKGROUND: Taraxacum species (commonly known as dandelion) used as herbal medicine have been reported to exhibit an antiproliferative effect on hepatoma cells and antitumor activity in non-small-cell lung cancer cells. Although several investigations have demonstrated the safety of Taraxacum officinale, the safety of tissue-cultured plants of T. formosanum has not been assessed so far. Therefore, the present study examines the safety of the water extract of the entire plant of tissue cultured T. formosanum based on acute and subacute toxicity tests in rats, as well as the Ames tests. RESULTS: No death or toxicity symptoms were observed in the acute and subacute tests. The results of the acute test revealed that the LD50 (50% of lethal dose) value of the T. formosanum water extract for rats exceeded 5â¯g/kg bw. No abnormal changes in the body weight, weekly food consumption, organ weight, or hematological, biochemical, and morphological parameters were observed in the subacute toxicity test. Thus, the no observed adverse effect level (NOAEL) of T. formosanum water extract was estimated to be higher than 2.0â¯g/kg. Finally, the results of the Ames test revealed that T. formosanum water extract was not genotoxic at any tested concentration to any of five Salmonella strains. CONCLUSIONS: The water extract of tissue-cultured T. formosanum was non-toxic to rats in acute and subacute tests and exhibited no genotoxicity to five Salmonella strains.
Subject(s)
Animals , Rats , Plant Extracts/toxicity , Taraxacum/toxicity , Tissue Culture Techniques/methods , Safety , Flavonoids/analysis , Chromatography, High Pressure Liquid , Urinalysis , Rats, Sprague-Dawley , Phenol/analysis , Toxicity Tests, Acute , Herbal Medicine , Taraxacum/chemistry , Serum , Cell Proliferation/drug effects , Toxicity Tests, Subacute , Mutagenicity TestsABSTRACT
BACKGROUND: Soil salinity causes huge economic losses to agriculture productivity in arid and semiarid areas world-wide. The affected plants face disturbances in osmotic adjustment, nutrient transport, ionic toxicity and reduced photosynthesis. Conventional breeding approaches produce little success in combating various stresses in plants. However, non-conventional approaches, such as in vitro tissue culturing, produce genetic variability in the development of salt-tolerant plants, particularly in woody trees. RESULTS: Embryogenic callus cultures of the date palm cultivar Khalas were subjected to various salt levels ranging from 0 to 300 mM in eight subcultures. The regenerants obtained from the salt-treated cultures were regenerated and evaluated using the same concentration of NaCl with which the calli were treated. All the salt-adapted (SA) regenerants showed improved growth characteristics, physiological performance, ion concentrations and K+/Na+ ratios than the salt non-adapted (SNA) regenerants and the control. Regression between the leaf Na+ concentration and net photosynthesis revealed an inverse nonlinear correlation in the SNA regenerants. Leaf K+ contents and stomatal conductance showed a strong linear relationship in SA regenerants compared with the inverse linear correlation, and a very poor coefficient of determination in SNA regenerants. The genetic fidelity of the selected SA regenerants was also tested using 36 random amplified polymorphic DNA (RAPD) primers, of which 26 produced scorable bands. The primers generated 1-10 bands, with an average of 5.4 bands per RAPD primer; there was no variation between SA regenerants and the negative control. CONCLUSION: This is the first report of the variants generated from salt-stressed cultures and their potential adaptation to salinity in date palm cv. Khalas. The massive production of salt stress-adapted date palm plants may be much easier using the salt adaptation approach. Such plants can perform better during exposure to salt stress compared to the non-treated date palm plants.
Subject(s)
Salt Tolerance/genetics , Phoeniceae/genetics , Acclimatization , Random Amplified Polymorphic DNA Technique , SalinityABSTRACT
ABSTRACT: The objective of the research was to evaluate cytokinin concentrations and agricultural residues as physical structuring agents of the culture medium in the in vitro propagation of highbush blueberry (Vaccinium corymbosum 'Duke'). To that end, three different concentrations of 2-isopentenyladenine (2iP) (0, 5, and 10 mg L-1)in woody plant medium (WPM) and five structuring agents (in natura rice husks, carbonized rice husks, coconut fiber, S-10 Beifort®, and agar)were tested. After 60days of culture, the following parameters were evaluated: survival (%),sprout and shoot lengths (cm),propagation rate, number of sprouts, and shoot dry mass (g). The substrates used as structuring agents were analyzed in terms of pH and electrical conductivity. The in natura rice husk, S-10 Beifort®, and carbonized rice husk did not differ from the agar in terms of the sprout length. The shoots were longer (1.64 cm) in cultures within natura rice husk than in other treatments. In the absence of 2iP, S-10 Beifort® resulted in the highest propagation rate (2.97). Concerning the number of sprouts, S-10 Beifort®, in natura rice husks, and carbonized rice husks did not differ from agar. However, when the regulator was added, the agar performed better in comparison to the other residues. Thus, in natura rice husks, carbonized rice husks, and S-10 Beifort®are potential physical structuring agents of the culture medium that can be used without 2iP.
RESUMO: O objetivo deste trabalho foi avaliar concentrações de citocinina e resíduos agrícolas como agentes físicos de estruturação do meio na multiplicação in vitro de mirtileiro (Vaccinium corymbosum) 'Duke'. Para isso, foram testadas três concentrações de 2-isopenteniladenina (2iP) (0; 5 e 10 mg L-1) no meio WPM (Wood Plant Media); e cinco agentes estruturantes (casca de arroz in natura, casca de arroz carbonizada, fibra de coco, S-10 Beifort® e ágar). Após 60 dias foram avaliados: sobrevivência (%); comprimento de brotações e de parte aérea (cm); taxa de multiplicação; número de brotações e massa seca de parte aérea (g). Analisou-se os substratos utilizados como agentes estruturantes quanto ao pH e condutividade elétrica. A casca de arroz in natura, o S-10 Beifort®, e a casca de arroz carbonizada não diferiram do ágar quanto ao comprimento de brotações. Na casca de arroz in natura obteve-se o maior comprimento de parte aérea (1,64 cm), diferindo dos demais tratamentos. Na ausência de 2iP o S-10 Beifort® apresentou a maior taxa de multiplicação (2,97). Para o número de brotações o S-10 Beifort®, a casca de arroz in natura e carbonizada não diferiram do ágar. Entretanto, quando acrescido de regulador, o ágar apresenta um desempenho maior em relação aos resíduos. Sendo assim, a casca de arroz in natura, a casca de arroz carbonizada, e o S-10 Beifort® apresentam-se como potencias estruturadores físicos do meio de cultura, podendo ser utilizados sem o uso de 2iP.
ABSTRACT
Objective@#To retrospectively analyze distribution characteristics of pathogenic bacteria and their antimicrobial susceptibility in patients with diabetic foot osteomyelitis(DFO).@*Methods@#Sixty cases of suspected DFO were collected from the Endocrinology Department of Henan Provincial People′s Hospital. After admission, bone biopsy was carried out to confirm the pathological diagnosis, and the pathogenic bacteria and drug sensitivity were determined by bone culture. In addition, bacterial culture was carried out in the basal tissue of the wound, and the results of bacterial culture were compared with those of bone culture.@*Results@#Sixty patients were diagnosed as DFO after bone biopsy. Among the 60 patients, 45 patients underwent bone culture and basal tissue culture. There are 24 patients of whom the results were consistent, accounting for 53.3%. The positive rate of bone culture was 55.0%, there were 16 strains of gram-positive bacteria and 22 strains of gram-negative bacteria. Staphylococcus aureus(9 strains) occurrence was the most, common finding, followed by Escherichia coli(6 strains). The course of diabetic foot, albumin(ALB), and antibiotic usage rate before admission were lower in bone culture positive group than those in bone culture negative group, while white blood cell(WBC) and C-reactive protein(CRP) were higher in bone culture negative group(P<0.05). There was no significant difference in gender, age, course of diabetes, HbA1C, and creatinine(CREA) levels between the two groups(P>0.05). The results of bone culture showed that Staphylococcus aureus was the main Gram-positive bacteria, which was more sensitive to vancomycin, tigecyclin, linezolid, etc. Escherichia coli was the main Gram-negative bacteria, which was more sensitive to tigecyclin, carbapenems, amikacin, etc.@*Conclusion@#Bone biopsy and bone culture should be carried out in cases for suspected DFO patients to identify the pathogenic bacteria, and the bone tissue should be preserved and obtained according to the operation specification before the application of antibiotics, and the appropriate antibiotics should be selected according to the drug sensitivity results.