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Abstract Background In ipecac (Carapichea ipecacuanha (Brot.) L. Andersson), adventitious shoots can be induced simply by placing internodal segments on phytohormone-free culture medium. The shoots form locally on the epidermis of the apical region of the segments, but not the basal region. Levels of endogenous auxin and cytokinin transiently increase in the segments after 1 week of culture. Results Here, we conducted RNA-seq analysis to compare gene expression patterns in apical and basal regions of segments before culture and after 1 week of culture for adventitious shoot formation. The results revealed 8987 differentially expressed genes in a de novo assembly of 76,684 genes. Among them, 276 genes were upregulated in the apical region after 1 week of culture relative to before culture and the basal region after 1 week of culture. These genes include 18 phytohormone-response genes and shoot-formation-related genes. Validation of the gene expression by quantitative real-time PCR assay confirmed that the expression patterns were similar to those of the RNA-seq data. Conclusions The transcriptome data show that expression of cytokinin biosynthesis genes is induced along with the acquisition of cellular pluripotency and the initiation of cell division by wounding in the apical region of internodal segments, that trigger adventitious shoot formation without callusing.

Background: In ipecac (Carapichea ipecacuanha (Brot.) L. Andersson), adventitious shoots can be induced simply by placing internodal segments on phytohormone-free culture medium. The shoots form locally on the epidermis of the apical region of the segments, but not the basal region. Levels of endogenous auxin and cytokinin transiently increase in the segments after 1 week of culture. Results: Here, we conducted RNA-seq analysis to compare gene expression patterns in apical and basal regions of segments before culture and after 1 week of culture for adventitious shoot formation. The results revealed 8987 differentially expressed genes in a de novo assembly of 76,684 genes. Among them, 276 genes were upregulated in the apical region after 1 week of culture relative to before culture and the basal region after 1 week of culture. These genes include 18 phytohormone-response genes and shoot-formation-related genes. Validation of the gene expression by quantitative real-time PCR assay confirmed that the expression patterns were similar to those of the RNA-seq data. Conclusions: The transcriptome data show that expression of cytokinin biosynthesis genes is induced along with the acquisition of cellular pluripotency and the initiation of cell division by wounding in the apical region of internodal segments, that trigger adventitious shoot formation without callusing. [ABSTRACT FROM AUTHOR]

For most plant species, auxins and cytokinins are added to the culture medium to induce adventitious shoot formation. In ipecac ( Carapichea ipecacuanha (Brot.) L. Andersson), however, shoots form on internodal segments without phytohormone treatment. This creates an opportunity to analyze the dynamics of endogenous phytohormones during adventitious shoot formation. Ipecac is a medicinal plant whose root extract is used as an expectorant and emetic. We explored the relationships among adventitious shoot formation and levels of an auxin (indole-3-acetic acid) and four cytokinins in ipecac. Adventitious shoots that formed in the apical region of internodal segments were derived from epidermal cells. One of the shoots grew vigorously with a vascular bundle connected to that of the segment, and the others were suppressed in the outgrowth. When the biggest shoot was removed, another began to grow. During adventitious shoot formation, endogenous auxin accumulated in the basal region of segments, whereas cytokinins accumulated in the middle region. Thus, the distribution of auxin, not cytokinins, determined where adventitious shoots formed. [ABSTRACT FROM AUTHOR]

In vitro organogenesis of Citrus was studied for the genotypes Citrus sinensis cv. 'Natal', C. limonia, C. volkameriana, and C. aurantium, with the use of epicotyl segments-derived explants, cultured in MT salts and vitamins medium supplemented with different concentrations of 6-benzylaminopurine (BAP - 0.0; 0.5; 1.0; 1.5 or 2.0 mg L-1). For the recalcitrant genotypes C. limonia and C. aurantium the in vitro organogenesis was also studied with internodal segments-derived explants, cultured in MT salts and vitamins medium supplemented with 0; 0.5; 1.0; 2.0, or 4.0 mg L-1 of BAP. The efficiency of culture medium supplementation with the combination of BAP (0.0; 1.0, or 2.0 mg L-1) and NAA (1-naphthaleneacetic acid - 0.0; 0.3, or 0.5 mg L-1) in the development of adventitious shoots was evaluated for C. aurantium. Culture medium supplementation with BAP is not essential for the adventitious shoots development in the four genotypes studied when epicotyl segments-derived explants are used. In general, culture media supplementation with BAP decreased the percentage of responsive explants excepted for C. sinensis cv. 'Natal' and C. limonia when the concentrations of 1.5 and 2.0 mg/L were used. The presence of cytokinin, in concentrations up to 2 mg/L, stimulated the in vitro organogenesis when internodal segments-derived explants were used for C. limonia and C. aurantium. For C. aurantium no adventitious shoots developed in explants (internodal segments) cultured in basal culture medium, without BAP supplementation. Although no statistic differences could be detected, culture media supplementation with the combination of BAP and NAA favored the development of adventitious shoots in C. aurantium. The best concentration of NAA varied according to BAP concentration. The results presented herein, show that Citrus in vitro organogenesis depends on the interaction of culture medium composition, explant differentiation level, and genotype.A organogênese in vitro de Cítrus foi estudada para os genótipos Citrus sinensis cv. 'Natal', C. limonia, C. volkameriana e C. aurantium, utilizando-se de segmentos de epicótilo como explante, cultivados em meio de cultura MT suplementado com diferentes concentrações de 6-benzilaminopurina (BAP - 0,0; 0,5; 1,0; 1,5 ou 2,0 mg L-1). Para os genótipos recalcitrantes C. limonia e C. aurantium, a organogênese in vitro também foi estudada, utilizando-se de segmento internodal como explante, cultivado em meio de cultura MT suplementado com 0; 0,5; 1,0; 2,0 ou 4,0 mg L-1 de BAP. A eficiência da suplementação do meio de cultura com a combinação de BAP (0,0; 1,0 ou 2,0 mg L-1) e NAA (0,0; 0,3 ou 0,5 mg L-1) no desenvolvimento de gemas adventícias foi avaliada para C. aurantium. Pode-se verificar que, nos quatro genótipos estudados, a suplementação do meio de cultura com BAP não é essencial para o desenvolvimento de gemas adventícias, quando segmentos de epicótilo são utilizados como explantes. De maneira geral, a suplementação do meio de cultura com BAP diminuiu a porcentagem de explantes responsivos, exceto para C. sinensis cv. 'Natal' e C. limonia, quando as concentrações de 1,5 e 2,0 mg L-1 foram utilizadas. Para C. limonia e C. aurantium, quando segmento internodal foi utilizado como explante, a presença de citocinina em concentrações até 2,0 mg L-1 estimulou a organogênese in vitro. No caso específico de C. aurantium, não ocorreu o desenvolvimento de gemas adventícias em explantes (segmento internodal) cultivados em meio de cultura basal, sem a presença de BAP. Embora não tenham sido detectadas diferenças estatísticas significativas, a suplementação do meio de cultura com a combinação BAP e NAA favoreceu o desenvolvimento de gemas adventícias em C. aurantium. A melhor concentração de NAA variou conforme a concentração de BAP utilizada. Pela análise dos resultados obtidos, pode-se verificar que a eficiência da organogênese in vitro de Cítrus depende da interação da composição do meio de cultura, do nível de diferenciação do explante e do genótipo utilizado.

In order to evaluate the formation of adventitious buds and in vitro regeneration of sour orange plants (Citrus aurantium L.) two organogenesis-inducing experiments were conducted. In the first experiment, the induction and in vitro regeneration of adventitious buds were tested on epicotyl and internodal segments under the influence of BAP or KIN associated with NAA. The second experiment evaluated the in vitro regeneration of sour orange plants related to different explant types (epicotyls segments, internodal segments of in vitro germinated plantlets and internodal segments of greenhouse cultivated plants). Data collected on both experiments included the percentage of responsive explants (explants that formed buds), and the number of buds per explant. The addition of BAP showed the best organogenic response. In vitro germinated epicotyl segments and internodal segments are recommended as explants for sour orange in vitro organogenesis. Rooting of regenerated shoots was achieved without the need of auxin in the medium.Com o objetivo de avaliar a formação de gemas adventícias e regeneração in vitro de plantas de laranja-azeda (Citrus aurantium L.), foram realizados dois experimentos de indução à organogênese. No primeiro experimento, a indução e a regeneração in vitro de gemas adventícias foram investigadas a partir de segmentos internodais e segmentos de epicótilo sob o efeito de BAP ou CIN associados com ANA. O segundo experimento avaliou a regeneração in vitro de plantas de laranja-azeda em função do tipo de explante (segmentos de epicótilo, segmentos internodais de plantas germinadas in vitro e segmentos internodais de plantas cultivadas em casa de vegetação). Os dados coletados em ambos os experimentos incluíram a porcentagem de explantes responsivos (explantes que formaram gemas) e número de gemas por explante. A adição de BAP revelou a melhor resposta organogenética. Segmentos de epicótilo e segmentos internodais são explantes recomendados para a indução de organogênese in vitro de laranja-azeda. Enraizamento das brotações foi alcançado sem a adição de auxinas ao meio de cultura.

No presente trabalho, avaliaram-se o efeito do tamanho e posição de explantes na planta de ipeca e diferentes volumes de meio de cultivo na multiplicação "in vitro" de Psychotria ipecacuanha (Brot.) Stokes. O experimento que avaliou o efeito dos diferentes tamanhos e posições de explantes foi conduzido em tubos de ensaio de 25x150 mm, contendo 20 mL de meio de cultivo por tubo, sendo avaliados os tamanhos de 0,5; 1,0 e 1,5 cm nas posições basal, mediana e apical. No segundo experimento, utilizaram-se frascos nos quais os tratamentos consistiam nos volumes de 10, 20, 30 e 40 mL de meio de cultivo por frasco. O meio de cultivo utilizado nos dois experimentos foi o MS (MURASHIGE e SKOOG, 1962) acrescido de 1,5 mg. L-1 de BAP (6-benzilaminopurina). A posição do segmento internodal não influenciou o desenvolvimento vegetativo; os tamanhos de 1,0 e 1,5 cm apresentaram maior número de brotações que os segmentos com 0,5 cm. No experimento com diferentes volumes de meio de cultivo, observou-se que 30 e 40 mL foram significativamente melhores na indução de brotações adventícias nessa espécie.In this work evaluated the effect of size and position of explant in plant and volume different culture medium on the multiplication of Psychotria ipecacuanha (Brot.) Stokes "in vitro" conditions. Experiment with different size and positions was conducted in test tube of 25x150 mm, content 20 mL of culture medium with 0,5; 1,0 and 1,5 cm of size and three different positions (basal, median and apical). In the second experiment were utilized differents volumes (10, 20, 30 and 40 mL) of culture medium. Culture medium utilized on two experiments was MS (MURASHIGE and SKOOG, 1962) supplemented with 1,5 mgL-1 of BAP (6-benzilaminopurine). The differents position not observed influenced, where size of 1,0 and 1,5 showed higher numbers of shoots segments. The different volumes of culture medium were observed that 30 and 40 mL showed higher shoot number.

The myelinated axon can be modeled by means of a distributed RGC circuit. The Green function of this model allows for a generic formulation of the internodal segment response to any kind of stimulus, and accounts for the delay of the action potential associated with this segment. The RGC model accuracy is comparable to that of a more complex electromagnetic model, and predicted delay agrees with experimental measurements. [ABSTRACT FROM AUTHOR]