RESUMO

One of the main objectives to achieve in plant tissue culture is the multiplication of the available plant material, taking full advantage of the regenerative capacities of plant cells. Somatic embryogenesis leverages cell totipotency to produce new explants from a cell, thus obtaining many propagules for scientific research, industrial, or exploitation purposes. Somatic embryogenesis (ES) characterizes by being one of the most efficient techniques in plant micropropagation. However, developing an efficient plant ES protocol requires several key factors to consider, as demonstrated throughout the chapters of this book. These chapters highlight the major drivers of the success of ES in different plant species: plant growth regulators, concentration of auxins and cytokines, water deficit, photoperiod, and type of culture medium; techniques such as the use of bioreactors and Thin Cell Layer (TCL); and the influence of stress on the formation of somatic embryos. Research has been conducted to address each phase of somatic embryogenesis, either individually or for all phases. The chapters of this book cover in detail the techniques used and provide guidance that will allow readers to successfully develop all the somatic embryogenesis phases in different cultures, from cell dedifferentiation to differentiation.

Assuntos

RESUMO

Taro is important for its nutritional content, medicinal use, and bioethanol production. The aim of the present study was to compare different semi-automated bioreactors (SABs) during in vitro multiplication of C. esculenta. The SABs used were temporary immersion bioreactors (TIBs), SETIS™ bioreactors and ebb-and-flow bioreactors; semi-solid culture medium was used as a control treatment. At 30 d of culture, different developmental variables, determination of chlorophyll, stomatal content, and survival percentage during acclimatization were evaluated. SABs increased the shoot multiplication rate relative to the semi-solid medium; however, the SETIS™ bioreactor showed the highest shoot production, with 36 shoots per explant, and the highest chlorophyll content. The stomatal index was higher in the semi-solid medium compared to the SABs, while the percentage of closed stomata was higher in the SABs than in the semi-solid culture medium. The survival rate during acclimatization showed no differences among the culture systems assessed, obtaining survival rates higher than 99%. In conclusion, the SETIS™ bioreactor showed the highest multiplication rate; however, other bioreactor alternatives are available for semi-automation and cost reduction for micropropagation of C. esculenta.

RESUMO

El objetivo del trabajo fue evaluar los efectos del carbón activado y las condiciones de oscuridad inicial sobre la propagación in vitro de banana (Musa spp.) variedad Nanicão. Fueron empleados 64 ápices provenientes de hijuelos de 20 - 30 cm. Los tratamientos consistieron en la combinación de dos concentraciones de carbón activado (2 y 3 g/l) y dos condiciones de oscuridad inicial (15 y 30 días). El diseño experimental utilizado fue completamente al azar con arreglo factorial 2 x 2 y cuatro repeticiones. Cada unidad experimental estuvo constituida por cuatro ápices. Las variables evaluadas fueron la tasa media de multiplicación, el porcentaje de supervivencia, oxidación, contaminación y aclimatización de plántulas. El análisis estadístico consistió en el ANAVA con un nivel de significancia del 5 %. Las medias fueron comparadas a través del test de Tukey al 5 % de probabilidad. Los resultados mostraron que la combinación de 2 g/l de carbón activado y 15 días de oscuridad inicial promovió el mayor porcentaje de supervivencia del primer subcultivo que alcanzo 57,30 %. En la fase de establecimiento el porcentaje de contaminación de los ápices de 39,06 % mientras que en el primer subcultivo de 22,39 %. La oxidación en la fase de establecimiento fue de 9,37 % y en el primer subcultivo fue de 8,85 %. Se concluye que la combinación entre la concentración de 2 g/l de carbón activado y 15 días de período de incubación inicial resulta efectiva en el aumento de la supervivencia de los ápices.

The objective of this work was to evaluate the effects of activated charcoal and the initial darkness conditions in the propagation in vitro of banana (Musa spp.) variety Nanicão. Were employed 64 apexes coming from shoots of approximately 20 to 30 cm. The treatments consisted in the combination of two concentrations of activated charcoal (2 and 3 g/l) and two initial dark conditions (15 and 30 days). The experimental design went completely randomized with 2 x 2 factorial arrangement and four replications. Each experimental unit consisted of four apexes. Statistical analysis consisted ANOVA with significance level of 5 %. The variables were the average rate of multiplication, survival percentage, oxidation, contamination and acclimatization of plantlets. Means were compared by Tukey test at 5 % of probability. The results showed significant difference in the interaction between the concentration of 2 g/l of activated charcoal and 15 days of initial darkness at the survival percentage from the first subculture which reached 57.30 %. In the establishment phase the percentage of contamination of the apexes was 39.06 % while that in the first subculture was 22.39 %. The oxidation in the establishment phase was 9.37 % and in the first subculture was 8.85 %. It is concluded the interaction between the concentration of 2 g/l of activated charcoal and 15 days of initial incubation period is effective in increasing survival.

RESUMO

O objetivo deste trabalho foi averiguar a melhor densidade de explantes e o melhor tipo de sistema de cultivo visando desenvolver um protocolo de micropropagação de baixo custo para a Carobinha. Foram realizados experimentos de multiplicação in vitro com quatro tipos de frascos: R.I.T.A. (50 explantes/frasco), erlenmayer, (50 explantes/frasco), potes tipo maionese (6 explantes/frasco) e cubetas (1 explante/frasco). O co-cultivo de explantes, tanto em meio sólido quanto em meio líquido (R.I.T.A.), promoveu maiores taxas de explantes com brotação e de sobrevivência. O sistema de imersão temporária proporcionou melhores índices de desenvolvimento, brotação, sobrevivência e altura dos explantes. Concluímos que biorreatores podem ser utilizados eficientemente para a micropropagação de carobinha.

The aim of this study was to identify the best explant density and the best cultivation system with the goal of developing a micropropagation protocol of low cost for "carobinha" (Jacaranda decurrens CHAM.). Experiments of in vitro multiplication were carried out using four flask types: R.I.T.A. (50 explants/flask), Erlenmeyer (50 explants/flask), mayonnaise pots (6 explants/flask) and cuvettes (1 explant/flask). The co-cultivation of explants, in both solid and liquid medium (R.I.T.A.), led explants to show higher sprouting and survival rates. The temporary immersion system provided better rates of development, sprouting, survival and height of explants. We concluded that bioreactors may be efficiently used for the micropropagation of "carobinha".

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RESUMO

The aim of this research was to evaluate the micropropagation potential of holly (Ilex paraguariensis Saint Hilaire) aseptic plantlets derived from immature zygotic embryos. Shoot tip were inoculated in a quarter-strength MS basal medium with different doses of 6-benzylaminopurine (BAP) to induce multiple shoots. Shoot tips were inoculated in a basal medium with different doses of indol-butiric acid (IBA) for in vitro rooting and to test their effects in plantlet acclimatization. The addition of 8.88µM BAP to the basal medium increased the number of adventitious shoots and shoot height. Shoot tip rooted in basal medium with 7.38µM IBA in 30 days of culture. The IBA doses of the rooting medium affect plantlet acclimatization. The described micropropagation protocol might be used to produce a high number of holly plants.

O objetivo deste trabalho foi avaliar o potencial de micropropagação de erva-mate (Ilex paraguariensis Saint Hilaire) a partir de plântulas assépticas oriundas de embriões zigóticos imaturos. Ápices caulinares foram inoculados em meio base, formado por » da concentração dos sais do meio MS, com diferentes doses de 6-benzilaminopurina (BAP). Ápices caulinares foram enraizados em meio base com diferentes doses de ácido indolbutírico (AIB) e avaliado o efeito do AIB do meio de enraizamento in vitro na aclimatização das plantas. O meio base com 8,88µM de BAP foi o mais efetivo para a multiplicação, promovendo o maior número e comprimento de brotações adventícias. Ápices caulinares enraizaram em 30 dias de cultivo em meio base, acrescido de 7,38µM de AIB. A dose de AIB utilizada no enraizamento in vitro interfere na aclimatização das plantas. A técnica de micropropagação descrita pode ser utilizada para a produção massal de plantas de erva-mate.

RESUMO

O objetivo deste trabalho foi avaliar o potencial de micropropagação de erva-mate (Ilex paraguariensis Saint Hilaire) a partir de plântulas assépticas oriundas de embriões zigóticos imaturos. Ápices caulinares foram inoculados em meio base, formado por ¼ da concentração dos sais do meio MS, com diferentes doses de 6-benzilaminopurina (BAP). Ápices caulinares foram enraizados em meio base com diferentes doses de ácido indolbutírico (AIB) e avaliado o efeito do AIB do meio de enraizamento in vitro na aclimatização das plantas. O meio base com 8,88µM de BAP foi o mais efetivo para a multiplicação, promovendo o maior número e comprimento de brotações adventícias. Ápices caulinares enraizaram em 30 dias de cultivo em meio base, acrescido de 7,38µM de AIB. A dose de AIB utilizada no enraizamento in vitro interfere na aclimatização das plantas. A técnica de micropropagação descrita pode ser utilizada para a produção massal de plantas de erva-mate.

The aim of this research was to evaluate the micropropagation potential of holly (Ilex paraguariensis Saint Hilaire) aseptic plantlets derived from immature zygotic embryos. Shoot tip were inoculated in a quarter-strength MS basal medium with different doses of 6-benzylaminopurine (BAP) to induce multiple shoots. Shoot tips were inoculated in a basal medium with different doses of indol-butiric acid (IBA) for in vitro rooting and to test their effects in plantlet acclimatization. The addition of 8.88µM BAP to the basal medium increased the number of adventitious shoots and shoot height. Shoot tip rooted in basal medium with 7.38µM IBA in 30 days of culture. The IBA doses of the rooting medium affect plantlet acclimatization. The described micropropagation protocol might be used to produce a high number of holly plants.

RESUMO

O gerânio (Pelargonium graveolens L.) é uma planta medicinal e aromática nativa do sul da África, cujo óleo essencial é amplamente empregado nas indústrias de perfumaria e cosmético, além de ser usado como terapêutico. Avaliou-se o efeito da luminosidade e dos reguladores de crescimento BAP e ANA na multiplicação in vitro e o uso de diferentes substratos na aclimatização de mudas micropropagadas. Foram avaliadas as concentrações 0; 0,5; 1,0 e 2,0 mg L-1 de BAP, 0; 0,1 e 0,5 mg L-1 de ANA e duas condições de luminosidade (ausência e presença de luz), em esquema fatorial 4x3x2; para a aclimatização foram utilizados quatro substratos, pó de coco + Biosafra® (3-12-6) (12 g L-1) + calcário (1 g L-1) (PCBC); pó de coco + Biosafra® (3-12-6) (12 g L-1) + calcário (1 g L-1) + vermiculita (1:1) (PCBCV 1:1); pó de coco + Biosafra® (3-12-6) (12 g L­-1) + calcário (1 g L-1) + vermiculita (PCBCV 2:1) e vermiculita e adição semanal de sais MS (VS). Para a micropropagação de P. graveolens a utilização de 1,3 mg L-1 de BAP e 0,5 mg L-1 de ANA é eficiente na regeneração direta de plantas de gerânio, sendo a condição de escuro a mais indicada por proporcionar o maior número de brotos por explante. Para a aclimatização das mudas de gerânio, os melhores resultados foram obtidos em mudas aclimatizadas no substrato vermiculita com adição semanal de sais de MS seguido do substrato PCBCV 1:1.

Geranium (Pelargonium graveolens L.) is a medicinal and aromatic plant native to the south of Africa and whose essential oil is widely used by perfume and cosmetic industries, as well as in therapeutics. The effect of luminosity and growth regulators BAP and NAA on in vitro multiplication was evaluated, together with the use of different substrates on the acclimatization of micropropagated seedlings. The evaluated concentrations were 0, 0.5, 1.0 and 2.0 mg L-1 BAP; 0, 0.1 and 0.5 mg L-1 NAA; and two luminosity conditions (absence and presence of light), in a 4 x 3 x 2 factorial arrangement. For acclimatization, four substrates were tested: coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) (CDBL); coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) + vermiculite (1:1) (CDBLV 1:1); coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) + vermiculite (CDBLV 2:1); and vermiculite with weekly addition of MS salts (VS). For P. graveolens micropropagation, the use of 1.3 mg L-1 BAP and 0.5 mg L-1 NAA is efficient for the direct regeneration of geranium plants, and the dark condition is the most suitable since it provides a larger number of shoots per explant. For the acclimatization of geranium seedlings, the best results were obtained with seedlings acclimatized in the substrate vermiculite with weekly addition of MS salts, followed by the substrate CDBLV 1:1.

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RESUMO

A banana (Musa spp.) é uma das frutas mais consumidas no mundo, e amplamente cultivada no Brasil, porém doenças como as sigatokas, negra e amarela, vêm reduzindo a sua produção. A disponibilização imediata de novas cultivares resistentes às principais doenças é limitada pela propagação convencional. A micropropagação é uma alternativa para a produção de mudas com qualidade fitossanitária e vegetativa, mas apresenta fatores que dificultam sua aplicação como a contaminação por fungos e bactérias, associada à oxidação dos explantes. O objetivo desse trabalho foi adaptar e/ou otimizar as etapas do processo de micropropagação para diferentes cultivares de bananeira, por meio do controle de oxidação, contaminação, e multiplicação de brotos, sendo utilizadas as cultivares Caipira (AAA), BRS Caprichosa (AAAB), Pacovan Ken (AAAB), Preciosa (AAAB), PV 03-76 (AAAB), Thap Maeo (AAB). No estudo foram utilizados o antibiótico sulfato de estreptomicina e o fungicida Opera® (BASF) visando reduzir a contaminação in vitro provocada por bactérias e fungos, além do anti-oxidante PVP (polivinilpirrolidona) para controlar a oxidação. Houve redução da contaminação com uso do sulfato de estreptomicina à concentração de 100 mg L-1 e da oxidação com PVP a 4 g L-1. Na fase de multiplicação de brotos, as cultivares apresentaram médias que variaram de 1,90 a 4,75 brotos/explante. A cultivar caipira (AAA) destacou-se das demais com a maior taxa de multiplicação de brotos após três subcultivos, média de 41,50 brotos por rizoma.

The banana (Musa spp) is one of the most consumed fruits in the world and is widely consumed in Brazil, but diseases such as yellow and black sigatoka have been reducing its production. The immediate availability of new cultivars resistant to major diseases is limited by conventional propagation. The micropropagation, is an alternative for the production of seedlings with phytosanitarium and vegetative quality, but presents factors that difficult its application such as contamination by fungi and bacteria, associated with oxidation of the explants. The objective of this work was to adapt and/or optimize the stages of the micropropagation of banana cultivars, through the control of oxidation, contamination, and multiplication of shoots. The cultivars used Caipira (AAA), BRS Caprichosa (AAAB), Pacovan Ken (AAAB), Preciosa (AAAB), PV 03-76 (AAAB), Thap Maeo (AAB) were subjected to different micropropagation stages. In the study we used the streptomycin sulfate antibiotic and fungicide Opera® (BASF) to reduce contamination in vitro caused by bacteria and fungi, besides the anti-oxidant PVP (polivinilpirrolidona) to control the oxidation. We found contamination reduction with the use of streptomycin sulfate in the concentration of 100 mg L-1 and of oxidation with PVP at 4 g L-1. At the stage of multiplication of shoots, the cultivates showed means ranging from 1,90 to 4,75 shoots / explant. The cultivate Caipira (AAA) stood out from the others with the highest rate of shoot multiplication after three subcultivations, 41,50 shoots per rhizome average.

Assuntos

RESUMO

The aim of this research was to evaluate the micropropagation potential of holly (Ilex paraguariensis Saint Hilaire) aseptic plantlets derived from immature zygotic embryos. Shoot tip were inoculated in a quarter-strength MS basal medium with different doses of 6-benzylaminopurine (BAP) to induce multiple shoots. Shoot tips were inoculated in a basal medium with different doses of indol-butiric acid (IBA) for in vitro rooting and to test their effects in plantlet acclimatization. The addition of 8.88µM BAP to the basal medium increased the number of adventitious shoots and shoot height. Shoot tip rooted in basal medium with 7.38µM IBA in 30 days of culture. The IBA doses of the rooting medium affect plantlet acclimatization. The described micropropagation protocol might be used to produce a high number of holly plants.

O objetivo deste trabalho foi avaliar o potencial de micropropagação de erva-mate (Ilex paraguariensis Saint Hilaire) a partir de plântulas assépticas oriundas de embriões zigóticos imaturos. Ápices caulinares foram inoculados em meio base, formado por » da concentração dos sais do meio MS, com diferentes doses de 6-benzilaminopurina (BAP). Ápices caulinares foram enraizados em meio base com diferentes doses de ácido indolbutírico (AIB) e avaliado o efeito do AIB do meio de enraizamento in vitro na aclimatização das plantas. O meio base com 8,88µM de BAP foi o mais efetivo para a multiplicação, promovendo o maior número e comprimento de brotações adventícias. Ápices caulinares enraizaram em 30 dias de cultivo em meio base, acrescido de 7,38µM de AIB. A dose de AIB utilizada no enraizamento in vitro interfere na aclimatização das plantas. A técnica de micropropagação descrita pode ser utilizada para a produção massal de plantas de erva-mate.

RESUMO

Objetivou-se com este experimento avaliar o rejuvenescimento do material vegetal através da técnica de micropropagação na produção de mudas de mirtilo, cv. Climax. Os tratamentos aplicados constituíram-se de dois tipos de citocininas (zeatina e 2iP), em quatro concentrações (0; 2,5; 5,0 e 7,5 mg.L-1) e duas fontes de explantes (plantas micropropagadas e plantas obtidas através da germinação de sementes in vitro). O delineamento experimental utilizado foi inteiramente casualizado, em esquema fatorial 2x4x2. Aos 60 dias após a instalação do experimento avaliou-se o número médio de brotos e de gemas por explante, o comprimento médio dos brotos e a taxa de multiplicação. Concluiu-se que plantas de mirtilo, micropropagadas na presença de citocinina e submetidas a sucessivas repicagens, demonstram elevada habilidade de rejuvenescimento in vitro do material adulto, podendo ser comparadas às plantas obtidas de semente, tanto na capacidade de emitir novas brotações, quanto no número de gemas e taxa de multiplicação.

This experiment was installed aiming to evaluate the plant rejuvenation capacity of rabbiteye blueberry seedlings cultivar Climax by using micropropagation technique. The treatments consisted of two cytokinin types (zeatin and 2iP), in four concentrations (0; 2.5; 5.0 and 7.5 mg.L-1) and two sources of explants (micropropagated plants and plants from in vitro seeds germination). The experiment was carried out in a 2x4x2 factorial arranged in a randomized complete design. After 60 days from experiment installation it was assessed the shoots average number and shoots average length, buds average number and the multiplication rate. In conclusion, the present results suggest that the micropropagated rabbiteye blueberry plants in the presence of cytokinin and submitted at successive cuttings showed a high in vitro rejuvenation ability of the adult material, which might be compared to the plants germinated in vitro, in the capacity of emitting new shoots as well as in the buds number and multiplication rate.