In vitro conservation of 'Florida Rough' lemon plants / Conservação in vitro de plantas de limoeiro 'Rugoso da Florida'

The establishment of minimum growth conditions is essential for in vitro germplasm conservation. Changes to the basic medium and carbon source concentrations are important factors for reducing plant growth in vitro. This study adjusted a protocol for the in vitro conservation of 'Florida Rough' lemon plants. Microcuttings (approximately 1 cm) from plants that were previously cultivated in vitro were inoculated into test tubes with 10 mL of woody plant medium (WPM) at different concentrations (1/1, 1/2 and 1/4) and supplemented with 0, 12.5, 25 and 50 g.L-1 of sucrose, solidified with 7 g.L-1 agar and adjusted to pH 5.8. The experiment was completely randomized in a 3 x 4 factorial design with 15 replications and was maintained under controlled conditions for 360 days. After this period the plant height in cm (PH), the plant dry mass in g (PDM) and the 21 numbers of green leaves (NGL), senescent leaves (NSL) and microcuttings (NM) were evaluated. The variables that best explained the observed behavior of the 'Florida Rough' lemon plants were NGL and PH, with values of 61.63 and 35.08%, respectively. The original concentration of the WPM with the addition of 25 g. L-1 of sucrose yielded the best 'Florida Rough' lemon plant growth reduction in vitro while maintaining the physiologically health of the plants.

O estabelecimento de condições de crescimento mínimo é fundamental para a conservação in vitro de germoplasma. Alterações nas concentrações do meio básico, na fonte de carbono, são fatores importantes na desaceleração do crescimento de plantas in vitro. Este trabalho teve como objetivo ajustar um protocolo de conservação in vitro de plantas do limoeiro 'Rugoso da Flórida'. Microestacas de plantas previamente cultivadas in vitro, com aproximadamente 1 cm, foram inoculadas em tubos de ensaio contendo 10 mL do meio de cultura WPM em diferentes concentrações (1/1, 1/2 e 1/4), suplementado com 0; 12,5; 25 e 50 g.L-1 de sacarose, solidificado com 7 g.L-1 de ágar e o pH ajustado em 5,8. O experimento foi instalado em delineamento inteiramente casualizado com 15 repetições, em esquema fatorial 3 x 4, e mantido sob condições controladas durante 360 dias. Após este período, foram avaliadas altura de planta em cm (AP), número de folhas verdes (NFV), número de folhas senescentes (NFS), número de microestacas (NM) e massa da planta seca em g (MPS). As variáveis que mais contribuíram para explicar a diferença existente no comportamento das plantas do limoeiro 'Rugoso da Flórida' no meio de cultura WPM foram NFV e AP com 61,63 e 35,08%, respectivamente. O meio de cultura WPM na sua concentração original com a adição de 25 g.L-1 de sacarose promoveu os melhores resultados na redução do crescimento das plantas in vitro do limoeiro 'Rugoso da Flórida', mantendo-as fisiologicamente sadias.

In vitro multiplication of wild Manihot species with different naphthaleneacetic acid and benzylaminopurine concentrations

In vitro multiplication is an important tissue culture technique that is capable of efficiently producing seedlings at any scale. It is a propagation method based on the aseptic culture of small propagules in a suitable culture medium to enable plant regeneration. Multiplication experiments conducted in vitro to set protocols adapted to wild Manihot species have used modified mineral salts and MS vitamins as basic culture medium. Here, 25 treatments based on combinations of the regulators benzylaminopurine (BAP) and naphthaleneacetic acid (NAA) at 0, 0.025, 0.05, 0.075, and 0.1 mg L-1 were used for in vitro multiplication of three genotypes of wild Manihot species (M. violaceae Pohl Müll. Arg., M. pseudoglaziovii Pax & Hoff., and M. flabellifolia Pohl). Plant height and the number of 1 cm minicuttings, number of roots, shoots, green leaves and senescent leaves were recorded 120 days after explant inoculation. M. violaceae Pohl. Müll. Arg. and M. flabellifolia Pohl. presented favorable results with 0.05 and 0.025 mg L-1 NAA, respectively. Culture medium lacking NAA and BAP favored the in vitro growth of M. pseudoglaziovii Pax & Hoff.

In vitro multiplication of wild Manihot species with different naphthaleneacetic acid and benzylaminopurine concentrations

In vitro multiplication is an important tissue culture technique that is capable of efficiently producing seedlings at any scale. It is a propagation method based on the aseptic culture of small propagules in a suitable culture medium to enable plant regeneration. Multiplication experiments conducted in vitro to set protocols adapted to wild Manihot species have used modified mineral salts and MS vitamins as basic culture medium. Here, 25 treatments based on combinations of the regulators benzylaminopurine (BAP) and naphthaleneacetic acid (NAA) at 0, 0.025, 0.05, 0.075, and 0.1 mg L-1 were used for in vitro multiplication of three genotypes of wild Manihot species (M. violaceae Pohl Müll. Arg., M. pseudoglaziovii Pax & Hoff., and M. flabellifolia Pohl). Plant height and the number of 1 cm minicuttings, number of roots, shoots, green leaves and senescent leaves were recorded 120 days after explant inoculation. M. violaceae Pohl. Müll. Arg. and M. flabellifolia Pohl. presented favorable results with 0.05 and 0.025 mg L-1 NAA, respectively. Culture medium lacking NAA and BAP favored the in vitro growth of M. pseudoglaziovii Pax & Hoff.(AU)

Vernonia condensata Baker: an alternative for large-scale seedling production / Uma alternativa para a produção de mudas de Vernonia condensata Baker em larga escala

The increasing use of Vernonia condensata Baker highlights the importance of developing strategies to reduce the impact of exploitation on nature reserves. The aim of this study was to establish a micropropagation protocol to produce homogenous plants with high phytosanitary quality. Apical, nodal, and internodal segments of plants grown in the field were used for in vitro growth. The segments were disinfected in sodium hypochlorite solution (1.0 and 2.0%) for 15 and 30 minutes and then transferred to Petri dishes containing MS culture medium for 30 days. A completely randomized factorial experiment (3 x 2 x 2) with five replicates was designed. After this period, a completely randomized in vitro multiplication experiment was carried out with six treatments (BAP - 0.0; 0.5; 1.0; 1.5; 2.0; 2.5 mg L-1) and six replicates. The shoots obtained in the best treatment were transferred to flasks with rooting medium (MS, MS/2 or MS/4). The experiment was completely randomized with 12 replicates. Microplants were acclimatized in polyethylene terephthalate (PET) bottles filled with autoclaved topsoil. Our results showed that 40.0% of the nodal segments (immersed in 1.0% sodium hypochlorite for 30 minutes) were adequately disinfected and survived. In the in vitro multiplication experiment, the 0.5 mg L-1 concentration of BAP yielded the highest number of shoots and the best vegetative growth. With regard to the assessed characteristics, MS/4 was the best rooting medium, with 100% survival during acclimatization. This study showed that V. condensata in vitro culture might produce 32,000 seedlings in 7 months.(AU)

A crescente utilização de Vernonia condensata Baker evidencia a importância de desenvolver estratégias que reduzam o extrativismo nos ambientes naturais. O estudo objetivou estabelecer protocolo de micropropagação produzindo plantas homogêneas e de alta qualidade fitossanitária. Foram utilizados, na etapa de estabelecimento in vitro, segmentos apicais, intermodais e nodais de plantas cultivadas no campo. Desinfestados em solução de hipoclorito de sódio (1,0 e 2,0%) no tempo de 15 e 30 minutos e em seguida introduzidos em placa de Petri, com meio de cultura MS, durante 30 dias. Foi utilizado o delineamento inteiramente casualizado no esquema fatorial (3 x 2 x 2), com cinco repetições. Após este período, foi montado o experimento de multiplicação in vitro, em delineamento inteiramente casualizado com seis tratamentos (BAP - 0,0; 0,5; 1,0; 1,5; 2,0; 2,5 mg L-1) e seis repetições. As brotações obtidas no melhor tratamento foram transferidas para frascos com meio de cultura de enraizamento (MS, MS/2 e MS/4). Esse experimento foi conduzido no delineamento inteiramente casualizado, com 12 repetições. A aclimatização foi realizada introduzindo as microplantas em garrafas do tipo PET contendo terra vegetal autoclavada. Os resultados mostraram que 40,0% dos segmentos nodais (imersos em 1,0% de hipoclorito de sódio, durante 30 minutos) foram desinfestados e sobreviveram. No experimento de multiplicação in vitro obteve-se a melhor resposta no número de brotos e no desenvolvimento vegetativo, na concentração de 0,5 mg L-1 de BAP. O meio de cultura de enraizamento que possibilitou a melhor resposta, nas características avaliadas, foi o MS/4. Durante a aclimatização obteve-se 100% de plantas sobreviventes, oriundas do meio MS/4. A realização desse trabalho permite estimar a obtenção de 32.000 mudas de V. condensata, após sete meses de cultivo in vitro.(AU)