Stomatal behaviour and endogenous phytohormones promotes intrinsic water use efficiency differently in cotton under drought

The coordination between phytohormones regulation, stomatal behaviour (stomatal index and opening/closing) and gas exchange are potent determinants of plant survival under drought stress. However, we found a knowledge gap in the mechanism regulating the fine-tuning of these features during drought. In order to address this we evaluated gas exchange, stomatal behaviour and endogenous phytohormones content in two cotton varieties (LRA-5166 and NBRI-67) differing in drought sensitivity during water deficit conditions. Variety specific differences were recorded in net photosynthesis rate (A), transpiration rate (E) and stomatal conductance (gs) with significantly less decrease in drought tolerant LRA-5166 than drought sensitive NBRI-67. The abscisic acid (ABA) accumulation was significantly increased in LRA-5166 while reduced in NBRI-67 under water deficit, which was accompanied by relatively less reduced 6-benzylaminopurine (6-BAP) level in LRA-5166 than NBRI-67.Thus, improved ABA/6-BAP ratio was observed in both the varieties of cotton. Critically, LRA- 5166 has reduced stomatal index, aperture size and significantly higher A and intrinsic water use efficiency (WUEi), thus higher drought tolerance than NBRI-67. Furthermore, we found that endogenous ABA predominantly maintains the stomatal behaviour and regulates its physiology either by antagonizing 6-BAP or alone to coordinate with water deficit signals. Overall, our findings describe a new insight as to how drought modulates endogenous ABA and 6-BAP homeostasis in cotton leaf and the mechanism of stomatal regulation by ABA and 6-BAP in cotton.

Multiplication, rooting in vitro, and acclimatization of Brassavola tuberculata Hook. (Orchidaceae), an orchid endemic to the brazilian atlantic rainforest / Multiplicação, enraizamento in vitro e aclimatização de Brassavola tuberculata Hook (Orchidaceae), uma orquídea endêmica da mata atlântica brasileira

The objective of this study was to promote the establishment of an in vitro culture of Brassavola tuberculata, testing different concentrations of naphthalene acetic acid (NAA) and 6-benzylaminopurine (BAP) on multiplication and rooting, evaluating different substrates during acclimatization, as well as the effect of in vitro treatments. After germination, the seedlings of B. tuberculata were subjected to culture on MS medium supplemented with different concentrations of NAA and BAP, and multiplication and rooting were assessed. During acclimatization, different substrates were tested: S1, Plantmax® and vermiculite (1: 1); S2, Plantmax® and grit (1: 1); and S3, dust fern. Also the effect of the in vitro culture treatments was evaluated: T1, control; T5, (2.5 µM NAA +5 µM BAP); and T7, (5 µM NAA + 0 µM BAP). The favorable balance of cytokinins promoted by treatment T5 yielded the largest number of shoots and leaves in B. tuberculata. The greatest length of leaves and roots, and highest root number were observed in the treatment T7, favored by the presence of auxin. This treatment had a positive effect with respect to plant acclimatization: T7 associated with substrate S1 provided the most suitable conditions for acclimatization of seedlings of B. tuberculata, providing greater number and length of leaves, and high survival rate.

Objetivou-se, com este trabalho, promover o estabelecimento do cultivo in vitro de Brassavola tuberculata, testando diferentes concentrações de ANA e BAP na multiplicação e no enraizamento, e avaliar diferentes substratos e o efeito dos tratamentos de cultivo in vitro na aclimatização. Após a germinação das sementes, as plântulas de B. tuberculata foram submetidas ao cultivo em meio MS suplementado com diferentes concentrações de ácido naftaleno acético (ANA) e 6-benzilaminopurina (BAP), sendo avaliados a multiplicação e o enraizamento. Foram testados diferentes substratos: S1 (Plantmax e vermiculita (1:1)); S2 (Plantmax e areia (1:1)) e S3 (pó de xaxim) na aclimatização e, posteriormente, o efeito dos tratamentos do cultivo in vitro: T1 (controle), T5 (2,5 ANA + 5 BAP) e T7 (5 ANA + 0 BAP), na aclimatização. O balanço favorável às citocininas promovido pelo tratamento T5 (2,5 µM ANA + 5 µM BAP) promoveu maior número de brotos e de folhas em B. tuberculata. O maior comprimento das folhas, das raízes e maior número de raízes foi observado no tratamento T7 (5 µM ANA e 0 µM BAP), favorável a auxina. Este tratamento apresentou efeito positivo com relação a aclimatização das plantas: T7 associado ao substrato S1, Plantmax e vermiculita (1:1) proporcionou melhores condições para a aclimatização das plântulas de B. tuberculata, propiciando maior número e comprimento das folhas, e elevada taxa de sobrevivência.

Micropropagation of Calophyllum brasiliense (Cambess) from nodal segments / Micropropagação de Calophyllum brasiliense (Cambess) a partir de segmentos nodais

Abstract Micropropagation of Calophyllum brasiliense Cambess. (Clusiaceae) is a way to overcome difficulties in achieving large-scale plant production, given the recalcitrant nature of the seeds, irregular fructification and absence of natural vegetative propagation of the species. Cultures were established using nodal segments 2 cm in length, obtained from 1-2 year old seedlings, maintained in a greenhouse. Mercury chloride and Plant Preservative Mixture™ were used in the surface sterilizing stage, better results being achieved with Plant Preservative Mixture™ incorporation in culture medium, at any concentration. Polyvinylpyrrolidone, activated charcoal, cysteine, ascorbic acid or citric acid were added to the culture medium to avoid oxidation. After 30 days of culture, polyvinylpirrolidone and ascorbic acid gave better results, eliminating oxidation in most explants. For shoot multiplication, benzylaminopurine was used in concentrations of 4.4 and 8.8 µM in Woody Plant Medium, resulting in an average of 4.43 and 4.68 shoots per explant, respectively, after 90 days. Indole-3-butyric acid and α-naphthalene acetic acid were used to induce root formation, reaching a maximum rooting rate of 24% with 20µM α-naphthalene acetic acid. For acclimatization. the rooted plants were transferred to Plantmax® substrate and cultured in a greenhouse, reaching 79% of survival after 30 days and 60% after one year.

Resumo A micropropagação de Calophyllum brasiliense Cambess. (Clusiaceae) é uma maneira de superar dificuldades para sua produção em larga escala, devido à natureza recalcitrante das sementes, frutificação irregular e ausência de propagação vegetativa natural da espécie. Culturas foram estabelecidas utilizando segmentos nodais com 2 cm de comprimento, obtidos de plantas com 1 a 2 anos de idade, mantidas em casa de vegetação. Cloreto de mercúrio e Plant Preservative Mixture™ foram utilizados durante a etapa de desinfestação, com melhores resultados alcançados com a incorporação de Plant Preservative Mixture™ ao meio de cultura. Polivinilpirrolidona, carvão ativado, cisteína, ácido ascórbico ou ácido cítrico foram adicionados ao meio de cultura para evitar a oxidação dos explantes. Após 30 dias de cultivo, o uso de polivinilpirrolidona ou ácido ascórbico proporcionou melhores resultados, eliminando a oxidação na maioria dos explantes. Para multiplicação das brotações, benzilaminopurina foi usada em concentrações de 4.4 e 8.8 µM em meio WPM, resultando em uma média de 4.43 e 4.68 brotações por explante, respectivamente, após 90 dias. Ácido indol-3-butírico e ácido α-naftaleno acético foram usados para a indução de raízes, alcançando um enraizamento máximo de 24% com o uso de 20µM de ácido α-naftaleno acético. As plantas enraizadas foram transferidas para substrato Plantmax® e cultivadas em casa de vegetação, alcançando 79% de sobrevivência após 30 dias e 60% após um ano.

ESTABLECIMIENTO DEL SISTEMA DE REGENERACIÓN POR EMBRIOGÉNESIS SOMÁTICA DE Azadirachta indica A. Juss. / Establishment of Regeneration System for Somatic Embryogenesis of Azadirachta Indica A. Juss.

Azadirachta indica, es una planta con múltiples aplicaciones tanto forestal como farmacológica. Por ende, el establecimiento del sistema de cultivo in vitro por embriogénesis somática ofrece diversas y variadas ventajas, tales como obtener plantas altamente productivas en metabolitos. En este estudio, se utilizaron secciones foliares y cotiledonares, inducidas en medios MS (1962) suplementados con: BAP sólo y combinado con ANA / 2,4-D, TDZ sólo y con ABA. La regeneración fue con MS sólo o con K + AIA y BAP + AIA. Como resultado se estableció un sistema eficiente con secciones de cotiledones, observándose organogénesis a bajas concentraciones de BAP, mientras a altos niveles de BAP (2,5 mg.L-1), así como con TDZ + ABA (0,02 + 1 mg.L-1) respectivamente favorecieron la embriogénesis somática primaria y secundaria en un 96 % y 71 % respectivamente. La regeneración fue 71 % con MS, mientras que el enraizamiento fue de 86,67 % con MS½, obteniéndose plantas completas a corto plazo.

Azadirachta indica, is a plant with multiple forest and pharmacological application. Therefore, the establishment of in vitro culture system for somatic embryogenesis offers several distinct advantages such as obtaining highly productive plant metabolites. In this study, were used sections cotyledon and leaf, induced on MS medium (1962) supplemented with: BAP alone and combined with NAA / 2,4-D, TDZ alone and ABA. Regeneration was with MS alone or with K + BAP + IAA and IAA. As a result was established an efficient system with cotyledon sections, being observed organogenesis at low concentrations of BAP, while high levels of BAP (2.5 mg.L-1) with 96 % and TDZ + ABA (0.02 + 1 mg.L-1) with 71 %, favoring the primary and secondary somatic embryogenesis. Regeneration was 71 % with MS; rooting was 86.67 % with MS½, presenting whole plants obtained short term.

FORMACIÓN DE EMBRIONES SOMÁTICOS A PARTIR DE SEMILLAS INMADURAS EN Sorghum bicolor VARIEDAD CIAP 132-R / Formation of Somatic Embryos from Immature Seeds of Sorghum bicolor Sorghum Variety CIAP 132-R

Existen varios protocolos de regeneración de plantas vía embriogénesis somática de Sorghum bicolor (L.) Moench, sin embargo los porcentajes de formación de callos con estructuras embriogénicas y regeneración de plantas son bajos. Es por ello que esta investigación tuvo como objetivo generar embriones somáticos en sorgo rojo variedad CIAP 132-R. Se ensayaron diferentes concentraciones de 2,4-D para la formación de callos, así como tres concentraciones de ácido ascórbico para eliminar la exudación de compuestos fenólicos por el explante. También para la formación de los embriones somáticos a partir de los callos se evaluaron diferentes concentraciones de 2,4-D y 6-BAP. El mayor porcentaje de formación de callos (57,5 %) se alcanzó con 18,1 µM de 2,4-D. Con la adición al medio de cultivo de 50,0 mg.l-1 de ácido ascórbico fue posible eliminar los compuestos fenólicos en el explante y en el medio de cultivo, además permitió incrementar el porcentaje de formación de callos con estructuras embriogénicas hasta un 95 %. El número mayor de embriones somáticos por callo se alcanzó en el medio de cultivo con concentraciones de 4,52 µM de 2,4- D, combinada con 2,22 µM de 6-BAP. Por primera vez, se logró la formación eficiente de embriones somáticos a partir de los callos obtenidos de semillas inmaduras germinadas como explante inicial en la variedad CIAP 132-R.

Several protocols of plant regeneration via somatic embryogenesis from Sorghum bicolor (L.) Moench have been development, however the percentage of calluses with embryogenic structures and plant regeneration are low. Therefore this study aimed to generate somatic embryos in red sorghum variety CIAP 132-R. Different concentrations of 2,4-D for callus formation, and three concentrations of ascorbic acid to remove phenolics exudation were assayed by explant. For the formation of embryos different concentrations of 2,4-D and 6-BAP were evaluated. The highest percentage of callus formation (57.5 %) was achieved with 18.1 µM 2,4-D. With the addition to the culture medium of 50.0 mg.l-1 of ascorbic acid was possible to eliminate the phenolic compounds in the explant and in the culture medium; also it allows increasing the percentage of calluses with embryogenic structures up to 95 %. The highest number of somatic embryos per callus was achieved with a reduction in the culture medium of 2,4-D to 4.52 µM in combination with 2.22 µM 6-BAP. For the first time, the efficiency of somatic embryo formation was obtained from the freshly germinated sprouts of immature seeds as initial explant CIAP 132-R.

Direct Multiple Shoot Induction and Plantlet Regeneration from Cotyledonary Explants of Sapindus emarginatus Vahl (Soapnut).

germination of S. emarginatus in vitro cotyledon explants in BAP/Kn/TDZ (1.0-3.0 mg/L) supplemented MS medium and (2) in plant treatment with BAP/Kn/TDZ (3.0 mg/ L) in combination of 1AA (0.5 mg/L) of the cotyledon explants of plants and maintained under sterile conditions. While the former method resulted in as many as (7.5±8.6 shoot buds) from the cotyledonary explants within four weeks, the latter yielded on average approximately 8 shoot buds from each treated node in eight weeks. The cytokinin treatment in plant consisted of placing sterile filter paper moistened with sterile distilled water over the node and adding different concentrations of 6- benzylaminopurine. The best results for shoot bud regeneration were obtained with cotyledons, when cultured in the presence of (0.5 mg/L) IAA in combination with (3.0 mg/L). The shoots elongated and rooted directly in vermiculite after a pulse treatment with IBA (2.5 mg/L) for 15 min. Fungus growth, a serious problem in S. emarginatus tissue culture, was controlled using a fungicide, Bavistin, along with elimination of organic nutrients from the growth medium.

Establecimiento in vitro de dos especies nativas de Costa Rica: Terminalia amazonia (Amarillón) y Vochysia allenii (Botarrama Blanco) / In vitro establishment of two species native to Costa Rica: Terminalia amazonia (Amarillón) and Vochysia allenii (Botarrama Blanco)

El objetivo de esta investigación fue lograr el establecimiento in vitro de la especie Terminalia amazonia y Vochysia allenii debido a la dificultad de propagarlas sexual y asexualmente con técnicas convencionales. Se logró establecer segmentos nodales de ambas especies en condiciones in vitro empleando el HgCl2 0,1 % con un tiempo de exposición de 5 minutos. El mejor medio de cultivo para nudos fue el WPM 100 % de sales para T.amazonia y para V. allenii fue el WPM 50 % de sales. Después de 28 días de cultivo se obtuvo un 42 % de nudos establecidos para T. amazonia y 10% para V. allenii. En ambas especies se evaluó el efecto sobre la brotación de cinco concentraciones de 6-bencilaminopurina (6-BAP) (0,0; 2.22, 4.44, 6.66, 8.88 μM L-1) y cinco de tidiazuron (TDZ) (0,0; 0.22, 0.45, 0.68, 0.90 μM L-1). Se obtuvó en promedio un brote por explante en los cinco tratamientos de BAP y TDZ utilizados.

The objective of this research was to achieve the in vitro establishment of the species Terminalia amazonia and Vochysia allenii due to the difficulty propagate sexually and asexually with conventional techniques. It was possible to establish the nodal segments of both species in in vitro conditions using 0.1 % HgCl2 with an exposure time of 5 minutes. The best nodal culture medium was 100 % WPM salts in T. amazonia and V. allenii was WPM 50% salts. After 28 days of culture 42 % of nodal segments to T. amazonia and V. allenii 10% was obtained. In both species, the effect on sprouting of five concentrations of 6-benzylaminopurine (6-BAP) (0.0, 2.22, 4.44, 6.66, 8.88 μM L-1) and five thidiazuron (TDZ) (0.0, 0.22, 0.45, 0.68, 0.90 μM L-1) was evaluated. Outbreak was scored on average per explant in the five treatments of BAP and TDZ used.

In vitro establishment and multiplication of the Normania triphylla (Lowe) Lowe

Normania triphylla is an endemic species from Madeira island (Portugal) extinct in the wild since 1991. The aim of this work was to culture the meristems of this species in vitro and to multiply its shoots in order to preserve this endangered species. The best results in terms of multiplication were obtained in Murashige and Skoog medium supplemented with 10 µM 6‑benzylaminopurine (BAP). The number of shoots, the number of nodes and the number of leaves were the most important in this medium. However, the best results concerning the total shoot length were obtained when BAP was not supplemented into the medium and in the presence of 5 or 7.5 µM 1-naphtalene acetic acid (NAA). This is the first report on the in vitro culture of N. triphylla which could brings new avenues for the development of this species.

Efeito do BAP no cultivo in vitro de Ocimum selloi Benth / Effect of BAP on in vitro culture of Ocimum selloi Benth

Visando promover a proliferação de brotações em segmentos apicais e nodais de Ocimum selloi em diferentes concentrações de BAP, plantas jovens de 60 dias serviram de doadoras de segmentos apicais e nodais. Os segmentos foram inoculados em meio MS preparado com a metade da concentração dos sais, e acrescido de 1,5% de sacarose e diferentes concentrações de BAP. O experimento foi conduzido no delineamento fatorial, 3 x 4, sendo 3 as posições dos segmentos de O. selloi (segmento apical, primeiro e segundo segmentos nodais) e 4 concentrações de BAP (0 - controle; 2; 4 e 6 mg L-1). Aos 30 dias, foram avaliados o número, comprimento e biomassa fresca e seca de brotos e raízes. Os primeiros e segundos segmentos apresentaram melhores resultados na indução de brotos de O. selloi, 7 e 8 brotos/explante, nas diferentes concentrações de BAP; porém, não houve formação de raízes na presença da citocinina. Nas condições testadas, recomenda-se o uso do primeiro e segundo segmento nodal suplementando o meio de cultivo com BAP para a proliferação in vitro de brotações de O. selloi.

The present study was undertaken to develop the proliferation of sprouts in apical and nodal segments of Ocimum selloi with different BAP levels. Young plants aged 60 days were used as donors of nodal and apical segments. The segments were inoculated in MS medium at half the concentration of salts supplemented with 1.5% of sucrose and different BAP levels. The experiment was in 3 x 4 factorial arrangement, 3 positions of O. selloi segments (apical segment; first and second nodal segment) and 3 BAP levels (0 - control; 2; 4 and 6 mg L-1). After 30 days, the number, the length, and the fresh and dry biomass of sprouts and roots were evaluated. The first and the second segments showed better results in inducing O. selloi sprouts, 7 and 8 sprouts/explant, at the different BAP levels, but there was not root formation in the presence of the cytokinin. Under the tested conditions, use of the first and the second nodal segments is recommended in addition to supplementing the culture medium with BAP for in vitro proliferation of O. selloi sprouts.

Optimización de un medio de cultivo para plantas micropropagadas de dioscorea alata L / Optimization of a culture medium for micropropagated plants of dioscorea alata L

La propagación de material de ñame de buena calidad es esencial para incrementar la producción sostenible de este cultivo. El presente trabajo tuvo como propósito optimizar el medio de cultivo de micropropagación de Dioscorea alata L. clon Caraqueño a través de los siguientes objetivos: determinar el efecto de diferentes antioxidantes (carbón activado 0,5 g/L-1 ; carbón activado 1,0 g/L-1; cisteína 10 mg/L-1, 20 mg/L-1 y 30 mg/L-1) y concentraciones de sales de Murashige y Skoog (MS) (25, 50, 75 y 100 %) en el medio de cultivo durante el establecimiento y la multiplicación de las plantas in vitro, y evaluar la utilización de distintas combinaciones de ácido naftalenacético (0,01; 0,1 mg/L-1) y bencilaminopurina (0,01; 0,1 mg/L-1) en el mejor medio de cultivo de multiplicación obtenido en el experimento anterior. A los 35 días se seleccionaron 40 plantas in vitro, a las cuales se les determinaron las siguientes variables: longitud en cm del vástago; número de nudos de novo por explantes; número de hojas por explante y porcentaje de fenolización. Se evaluó además, en el experimento con los reguladores de crecimiento, el número de raíces y longitud de la raíz de mayor tamaño. Se aplicó un diseño experimental completamente aleatorio con análisis de varianza bifactorial y clasificación simple. Se realizó la prueba de comparación de medias de Tukey para un nivel de significación del 5%. Los resultados obtenidos mostraron que las sales MS al 75% de su concentración, el carbón activado (0,5 g/L-1) o la cisteína (10 mg/L-1), en combinación con los reguladores de crecimiento ANA/BAP (0,01/0,01 mg/L-1) en el medio de cultivo MS, incrementaron los indicadores de desarrollo de las plantas in vitro tales como número de nudos de novo (3,5), longitud del vástago (4,1 cm), número de hojas (3,8), número de raíces (5,7) y longitud de las raíces (6 cm).

The material propagation of good quality yam is essential to increase the sustainable production of this cultivation. In the present work the optimization of culture medium of Dioscorea alata L. clone Caraqueño micropropagation was carried out through the following objectives: to determine the effect of different anti-oxydants (activated charcoal 0,5 g.L-1; activated charcoal 1,0 g.L-1; cysteine 10 mg.L-1, 20 mg.L-1 and 30 mg.L-1) and Murashige and Skoog salts concentrations (25, 50, 75 and 100%) in the culture medium during the in vitro plants establishment and the multiplication, and to evaluate the use of different combinations of naftalenacetic acid (0,01; 0,1 mg.L-1) and benzylaminopurine (0,01; 0,1 mg.L-1) in the best multiplication medium obtained in the above experiment. At 35 days, 40 in vitro plants were selected. The following variables were determined in these plants: shoot length, cm; leaf and bud explant number; and oxydation phenolic percentage. In the experiment of plant growth regulators was also evaluated, the roots number and greater size root length. A totally randomized experimental design with one and two factor variance analysis and the Tukey test for means comparison at 5% significance level were applied. The obtained results showed that the salts MS at 75% concentration, the activated charcoal (0,5 g. L-1) or the cysteine (10 mg. L-1), in combination with the growth regulators ANA/BAP (0,01/0,01 mg.L-1) in the MS culture medium, increase the development of the in vitro plants, number of novo buds (3,5), shoot length (4,1 cm), number of leaves (3,8), number of roots (5,7) and greater size root length (6 cm).

Micro propagation and tissue culture of the endangered medicinal plant withania somnifera by the direct shoot and root initiation method.

Leaf and Cotyledon explants of Withania somnifera (L). Dunal were used to evaluate the effect of different growth regulators on the in vitro direct shoot and root initiation methods. Four different explants were used to establish callus shoot and root direct regeneration. In the first experiment leaf segments were cultured on MS basal supplemented with 2,4 – Dichlorophenoxyacetic acid (2,4 – D, 0.1-20.0 mg/L), with combination of Naphthalene acetic acid (NAA 0.1-20 mg/L) and Benzylaminopurin (0.1-20 mg/L). This new protocol was standardized for easy mass propagation of W. somnifera medicinal plant. Callus initiation was observed best in MS media with (2,4- D 1.0-5.0 mg/L) after 16-20 days (93%). Highest maximum number of multiple shoots was obtained on MS medium (BAP 3.0 – 5.0 mg/L). The shoots were seaperated from the multiple-shoots, transferred to MS medium supplemented with 1.5 – 20 mg/L NAA favored roots formation occurred in most of the shoot let 88% were successfully achieved in the MS media. The rooted plantlets were transferred to polythene bags which was containing vermi compost, sand and red soil in the ratio of 1:2:2 and kept in a mist house. After acclimatization in the mist house for 2-months, it transferred to greenhouse. The plantlets were successfully planted in the field.

In vitro shoot development in blepharis repnes.

An herb is a plant or plant part valued for its medicinal aromatic or savory qualities. In vitro shoot development of Blepharis repnes was achieved using thin sections of nodes and stems (1.0-1.5 mm) were cultured in MS medium containing varying concentrations of different combination of phytohormones Indole-3-butyric acid (IBA), Benzylaminopurine (BAP) & Gibberlic acid. The combination of BAP, IBA & Gibberellic acid at different concentrations showed the length of shoot development 0.6 -1.8cm with in three weeks in suitable laboratory condition.

Diodos emissores de luz e concentrações de BAP na multiplicação in vitro de morangueiro / Light-emitting diodes and BAP concentrations in the in vitro strawberry multiplication

Realizou-se um estudo para avaliar o efeito de fontes de luz na micropropagação de morangueiro, com níveis crescentes de BAP no meio de cultivo. Para tanto, inocularam-se gemas de brotações da cultivar 'Sabrosa' em meio MS com 30g L-1 de sacarose, 100mg L-1 de mio-inositol, 7g L-1 de ágar e BAP (0; 0,3; 0,6; 0,9; e 1,5mg L-1), em pH 5,8. Os explantes foram cultivados a 25+2°C, com 16 horas de fotoperíodo e luminosidade de 20µmol m-2 s-1, esta última fornecida por diferentes fontes de luz (LED azul-EDEB 3LA1, LED verde-EDET 3LA1, LED vermelho-EDER 3LA3, lâmpada fluorescente Growlux e lâmpada fluorescente branca). Os tratamentos foram dispostos em delineamento inteiramente ao acaso em um fatorial 5x5 (concentrações de BAP x fontes de luz), com seis repetições. O experimento foi repetido em três subcultivos sucessivos de 35 dias cada. Nestes avaliaram-se o número de brotações por explante e a altura das brotações. Ao final do terceiro subcultivo, determinaram-se, ainda, as concentrações de carotenoides e de clorofilas a e b, independentemente do nível de BAP. Maior número de brotações por explante foi obtido sob LEDs vermelhos e verdes. Concentrações de BAP no meio de cultura entre 0,82 e 1,22mg L-1, dependendo da fonte de luz, proporcionaram maior multiplicação in vitro de brotações. Sob todas as fontes de luz foram obtidas brotações de maior comprimento em meio isento de BAP. Brotações cultivadas sob LEDs vermelhos apresentaram maior quantidade de pigmentos fotossintéticos, enquanto aquelas sob LEDs verdes e lâmpadas Growlux apresentaram a menor.

The objective of this research was to evaluate the effect of different sources of light in strawberry micropropagation, under increasing levels of BAP in culture medium. 'Sabrosa' shoots were inoculated in MS medium supplemented with 30g L-1 sucrose, 100mg L-1 myo-inositol, 7 agar g L-1 and BAP (0; 0.3; 0.6; 0.9; e 1.5mg L-1), pH 5.8. The explants were cultivated at 25+2°C, 16 hours photoperiod and 20µmol m-2 s-1. The luminosity was supplied by different sources of light (blue-EDEB 3LA1 LED, green-EDET 3LA1 LED, red-EDER 3LA3 LED, Growlux fluorescent lamp and white fluorescent lamp). The experimental design was a factorial entirely randomized (5 concentrations of BAP x 5 light sources) with six replications. The experiment was repeated in three successive subcultures of 35 days each, being evaluated the shoot number per explant and shoot height. The carotenoids and chlorophyll a and b determinations were carried out after the third subculture, independently of BAP concentration. Shoot number per explant was higher under red and green LEDs. BAP concentrations between 0.82 and 1.22mg L-1 in culture medium showed higher multiplication rate depending on the light source. Shoot length was highest in culture medium without BAP under all light sources. Shoots cultivated under red LEDs showed higher concentration of photosynthetic pigments, while those under green LEDs and Growlux light bulbs showed the lowest.

Crescimento de plantas de Salvia officinalis sob ação de reguladores de crescimento vegetal / Growth of Salvia officinalis plants under action of plant growth regulators

O objetivo deste trabalho foi estimar os parâmetros da análise de crescimento em função de diferentes reguladores vegetais aplicados na parte aérea de plantas de Salvia officinalis L. Para tanto,o experimento foi instalado em casa de vegetação do Departamento de Botânica, Instituto de Biociências, da Universidade Estadual Paulista, Botucatu, SP. Os tratamentos consistiram na pulverização da solução de 100mg L-1 de ácido giberélico (GA3); 100mg L-1 de benzilaminopurina (BAP); 100mg L-1 de ácido 2-cloroetil-fosfônico (ethephon); Stimulate® a 2 por cento (90mg L-1 de cinetina, 50mg L-1 de ácido giberélico e 50mg L-1 de ácido indolilbutírico) e água (testemunha). As aplicações foram realizadas em três épocas, aos 15, 25 e 35 dias após o transplante (d.a.t.) e o crescimento foi avaliado em cinco épocas de coletas a intervalos de 21 dias, sendo a primeira realizada aos 47 (d.a.t.). Foram determinados os parâmetros fisiológicos da análise de crescimento: razão de área foliar (RAF), área foliar específica (AFE), taxa assimilatória líquida (TAL) e taxa de crescimento relativo (TCR). Os resultados mostram que os reguladores de crescimento vegetal influenciaram os parâmetros fisiológicos da análise de crescimento. As plantas tratadas com BAP apresentaram maiores valores de RAF aos 47 d.a.t., já as plantas tratadas com GA3, a TAL apresentou aumento até o 131 d.a.t. A TCR é decrescente para todos os tratamentos com reguladores de crescimento vegetal testados e a testemunha.

The objective of this study was to estimate the effects of different plant regulators on the index growth analysis application of sage plants. An experiment was conducted in a greenhouse with controlled temperature and relative humidity, at the Departmento de Botânica, Instituto de Biociências, Universidade Estadual Paulista, UNESP, Botucatu, SP, Brazil. The experimental design was completely randomized, with five treatments containing three replicates. Treatments consisted of the spraying of the solution of 100mg L-1 of gibberellic acid (GA3); 100mg L-1 of benzylaminopurine (BAP); 100mg L-1 of 2-chloroethyl phosphonic acid (ethephon); Stimulate® at 2 percent (90mg L-1 of kinetin, 50mg L-1 of gibberellic acid and 50mg L-1 of indolylbutyric acid) and water (control). Applications of plant growth regulators were performed in 3 times, at 15, 25, and 35 days after transplanting, and growth was evaluated in five successive harvests at 21-day intervals, performed at 47, 68, 89, 110, and 131 days after transplanting. The following growth index were determined: leaf area ratio (LAR), specific leaf area (SLA), net assimilation rate (NAR), and relative growth rate (RGR). These results suggest that the plant growth regulators influence the index growth analysis. The plants treated with BAP presented increase values of LAR to the 47 days after transplanting. The plants treated with GA3 to NAR presented increase until the 131 days after transplanting. The RGR is decreasing for all the treatments with vegetal regulators tested and the control.

Influência da benzilaminopurina (BAP) NA multiplicação in vitro de kiwi (Actinidia deliciosa) / The influence of benzylaminopurine in the in vitro multiplication of kiwi (Actinidia deliciosa)

Este experimento foi conduzido no Laboratório de Cultura de Tecidos da EMBRAPA/CPACT, Pelotas, RS, com o objetivo de estudar a influência de diferentes concentrações de 6-benzilaminopurina (BAP) na multiplicação in vitro de kiwi (Actinidia deliciosa), cv. Tomun. Utilizou-se o meio de cultura MS, acrescido de sacarose, mio-inositol, ágar e das seguintes concentrações de BAP: 0,0; 0,5; 1,0; 1,5; 2,0 e 2,5 mg/l. Os explantes foram constituídos de microestacas provenientes da coleção in vitro do Laboratório, com aproximadamente 10mm de comprimento. Verificou-se que, para o número e comprimento de brotações, número de gemas e folhas a concentração em torno de 1,5mg/l de BAP proporcionou os memores resultados. Nas condições em que o trabalho foi realizado, pode-se concluir que o BAP foi eficiente na multiplicação in vitro de kiwi, cv. Tomuri.

This trial was carried out in the Tissue Culture Laboratory at EMBRAPA/CPACT, Pelotas, RS, Brazil, aiming to study the influence of different 6-benzylaminopurine (BAP) concentrations in the in vitro multiplication of kiwi, cv. Tomuri. A MS medium was used adding sucrose, myo-inositol, agar and concentrations of BAP as follows: 0.0; 0.5; 1.0; 1.5; 2.0 and 2.5mg/l. The explants consisting of microshoots of 10mm length were obtained from the in vitro collection at the laboratory. The best results of bud and shoot numbers, shoot length and number of leaves were obtained using about 1.5mg/l BAP. Results of this experiment demonstrated that BAP is efficient in the in vitro multiplication of kiwi, cv. Tomuri.