ACTIVATED CHARCOAL APPLICATION FOR THE MICROPROPAGATION OF Cattleya crispata (Thunb.) Van den Berg

 Micropropagation is an alternative for the genetic conservation and propagation of endemic species from “Campo Rupestre Ferruginoso”, such as the orchid Cattleya crispata. The aim of the present study is to assess the influence of activated charcoal on the in vitro germination, multiplication and elongation phases of C. crispata. Seeds extracted from mature capsules were used for inoculation in the culture medium that was adopted to assess the effect of supplementation, or not, with activated charcoal. Data about germination speed, seedling number, length, vigor, oxidation and contamination (bacterial and/or fungal) were assessed through these phases. Based on the results obtained, the use of activated charcoal was efficient in the in vitro germination and multiplication phases of C. crispata, providing greater speed and percentage of germination, less contamination and oxidation of the tissues, greater number, length and vigor of shoots, being effective for the genetic conservation and production of seedlings of the species. Culture medium without the supplementation of activated charcoal provided the best results for the in vitro elongation, with greater length, vigor and less oxidation of shoots.Aplicação de carvão ativado para micropropagação de Cattleya crispata (Thunb.) Van den Berg  RESUMO: A micropropagação é uma alternativa para a conservação genética e propagação de espécies endêmicas do “Campo Rupestre Ferruginoso”, como a orquídea Cattleya crispata. O objetivo do presente estudo é avaliar a influência do carvão ativado nas fases de germinação, multiplicação e alongamento in vitro de C. crispata. Sementes extraídas de cápsulas maduras foram utilizadas para inoculação no meio de cultura adotado para avaliar o efeito da suplementação, ou não, com carvão ativado. Dados sobre velocidade de germinação, número de brotos por explante, comprimento, vigor, oxidação e contaminação (bacteriana e / ou fúngica) foram avaliados por meio dessas fases. Com base nos resultados obtidos, o uso do carvão ativado foi eficiente nas fases de germinação e multiplicação in vitro de C. crispata, proporcionando maior velocidade e porcentagem de germinação, menor contaminação e oxidação dos tecidos, maior número, comprimento e vigor dos brotos, sendo eficaz para a conservação genética e produção de mudas da espécie. O meio de cultura sem a suplementação de carvão ativado proporcionou os melhores resultados para o alongamento in vitro, com maior comprimento, vigor e menor oxidação dos brotos. Palavras-chave: antioxidante; campos rupestres; orquídea; propagação in vitro.   ABSTRACT: Micropropagation is an alternative for the genetic conservation and propagation of endemic species from rupestrian grasslands, such as the orchid Cattleya crispata. The aim of the present study is to assess the influence of activated charcoal on the in vitro germination, multiplication and elongation phases of C. crispata. Seeds extracted from mature capsules were used for inoculation in the culture medium that was adopted to assess the effect of supplementation, or not, with activated charcoal. Data about germination speed, number of shoots per explant, length, vigor, oxidation and contamination (bacterial and/or fungal) were assessed through these phases. Based on the results obtained, the use of activated charcoal was efficient in the in vitro germination and multiplication phases of C. crispata, providing greater speed and percentage of germination, less contamination and oxidation of the tissues, greater number, length and vigor of shoots, being effective for the genetic conservation and production of plants of the species. Culture medium without the supplementation of activated charcoal provided the best results for the in vitro elongation, with greater length, vigor and less oxidation of shoots. Keywords: antioxidant; orchid; propagation in vitro; rupestrian grasslands

CLONAL MICROPLANT PRODUCTION, MORPHOLOGICAL EVALUATION AND GENETIC STABILITY OF Dendrocalamus asper (Schult. & Schult.) Backer ex. K. Heyneke

ABSTRACT: Bamboo species have many commercial applications, considering that homogeneous plantations (formed from clonal plants) are essential to high sustainable biomass production. The cloning of selected plants on an industrial scale through in vitro cultivation has many advantages, being important for the supply of plants in sufficient quantity and quality to meet commercial demand. The control of the cloning is the basis for an industrial scale, and its knowledge can optimize the process. This work aimed to evaluate the cloning of Dendrocalamus asper selected plant through micropropagation. Morphological features by scanning electron microscopy and genetic stability with ISSR molecular markers were evaluated. Four times of immersion in sodium hypochlorite (NaOCl) on in vitro establishment of nodal segments were evaluated. The established explants were transferred to a culture medium that was supplemented with three concentrations of 6-benzylaminopurine (BAP). Three concentrations of indole-3-butyric acid (IBA) to the in vitro adventitious rooting were evaluated. NaOCl application for 10 min resulted in 71.4 % of establishment in 30 d. Supplementation of the culture medium with 2.0 and 3.0 mg L-1 BAP de resulted in the highest averages for multiplication and elongation stages. The formation of adventitious roots occurred with 4.0 mg L-1 IBA of supplementation. Micropropagated plants showed normal morphological features and genetic stability, confirming the cloning of selected plant. Keywords: bamboo; micropropagation; vegetative propagation; In vitro culture; ISSR; plant growth regulator.   Produção de microplantas clonais, avaliação morfofisiológica e estabilidade genética de Dendrocalamus asper (Schult. & Schult.) Backer ex. K. Heyneke   RESUMO: Espécies de bambus apresentam diversas aplicações comerciais, visto que os plantios homogêneos (formados a partir de plantas clonais) são essenciais para a alta produção de biomassa sustentável. A clonagem de plantas selecionadas em escala industrial por meio do cultivo in vitro apresenta muitas vantagens, sendo uma importante ferramenta para o fornecimento de plantas em quantidade e qualidade suficientes para atender a demanda comercial. O controle da clonagem é a base para escala industrial, e seu conhecimento pode otimizar os processos. O trabalho teve como objetivo avaliar a clonagem de planta selecionada de Dendrocalamus asper por meio da técnica de micropropagação. Foram avaliadas as características morfológicas por microscopia eletrônica de varredura e estabilidade genética por meio de marcadores moleculares ISSR. Além disso, foram avaliados quatro tempos de imersão em hipoclorito de sódio (NaOCl) no estabelecimento in vitro de segmentos nodais. Os explantes estabelecidos foram transferidos para um meio de cultura que foi suplementado com três concentrações de benzilaminopurina (BAP). Por fim, foram avaliadas três concentrações de ácido indolbutírico (AIB) durante o enraizamento adventício in vitro. A adição de NaOCl por 10 min resultou em 71,4 % de estabelecimento em 30 d. A suplementação do meio de cultura com 2,0 e 3,0 mg L-1 BAP resultou nas maiores médias para as fases de multiplicação e alongamento. A formação de raízes adventícias ocorreu com a suplementação de 4,0 mg L-1 de AIB. Plantas micropropagadas apresentaram características morfológicas normais e estabilidade genética, confirmando a clonagem da planta selecionada. Palavras-chave: bambu; micropropagação; propagação vegetativa; cultivo in vitro; ISSR; regulador de crescimento vegetal.ABSTRACT: Bamboo species have many commercial applications, considering that homogeneous plantations (formed from clonal plants) are essential to high sustainable biomass production. The cloning of selected plants on an industrial scale through in vitro cultivation has many advantages, being important for the supply of plants in sufficient quantity and quality to meet commercial demand. The control of the cloning is the basis for an industrial scale, and its knowledge can optimize the process. This work aimed to evaluate the cloning of Dendrocalamus asper selected plant through micropropagation. Morphological features by scanning electron microscopy and genetic stability with ISSR molecular markers were evaluated. Four times of immersion in sodium hypochlorite (NaOCl) on in vitro establishment of nodal segments were evaluated. The established explants were transferred to a culture medium that was supplemented with three concentrations of 6-benzylaminopurine (BAP). Three concentrations of indole-3-butyric acid (IBA) to the in vitro adventitious rooting were evaluated. NaOCl application for 10 min resulted in 71.4 % of establishment in 30 d. Supplementation of the culture medium with 2.0 and 3.0 mg L-1 BAP de resulted in the highest averages for multiplication and elongation stages. The formation of adventitious roots occurred with 4.0 mg L-1 IBA of supplementation. Micropropagated plants showed normal morphological features and genetic stability, confirming the cloning of selected plant. Keywords: bamboo; micropropagation; vegetative propagation; In vitro culture; ISSR; plant growth regulator.   Produção de microplantas clonais, avaliação morfofisiológica e estabilidade genética de Dendrocalamus asper (Schult. & Schult.) Backer ex. K. Heyneke   RESUMO: Espécies de bambus apresentam diversas aplicações comerciais, visto que os plantios homogêneos (formados a partir de plantas clonais) são essenciais para a alta produção de biomassa sustentável. A clonagem de plantas selecionadas em escala industrial por meio do cultivo in vitro apresenta muitas vantagens, sendo uma importante ferramenta para o fornecimento de plantas em quantidade e qualidade suficientes para atender a demanda comercial. O controle da clonagem é a base para escala industrial, e seu conhecimento pode otimizar os processos. O trabalho teve como objetivo avaliar a clonagem de planta selecionada de Dendrocalamus asper por meio da técnica de micropropagação. Foram avaliadas as características morfológicas por microscopia eletrônica de varredura e estabilidade genética por meio de marcadores moleculares ISSR. Além disso, foram avaliados quatro tempos de imersão em hipoclorito de sódio (NaOCl) no estabelecimento in vitro de segmentos nodais. Os explantes estabelecidos foram transferidos para um meio de cultura que foi suplementado com três concentrações de benzilaminopurina (BAP). Por fim, foram avaliadas três concentrações de ácido indolbutírico (AIB) durante o enraizamento adventício in vitro. A adição de NaOCl por 10 min resultou em 71,4 % de estabelecimento em 30 d. A suplementação do meio de cultura com 2,0 e 3,0 mg L-1 BAP resultou nas maiores médias para as fases de multiplicação e alongamento. A formação de raízes adventícias ocorreu com a suplementação de 4,0 mg L-1 de AIB. Plantas micropropagadas apresentaram características morfológicas normais e estabilidade genética, confirmando a clonagem da planta selecionada. Palavras-chave: bambu; micropropagação; propagação vegetativa; cultivo in vitro; ISSR; regulador de crescimento vegetal

Sewage sludge and rice husk as potential substrate to produce Mimosa setosa seedlings

In the production of forest seedlings one of the main factors to be analyzed is the formulation of the substrate that meets the needs of the plants that will be produced. For this, the choice of substrate must consider the physical characteristics of the material, its chemical composition, as well as its availability, quality, easy handling and cost. This study aimed to investigate the potential of sewage sludge and rice husk for the production of seedlings of Mimosa setosa. The experiment was conducted in structures of a forest nursery, localized in the municipality of Alegre-ES, in 110 cm³ tubes arranged in a completely randomized design constituted by ten treatments with four replications of 54 seedlings per plot. The treatments were formulated using sewage sludge, raw rice husks and carbonized rice husk. At five months after sowing we measured plant height, root collar diameter, height/diameter ratio, shoot dry weight, root dry weight, total dry matter, shoot to root dry weight ratio and Dickson Quality Index. The substrates formulated with carbonized rice husk promoted the best results for all morphological characteristics analyzed. The multivariate analysis showed higher correlation of treatments with carbonized rice husk (T6, T7, T8 and T9) regarding height, root collar diameter, root dry weight, shoot dry weight and total dry matter. For the production of Mimosa setosa seedlings the best responses resulted from the substrate formulated with 60% sewage sludge + 40% carbonized rice husk

ACTIVATED CHARCOAL APPLICATION FOR THE MICROPROPAGATION OF Cattleya crispata (Thunb.) Van den Berg

 Micropropagation is an alternative for the genetic conservation and propagation of endemic species from “Campo Rupestre Ferruginoso”, such as the orchid Cattleya crispata. The aim of the present study is to assess the influence of activated charcoal on the in vitro germination, multiplication and elongation phases of C. crispata. Seeds extracted from mature capsules were used for inoculation in the culture medium that was adopted to assess the effect of supplementation, or not, with activated charcoal. Data about germination speed, seedling number, length, vigor, oxidation and contamination (bacterial and/or fungal) were assessed through these phases. Based on the results obtained, the use of activated charcoal was efficient in the in vitro germination and multiplication phases of C. crispata, providing greater speed and percentage of germination, less contamination and oxidation of the tissues, greater number, length and vigor of shoots, being effective for the genetic conservation and production of seedlings of the species. Culture medium without the supplementation of activated charcoal provided the best results for the in vitro elongation, with greater length, vigor and less oxidation of shoots.Aplicação de carvão ativado para micropropagação de Cattleya crispata (Thunb.) Van den Berg  RESUMO: A micropropagação é uma alternativa para a conservação genética e propagação de espécies endêmicas do “Campo Rupestre Ferruginoso”, como a orquídea Cattleya crispata. O objetivo do presente estudo é avaliar a influência do carvão ativado nas fases de germinação, multiplicação e alongamento in vitro de C. crispata. Sementes extraídas de cápsulas maduras foram utilizadas para inoculação no meio de cultura adotado para avaliar o efeito da suplementação, ou não, com carvão ativado. Dados sobre velocidade de germinação, número de brotos por explante, comprimento, vigor, oxidação e contaminação (bacteriana e / ou fúngica) foram avaliados por meio dessas fases. Com base nos resultados obtidos, o uso do carvão ativado foi eficiente nas fases de germinação e multiplicação in vitro de C. crispata, proporcionando maior velocidade e porcentagem de germinação, menor contaminação e oxidação dos tecidos, maior número, comprimento e vigor dos brotos, sendo eficaz para a conservação genética e produção de mudas da espécie. O meio de cultura sem a suplementação de carvão ativado proporcionou os melhores resultados para o alongamento in vitro, com maior comprimento, vigor e menor oxidação dos brotos. Palavras-chave: antioxidante; campos rupestres; orquídea; propagação in vitro.   ABSTRACT: Micropropagation is an alternative for the genetic conservation and propagation of endemic species from rupestrian grasslands, such as the orchid Cattleya crispata. The aim of the present study is to assess the influence of activated charcoal on the in vitro germination, multiplication and elongation phases of C. crispata. Seeds extracted from mature capsules were used for inoculation in the culture medium that was adopted to assess the effect of supplementation, or not, with activated charcoal. Data about germination speed, number of shoots per explant, length, vigor, oxidation and contamination (bacterial and/or fungal) were assessed through these phases. Based on the results obtained, the use of activated charcoal was efficient in the in vitro germination and multiplication phases of C. crispata, providing greater speed and percentage of germination, less contamination and oxidation of the tissues, greater number, length and vigor of shoots, being effective for the genetic conservation and production of plants of the species. Culture medium without the supplementation of activated charcoal provided the best results for the in vitro elongation, with greater length, vigor and less oxidation of shoots. Keywords: antioxidant; orchid; propagation in vitro; rupestrian grasslands

ONTOGENETIC AGE AND INOCULATION METHODS FOR THE IN VITRO ESTABLISHMENT OF Eucalyptus pilularis Smith

We aimed to evaluate the in vitro establishment of nodal segments of Eucalyptus pilularis Smith considering two origins of tissues (Or1 - epicormic shoots collected from pruned branches of selected adult trees; Or2 - shoots collected from seminal mini-stumps) and four inoculation methods (Me1 - culture medium supplemented with 0.5 g L-1 activated charcoal; Me2 - culture medium supplemented with 800 mg L-1 PVP30; Me3 - exposure to light for 30 days; Me4 - exposure to dark for 7 days). At 30 days after the in vitro inoculation of tissues, there was no establishment of tissues from epicormic shoots (Or1). Or2 resulted in lower percentages of tissue oxidation and contamination by microorganisms, in addition to having presented establishment and formation of shoots. Me1 resulted in a lower mean tissue oxidation, although it differed statistically only from Me4. An origin of the tissues of ontogenetic age was a determining factor for the successful in vitro establishment of E. pilularis. The use of the Or2 origin and the Me1, Me2, and Me3 methods are recommended to reduce phenolic oxidation of tissues in the in vitro establishment. Keywords: antioxidant agents; exposure to light; epicormic shoots; tissue oxidation.   Idade ontogenética e métodos de inoculação para o estabelecimento in vitro de Eucalyptus pilularis Smith   RESUMO: Objetivou-se avaliar o estabelecimento in vitro de segmentos nodais de Eucalyptus pilularis considerando duas origens de tecidos (Or1 - brotos epicórmicos coletados em galhos podados de matrizes adultas e Or2 - brotos coletados de minicepas seminais) e quatro métodos de inoculação (Me1 - meio de cultura suplementado com 0,5 g L-1 de carvão ativado, Me2 - meio de cultura suplementado com 800 mg L-1 de PVP30, Me3 - exposição à luminosidade por trinta dias e Me4 - exposição em ambiente com ausência de luminosidade por sete dias). Aos 30 dias após a inoculação in vitro dos tecidos, constatou-se que não houve estabelecimento de tecidos oriundos da Or1. A utilização de segmentos nodais provenientes da Or2 resultou em menores percentuais de oxidação e de contaminação por microrganismos, além de ter apresentado estabelecimento e emissão de brotos. O uso do Me1 resultou em menor média de oxidação, embora tenha diferido estatisticamente somente do Me4. A origem dos tecidos associada à idade ontogenética foi um fator determinante para o sucesso do estabelecimento in vitro de E. pilularis. Recomenda-se a utilização da Or2 e dos métodos Me1, Me2 e Me3 a fim de reduzir a oxidação fenólica dos tecidos durante o estabelecimento in vitro. Palavras-chave: agentes antioxidantes; exposição à luminosidade; brotos epicórmicos; oxidação de tecidos.We aimed to evaluate the in vitro establishment of nodal segments of Eucalyptus pilularis Smith considering two origins of tissues (Or1 - epicormic shoots collected from pruned branches of selected adult trees; Or2 - shoots collected from seminal mini-stumps) and four inoculation methods (Me1 - culture medium supplemented with 0.5 g L-1 activated charcoal; Me2 - culture medium supplemented with 800 mg L-1 PVP30; Me3 - exposure to light for 30 days; Me4 - exposure to dark for 7 days). At 30 days after the in vitro inoculation of tissues, there was no establishment of tissues from epicormic shoots (Or1). Or2 resulted in lower percentages of tissue oxidation and contamination by microorganisms, in addition to having presented establishment and formation of shoots. Me1 resulted in a lower mean tissue oxidation, although it differed statistically only from Me4. An origin of the tissues of ontogenetic age was a determining factor for the successful in vitro establishment of E. pilularis. The use of the Or2 origin and the Me1, Me2, and Me3 methods are recommended to reduce phenolic oxidation of tissues in the in vitro establishment. Keywords: antioxidant agents; exposure to light; epicormic shoots; tissue oxidation.   Idade ontogenética e métodos de inoculação para o estabelecimento in vitro de Eucalyptus pilularis Smith   RESUMO: Objetivou-se avaliar o estabelecimento in vitro de segmentos nodais de Eucalyptus pilularis considerando duas origens de tecidos (Or1 - brotos epicórmicos coletados em galhos podados de matrizes adultas e Or2 - brotos coletados de minicepas seminais) e quatro métodos de inoculação (Me1 - meio de cultura suplementado com 0,5 g L-1 de carvão ativado, Me2 - meio de cultura suplementado com 800 mg L-1 de PVP30, Me3 - exposição à luminosidade por trinta dias e Me4 - exposição em ambiente com ausência de luminosidade por sete dias). Aos 30 dias após a inoculação in vitro dos tecidos, constatou-se que não houve estabelecimento de tecidos oriundos da Or1. A utilização de segmentos nodais provenientes da Or2 resultou em menores percentuais de oxidação e de contaminação por microrganismos, além de ter apresentado estabelecimento e emissão de brotos. O uso do Me1 resultou em menor média de oxidação, embora tenha diferido estatisticamente somente do Me4. A origem dos tecidos associada à idade ontogenética foi um fator determinante para o sucesso do estabelecimento in vitro de E. pilularis. Recomenda-se a utilização da Or2 e dos métodos Me1, Me2 e Me3 a fim de reduzir a oxidação fenólica dos tecidos durante o estabelecimento in vitro. Palavras-chave: agentes antioxidantes; exposição à luminosidade; brotos epicórmicos; oxidação de tecidos

Effect of light spectra on in vitro multiplication, elongation and adventitious rooting stages of Bambusa vulgaris Schrad. ex J. C. Wendl.

Bamboos occur throughout much of the temperate and tropical world, have rapid growth, and have various commercial and environmental applications. Clonal production of selected plants on a industrial scale is an important strategy for the bamboo sector. This study aimed to evaluate the effects of the light spectrum on in vitro multiplication, elongation, adventitious rooting, and anatomical features of the leaf surface of Bambusa vulgaris. In the multiplication and elongation stages, in vitro–established explants were transferred to a culture medium supplemented with 8.88 µmol of 6-benzylaminopurine (BAP) and 2.69 µmol of α-naphthalene acetic acid (NAA), and subjected to four light spectra (i.e., white, blue, green, and red). At the adventitious rooting stage, the culture medium was supplemented with 9.84 µmol of indole-3-butyric acid (IBA), 5.37 µmol NAA, and 2.22 µmol BAP under identical light spectra. Explant survival was not influenced by light spectra in the multiplication and elongation stages. White (2.2 shoots) and blue (1.8 shoots) light spectra were the most suitable for the number of shoots per explant. The white spectrum was associated with the highest average length of shoots (7.4 cm) and number of leaves per explant (3.0 leaves). The white light spectrum resulted in the highest average chlorophyll a contents (12.60 µg mg−1), total chlorophyll (16.60 µg mg−1), and carotenoids (10.10 µg mg−1). White and blue light spectra resulted in the best responses for vigor, and least senescence and tissue oxidation. White and blue light spectra favored the chlorophyll b content, resulting in 4.60 and 3.60 µg mg−1, respectively. Survival (80.0 %), adventitious rooting (50.0 %), vigor, senescence, and tissue oxidation were favored in the white light spectrum in the adventitious rooting stage. Scanning electron microscopy of leaves exposed to the white light spectrum revealed microtrichomes and spines on the adaxial surface of the leaf blade, papillae and stomata; on the abaxial surface, there were many unicellular trichomes arranged in rows, denoting normal growth and development. These results may help the production of micropropagated plants of Bambusa vulgaris on an industrial scale

THE INFLUENCE OF LIGHT QUALITY IN THE in vitro CULTIVATION OF Cattleya crispata Van den Berg

ABSTRACT: Micropropagation technique is a valuable alternative for high quality genetic preservation of endemic species such as the orchid Cattleya crispata from “Campo Rupestre Ferruginoso”. This study aims to evaluate the influence of light quality on in vitro multiplication and elongation phases, offering new insights on the limiting factors of C. crispata. Seeds extracted from capsules were used for inoculation in the culture medium. Four light sources were evaluated for in vitro culture, namely: fluorescent lamp, white LEDs, red LEDs and red/blue LEDs. Data about the number of shoots, shoot length, shooting vigor and pigment content were assessed at 90 days of in vitro culture. Based on the recorded results, white LEDs are the most suitable ones for in vitro multiplication and elongation phases of C. crispata. It offers higher quality for seedling production and increases the chances of genetic conservation of the species. Keywords: ‘Campo Rupestre Ferruginoso’; in vitro propagation; wavelength; LEDs.ABSTRACT: Micropropagation technique is a valuable alternative for high quality genetic preservation of endemic species such as the orchid Cattleya crispata from “Campo Rupestre Ferruginoso”. This study aims to evaluate the influence of light quality on in vitro multiplication and elongation phases, offering new insights on the limiting factors of C. crispata. Seeds extracted from capsules were used for inoculation in the culture medium. Four light sources were evaluated for in vitro culture, namely: fluorescent lamp, white LEDs, red LEDs and red/blue LEDs. Data about the number of shoots, shoot length, shooting vigor and pigment content were assessed at 90 days of in vitro culture. Based on the recorded results, white LEDs are the most suitable ones for in vitro multiplication and elongation phases of C. crispata. It offers higher quality for seedling production and increases the chances of genetic conservation of the species.Keywords: ‘Campo Rupestre Ferruginoso’; in vitro propagation; wavelength; LEDs.   A qualidade da luminosidade influencia o cultivo in vitro de Cattleya crispata (Thunb.) Van den Berg   RESUMO: A micropropagação é uma alternativa para a conservação genética de espécies endêmicas do Campo Rupestre Ferruginoso, como a orquídea Cattleya crispata, tendo vantagem a maximização da propagação em elevada qualidade. Para solucionar os fatores limitantes ao cultivo in vitro de C. crispata, o presente trabalho teve como objetivo avaliar a influência da qualidade de luz nas fases de multiplicação e alongamento in vitro. Sementes extraídas de cápsulas foram utilizadas para a inoculação em meio de cultura. Avaliaram-se quatro fontes de luz (Lâmpada fluorescente, LEDs branco, LEDs vermelho e LEDs vermelho/azul). Dados referentes ao número, comprimento de brotos, vigor das brotações e conteúdo de pigmentos foram avaliados aos 90 dias de cultivo. Com base nos resultados obtidos, pode-se inferir que LEDs branco é mais adequada para ser utilizada nas fases de multiplicação e alongamento in vitro de C. crispata, sendo uma alternativa para a produção de mudas e conservação genética. Palavras-chave: Campo Rupestre Ferruginoso; propagação in vitro; comprimento de onda; LEDs