Catasetum fimbriatum (Morren) Lindl. é uma orquídea estudada no Laboratório de Fisiologia Vegetal do IBUSP desde 1990. As plantas deste gênero, quando incubadas in vitro no escuro, apresentam crescimento contínuo do caule na ausência de qualquer regulador de crescimento. Quando as gemas laterais dos caules estiolados são isoladas e incubadas na presença de luz, originam novas plantas, constituindo-se, desta forma, uma técnica relativamente simples e geneticamente segura de clonagem de plantas. Baseando-se no conhecimento consolidado com esta planta, procurou-se por um lado aprofundar conhecimentos básicos a respeito dos processos envolvidos na indução do crescimento caulinar em C. fimbriatum, e por outro, tomando-a como parâmetro de referência, estendê-los às plantas de Cymbidium e Cattleya labiata tratadas com etileno, giberelina (GA) e óxido nítrico (NO). Foram utilizadas plantas micropropagadas de C. fimbriatum e Cymbidium, enquanto as de Cattleya foram obtidas por meio de germinação assimbiótica. Após 120 dias de incubação, as plantas foram transferidas para o escuro e tratadas com diferentes concentrações de GA, paclobutrazol (PA), um inibidor de biossíntese de GA, etileno, 1-metilciclopropeno (1-MCP), um inibidor da ação do etileno, e NO. No 30º e 60º dia de incubação no escuro, foram mensurados o número de nós formados e de gemas laterais liberadas, bem como o tamanho e as massas fresca e seca dos estolões. Análises dos teores de etileno e CO2 acumulados nos frascos foram determinadas por meio de cromatografia gasosa. Os três gêneros estudados apresentaram respostas diferentes quanto ao estiolamento. Nitidamente, sob as condições utilizadas, as plantas de Cattleya labiata mostraram-se recalcitrantes à formação de estolões e à quebra da dominância apical, mantendo inalterado o crescimento foliar ao longo de todo o período de tratamento. Não obstante os caules de C. fimbriatum e Cymbidium tenham estiolado conspicuamente, na primeira planta eles se formaram na base do pseudobulbo, enquanto na segunda houve a retomada da atividade meristemática apical já nos primeiros 30 dias de incubação. Os tratamentos com GA promoveram significativamente o alongamento caulinar em C. fimbriatum, enquanto que nas plantas de Cymbidium a concentração mais elevada inibiu esse processo. Os tratamentos com PA e 1-MCP reduziram o alongamento caulinar em ambos os gêneros. O etileno aumentou a liberação de gemas laterais e a ramificação das mesmas em C. fimbriatum e promoveu incremento no tamanho do caule principal e no número de segmentos nodais em Cymbidium. Os efeitos mais marcantes da aplicação de NO ocorreram após 30 dias de tratamento, refletindo positivamente sobre o alongamento caulinar e o número de segmentos nodais em C. fimbriatum. Nas plantas de Cymbidium, contudo, a presença de NO inibiu esse processo. No que se refere à emissão de etileno e CO2, observou-se uma variação acentuada entre os gêneros e os tratamentos realizados. Quanto ao etileno, a liberação mais elevada deu-se nos tratamentos com etileno nas três plantas estudadas. A liberação deste hormônio foi mais proeminente nas plantas de C. fimbriatum do que nas de Cymbidium, o que, em princípio, poderia ser relacionado à rápida retomada da atividade meristemática nestas últimas e a manutenção da dominância apical, tendo aparentemente o oposto ocorrido com plantas de C. fimbriatum. Embora ambas originem estolões, os meristemas envolvidos comportam-se de forma distintas, podendo ser separados em três grupos. Nos primeiros dois se enquadrariam os gêneros Catasetum e Cymbidium, com inibição do crescimento foliar e formação de estolões a partir apenas da atividade do MAC (Cymbidium), ou a partir da liberação de gemas laterais (Catasetum), enquanto no terceiro grupo se enquadraria o gênero Cattleya, com manutenção do crescimento foliar e não formação de caules estiolados. / Catasetum fimbriatum (Morren) Lindl. is an epiphytic orchid that has been studied since 1990 in our Laboratory. Plants of this genus have the peculiarity of being easily propagated, using the lateral buds of its etiolated shoot. This is possible because the shoot apical meristem shows a sustained activity when kept in darkness, resulting in an expressive shoot elongation. When the etiolated nodal segments are incubated under light, they rapidly form new plants. This micropropagation process dismisses the application of any plant growth regulators, representing an efficient in vitro multiplication method. The aim of this study was to apply the existing knowledge about the physiology of dark-grown C. fimbriatum plants, to other two genera, Cymbidium and Cattleya, intending to characterize the role of ethylene, gibberellin and nitric oxide in the signaling of shoot elongation process of these plants. C. fimbriatum and Cymbidium plants were obtained by micropropagation technique, using etiolated nodal segments, whereas Cattleya plants were obtained by assimbiotic germination. After 3 months, these plants were incubated in the dark under different levels of concentration of gibberellin (GA), ethylene, nitric oxid (NO), paclobutrazol (PA, inhibitor of gibberellin biosynthesis) and 1-methylcyclopropene (1-MCP, inhibitor of ethylene perception),. After 30 and 60 days, the treatments effects were measured through the number of nodal segments of the stolons, shoot size, and wet and dry mass. Ethylene and CO2 contents were determined using gas chromatography. The three studied genera showed different responses to dark incubation and treatments. Cattleya was not able to recover shoot apical meristem (SAM) activity and its phenotype remained unaltered, maintaining foliar growth until the end of the treatment. C. fimbriatum originated stolons from the base of the pseudobulb, forming etiolated shoots. On the other hand, Cymbidium showed a different meristematic activity, giving rise to an elongated stem from the apical end of the pseudobulb within the first 30 days of the experiment. GA promoted a significant increase in stem elongation in C. fimbriatum whereas in Cymbidium plants this parameter was inhibited at the highest level of hormone. PA and 1-MCP reduced stem elongation in both C. fimbriatum and Cymbidium plants. Ethylene treatment increased the development of lateral buds and their branching in C. fimbriatum and increased the size of Cymbidium etiolated stem and the number of node segments. The most conspicuous effects of the application of NO occurred after 30 days of treatment, increasing shoot elongation and number of node segments in C. fimbriatum and reducing these parameters in Cymbidium plants. With regard to ethylene and CO2 emission the different genera presented different responses. C. fimbriatum, Cattleya and Cymbidium showed increasing hormone liberation within the ethylene treatments. Furthermore, C. fimbriatum showed conspicuously higher ethylene and CO2 emission than Cymbidium plants. When incubated in the absence of light, the three orchid genera showed distinct behavior, separated into three groups: (1) etiolated stem formation only from SAM activity, and development of reduced leaves (Cymbidium) (2) etiolated stem formation from lateral buds and development of reduced leaves (C. fimbriatum), (3) absence of etiolated stems, with maintenance of foliar growth (Cattleya).

A multiplicação de orquídeas in vitro vem sendo utilizada há algum tempo com objetivo de elevar a taxa de multiplicação, além de eliminar patógenos e reduzir gastos na produção. Esta ferramenta de trabalho vem sendo rotineiramente utilizada no nosso laboratório, ao longo de mais de duas décadas em nosso laboratório, em pesquisas básicas de fisiologia e de aprimoramento da técnica de clonagem, principalmente de orquídeas. Neste caso,o uso da técnica visa a obtenção de maior estabilidade genética dos regenerantes em cultivos de longa duração. Plantas do gênero Catasetum apresentam atividade indeterminada do meristema apical caulinar quando incubadas no escuro, originando em pouco tempo longos estolões com crescimento indeterminado, comportamento raro no reino vegetal. Cada nó do caule estiolado possui uma gema lateral que, quando isolada e incubada no claro, forma rapidamente uma planta completa, facilitando a micropropagação. Outras espécies de orquídeas valorizadas na floricultura não apresentam tal facilidade na multiplicação, mostrando-se recalcitrantes à micropropagação, como é o caso do gênero Dendrobium (Orchidaceae). O objetivo deste estudo foi obter uma melhor compreensão dos mecanismos fisiológicos envolvidos no estiolamento de plantas Dendrobium \"Second Love\", que apresenta crescimento caulinar limitado quando sob ausência de luz, buscando compreender os efeitos do escuro e dos hormônios etileno e giberelina, bem como do radical livre óxido nítrico e gás carbônico na atividade dos meristemas apicais e laterais dessa orquídea. Como objetivo complementar, buscou-se estimular um estiolamento mais pronunciado, visando com isto um aumento potencial na formação de gemas laterais, paralelamente à quebra da dominância apical e o crescimento subsequente dos estolões. As plantas de Dendrobium utilizadas faziam parte do nosso estoque de germoplasma in vitro. Após 120 dias de incubação no claro, as plantas foram transferidas para o escuro e tratadas com diferentes concentrações de ácido giberélico (GA), paclobutrazol (PA - inibidor de biossíntese de giberelina), etileno, 1-metilciclopropeno (1-MCP - inibidor da ação do etileno) e óxido nítrico (NO). Análises mensais dos teores de etileno e CO2 acumulados nos frascos foram realizadas por meio de cromatografia gasosa durante três meses. Após 30, 60 e 90 dias de tratamento no escuro, quantificou-se o número de gemas laterais presentes nos estolões, o número de gemas laterais e apicais que se desenvolviam, o tamanho dos estolões formados, bem como os respectivos valores de massas fresca e seca destes. Por fim, buscou-se avaliar ainda a importância da incubação na penumbra e no escuro sobre o crescimento caulinar, o número de gemas laterais e o desenvolvimento destas após três meses de cultivo. O crescimento no escuro dos caules das plantas de Dendrobium \"Second Love\" mostrou-se extremamente lento e limitado quando comparado ao das plantas de Catasetum fimbriatum. No entanto, quando tratadas com 1.000 μM de óxido nítrico, verificou-se ao final do terceiro mês que o número de gemas laterais era cinco vezes maior do que nas respectivas plantas controle. O tratamento com 10 ppm de etileno apresentou um aumento significativo no número de gemas e de estolões laterais, quando comparados ao controle a partir do segundo mês de incubação. Quanto ao tamanho do estolão apical, os tratamentos com 5 e 50 μM de GA não apresentaram nenhum efeito promotor sobre alongamento caulinar. Mesmo não apresentando a retomada da atividade meristemática apical, o tratamento com 5 μM de PA liberou um número maior de estolões laterais que o controle. Plantas tratadas com 1.000 ppm de NO, a partir do segundo mês de incubação, apresentaram um número elevado de estolões laterais, além dos mesmos apresentarem-se significativamente maiores. O tratamento com 100 ppm de 1-MCP apresentou o mesmo fenótipo das plantas tratadas no claro, ou seja, não estiolaram mesmo sob a ausência de luz. Quanto à emissão de etileno, observou-se que o tratamento com 1-MCP acarretou um aumento significativo na emissão deste gás pela planta, alcançando valores vinte vezes maiores do que no tratamento controle. Já a emissão de CO2 foi menor no tratamento claro quando comparada a maioria dos outros tratamentos no escuro. Os tratamentos em maiores concentrações de GA e NO pareceram promover algum tipo de estresse na planta (evidenciado pela necrose dos tecidos), demonstrando que a espécie em questão pode ser sensível à níveis elevados destas substâncias / In vitro multiplication has been used for some time in order to improve multiplication rate, eliminate pathogens and reduce the production costs. This working tool has been routinely used in our laboratory for over than two decades of basic research in plant physiology and enhancement of cloning technique, especially orchids, aiming to obtain greater genetic stability of regenerants in long term crops. Genus Catasetum present indeterminate shoot apical meristem activity when incubated in the dark, resulting, in a short period of time, long stolons with indeterminate growth: rare behavior in the plant kingdom. Each etiolated steam node has a lateral bud that, when isolated and incubated in light, quickly forms a complete plant, facilitating micropropagation. Other species of valued orchids in floriculture, such as genus Dendrobium (Orchidaceae), have no such facility in multiplication, being recalcitrant to micropropagation. The goals of this study were to gain a better understanding of the physiological mechanisms involved in plant etiolation in Dendrobium \"Second Love\", which has limited stem growth when in dark: and to understand the effects of dark, gibberellin and ethylene (plant hormones), as well as the free radical nitric oxide and carbon dioxide in the activity of apical and lateral meristems of the orchid. As a complementary objective, we tried to stimulate etiolation, aiming to potentially increase lateral buds formation and to break apical dominance with a subsequent stolons growth. Dendrobium plants used in this work were part of our in vitro germplasm stock. After 120 days of incubation in light, the plants were transferred to dark and treated with different concentrations of gibberellic acid (GA), paclobutrazol (PA - gibberellin biosynthesis inhibitor), ethylene, 1-methylcyclopropene (1-MCP - ethylene action inhibitor) and nitric oxide (NO). During a three months period, monthly analyzes of the accumulated levels of ethylene and CO2 in the flasks were performed using gas chromatography. After 30, 60 and 90 days of dark treatment the number of lateral buds presented in stolons, the number of developed lateral and apical buds, the size of formed stolons, and the respective amounts of fresh and dry mass were quantified. Finally, we evaluated the importance of incubation to steam growth in low light and in the dark, and the number of lateral buds and their development after three months of incubation. The Dendrobium \"Second Love\" steam growth in dark is extremely slow and limited when compared to Catasetum fimbriatum plants. However, after three months of treatment with 1.000 μM nitric oxide it was found to have five times more lateral buds than the respective control treatment plants. The treatment with 10 ppm ethylene showed a significant increase in the number of buds and lateral stolons compared to the control treatment from the second month of incubation. Treatments with 5 and 50 μM of GA had no promoting effect on the apical stolon stem elongation. Although not presenting the resumption of apical meristem activity, 5 μM of PA treatment has released a greater number of lateral stolons than the control treatment. Plants treated with 1000 ppm of NO, from the second month of incubation, showed a higher number of lateral stolons, moreover they were significantly larger. Treatment with 100 ppm 1-MCP had the same phenotype as plants treated in light: in other words, they did not etiolate even in light absence. Regarding the ethylene emission, we observed that the treatment with 1-MCP caused a significant increase in the emission of this gas by the plant, reaching values twenty times higher than the control treatment. CO2 emission was lower in light treatment when compared to most of the other treatments in dark. Treatments at higher concentrations of NO and GA seemed to foster some sort of plant stress (evidenced by tissue necrosis), demonstrating that the specie in question may be sensitive to high levels of these substances

Lowther and Boll in the study of the effect of 2,4,5-TCPAA on etiolated bean leaf discs observed that inhibition of expansion at low concentrations was relieved at high concentrations where growth was equal to that of the light plus Co control. This means that at appropriate concentrations, TCPAA completely replaced Co in the expansion of light-treated leaf discs. [...]

A multiplicação de orquídeas in vitro vem sendo utilizada há algum tempo com objetivo de elevar a taxa de multiplicação, além de eliminar patógenos e reduzir gastos na produção. Esta ferramenta de trabalho vem sendo rotineiramente utilizada no nosso laboratório, ao longo de mais de duas décadas em nosso laboratório, em pesquisas básicas de fisiologia e de aprimoramento da técnica de clonagem, principalmente de orquídeas. Neste caso,o uso da técnica visa a obtenção de maior estabilidade genética dos regenerantes em cultivos de longa duração. Plantas do gênero Catasetum apresentam atividade indeterminada do meristema apical caulinar quando incubadas no escuro, originando em pouco tempo longos estolões com crescimento indeterminado, comportamento raro no reino vegetal. Cada nó do caule estiolado possui uma gema lateral que, quando isolada e incubada no claro, forma rapidamente uma planta completa, facilitando a micropropagação. Outras espécies de orquídeas valorizadas na floricultura não apresentam tal facilidade na multiplicação, mostrando-se recalcitrantes à micropropagação, como é o caso do gênero Dendrobium (Orchidaceae). O objetivo deste estudo foi obter uma melhor compreensão dos mecanismos fisiológicos envolvidos no estiolamento de plantas Dendrobium \"Second Love\", que apresenta crescimento caulinar limitado quando sob ausência de luz, buscando compreender os efeitos do escuro e dos hormônios etileno e giberelina, bem como do radical livre óxido nítrico e gás carbônico na atividade dos meristemas apicais e laterais dessa orquídea. Como objetivo complementar, buscou-se estimular um estiolamento mais pronunciado, visando com isto um aumento potencial na formação de gemas laterais, paralelamente à quebra da dominância apical e o crescimento subsequente dos estolões. As plantas de Dendrobium utilizadas faziam parte do nosso estoque de germoplasma in vitro. Após 120 dias de incubação no claro, as plantas foram transferidas para o escuro e tratadas com diferentes concentrações de ácido giberélico (GA), paclobutrazol (PA - inibidor de biossíntese de giberelina), etileno, 1-metilciclopropeno (1-MCP - inibidor da ação do etileno) e óxido nítrico (NO). Análises mensais dos teores de etileno e CO2 acumulados nos frascos foram realizadas por meio de cromatografia gasosa durante três meses. Após 30, 60 e 90 dias de tratamento no escuro, quantificou-se o número de gemas laterais presentes nos estolões, o número de gemas laterais e apicais que se desenvolviam, o tamanho dos estolões formados, bem como os respectivos valores de massas fresca e seca destes. Por fim, buscou-se avaliar ainda a importância da incubação na penumbra e no escuro sobre o crescimento caulinar, o número de gemas laterais e o desenvolvimento destas após três meses de cultivo. O crescimento no escuro dos caules das plantas de Dendrobium \"Second Love\" mostrou-se extremamente lento e limitado quando comparado ao das plantas de Catasetum fimbriatum. No entanto, quando tratadas com 1.000 μM de óxido nítrico, verificou-se ao final do terceiro mês que o número de gemas laterais era cinco vezes maior do que nas respectivas plantas controle. O tratamento com 10 ppm de etileno apresentou um aumento significativo no número de gemas e de estolões laterais, quando comparados ao controle a partir do segundo mês de incubação. Quanto ao tamanho do estolão apical, os tratamentos com 5 e 50 μM de GA não apresentaram nenhum efeito promotor sobre alongamento caulinar. Mesmo não apresentando a retomada da atividade meristemática apical, o tratamento com 5 μM de PA liberou um número maior de estolões laterais que o controle. Plantas tratadas com 1.000 ppm de NO, a partir do segundo mês de incubação, apresentaram um número elevado de estolões laterais, além dos mesmos apresentarem-se significativamente maiores. O tratamento com 100 ppm de 1-MCP apresentou o mesmo fenótipo das plantas tratadas no claro, ou seja, não estiolaram mesmo sob a ausência de luz. Quanto à emissão de etileno, observou-se que o tratamento com 1-MCP acarretou um aumento significativo na emissão deste gás pela planta, alcançando valores vinte vezes maiores do que no tratamento controle. Já a emissão de CO2 foi menor no tratamento claro quando comparada a maioria dos outros tratamentos no escuro. Os tratamentos em maiores concentrações de GA e NO pareceram promover algum tipo de estresse na planta (evidenciado pela necrose dos tecidos), demonstrando que a espécie em questão pode ser sensível à níveis elevados destas substâncias / In vitro multiplication has been used for some time in order to improve multiplication rate, eliminate pathogens and reduce the production costs. This working tool has been routinely used in our laboratory for over than two decades of basic research in plant physiology and enhancement of cloning technique, especially orchids, aiming to obtain greater genetic stability of regenerants in long term crops. Genus Catasetum present indeterminate shoot apical meristem activity when incubated in the dark, resulting, in a short period of time, long stolons with indeterminate growth: rare behavior in the plant kingdom. Each etiolated steam node has a lateral bud that, when isolated and incubated in light, quickly forms a complete plant, facilitating micropropagation. Other species of valued orchids in floriculture, such as genus Dendrobium (Orchidaceae), have no such facility in multiplication, being recalcitrant to micropropagation. The goals of this study were to gain a better understanding of the physiological mechanisms involved in plant etiolation in Dendrobium \"Second Love\", which has limited stem growth when in dark: and to understand the effects of dark, gibberellin and ethylene (plant hormones), as well as the free radical nitric oxide and carbon dioxide in the activity of apical and lateral meristems of the orchid. As a complementary objective, we tried to stimulate etiolation, aiming to potentially increase lateral buds formation and to break apical dominance with a subsequent stolons growth. Dendrobium plants used in this work were part of our in vitro germplasm stock. After 120 days of incubation in light, the plants were transferred to dark and treated with different concentrations of gibberellic acid (GA), paclobutrazol (PA - gibberellin biosynthesis inhibitor), ethylene, 1-methylcyclopropene (1-MCP - ethylene action inhibitor) and nitric oxide (NO). During a three months period, monthly analyzes of the accumulated levels of ethylene and CO2 in the flasks were performed using gas chromatography. After 30, 60 and 90 days of dark treatment the number of lateral buds presented in stolons, the number of developed lateral and apical buds, the size of formed stolons, and the respective amounts of fresh and dry mass were quantified. Finally, we evaluated the importance of incubation to steam growth in low light and in the dark, and the number of lateral buds and their development after three months of incubation. The Dendrobium \"Second Love\" steam growth in dark is extremely slow and limited when compared to Catasetum fimbriatum plants. However, after three months of treatment with 1.000 μM nitric oxide it was found to have five times more lateral buds than the respective control treatment plants. The treatment with 10 ppm ethylene showed a significant increase in the number of buds and lateral stolons compared to the control treatment from the second month of incubation. Treatments with 5 and 50 μM of GA had no promoting effect on the apical stolon stem elongation. Although not presenting the resumption of apical meristem activity, 5 μM of PA treatment has released a greater number of lateral stolons than the control treatment. Plants treated with 1000 ppm of NO, from the second month of incubation, showed a higher number of lateral stolons, moreover they were significantly larger. Treatment with 100 ppm 1-MCP had the same phenotype as plants treated in light: in other words, they did not etiolate even in light absence. Regarding the ethylene emission, we observed that the treatment with 1-MCP caused a significant increase in the emission of this gas by the plant, reaching values twenty times higher than the control treatment. CO2 emission was lower in light treatment when compared to most of the other treatments in dark. Treatments at higher concentrations of NO and GA seemed to foster some sort of plant stress (evidenced by tissue necrosis), demonstrating that the specie in question may be sensitive to high levels of these substances

The C4 cycle represents a series of biochemical and anatomical modifications that are targeted to overcome the effects of photorespiration caused by the oxygenase capability of Ribulose Bisphosphate Carboxylase/Oxygenase (RuBisCO). The cycle has evolved independently in over 60 lineages, which suggests that recruitment of genes into the C4 cycle is a relatively easy process. However, the mechanisms by which the anatomy and cell-specificity of the components of the C4 cycle is achieved is poorly understood. Preliminary work in maize indicated several components of the C4 cycle may be targeted by microRNAs (miRNAs). To explore this, a library of sRNA sequences from mature leaf tissue of the model C4 species Cleome gynandra L. was generated and then searched against a list of expressed sequence tag sequences for candidate genes of the C4 cycle. To complement this, transgenic C. gynandra containing the viral p19 protein, which is capable of suppressing miRNA activity, were produced. A limited subset of the candidate C4 genes showed a high level of sRNA read alignment. In C. gynandra plants expressing p19 photosynthesis was compromised and transcripts of several genes (most notably RbcS and RCA) were upregulated. These data were complemented by examining the effect of illumination on developing C. gynandra cotyledons, and attempts to generate a hybrid between C. gynandra and the C3 C. hassleriana Chodat. RbcS also showed elevated abundance in etiolated cotyledons, although this rapidly declined after illumination. The remainder of the C4 genes profiled accumulated in etiolated tissue, but were upregulated within 6 hours of illumination. Therefore, this study has illustrated that miRNA activity may play a role in maintaining the C4 photosynthetic cycle at optimum efficiency, although it has not been possible to identify at which point(s) this regulation is applied. Secondly, RbcS appears to be subject to multiple regulatory mechanisms in C. gynandra, and it is possible that miRNAs have a role in negatively regulating expression of RbcS.

Catasetum fimbriatum (Morren) Lindl. é uma orquídea estudada no Laboratório de Fisiologia Vegetal do IBUSP desde 1990. As plantas deste gênero, quando incubadas in vitro no escuro, apresentam crescimento contínuo do caule na ausência de qualquer regulador de crescimento. Quando as gemas laterais dos caules estiolados são isoladas e incubadas na presença de luz, originam novas plantas, constituindo-se, desta forma, uma técnica relativamente simples e geneticamente segura de clonagem de plantas. Baseando-se no conhecimento consolidado com esta planta, procurou-se por um lado aprofundar conhecimentos básicos a respeito dos processos envolvidos na indução do crescimento caulinar em C. fimbriatum, e por outro, tomando-a como parâmetro de referência, estendê-los às plantas de Cymbidium e Cattleya labiata tratadas com etileno, giberelina (GA) e óxido nítrico (NO). Foram utilizadas plantas micropropagadas de C. fimbriatum e Cymbidium, enquanto as de Cattleya foram obtidas por meio de germinação assimbiótica. Após 120 dias de incubação, as plantas foram transferidas para o escuro e tratadas com diferentes concentrações de GA, paclobutrazol (PA), um inibidor de biossíntese de GA, etileno, 1-metilciclopropeno (1-MCP), um inibidor da ação do etileno, e NO. No 30º e 60º dia de incubação no escuro, foram mensurados o número de nós formados e de gemas laterais liberadas, bem como o tamanho e as massas fresca e seca dos estolões. Análises dos teores de etileno e CO2 acumulados nos frascos foram determinadas por meio de cromatografia gasosa. Os três gêneros estudados apresentaram respostas diferentes quanto ao estiolamento. Nitidamente, sob as condições utilizadas, as plantas de Cattleya labiata mostraram-se recalcitrantes à formação de estolões e à quebra da dominância apical, mantendo inalterado o crescimento foliar ao longo de todo o período de tratamento. Não obstante os caules de C. fimbriatum e Cymbidium tenham estiolado conspicuamente, na primeira planta eles se formaram na base do pseudobulbo, enquanto na segunda houve a retomada da atividade meristemática apical já nos primeiros 30 dias de incubação. Os tratamentos com GA promoveram significativamente o alongamento caulinar em C. fimbriatum, enquanto que nas plantas de Cymbidium a concentração mais elevada inibiu esse processo. Os tratamentos com PA e 1-MCP reduziram o alongamento caulinar em ambos os gêneros. O etileno aumentou a liberação de gemas laterais e a ramificação das mesmas em C. fimbriatum e promoveu incremento no tamanho do caule principal e no número de segmentos nodais em Cymbidium. Os efeitos mais marcantes da aplicação de NO ocorreram após 30 dias de tratamento, refletindo positivamente sobre o alongamento caulinar e o número de segmentos nodais em C. fimbriatum. Nas plantas de Cymbidium, contudo, a presença de NO inibiu esse processo. No que se refere à emissão de etileno e CO2, observou-se uma variação acentuada entre os gêneros e os tratamentos realizados. Quanto ao etileno, a liberação mais elevada deu-se nos tratamentos com etileno nas três plantas estudadas. A liberação deste hormônio foi mais proeminente nas plantas de C. fimbriatum do que nas de Cymbidium, o que, em princípio, poderia ser relacionado à rápida retomada da atividade meristemática nestas últimas e a manutenção da dominância apical, tendo aparentemente o oposto ocorrido com plantas de C. fimbriatum. Embora ambas originem estolões, os meristemas envolvidos comportam-se de forma distintas, podendo ser separados em três grupos. Nos primeiros dois se enquadrariam os gêneros Catasetum e Cymbidium, com inibição do crescimento foliar e formação de estolões a partir apenas da atividade do MAC (Cymbidium), ou a partir da liberação de gemas laterais (Catasetum), enquanto no terceiro grupo se enquadraria o gênero Cattleya, com manutenção do crescimento foliar e não formação de caules estiolados. / Catasetum fimbriatum (Morren) Lindl. is an epiphytic orchid that has been studied since 1990 in our Laboratory. Plants of this genus have the peculiarity of being easily propagated, using the lateral buds of its etiolated shoot. This is possible because the shoot apical meristem shows a sustained activity when kept in darkness, resulting in an expressive shoot elongation. When the etiolated nodal segments are incubated under light, they rapidly form new plants. This micropropagation process dismisses the application of any plant growth regulators, representing an efficient in vitro multiplication method. The aim of this study was to apply the existing knowledge about the physiology of dark-grown C. fimbriatum plants, to other two genera, Cymbidium and Cattleya, intending to characterize the role of ethylene, gibberellin and nitric oxide in the signaling of shoot elongation process of these plants. C. fimbriatum and Cymbidium plants were obtained by micropropagation technique, using etiolated nodal segments, whereas Cattleya plants were obtained by assimbiotic germination. After 3 months, these plants were incubated in the dark under different levels of concentration of gibberellin (GA), ethylene, nitric oxid (NO), paclobutrazol (PA, inhibitor of gibberellin biosynthesis) and 1-methylcyclopropene (1-MCP, inhibitor of ethylene perception),. After 30 and 60 days, the treatments effects were measured through the number of nodal segments of the stolons, shoot size, and wet and dry mass. Ethylene and CO2 contents were determined using gas chromatography. The three studied genera showed different responses to dark incubation and treatments. Cattleya was not able to recover shoot apical meristem (SAM) activity and its phenotype remained unaltered, maintaining foliar growth until the end of the treatment. C. fimbriatum originated stolons from the base of the pseudobulb, forming etiolated shoots. On the other hand, Cymbidium showed a different meristematic activity, giving rise to an elongated stem from the apical end of the pseudobulb within the first 30 days of the experiment. GA promoted a significant increase in stem elongation in C. fimbriatum whereas in Cymbidium plants this parameter was inhibited at the highest level of hormone. PA and 1-MCP reduced stem elongation in both C. fimbriatum and Cymbidium plants. Ethylene treatment increased the development of lateral buds and their branching in C. fimbriatum and increased the size of Cymbidium etiolated stem and the number of node segments. The most conspicuous effects of the application of NO occurred after 30 days of treatment, increasing shoot elongation and number of node segments in C. fimbriatum and reducing these parameters in Cymbidium plants. With regard to ethylene and CO2 emission the different genera presented different responses. C. fimbriatum, Cattleya and Cymbidium showed increasing hormone liberation within the ethylene treatments. Furthermore, C. fimbriatum showed conspicuously higher ethylene and CO2 emission than Cymbidium plants. When incubated in the absence of light, the three orchid genera showed distinct behavior, separated into three groups: (1) etiolated stem formation only from SAM activity, and development of reduced leaves (Cymbidium) (2) etiolated stem formation from lateral buds and development of reduced leaves (C. fimbriatum), (3) absence of etiolated stems, with maintenance of foliar growth (Cattleya).

No description available.

Numerous wild bigtooth maple (Acer grandidentatum Nutt.) specimens in northern Utah have potential for use in landscapes, but improvements in selection and propagation need to be developed before these specimens can be introduced to the green industry. Criteria-based evaluations centered on aesthetics, function, and fall color were performed to objectively select superior bigtooth maple specimens. Out of 56 trees initially selected for red fall color, six were selected for propagation based on all three criteria. Five of the six selected trees yielded viable bud take via chip budding. Optimum time for chip budding propagation was determined by four experiments. Coppiced seedling rootstocks were used with the "return budding" of excised buds as scions to parent stock (2006) and grafting buds from wild trees as scions (2007 and 2009). A fourth experiment examined chip budding of wild scions on 2-year-old, containerized, seedling rootstocks. The general time period identified as the optimum time for budding bigtooth maple was July through mid-August. Propagation by cuttings was also explored as an alternative production method among bigtooth maple selections. Softwood cuttings were taken from six selections of wild bigtooth maples grafted on seedling rootstocks growing in a coppiced stool bed environment. Open-ended, black, velour, drawstring bags were placed over the end of pruned shoots at bud swell to initiate etiolation of the cuttings. The bags were left in place during shoot elongation to insure etiolation of the shoot base. Cuttings were harvested after 3 to 4 weeks, wounded, dipped in auxin, and placed on heating mats under an intermittent mist system. Rooting was evaluated on the cuttings after four weeks. Results showed the effects of etiolation to significantly increase the percentage of rooted cuttings and the number of roots per cutting.

Abstract The elongation response of Scots pine (Pinus sylvestris L.) seedlings to the removal of blue light (400–500 nm) was studied in field experiments in northern Finland. The seedlings were grown in orange or transparent plexiglass chambers or in ambient control plots. The orange plexiglass removed the blue wavelengths from sunlight, while the others served as controls. The experiment was conducted at sub-arctic (69°N) and mid-boreal (64°N) latitudes with three- and two-year-old seedlings originating from 67°N latitude. The response to blue light depletion was also investigated at the 69°N latitude in the following plant subjects: one-year-old Scots pine seedlings of northern (67°N) and southern (62°N) provenances, deciduous Betula pubescens ssp. czerepanovii and Betula pubescens f. rubra seedlings and herbaceous Epilobium angustifolium and Glechoma hederacea plants. Additionally, diurnal change in light quality at the 69°N latitude during the summer was measured. The elongation of Scots pine seedlings was increased by the removal of blue wavelengths. The increase was more pronounced at the 69°N latitude, while at the 64°N latitude the response was smaller or absent. This is due to increased amount of scattered growth-inhibiting blue light during the nights at the high latitude. The removal of blue light increased stem elongation in northern origin Scots pine seedlings much more compared to the southern origin seedlings, which suggests that the northern provenance is more sensitive to blue light. Irrespective of that, southern origins also suffer from reduced elongation in the north as they migrate according to climatic change scenarios. However, it is obvious that they grow longer than local origins in the north. Morphological variables and photosynthetic pigments confirm that the increased elongation of Scots pine seedlings under blue light depletion is not a result of etiolation or it is only a marginal factor. Also, it was neither dependent on temperature nor photosynthesis and growth resources. Instead, the increased elongation is probably a photomorphogenic regulation response of metabolism. In addition, shade intolerant Scots pine, Betula seedlings and herbaceous Epilobium angustifolium responded stronger to blue light removal compared to the more shade-tolerant herbaceous Glechoma hederacea.

Plants absorb light for photosynthesis, but not all is used. Excess light energy may lead to photoinhibition of photosynthesis and irreversible photooxidative damage. Plants have evolved mechanisms for energy dissipation under high light stress. One such response may involve production of ELIP. It is of interest to know what signal(s) may be involved in ELIP expression. My hypothesis is that redox status of the chloroplast photosynthetic electron transport Chain (PETC) and/or chlororespiration may induce ELIP expression. Using the Arabidopsis thaliana immutans (im) chlororespiratory mutant, this hypothesis was tested. Etiolated seedlings of this variegated mutant were subjected to various light intensities over 0-24 hr period and ELIP mRNA levels were analyzed. These were compared with the wild type plants treated in the same manner. It was found that mature thylakoids may not be required for ELIP expression, and that both photoreceptor-dependent and independent components may be involved in ELIP expression.