ABSTRACT
RESUMEN La inmovilización de las algas tiene múltiples aplicaciones, tales como la biorremediación del agua y la producción de metabolitos. Una de las variables que se puede determinar en las algas inmovilizadas es la fluorescencia de la clorofila a, debido a que este parámetro está relacionado con la respuesta fisiológica de estos organismos. Por ello, el objetivo de este estudio fue explorar un método para la medición de la fluorescencia de la clorofila a en algas encapsuladas en esferas de alginato de calcio. Con este fin, se cultivaron dos especies de microalgas (Scenedesmus ovalternus LAUN 001 y Parachlorella kessleri LAUN 002) en monocultivos, tanto en condiciones de cultivo libres (10 mL de la preparación algal en 250 mL de Medio Básico de Bold), como encapsuladas (250 esferas de alginato de calcio en 250 mL de Medio Básico de Bold). Se realizaron diferentes protocolos de medición de la fluorescencia de la clorofila a del fotosistema II (PSII) variando a) el tiempo de preadaptación a la oscuridad (10, 15 y 30 min), b) la intensidad de luz generada por fluorómetro no modulado (entre 1000 y 3500 -jmoles m-2 s-1), y c) el tiempo de exposición a la luz actínica (1, 2 y 5 s). Se lograron establecer como condiciones óptimas para la medición de la eficiencia cuántica potencial del fotosistema II (Fv/Fm) en las algas encapsuladas las siguientes: a) 30 min de preadaptación a la oscuridad; b) 3000 -jmoles m-2 s-1, de intensidad lumínica generada desde el fluorómetro; y c) 1 o 2 s de exposición a la luz actínica. Se encontraron los siguientes valores en la Fv/Fm en condiciones no estresantes: 0,760 a 0,764 para S. ovalternus y 0,732 a 0,748 para P. kessleri. Esta metodología permite observar algunos cambios en la actividad fotoquímica relacionados con variaciones de los factores bajo los cuales se encuentran las algas inmovilizadas.
ABSTRACT Immobilization of algae has many applications, such as water bioremediation and production of metabolites. One of the variables that can be determined in the immobilized algae is chlorophyll a fluorescence, because this parameter is related to the physiological response ofthese organisms. Therefore, the objective ofthis study was to explore a method for measuring the chlorophyll a fluorescence in calcium alginate-encapsulated algae. To do this, two species of microalgae (Scenedesmus ovalternus LAUN 001 and Parachlorella kessleri LAUN 002) were grown in monocultures in both free culture conditions (10 mL of algae preparation in 250 mL of Basal Bold Medium) and encapsulated (250 spheres in 250 mL of Basal Bold Medium). Different measurement protocols of chlorophyll a fluorescence of photosystem II (PSII) were performed by varying a) the preadaptation time to darkness (10, 15 and 30 min), b) the light intensity of the non-modulated fluorometer (between 1000 and 3500 Llmoles m-2s-1), and c) the time of exposure to actinic light (1, 2 and 5 s). The optimal conditions for the measurement of the maximum quantum yield ofPSII (Fv/Fm) in encapsulated algae were established as follow: a) 30 min of preadaptation time; b) 3000 Llmoles m-2s-1 of the fluorometer light intensity; and c) 1 to 2 s of exposure to actinic light. The following values in the photochemical activity of algae in non-stressful conditions were found: 0.760 - 0.764 for S. ovalternus, and 0.732 - 0.748 for P. kessleri. This methodology allows to observe some changes in the photochemical activity related with variations in the factors under which are the immobilized algae.
ABSTRACT
The standard physiological parameters for assessing stress physiology, of plants, such as chlorophyll index and photosystem II fluorescence, are essential for measuring reactions of plants to stress conditions. To help standardize the parameters for chlorophyll indices of chlorophylls a, b, and total and the fluorescence of photosystem II (Fv/Fm), which are physiological indicators of stress conditions, 6-mo-old seedlings of Persea americana Mill. cv. 'Duke7' and 'Toro canyon', were evaluated under shade house conditions. For each plant, chlorophyll indices were measured from the second through the fourteenth fully expanded leaves. Fluorescence was measured in the third, fifth, seventh, and ninth fully expanded leaf, and determined in function of both, time and intensity of the exposure light source, and the time for dark pre-acclimation of the leaf; was also compared right and left sides of the leaves. Chlorophyll indices were not different between the left and right sides of the leaves, but were different between varieties with 'Duke 7' having the highest value from the eighth leaf, while leaves tested for 'Toro canyon' did not show a difference. For Fv/Fm, there was an interaction among the three factors in both cultivars. A prolonged exposure time (nine seconds), short time for acclimation to darkness, and low intensity of exposure did not induce maximum fluorescence levels. On both avocado cultivars, exposure to maximum light intensity for 7 seconds after 25 to 30 minutes of acclimation to darkness, was the most favorable combination allowing the measurement of photosynthetic efficiency.
Os parâmetros padronizados para avaliação do estresse fisiológico de plantas, tais como índice de clorofila: fluorescência do fotossistema II, são essenciais para medir as reações de plantas a condições de estresse. Para auxiliar na padronização dos parâmetros para os índices de clorofila de clorofilas a, b e total e a fluorescência do fotossistema II (Fv / Fm), que são indicadores fisiológicos de condições de estresse, mudas de 6 meses de idade, de Persea americana Mill. Cv. 'Duke7' e 'Toro canyon' foram avaliadas sob condições casa de vegetação. Para cada planta, índices de clorofila foram medidos da segunda até a décima quarta folha completamente expandida. A fluorescência foi medida na terceira, quinta, sétima e nona folha totalmente expandida, e determinada em função do tempo e intensidade de exposição à fonte de luz, assim como do tempo para pré-adaptação da folha no escuro; também foram comparados entre o lado direito e esquerdo das folhas. Os índices de clorofila não foram diferentes entre os lados direito e esquerdo das folhas, mas foram entre diferentes variedades com 'Duke 7' tendo o valor mais alto a partir da oitava folha, enquanto que as de 'Toro canyon' não mostraram diferença. Para Fv/Fm, houve interação entre os três fatores em ambas as cultivares. O tempo prolongado de exposição (nove segundos), tempo curto para aclimatação no escuro, e baixa intensidade de exposição não induziu níveis máximos de fluorescência. Em ambas as cultivares de abacate, 100% de exposição à intensidade máxima de luz durante 7 segundos, após 25 a 30 minutos de adaptação no escuro, foi a combinação mais adequada para a medição da eficiência fotossintética.
Subject(s)
Stress, Physiological , Chlorophyll , Persea , FluorescenceABSTRACT
Enrichment planting in naturally recovering secondary forests or in tree plantations is increasingly being used as strategy to restore later-successional, large-seeded tropical forest trees. We seeded two tree species (Otoba novogranatensis and Ruagea glabra) in three agricultural sites in Southern Costa Rica: abandoned pastures, eight to ten year old secondary forests and three year old tree plantations (containing two N-fixing of four total tree species). We measured micrometeorological conditions, soil water content, plant water potential, leaf area, foliar C and N, and photosynthesis to better understand mechanistic responses of seedlings to conditions in the different successional habitats. Micrometeorological conditions, soil water content, and plant water potential were generally similar across habitats. Certain aspects of leaves (such as Specific Leaf Area and foliar N content), and photosynthesis (e.g. quantum yield and electron transport rate) were highest in the plantations, intermediate in the secondary forests, and lowest in abandoned pastures. Enhanced rates of photosynthetic biochemistry (such as Vcmax and Jmax) and Photosystem II efficiency (e.g. thermal energy dissipation) occurred in leaves from the plantations compared to the abandoned pastures, which may be related to higher leaf %N content. Results suggest that foliar N may be of greater importance than soil water content and micrometeorological factors in driving differences in photosynthetic processes across planting habitats. Planting seeds of these two species in plantations containing three year old trees (including two N-fixing species) enhances certain aspects of their photosynthesis and growth, compared to seedlings in abandoned pastures with non-native grasses, and thus can help increase forest recovery on abandoned agricultural lands.
El enriquecimiento de bosques secundarios o plantaciones forestales en proceso de regeneración natural por medio de la siembra de plántulas es una práctica cada vez más utilizada para restaurar bosques tropicales en estado de sucesión tardía. Sembramos dos especies de árboles (Otoba novogranatensis y Ruagea glabra) en pastizales abandonados, bosques secundarios de ocho a diez años de edad y plantaciones forestales de tres años de edad (con dos especies fijadoras de Nitrógeno de un total de cuatro especies) en tres sitios agrícolas en el Sur de Costa Rica. Medimos condiciones micrometeorológicas, contenido de agua del suelo, potencial hídrico de las plantas, área foliar, C y N foliar, y fotosíntesis para entender de una mejor manera las respuestas funcionales de las plántulas ante condiciones de distintos estadíos sucesionales. Las condiciones micrometeorológicas, contenido hídrico del suelo y el potencial hídrico de las plantas fueron mayoritariamente similares entre hábitats. Algunos aspectos de las hojas (como Área Foliar Específica y contenido de N foliar) y fotosíntesis (ej.: rendimiento cuántico y tasa de transporte de electrones) presentaron valores mayores en las plantaciones, intermedios en los bosques secundarios y menores en los pastizales abandonados. Se obtuvo un aumento en las tasas fotosintéticas bioquímicas (como Vcmax, Jmax) y la eficiencia del Fotosistema II (ej.: disipación de energía térmica) en hojas provenientes de las plantaciones comparado a las de los pastizales, posiblemente relacionado a un mayor %N foliar. Los resultados sugieren que el N foliar puede ser más importante que el contenido de agua del suelo y que los factores micrometeorológicos para marcar diferencias en los procesos fotosintéticos entre hábitats. Las plántulas de estas dos especies en las plantaciones con árboles de tres años de edad (incluyendo dos fijadoras de N) incrementaron ciertos aspectos de su fotosíntesis y crecimiento comparado a las plántulas en los pastizales abandonados de especies exóticas, por lo tanto, esta práctica puede ayudar a incrementar la recuperación de los bosques en áreas agrícolas abandonadas.