Résumé
The physical dormancy of seeds has been poorly studied in species from tropical forests, such as the Atlantic Forest. This study aimed to examine the effect of moderate alternating temperatures on breaking the physical dormancy of seeds, the morphoanatomy and histochemistry of seed coats, and to locate the structure/region responsible for water entrance into the seed, after breaking the physical dormancy of seeds of two woody Fabaceae (subfamily Faboideae) species that occur in the Brazilian Atlantic Forest: Sophora tomentosa and Erythrina speciosa. To assess temperature effect, seeds were incubated in several temperature values that occur in the Atlantic Forest. For morphological and histochemical studies, sections of fixed seeds were subjected to different reagents, and were observed using light or epifluorescence microscopy, to analyze the anatomy and histochemistry of the seed coat. Treated and non-treated seeds were also analyzed using a scanning electron microscope (SEM) to observe the morphology of the seed coat. To localize the specific site of water entrance, the seeds were blocked with glue in different regions and also immersed in ink. In the present work a maximum temperature fluctuation of 15ºC was applied during a period of 20 days and these conditions did not increase the germination of S. tomentosa or E. speciosa. These results may indicate that these seeds require larger fluctuation of temperature than the applied or/and longer period of exposition to the temperature fluctuation. Blocking experiments water inlet combined with SEM analysis of the structures of seed coat for both species showed that besides the lens, the hilum and micropyle are involved in water absorption in seeds scarified with hot water. In seeds of E. speciosa the immersion of scarified seeds into an aniline aqueous solution showed that the solution first entered the seed through the hilum. Both species showed seed morphological and anatomical features for seed coats of the subfamily Faboideae. Lignin and callose were found around all palisade layers and the water impermeability and ecological role of these substances are discussed in the work.
Requisitos para romper la latencia en semillas de Sophora tomentosa y Erythrina speciosa (Fabaceae). La latencia física de las semillas ha sido poco estudiada en las especies de los bosques tropicales, como el bosque atlántico. Este estudio tuvo como objetivo examinar el efecto de las temperaturas moderadas alternantes en romper la latencia física de las semillas, la anatomía y la histoquímica de la cubierta de las semillas, y la localización de la estructura o región responsable de la entrada de agua, después de romper la latencia física de las semillas de Sophora tomentosa y Eythrina speciosa, dos especies leñosas de Fabaceae (subfamilia Faboideae) que presentes en el bosque atlántico de Brasil. Para cumplir con el primer objetivo se incubaron las semillas a varias temperaturas que se dan en el bosque atlántico. Para los estudios morfológicos e histoquímicos se fijaron secciones de semillas sometidos a diferentes reactivos y se observaron usando luz o microscopía de epifluorescencia para analizar la anatomía y la histoquímica de la cubierta de la semilla. Semillas tratadas y no tratadas se analizaron también usando un microscopio electrónico de barrido (MDB) o microscopio estereoscópico (ME) para observar la morfología de la cubierta de la semilla. Para localizar el sitio específico de la entrada de agua, las semillas fueron bloqueadas con pegamento en diferentes regiones y también sumergidas en tinta. En el presente trabajo se aplicó una fluctuación de temperatura máxima de 15°C durante un período de 20 días y estas condiciones no aumentó la germinación de S. tomentosa o E. speciosa. Estos resultados pueden indicar que estas semillas requieren mayor fluctuación de la temperatura que la aplicada y/o un período más largo de exposición a la fluctuación de la temperatura. Experimentos de bloqueo de entrada de agua combinada con el análisis de las estructuras de la cubierta de la semilla para ambas especies (SEM) mostró que, a pesar de la lente, el hilio y micropilo están implicados en la absorción de agua en las semillas escarificadas con agua caliente. En las semillas de E. speciosa la inmersión de semillas escarificadas en una solución acuosa de anilina mostró que la solución entró por primera vez a la semilla a través del hilio. Ambas especies mostraron características morfológicas y anatómicas de semillas con cubierta de la subfamilia Faboideae. La lignina y callosa se encontraron alrededor de todas las capas de empalizada y la impermeabilidad al agua y en el trabajo se discute el papel ecológico de estas sustancias.
Sujets)
Erythrina/croissance et développement , Germination/physiologie , Dormance des plantes/physiologie , Graines/croissance et développement , Sophora/croissance et développement , Microscopie électronique à balayageRésumé
The endophytic strain Zong1 isolated from root nodules of the legume Sophora alopecuroides was characterized by conducting physiological and biochemical tests employing gfp-marking, observing their plant growth promoting characteristics (PGPC) and detecting plant growth parameters of inoculation assays under greenhouse conditions. Results showed that strain Zong1 had an effective growth at 28 ºC after placed at 4-60 ºC for 15 min, had a wide range pH tolerance of 6.0-11.0 and salt tolerance up to 5% of NaCl. Zong1 was resistant to the following antibiotics (µg/mL): Phosphonomycin (100), Penicillin (100) and Ampicillin (100). It could grow in the medium supplemented with 1.2 mmol/L Cu, 0.1% (w/v) methylene blue and 0.1-0.2% (w/v) methyl red, respectively. Zong1 is closely related to Pseudomonas chlororaphis based on analysis the sequence of 16S rRNA gene. Its expression of the gfp gene indicated that strain Zong1 may colonize in root or root nodules and verified by microscopic observation. Furthermore, co-inoculation with Zong1 and SQ1 (Mesorhizobium sp.) showed significant effects compared to single inoculation for the following PGPC parameters: siderophore production, phosphate solubilization, organic acid production, IAA production and antifungal activity in vitro. These results suggest strains P. chlororaphi Zong1 and Mesorhizobium sp. SQ1 have better synergistic or addictive effect. It was noteworthy that each growth index of co-inoculated Zong1+SQ1 in growth assays under greenhouse conditions is higher than those of single inoculation, and showed a significant difference (p < 0.05) when compared to a negative control. Therefore, as an endophyte P. chlororaphis Zong1 may play important roles as a potential plantgrowth promoting agent.