Subcellular distribution and tolerance of cadmium in Canna indica L.

Canna indica L. is a promising species for heavy metal phytoremediation due to its fast growth rate and large biomass. However, few studies have investigated cadmium (Cd) tolerance mechanisms. In the present study, Canna plants were cultivated under hydroponic conditions with increasing Cd concentrations (0, 5, 10, 15 mg/L). We found that the plants performed well under 5 mg/L Cd2+ stress, but damage was observed under higher Cd exposure, such as leaf chlorosis, growth inhibition, a decreased chlorophyll content, and destruction of the ultrastructure of leaf cells. Additionally, Canna alleviated Cd toxicity to a certain extent. After Canna was exposed to 5, 10 and 15 mg/L Cd2+ for 45 d, the highest Cd concentration was exhibited in roots, which was almost 17-47 times the Cd concentration in leaves and 8-20 times that in stems. At the subcellular level, cellular debris and heat-stable proteins (HSPs) were the main binding sites for Cd, and the proportion of Cd in the two subcellular fractions accounted for 71.4-94.2% of the total Cd. Furthermore, we found that granules could participate in the detoxification process when Cd stress was enhanced. Our results indicated that Canna indica L. can tolerate Cd toxicity by sequestering heavy metals in root tissues, fencing out by cell wall, and binding with biologically detoxified fractions (granules and HSPs).

[Pollen morphology in species of Canna (Cannaceae), and systematics implications]. / Morfología del polen en especies de Canna (Cannaceae) y su implicancia sistemática.

The morphology of pollen grains of eight taxa of Canna, C. ascendens, C. coccinea, C compacta, C. glauca, C. indica, C. paniculata, C. variegatifolia and C. fuchsina, an unpublished new species, were studied using light and scanning electronic microscopes. We used the Wodehouse technique on samples of 20 grains per specimen to measure the intine with a light microscope; and the density of spines (in 400 microm2 fields) with scanning electronic microscopy. Pollen grains are spherical, echinate, omniaperturate. The sporoderm presents a very thin exine covering a thicker intine. Corrugate micro-perforate, sub-reticulate, rugate, rugulate, striate to folded, micro-striate, micro-granulate, and smooth types of the external surface of the sporoderm were found. The spines consist of exine, partially to completely covered by tryphine. The two-layered intine is the thicker part of the wall. Echinate ornamentation is a generic character in Canna, but size, surface and color of pollen walls, and density and shape of spines, are diagnostic for species. Pollen morphology supports the view of C. indica and C. coccinea as different species. Canna fuchsina grows in wild, dense colonies, in humid riverside forests from Buenos Aires and Santa Fe Provinces, Argentina; its characters suggest relationships with a not well known group of taxa, some of them hybrids, such as C. x generalis. However, as these plants showed normal, well formed grains, close to those of C. coccinea, that germinate over the stigmatic surfaces in fresh flowers, we decided to include their pollen in this study.

Characterization of biominerals in species of Canna (Cannaceae).

Plant biominerals are not always well characterized, although this information is important for plant physiology and can be useful for taxonomic purposes. In this work, fresh plant material of seven wild neotropical species of genus Canna, C. ascendens, C. coccinea, C. indica, C. glauca, C. plurituberosa, C. variegatifolia and C. fuchsina sp. ined., taken from different habitats, were studied to characterize the biominerals in their internal tissues. For the first time, samples from primary and secondary veins of leaves were investigated by means of infrared spectroscopy, complemented with X-ray powder diffractometry and scanning electron microscopy. The spectroscopic results, supported by X-ray powder diffractometry, suggest that the calcium oxalate is present in the form of whewellite (CaC2O4 x H2O) in all the investigated samples. It is interesting to emphasize that all IR spectra obtained were strongly similar in all species studied, thus indicating an identical chemical composition in terms of the biominerals found. In this sense, the results suggest that the species of Canna show similar ability to produce biogenic silica and produce an identical type of calcium oxalate within their tissues. These results can be an additional trait to support the relationship among the families of Zingiberales.

Organogenesis and Ultrastructural Features of In Vitro Grown Canna indica L.

An efficient protocol for micropropagation of Canna indica L., an economically and pharmaceutically important plant, was standardized using rhizome explants, excised from two-month-old aseptic seedlings. Complete plant regeneration was induced on MS medium supplemented with 3.0 mg/L BAP plus 1.5 mg/L NAA, which produced the highest number of shoots (73.3 ± 0.5%) and roots (86.7 ± 0.4%) after 2 weeks. Furthermore, the optimum media for multiple shoots regeneration were recorded on MS enriched with 7.0 mg/L BAP (33.0 ± 0.5%). Plantlets obtained were transplanted to pots after two months and acclimatized in the greenhouse, with 75% survival. In addition, ultrastructural studies showed that rhizomes of in vitro grown specimens were underdeveloped compared to the in vivo specimens, possibly due to the presence of wide spaces. Meanwhile, the leaves of in vivo specimens had more open stomata compared to in vitro specimens, yet their paracytic stomata structures were similar. Hence, there were no abnormalities or major differences between in vitro regenerants and mother plants.

Morfología del polen en especies de Canna (Cannaceae) y su implicancia sistemática

Pollen morphology in species of Canna (Cannaceae), and systematics implications. The morphology of pollen grains of eight taxa of Canna, C. ascendens, C. coccinea, C. compacta, C. glauca, C. indica, C. paniculata, C. variegatifolia and C. fuchsina, an unpublished new species, were studied using light and scanning electronic microscopes. We used the Wodehouse technique on samples of 20 grains per specimen to measure the intine with a light microscope; and the density of spines (in 400μm2 fields) with scanning electronic microscopy. Pollen grains are spherical, echinate, omniaperturate. The sporoderm presents a very thin exine covering a thicker intine. Corrugate micro-perforate, sub-reticulate, rugate, rugulate, striate to folded, micro-striate, micro-granulate, and smooth types of the external surface of the sporoderm were found. The spines consist of exine, partially to completely covered by tryphine. The two-layered intine is the thicker part of the wall. Echinate ornamentation is a generic character in Canna, but size, surface and color of pollen walls, and density and shape of spines, are diagnostic for species. Pollen morphology supports the view of C. indica and C. coccinea as different species. Canna fuchsina grows in wild, dense colonies, in humid riverside forests from Buenos Aires and Santa Fe Provinces, Argentina; its characters suggest relationships with a not well known group of taxa, some of them hybrids, such as C. x generalis. However, as these plants showed normal, well formed grains, close to those of C. coccinea, that germinate over the stigmatic surfaces in fresh flowers, we decided to include their pollen in this study. Rev. Biol. Trop. 58 (1): 63-79. Epub 2010 March 01.

Se estudió la morfología de los granos de polen de ocho táxones de Canna, C. ascendens, C. coccinea, C. compacta, C. glauca, C. indica, C. paniculata, C. variegatifolia y C. fuchsina, nueva especie aún no descrita, que fueron estudiadas usando microscopio de luz y microscopio electrónico de barrido. Nosotros utilizamos la técnica de Wodehouse en muestras de 20 granos por espécimen para medir la intina con el microscopio de luz; y la densidad de espinas (en campos de 400μm2) con el microscopio electrσnico de barrido. Los granos de polen son esfιricos, equinados y omniaberturados. El esporodermo presenta una exina muy delgada cubriendo una intina gruesa. La superficie del esporodermo puede ser corrugada, microperforada, sub-reticulada, rugada, rugulada, plegada-estriada, micro-estriada, microgranulada o lisa. Las espinas están formadas por exina, cubiertas total o parcialmente por trifina. La intina esta formada por dos capas, es la parte más gruesa de la pared. La ornamentación equinada es un rasgo genérico en Canna, pero el tamaño, la superficie y el color del polen, y la densidad y forma de las espinas, son rasgos diagnósticos de las especies. La morfología del polen apoya el tratamiento de C. coccinea y C. indica como especies diferentes. Canna fuchsina crece formando densas colonias silvestres en selvas ribereñas húmedas de las provincias de Buenos Aires y Santa Fe, Argentina; sus características sugieren relaciones no muy bien entendidas en el grupo de taxones, algunos son híbridos tales como C. x generalis. Sin embargo, estas plantas muestran granos normales, bien formados, cercanos a los de C. coccinea, que germinan sobre el estigma de flores frescas, nosotros decidimos incluir su polen en este estudio.

Characterization of biominerals in species of Canna (Cannaceae)

Plant biominerals are not always well characterized, although this information is important for plant physiology and can be useful for taxonomic purposes. In this work, fresh plant material of seven wild neotropical species of genus Canna, C. ascendens, C. coccinea, C. indica, C. glauca, C. plurituberosa, C. variegatifolia and C. fuchsina sp. ined., taken from different habitats, were studied to characterize the biominerals in their internal tissues. For the first time, samples from primary and secondary veins of leaves were investigated by means of infrared spectroscopy, complemented with X-ray powder diffractometry and scanning electron microscopy. The spectroscopic results, supported by X-ray powder diffractometry, suggest that the calcium oxalate is present in the form of whewellite (CaC2O4×H2O) in all the investigated samples. It is interesting to emphasize that all IR spectra obtained were strongly similar in all species studied, thus indicating an identical chemical composition in terms of the biominerals found. In this sense, the results suggest that the species of Canna show similar ability to produce biogenic silica and produce an identical type of calcium oxalate within their tissues. These results can be an additional trait to support the relationship among the families of Zingiberales. Rev. Biol. Trop. 58 (4): 1507-1515. Epub 2010 December 01.

Los biominerales de las plantas no siempre han sido bien caracterizados aunque esta información es importante en fisiología vegetal y puede ser de utilidad para fines taxonómicos. En este trabajo se estudió material vegetal fresco de siete especies silvestres neotropicales: Canna, C. ascendens, C. coccinea, C. indica, C. glauca, C. plurituberosa, C. variegatifolia and C. fuchsina sp. ined., provenientes de diferentes localidades, con el fin de caracterizar los biominerales presentes en sus tejidos foliares internos. Por vez primera, muestras de venas primarias (ejes foliares) y secundarias de hojas de estas especies se investigaron por medio de espectroscopia de infarrojo, complementada con estudios por difracción de rayos X de polvos y microscopía electrónica de barrido. Los resultados indicaron la presencia de ópalo (sílice biogénica) y oxalato de calcio en los tejidos vegetales analizados. Además, se determinó que el oxalato de calcio está presente en forma de whewellita (CaC2O4×H2O), información nueva para el género. Tanto el ópalo como la whewellita están presentes en todas las especies analizadas, que representan aproximadamente un tercio de las especies silvestres del género. La capacidad de biomineralizar SiO2 en forma de ópalo en especies de Canna de diversos ambientes resulta también un rasgo altamente sugerente para futuros estudios.