Byrsonima intermedia A Juss. is a species with pharmacological properties from the Brazilian Cerrado that shows difficulties related to sexual propagation. Research on cell viability may provide useful information for...Byrsonima intermedia A Juss. is a species with pharmacological properties from the Brazilian Cerrado that shows difficulties related to sexual propagation. Research on cell viability may provide useful information for the selection of cells with embryogenic potential during the callus culture, Within this context, our research is aimed at establishing the cell viability of calli from Byrsonima intermedia leaf segments. The calli went through three subculture phases, of 60 days each, in MS medium with 0.09 M sucrose, 0.6% agar, pH 5.8 and 4.52 laM 2,4-D. The calli were stored in dark conditions and samples were collected every 10 days from each subculture for viability tests with fluorescein 3,6-diacetate （FDA） and 2,3,5-triphenyltetrazolium chloride （TTC）. The staining methods allowed quantifying cell viability in each subculture. The best results from the FDA tests were obtained at 21, 25 and 29 days for the first, second and third subcultures respectively, with 53,86%, 61.88% and 53.73% viable cells. Regarding the TTC test, the largest absorbance values were obtained at 21, 27 and 28 days for the first, second and third subcultures respectively. Fluorescence and spectrophotometry analyses were efficient for determination of cell viability during callus cultivation period.展开更多
Byrsonima intermedia A. Juss. is a species from the Brazilian Cerrado that produces edible fruits and, in common with other species from the Byrsonima genus, has pharmacological potential. Previous attempts to propaga...Byrsonima intermedia A. Juss. is a species from the Brazilian Cerrado that produces edible fruits and, in common with other species from the Byrsonima genus, has pharmacological potential. Previous attempts to propagate the species through conventional methods showed difficulties. Thus, the purpose of this work was to characterize pro-embryogenic masses of Byrsonima intermedia callus, aiming for their in vitro propagation through somatic embryogenesis. Leaf segments from in vitro germinated seedlings were employed as explants for callus production. The calli were then subcultured and exposed to dyes to fulfill their embryogenic potential. Digitalizations of the cytological preparations were made in order to measure the area that was stained by both Aceto-Carmine and Evans-Blue, using image tool software. Somatic embryos were induced after treatments with l-naphthaleneacetic acid （NAA）. The percentages of double-colored areas （by Aceto-Carmine and Evans-Blue） were calculated and the data were analyzed by using the Skott-Knott test （P ≤ 0.05） and, the embryogenic callus, as well as the formation of somatic embryos were analyzed by using the Krsuskal-Wallis rank test （P ≤0.05）. The results show that double coloration is effective at identifying cells showing embryogenic potential. Early callus subculture phases show a larger percentage ofembryogenic area （83%） Somatic embryos were induced by using high auxin level.展开更多
文摘Byrsonima intermedia A Juss. is a species with pharmacological properties from the Brazilian Cerrado that shows difficulties related to sexual propagation. Research on cell viability may provide useful information for the selection of cells with embryogenic potential during the callus culture, Within this context, our research is aimed at establishing the cell viability of calli from Byrsonima intermedia leaf segments. The calli went through three subculture phases, of 60 days each, in MS medium with 0.09 M sucrose, 0.6% agar, pH 5.8 and 4.52 laM 2,4-D. The calli were stored in dark conditions and samples were collected every 10 days from each subculture for viability tests with fluorescein 3,6-diacetate （FDA） and 2,3,5-triphenyltetrazolium chloride （TTC）. The staining methods allowed quantifying cell viability in each subculture. The best results from the FDA tests were obtained at 21, 25 and 29 days for the first, second and third subcultures respectively, with 53,86%, 61.88% and 53.73% viable cells. Regarding the TTC test, the largest absorbance values were obtained at 21, 27 and 28 days for the first, second and third subcultures respectively. Fluorescence and spectrophotometry analyses were efficient for determination of cell viability during callus cultivation period.
文摘Byrsonima intermedia A. Juss. is a species from the Brazilian Cerrado that produces edible fruits and, in common with other species from the Byrsonima genus, has pharmacological potential. Previous attempts to propagate the species through conventional methods showed difficulties. Thus, the purpose of this work was to characterize pro-embryogenic masses of Byrsonima intermedia callus, aiming for their in vitro propagation through somatic embryogenesis. Leaf segments from in vitro germinated seedlings were employed as explants for callus production. The calli were then subcultured and exposed to dyes to fulfill their embryogenic potential. Digitalizations of the cytological preparations were made in order to measure the area that was stained by both Aceto-Carmine and Evans-Blue, using image tool software. Somatic embryos were induced after treatments with l-naphthaleneacetic acid （NAA）. The percentages of double-colored areas （by Aceto-Carmine and Evans-Blue） were calculated and the data were analyzed by using the Skott-Knott test （P ≤ 0.05） and, the embryogenic callus, as well as the formation of somatic embryos were analyzed by using the Krsuskal-Wallis rank test （P ≤0.05）. The results show that double coloration is effective at identifying cells showing embryogenic potential. Early callus subculture phases show a larger percentage ofembryogenic area （83%） Somatic embryos were induced by using high auxin level.
