Abstract: Quercetin and (+)-catechin are metabolites present in Phyllanthus niruri plant, have potential in medicinal uses as anticancer and antioxidant agents. Studies on production of quercetin and (+)-catechin from P. niruri callus culture via in vitro technique were carried out and the results were compared to the intact plant. P. niruri explants were cultured on Murashige and Skoog (MS) solidified media supplemented with several phytohormone combinations for one month. The metabolites were extracted from P. niruri callus and intact plant by using carbon dioxide supercritical fluid extraction (SFE) with ethanol as modifier and solvent extraction techniques. The extracts were analyzed by means of HPLC method. Results showed that P. niruri callus culture was successfully established. The highest content of quercetin (1.72%) was found from P. niruri callus grown in media supplemented with 0.8mg/L kinetin and 0.2mg/L 2,4-dicholophenoxyacetic acid (2,4-D), which was 1.2 fold higher than intact plant. Meanwhile, the highest amounts of (+)-catechin (0.63%) was found from P. niruri callus grown in media with addition of 0.2mg/L 1-naphthalene acetic acid (NAA) and 0.8mg/L 2,4-D. The SFE condition in this study showed better extraction efficiency when higher contents of selected metabolites were found in all SFE extracts compared to the common solvent extracts.
Abstract: Agropyron cristatum L. Gaertn. is a native grass of
semiarid region in Iran which is quit resistant to cool and drought
climate and withstand heavy grazing. This species has close
phylogenetic relationship with Triticum and Hordeum. In this
research, the effect of seven different concentrations of growth
regulator 2,4-D on callus production and somatic embryogenesis of
A. cristatum was investigated on Murashige and Skoog medium. The
results showed that the rate of callus, embryo and neomorph were
highest in 1 mg L-1 2,4-D. Callus production was increased in 1 mg
L-1 2,4-D but dramatically decreased at 5.5 and 9 mg L-1 2,4-D. The
somatic embryos were observed at 1 and 4 mg L-1 2,4-D but matured
embryos and plantlet were only occurred at 1 mg L-1 2,4-D. There
were significant differences between 1 mg L-1 2,4-D and other
treatments for producing globular and torpedo embryos, plantlet,
rooted callus and number of roots (p
Abstract: The project was undertaken to determine the effects of modified tissue culture protocols e.g. age of culture and hormone levels (2,4-D) in generating somaclonal variation. Moreover, the utility of molecular markers (SSR and MSAP) in sorting off types/somaclones were investigated.
Results show that somaclonal variation is in effect due to prolonged subculture and high 2,4-D concentration. The resultant variation was observed to be due to high level of methylation events specifically cytosine methylation either at the internal or external cytosine and was identified by methylation sensitive amplification polymorphism (MSAP).Simple sequence repeats (SSR) on the other hand, was able to associate a marker to a trait of interest.
These therefore, show that molecular markers can be an important tool in sorting out variation/mutants at an early stage.
Abstract: Stevia rebaudiana Bertoni (natural sweetener) belongs
to Asteraceae family and can be used as substitute of artificial
sweeteners for diabetic patients. Conventionally, it is cultivated by
seeds or stem cutting, but seed viability rate is poor. A protocol for
callus induction and multiplication was developed to produce large
no. of calli in short period. Surface sterilized nodal, leaf and root
explants were cultured on Murashige and Skoog (MS) medium with
different concentrations of plant hormone like, IBA, kinetin, NAA,
2,4-D, and NAA in combination with 2,4-D. 100% callusing was
observed from leaf explants cultured on combination of NAA and
2,4-D after three weeks while with 2,4-D, only 10% callusing was
observed. Calli obtained from leaf and root explants were shiny green
while with nodal explants it was hard and brown. The present
findings deal with induction of callusing in Stevia to achieve the
rapid callus multiplication for study of steviol glycosides in callus
culture.