Triterpene Production in Centella Asiatica (L.) Urban (Pegaga) Callus and Cell Suspension Cultures

Centella asiatica or locally known as ‘Pegaga’ is one of the most common medicinal plants used by diverse ancient cultures and tribal groups. Its medicinal values are mainly attributed to the presence of the triterpene constituents. As there is still no information available on the triterpene production in cultured tissues, studies were carried out in determining the triterpene distribution particularly asiatic acid, madecassic acid, asiaticoside and madecassoside in intact plants of the twelve accessions of C. asiatica collected throughout Malaysia as well as in the callus and cell suspension cultures. Results obtained from the studies revealed that twelve accessions of C. asiatica differed both in their morphologies and their triterpene contents. The triterpenes constituents were detected at a range of 0.134 to 1.655 mg/g dry weight in the whole plant intact tissues. Triterpenes were also successfully detected in the callus (0.014 to 0.773 mg/g dry weight) and cell suspension cultures (0.005 to 0.084 mg/g dry weight), the amount that were lower than that produced in the intact tissues. However, manipulating the physical culture conditions, feeding of precursor, elicitation as well as amino acid addition managed to increase the triterpenes content in cultured tissues. Studies on the effects of the medium composition show that full strength of the basal Murashige and Skoog medium supplemented with B5 vitamins and sucrose (3-4%) increased the triterpenes content in both callus and cell suspension cultures. An interaction of auxincytokinin has observed being important for both callus and cell suspension cultures in enhancing triterpenes production. Higher triterpenes content was obtained in callus treated with 2,4-D and kinetin while the combination of kinetin and dicamba enhanced the triterpenes production in cell suspension cultures. The precursor-feeding studies revealed that lower concentrations of squalene (0.16 mg/L in callus and 0.8 mg/L in cells) were preferred for triterpenes production. Squalene at 0.16 mg/L had successfully triggered the production of madecassoside, asiaticoside and madecassic acid in callus cultures while asiatic acid and madecassic acid content was increased in cells treated with 0.8 mg/L squalene. The elicitor studies exhibited that the different elicitors showed distinctive effects on triterpenes production. Nevertheless, supplementation of succinic acid at 3 and 4 mg/L was found the best in increasing the triterpenes production in callus and cell suspension cultures, respectively. Addition of amino acid into the culture media was also found to promote the triterpenes production in in vitro cultures. The study further concluded that the combinations of the optimized factors namely medium composition, precursor feeding, elicitation and amino acid addition is a very useful strategy in enhancing the triterpenes production particularly the asiatic acid and madecassic acid in in vitro cultures of C. asiatica

The modified Gompertz model demonstrates a variable growth rate between two Centella asiatica phenotypes

Centella asiatica, a weakly aromatic plant that flourishes in wet tropical and sub-tropical areas as a medicinal species since ancient times. It contained important terpenoids that impart important medicinal values. Currently, studies on the terpenoid content of various Centella asiatica phenotypes have shown not only variable content but variable growth rates of different phenotypes that can affect future selection of phenotypes. The use of mathematical growth modelling can reveal important growth constants and discriminate between faster and slower growth phenotypes. Two Centella asiatica phenotypes from South Africa is modelled using the modified Gompertz model and the results showed that the C. asiatica Type-1 exhibited a faster growth rates and a shorter lag period at 0.152 day-1 and 2.313 day than another phenotype; C. asiatica Type 2 with a growth rate and a lag period of 0.067 day-1 and 3.363 day, respectively. The data indicates that different phenotypes of C. asiatica can have different growth rates and lag period and this can be important for selection of phenotypes to be used as the best bioactive peptides producer

Modelling the growth kinetics of callus cultures from the seedling of Jatropha curcas L. according to the modified Gompertz model

One of the most important preliminary investigation of callus attributes is the growth characteristics. Most often than not, callus growth curve is sigmoidal in characteristics. In this work, we model callus growth from the seedling of Jatropha curcas L. according to the modified Gompertz model from published literature to acquire essential growth constants. These growth constants can be obtained with better precision using model such as the modified Gompertz. Parameters obtained from the fitting exercise were maximum callus growth rate (μm), lag time (l) and maximal callus production (Ymax) of 0.193 d-1, 2.91 days and 0.38 g callus/25 mL culture, respectively. Growth parameter constants extracted from the modeling exercise will be helpful for additional secondary modelling implicating the consequence of media conditions as well as other factors on the growth of callus from this plant

Modelling the effect of picloram on the growth kinetics of cell suspension cultures of Ficus deltoidea L.

Ficus deltoidea (or commonly known as mistletoe fig) in various parts of the world mainly serves as an ornamental shrub or houseplant and found native mainly in Asia tropical region, for example, Malaysia, Indonesia, Philippines and Thailand. Studies on the effect of plant growth regulators on cells production from this plant is important as optimization of cells production can lead to efficient production of secondary products characterization and production. An important aspect of the sigmoidal cells growth curve is the growth parameters. In this work, we model the effect of picloram (4-amino-3,5,6-trichloropicolinic acid) on the growth kinetics of the cell suspension cultures of Ficus deltoidea according to the modified Gompertz model. The adjusted coefficient of determination showed good agreement between experimental and predicted data with values ranging from 0.97-0.99. Parameters obtained from the fitting exercise were maximum cells growth rate (µm), lag time (λ) and maximal cells production (Ymax). The results showed that picloram at concentrations of 3 mg/L and above were optimal for giving the highest cells growth rate measured as PCV (packed cell volume). The parameter growth rate obtained from the modelling exercise will be helpful for additional secondary modelling implicating the consequence of media conditions as well as other factors on the effect of picloram on the growth rate of cell suspension from this plant

Induction and maintenance of callus from leaf explants of Mirabilis jalapa

Mirabilis jalapa L., commonly known as ‘four o’clock plant’ produces a strong, sweet smelling fragrance after the flowers open at late afternoon. It is a well known ornamental plant as the flowers of different colours can be found simultaneously on the same plant or an individual flower can be splashed with different colours. The colour-changing phenomenon is one of the unique characteristics of M. jalapa as it can display flowers with different colour when it matures. Apart from its ornamental value, it has also earned its place in herbal medicine practices around the world. Its array of biological activities continues to support its use worldwide for control of viruses, fungi and yeast. In this study, callus culture was initiated from the leaf explants of M. jalapa. The suitable strength of MS (Murashige and Skoog) nutrient media was determined and the effects of different types of auxins [4-amino-3,5,6-trichloro picolinic acid (picloram), 2,4-dichlorophenoxyacetic acid (2,4-D), -naphthaleneacetic acid (NAA)] and cytokinin [6-benzyl amino purine (BAP)] at 0.0 2.5, 5.0, 7.5, 10.0, and 20.0 M were investigated in order to determine the suitable callus induction and maintenance media. The establishment of callus culture was greatly influenced by the strength of MS medium, type as well as the concentration of plant growth regulators (PGRs) used. The best callus induction response was obtained on half strength (½) MS media supplemented with 20.0 M picloram which produced healthy and friable callus. Meanwhile, ½ MS supplemented with NAA or BAP as well as PGR-free medium did not produce any callus; rather, explants became necrotic after 3 to 4 weeks of culture. Calli were successfully maintained in ½ MS supplemented with 10.0 M picloram. Callus in maintenance medium showed a sigmoid growth pattern and reached a maximum growth rate between weeks 1 and 3

Multiple shoots formation of an important tropical medicinal plant, Eurycoma longifolia Jack

Eurycoma longifolia Jack is well known among the communities in Southeast Asia because of its aphrodisiac properties and its effectiveness as the cytotoxic, anti-malarial, anti-ulcer, anti-tumor promoting and anti-parasitic agent. Micropropagation through direct plant regeneration from in vivo shoot tip explants was carried out. The highest regeneration percentage (90%) and multiple shoots formation were obtained with the basal Murashige and Skoog (MS) medium supplemented with 5.0 mg l-1 kinetin. Roots were induced after 14 days of culture in the basal MS medium supplemented with 0.5 mg l -1 of indole-3-butyric acid. Plantlets regenerated from shoot tip explants survived well with no morphological differences from parent plants after two months of transplantation to soil. Copyright © 2005 The Japanese Society for Plant Cell and Molecular Biology

A Review on Medicinal Properties of Orientin

Medicinal plants continue to play an important role in modern medications and healthcare as consumers generally believe that most of them cause fewer or milder adverse effects than the conventional modern medicines. In order to use the plants as a source of medicinal agents, the bioactive compounds are usually extracted from plants. Therefore, the extraction of bioactive compounds from medicinal plants is a crucial step in producing plant-derived drugs. One of the bioactive compounds isolable from medicinal plants, orientin, is often used in various bioactivity studies due to its extensive beneficial properties. The extraction of orientin in different medicinal plants and its medicinal properties, which include antioxidant, antiaging, antiviral, antibacterial, anti-inflammation, vasodilatation and cardioprotective, radiation protective, neuroprotective, antidepressant-like, antiadipogenesis, and antinociceptive effects, are discussed in detail in this review

Adventitious roots induction of recalcitrant tropical woody plant, Eurycoma longifolia

Abstract Eurycoma longifolia is well known for its aphrodisiac and energy enhancing properties especially among the communities in Southeast Asi

Physicochemical characteristics and nutritional compositions of MR219 mutant rice and their effects on glycaemic responses in BALB/c mice

The awareness of the general public on healthy foods has been a major concern and people are looking for the right variety of rice for diabetic patients. High amylose content rice with low glycaemic index (GI), which is an indicator of sugar release in the blood, is beneficial for human health. The present work was aimed to determine the physicochemical characteristics and nutritional compositions of MR219 mutant rice, and the effects of amylose content to blood glucose response and glycaemic index in field condition. A total of 31 M4 mutant lines (ML1 to ML31) were evaluated for physicochemical characteristics and nutritional compositions in comparison with the parental variety, MR219. In glycaemic response study, 48 female BALB/c mice were fed with glucose (a baseline), saline water, two check varieties (MR219 and MRQ74) and four selected mutant lines with different amylose contents. The physicochemical and proximate analysis revealed highly significant differences among the mutant lines. Some mutant lines improved amylose content and nutritional composition. Mutant ML3 had slightly higher amylose content than the parental variety and was recommended for glycaemic responses. However, the field experiment results showed two mutant lines namely; ML3 and ML30, having significantly lower glucose reading (5.49 mmol/L and 5.47 mmol/L, respectively) as compared to the parental variety and other mutant lines. The glucose level was found highest at 60 min after feeding but significantly dropped at 120 min. The normal glucose reading in ML3 and ML30 also resulted in moderate GI values (65% and 66%, respectively). As low and moderate GI foods are recommended for diabetic patients, ML3 and ML30 had high potential for their consumption, and can be suggested for further breeding program to develop low GI rice