A novel cost-effective medium for in vitro cultivation of Gentiana kurroo Royle using sugarcane bagasse

Objective: To develop a simple, cost-effective, and efficient medium by using sugarcane bagasse (SB) as abase material to replace the conventional Murashige and Skoog (MS) medium.Materials and Methods: Water extracts of SB along with some macronutrients and plant growth regulators weregelled with 0.7% agar-agar powder. Nodal segments of Gentiana kurroo were used as explants and inoculatedin the medium and placed in a growth chamber under standard conditions of light and temperature. Out of thetested combinations of plant growth regulators, 0.5 mg/l each of kinetin (KN) and 6-Benzylaminopurine (BAP)showed the excellent shoot multiplication and proliferation rate on the bagasse medium with the same potentialas on the MS medium with an average of 5–6 shoots/explant. In vitro rooting was obtained on half strength MSmedium supplemented with IBA (0.5 mg/l) with an average length of 7–8 cm and 20–25 roots/explant. Theplants were hardened in a mixture of clay loam and farmyard manure in 1:1(w/w) with 70%–80% survival ratewithout any phenotypic aberrations.Conclusion: The results from the present investigation indicate that SB can be used as a cost-effective substituteof MS medium for in vitro propagation of G. kurroo.

Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L

Objective: To induce callus from the medicinally valuable species, Barringtonia racemosa L. (B. racemosa) whereby the formation of callus is essential for micropropagation studies and in vitro plant secondary metabolites production. Methods: The callus induction potential in B. racemosa was assessed from endosperm explant cultured on different culture media and plant hormonal treatments. Lloyd and McCown's woody plant medium and Murashige and Skoog's medium were used in the study as culture media. On the other hand, various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (1.0-2.0 mg/L) and kinetin (0.5-2.5 mg/L) had been incorporated in the culture media to exert the effects of auxin and cytokinin on callus induction. Results: From the present study, it was found that the profuse [(1.681 ± 0.770) g fresh weight, (0.239 ± 0.239) g dry weight] and friable callus formation was optimally produced with desirable morphology and considerable percentage of callus induction (56.70%) in endosperm explants cultured on 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.5 mg/L kinetin in Murashige and Skoog's medium. Conclusions: A reliable protocol for inducing callus formation of profuse and friable morphology in endosperm explant of B. racemosa had therefore been successfully established.

Studies on Influence of Growth Regulators in Micropropagation of Lavandula Angustifolia.

Lavandula angustifolia (Family Labiates) is a medicinal herb found in Mediterranean area. It is a well known herb in ayurvedic system of medicines and has traditionally been used to treat disorder of liver, fever and several conditions including infertility, infection and anxiety. There are few reports on tissue culture of Lavandula angustifolia that too mainly on micropropagation. Present study explored an in vitro micropropagation of Lavandula angustifolia. In vitro callus formation was established by using nodal segments on Murashique and Skoog, (1962) medium (MS) supplemented with IAA at 0.1mg/l, l BAP at 0.002mg/l and 2-4D at 0.2mg/l, significantly recorded complete callus formation after 6 weeks of incubation at 25±1ºC. The callus was allowed for organogenesis and then shoot multiplication was carried out at 4 concentrations of BAP (0.5, 1, 2 and 1mg/L) and IAA (0.5, 0.5, 0.5 and 1mg/L) on MS medium. The shoot regeneration medium for shoot multiplication and proliferation with higher number of shoots was recorded at 0.5mg/l of IAA and 2.0mg/l BAP. However, the growth was very steady and originating from the base. MS medium without any growth regulators tabulated the tallest shoot length of 35 mm, but the shoots were clustered not properly differentiated.

In Vitro Propagation of Chamomilla Recutita From Capitulum Inflorescence: A Medicinal Plant With Multiple Therapeutic Applications

A protocol has been developed for induction of somatic embryogenesis from whole inflorescence explants of Chamomilla recutita L. (chamomile). Chamomile is a well-known medicinal plant from the Asteraceae family often referred to as the “star among medicinal species.” Nowadays, it is a highly favoured medicinal plant in folk and traditional medicine. Its multitherapeutic, cosmetic and nutritional values have been established through the years of traditional and scientific use and research. Chamomile has an established domestic (Indian) and international market, which is increasing day by day. Among the various major constituents, α-bisabolol and chamazulene have been reported to be more useful than others. Chamazulene occurs in the capitula of the flowers in minute quantities and has been demonstrated to exert antiinflammatory activity in-vivo. Moreover, chamomile is a seasonal 4-5 months winter crop in India but is extensively required in various medicinal applications. Therefore, to increase the overall yield of this plant, its in-vitro propagation is needed. In the present study, somatic embryos were developed from capitulum explants after 2-4 weeks of culture on MS medium supplemented with 26.8 µM NAA and 11.5 µM Kin. The somatic embryos were further subcultured in-vitro, where new plantlets regenerated from embryos. It is concluded that in-vitro propagation is possible in case of chamomile and can be used to increase the overall yield of chamazulene present in the capitula of flowers as well as augment the overall yield of this important plant, which is conventionally propagated by seeds.

Optimization of Nutritive Factors in Culture Media for Growth and Tropane Alkaloid Production from Anisodus acutangulus Hairy Roots.

Anisodus acutangulus hairy roots were grown in N6 medium which was optimal for growth and alkaloid production. The cell biomass and alkaloid yield reached 2.7 g l-1 and 3.9 mg l-1 (dry weight) respectively in sucrose medium, higher than those obtained in other carbon source media. 90 mM nitrogen (NH4 +/NO3 – = 4:1) gave the highest cell yield (4.5 g l-1) and the maximum alkaloid production (9.9 mg l-1). The cell yield (4.1 g l-1) of hairy roots grown at pH 6.5 was 2 times higher than that at pH 4.5. However, the maximum alkaloid production (7.2 mg l-1) was yielded at pH 4.5.

Micropropagation and Phenolic exudation protocol for Excoecaria agallocha-an important mangrove

Objective: To develop a standard micropropagation protocol for an important vulnerable mangrove Excoecaria agallocha. Methods: Collection of explants, surface sterilization, phenolic exudation and medium was standardized. Shoot induction, shoot multiplication and rooting were carried out in MMS medium supplemented with BAP, Kinetin, Zeatin, 2ip, NAA, IAA and IBA. Hardening was carried out after root well established. Results: The best phenolic exudation removal was resulted in 4 g/L activated charcoal. The maximum shoot induction response showed in MMS medium and better shoot induction was performed in the concentration of BAP (3.9 μmol) and NAA (1.34 μmol). Rooting induction was performed high range at 5.02 μmol of IAA. Well rooted micro-shoots were hardened and acclimatized. Conclusions: From the present investigation, it can be concluded that a standard micropropagation protocol was developed for an important vulnerable mangrove species.

Micro propagation and tissue culture of the endangered medicinal plant withania somnifera by the direct shoot and root initiation method.

Leaf and Cotyledon explants of Withania somnifera (L). Dunal were used to evaluate the effect of different growth regulators on the in vitro direct shoot and root initiation methods. Four different explants were used to establish callus shoot and root direct regeneration. In the first experiment leaf segments were cultured on MS basal supplemented with 2,4 – Dichlorophenoxyacetic acid (2,4 – D, 0.1-20.0 mg/L), with combination of Naphthalene acetic acid (NAA 0.1-20 mg/L) and Benzylaminopurin (0.1-20 mg/L). This new protocol was standardized for easy mass propagation of W. somnifera medicinal plant. Callus initiation was observed best in MS media with (2,4- D 1.0-5.0 mg/L) after 16-20 days (93%). Highest maximum number of multiple shoots was obtained on MS medium (BAP 3.0 – 5.0 mg/L). The shoots were seaperated from the multiple-shoots, transferred to MS medium supplemented with 1.5 – 20 mg/L NAA favored roots formation occurred in most of the shoot let 88% were successfully achieved in the MS media. The rooted plantlets were transferred to polythene bags which was containing vermi compost, sand and red soil in the ratio of 1:2:2 and kept in a mist house. After acclimatization in the mist house for 2-months, it transferred to greenhouse. The plantlets were successfully planted in the field.

Effect of picloram, additives and plant growth regulators on somatic embryogenesis of Phyla nodiflora (L.) Greene

The present study describes the plant regeneration via somatic embryogenesis in suspension culture derived from the leaf and stem explants of Phyla nodiflora. The medium type, plant growth regulators, complex extract (coconut milk and malt extract) and anti-oxidant (activated charcoal, ascorbic acid, Polyvinylpyrrolidone and citric acid) markedly influenced the embryo regeneration of P. nodiflora. MS with 2,4-D and activated charcoal (10 mg/L) gave the highest stimulation of embryogenic callus growth. Optimized callus was transfered into suspension culture, which showed the globular, heart shaped embryos in MS with 2,4-D + BA + picloram (0.1 mg/L), coconut milk (10 ml/L), citric acid (100 mg/L) on 6th subcultures. Further development stages such as torpedo and cotyledonary stage embryos and fostered maturation of embryos were observed at 8th and 10th subculture. However, the high frequency embryo germination and plantlet (45 plants/20 mg cotyledonary stages embryos) formation was obtained in half-strength MS medium without growth regulators from cotyledonary embryos. All the plantlets established in the field exhibited morphological characters similar to those of the mother plant.