In vitro approaches for conservation of Asparagus racemosus Willd.

Abstract

Asparagus racemosus is a commercially important medicinal plant, traditionally used for combating gynecological problems in India. The majority of plants used by the pharmaceutical industry come from wild sources, endangering the natural population of the species. The plants are being overharvested, so this species faces a real danger of becoming vulnerable in its natural habitat. Ex situ conservation using in vitro tools is a possible solution to this problem. Ex situ conservation of plants involving in vitro tools has been initiated through axillary branching using nodal explants. Studies on in vitro storage under slow-growth conditions were carried out to develop an efficient protocol for conservation of A. racemosus germplasm. In vitro shoot cultures generally require a 4-wk subculture onto fresh medium when grown at 25±2°C under a 16-h photoperiod. In this research, the use of mannitol or sorbitol as an osmoticum and reduction of sucrose to 1.5% (w/v) in half-strength MS medium led to maintenance of the cultures for 6 mo at 25±2°C with no subculture. Surviving shoots from the slow-growth cultures could be regenerated with 100% efficiency, indicating that the subculture interval was successfully extended by this method. Temperature and medium modification both had significant effects on the growth of stored shoots, and the two factors showed significant interaction. In experiments designed to test encapsulation as a storage method, micropropagated shoot clusters encapsulated in calcium alginate beads were successfully stored up to 75 d at 25±2°C under a 16-h photoperiod. Stored shoots from both storage methods were subsequently recovered and multiplied on MS medium with 3% sucrose and 1.11 μM benzylaminopurine at 25 ± 2°C. Welldeveloped shoots were rooted and acclimatized successfully.

Enhanced production of berberine in In vitro regenerated cell of Tinospora cordifolia and its analysis through LCMS QToF

“…Number of shoots as well as shoot length decreased as the concentration of BAP decreased/increased beyond 1.0 mg L −1 (Table 1). The multiplication of shoots using BAP has also been reported in T. cordifolia (Raghu et al 2006) as well as in other medicinal plants such as Portulaca oleracea , Asparagus racemosus and Cedrela fissilis (Sharma et al 2011; Thakur et al 2015; Aragão et al 2016) which supports the present results. In contrast to the above results, Faisal and Anis (2003) reported that Kn was optimum for in vitro shooting in Tylophora indica .

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In vitro conservation of Poincianella pyramidalis (Tul.) L.P. Queiroz under minimal growth conditions

“…However, when these osmotic agents were combined with high concentrations of sucrose, the plants after culture for 240 days presented lack of vigor, poor development and irregular growth of the aerial part (Figure 2b) Analogously to the observation in this study, other researchers have observed that the addition of the osmotic agents mannitol or sorbitol together with sucrose was efficient in reducing the growth of other plant species, such as Passiflora gibertii N.E. Brown (Faria et al, 2006), Glycyrrhiza glabra L. (Srivastava et al, 2013) and A. racemosus (Thakur;Tiwari;Jadhav, 2015).…”

“…However, the authors observed a decline in the survival rate and growth of the plants when sorbitol and mannitol were both added to the medium at concentration of 87 mM. Analogously, other authors have reported a phytotoxic effect of these osmotic agents on various plant species, such as A. racemosus Willd (Thakur;Tiwari;Jadhav, 2015) and Piper aduncum L. and Piper hispidinervum C. DC. Scherwinski-Pereira, 2011).…”

“…The slow growth storage technique consists of slowing down or stopping the physiological metabolism of plants, to minimize the in vitro development and increase the in vitro period of the explant, without altering the genetic uniformity (or standard) of micropropagated plants (Kamińska et al, 2016). When reducing the in vitro metabolism of plants, the culture conditions such as temperature, light period and intensity and components of the culture medium (organic and inorganic nutrients, osmotic and growth regulators) can be altered during the incubation period (Singh;Kumar;Singh, 2015;Thakur;Tiwari;Jadhav, 2015;Kaur et al, 2012).…”

Slow-grown in vitro conservation of Heliconia champneiana cv. Splash under different light spectra

“…In effective reduction in the metabolism of plants in vitro, the cultivation conditions such as temperature, light period, and light intensity may be changed, and so may the components of the cultivation medium (organic and inorganic under different light spectra nutrients, osmotic and growth regulators) during the period of incubation (Thakur et al, 2015;Carvalho et al, 2014;Kaur et al, 2012).…”