Influence of Additives on Enhanced in Vitro Shoot Multiplication of <i>Stevia rebaudiana</i> (Bert.)—An Important Anti Diabetic Medicinal Plant

Abstract

The present study was designed to develop an efficient protocol for micro propagation of S. rebaudiana from nodal explants and study the influence of additives on enhancement of shoot proliferation. A two-step protocol has been standardized in which, first step comprising growth hormones concentration is optimized and it was found that MS medium supplemented with 2.0 mg/l BAP + 0.5 mg/l Kin + 0.1 mg/l NAA turned out to be the best treatment for shoot induction. In the second step, the best treatment for shoot induction was fortified with different growth additives for further shoot proliferation. Among the different types of additives used, casein hydrolysate at 0.05% (w/v) was found to be most effective, resulted with maximum of 15.0 shoots. 90% regeneration frequency and shoot length of 6.0 cm were recorded per explant. Thus, the procedure described is a quick and reliable method which could be applied for efficient large scale propagation, genetic transformation assays and secondary metabolite production of Stevia.

Mass propagation of Plectranthus bourneae Gamble through indirect organogenesis from leaf and internode explants

“…CH is an intricate assortment of amino acids and ammonium salts, which together with the combination of KN (1.0 mg/l) and NAA (0.7 mg/l) supported to increase the shoot multiplication rate in P. bourneae. Similar effect of CH have also been reported in Stevia rebaudiana (Sridhar and Aswath 2014), Digitalis lanata (Fatima et al 2009). The shoots cultured on medium containing AC, failed to develop more number of multiple shoots, because AC is known to absorb high concentrations of growth regulators and may reduce shoot regenerative response (Constantin et al 1977;Weatherhead et al 1978).…”

RETRACTED: Review on Medicinal Plants Propagation: A Comprehensive Study on Role of Natural Organic Extracts in Tissue Culture Medium

“…In Rawolfia serpentina [46] a simple regeneration protocol has been developed using cut ovarian peices cultured on MS medium supplemented with CW at 10% (v/v). In a recent report on Stevia [47] CW supplementation as growth additive highly influenced the shoot bud initiation and multiplication, in which two fold increase in shoot multiplication was achieved compared to without CW supplemented cultures. In vitro regeneration studies on Cyamopsis tetragonolobust [48], it was observed that both 2,4-D and coconut water had a synergistic effect, either of them was not able to produce the same result when used separately.…”

“…Synergistic effect of complex extracts was studied after determining the optimum cytokinin and auxin levels for shoot sprouting to improve the quality of shoots was extensively studied earlier in a number of medicinal plants regeneration. In a recent report on stevia [47] regeneration fortification of CH resulted with good regeneration responses and multiplication of shoots was increased two fold. Similar research findings were reported on Oroxyllum indicum [50] regeneration using CH as growth additive.…”

“…In the present review, reports on malt extract supplementation to the culture medium for regeneration is very limited compare to other additives such as coconut water, casein hydrolysate and yeast extract. Though the usage of malt extract was limited, but in some medicinal crops such as Gymnema sylvestre [61], Anogeissus pendula, A. latifolia [56] and Stevia rebaudiana [47] enhanced regeneration frequency and shoot multiplication were recorded on malt extract supplementation. ME at a concentration of 100 mg/L enhanced the shoot number, prevented yellowing of leaves and reduced the callus formation at the cut end of the axillary node explants.…”

“…Supplementation of yeast extract as growth adjuvant in plant tissue culture medium is a regular practice and it has been well established in many medicinal plants in vitro studies, such as Eryngium foetidum [44], Stevia rebaudiana [47], Gymnema sylvestre [61], Asclepias curassavica [51], Lavandula latifolia [63]. Majority of medicinal plants shows good regeneration response on supplementing YE to the culture medium at different concentrations.…”