EfficientIn-VitroRegeneration from Mature Leaf Explants ofScoparia dulcisL., an Ethnomedicinal Plant

Abstract

A plant regeneration protocol via multiple-shoot induction using leaf explants from field-grown mature plants of Scoparia dulcis L. (Scrophulariaceae), an ethnomedicinal plant, was established. Explants were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP, 13.2, 17.6 and 22.2 μM) in combination with indole-3-acetic acid (IAA, 0.5 and 2.8 μM) or napthalene-3-acetic acid (NAA, 0.5 and 2.6 μM). The maximum number of shoots (14.0 ± 1.14) with longest shoot length (2.97 ± 0.18) were obtained directly (without an intervening callus phase) from the leaf explants using a combination of BAP (22.2 μM) and IAA (0.5 μM). BAP used in combination with NAA produced fewer shoots per explant as compared with the use of IAA and the regeneration was mediated by callus formation. For root induction, the elongated shoots were separated and transferred onto MS medium supplemented with indole-3-butyric acid (IBA, 4.9 μM). Rooted Aileni et al. 201plantlets (90%) were successfully transferred to soil in the greenhouse with a 75% survival rate.

Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

“…Different from our previous regeneration studies of S. dulcis in which we had used basal medium for elongation of shoots after shoot induction (Aileni et al 2008), in the present report the transformed shoot primordia were transferred onto the medium with lower concentration of BAP for promoting shoot elongation (Fig. 1g).…”

“…Based on our prior report in regenerating 14 ± 1.14 plantlets per mature leaf explant (Aileni et al 2008), in the present investigation we have also used the optimal combination and concentrations of plant growth regulators for regeneration of shoot primordia after cocultivation. Different from our previous regeneration studies of S. dulcis in which we had used basal medium for elongation of shoots after shoot induction (Aileni et al 2008), in the present report the transformed shoot primordia were transferred onto the medium with lower concentration of BAP for promoting shoot elongation (Fig.…”

“…In comparison with the abundant pharmacological data and clinical studies using S. dulcis, information concerning its in vitro plant regeneration and genetic engineering is rather sparse (Aileni et al 2008). So far, there is only one report of production of transgenic herbicide-resistant S. dulcis plants using Agrobacterium rhizogenes (Yamazaki et al 1996), and there are no reports available on Agrobacterium tumefaciens-mediated transformation for this valuable medicinal plant.…”

Cytokinin induced shoot regeneration and flowering of Scoparia dulcis L. (Scrophulariaceae)–an ethnomedicinal herb

“…Although in vitro propagation of S. dulcis with leaf explants [14] and with nodal segment [13] were reported, in this report, the total time schedule was compressed as well as produced 59 shoots from a single explants. About three fold increased in direct shoot formation comparing with the previous report.…”

“…Traditionally the plant is used to treat kidney stone and urinary troubles [6][7][8][9][10][11][12] . Though micropropagation has been performed in S. dulcis [13,14] further studies on factors that may enhance the process are needed. The objective of the present study was to develop an improved protocol for micropropagation of S. dulcis.…”