An improved protocol for in vitro propagation of the medicinal plant Mimosa pudica L.

Abstract

This work aimed to develop a protocol for the in vitro establishment, multiplication, rooting and ex vitro acclimatization of Mimosa pudica L., a species used in folk medicine and with pharmacological activity. Aseptic cultures were established from seeds inoculated in MS medium, without growth regulators, followed by an in vitro stabilization phase in culture medium supplemented with 2.22 μM BAP. The cultures were transferred to MS medium supplemented with different cytokinins, combined or not with an auxin, aiming its large-scale propagation. The culture medium supplemented with 5 µM BAP plus 0.5 µM NAA provided the highest multiplication rate and top quality plantlets. The combination of 0.6 µM TDZ plus 0.05 µM NAA resulted in higher multiplication rates than in response to combination of BAP plus NAA, although the subsequent maintenance of the cultures in a medium without growth regulators has resulted in low regenerative response. In vitro rooting of micro-cuttings was high even in the absence of auxins. Over 90% of plantlets transferred to the greenhouse survived after the acclimatization phase. Acclimatized plants presented normal vegetative and reproductive development. The procedures established in the present study allow a massive production of M. pudica plants for further pharmacological studies.

Revisiting In vitro Micropropagation Protocols of Mimosa pudica for Enhanced Seed Germination, Shoot Multiplication, and Root Initiation

“…Above all, MS basal medium fortified with 17.6 μM KIN (Figure 3A) was selected as the best media for shoot multiplication of M. pudica due to the satisfactory shoot number and shoot length, a moderate amount of leaves with low basal callus formation. Consistent with the previous observation by Bianchetti et al (2017), the highest number of shoots recorded in 5 μM BAP was 5 shoots/explant, which was just marginally higher than those in the current study (4.87 shoots/explant in 35.2 μM KIN). It was also reported that increasing BAP concentration hindered shoot number and elongation, with plantlets reaching an average height of 2.5 cm, which was lower than the shoot length achieved in this study, which was over 3 cm.…”

“…In general, the medium composition is one of the many factors that influence the success of an in vitro micropropagation protocol. The result from the most recent micropropagation study of M. pudica by Bianchetti et al (2017) in the germination part was not reproducible in this present study. To investigate this issue, other germination media components not addressed in the previous study, such as the type and strength of the basal medium and sucrose concentration, were assessed to establish a reproducible in vitro germination media for M. pudica seeds.…”

“…These results reflect those of Bianchetti et al (2017), who found that no significant difference was observed between control and NAA-fortified treatments of M. pudica, indicating that this PGR did not promote rooting regardless of the concentration. Since there has been little research on 2,4-D as a source of auxin in M. pudica rooting, it was examined in this study.…”

An efficient high-frequency direct organogenesis protocol of Mimosa pudica, suitable for mass multiplication and transformation experiments

“…Plant tissue culture is the most popular method of ex-situ conservation strategy, which is user-friendly, low-cost; requires less space, energy and advanced tools. A few attempts have been tried to develop a tissue culture protocol for M. pudica using nodal explants and shoot tips as explants (Gharyal and Maheshwari 1982;Hassan et al 2010;Ramesh et al 2013, Bianchetti et al 2017 and hardened the plantlets as well. All the described protocols are similar to shoot multiplication methods and direct organogenesis protocol for the plant has been not reported so far.…”

In vitro propagation from axillary buds of the endangered tree <i>Dalbergia congestiflora</i> Pittier (Fabaceae)

“…This auxin is widely used to induce rooting in shoots of woody legumes, such as Abrus precatorius (Biswas et al 2007), Bauhinia racemosa (Rajanna et al 2011), Cassia angustifolia (Siddique et al 2015), Dalbergia latifolia (Gulati and Jaiwal 1996;Manikandan et al 2017;Swamy et al 1992), D. retusa (Valverde-Cerdas andAlvarado-Guzmán 2004), D. sissoo (Ali et al 2012;Rehman et al 2012;Singh et al 2002), Pithecellobium dulce (Goyal et al 2012), and Pterocarpus santalinum (Prakash et al 2006). However, in some tree species, such as Alnus glutinosa (San José et al 2012) and Mimosa pudica (Bianchetti et al 2017), optimal in vitro rooting has been shown without the addition of auxins, which also promoted the development of secondary roots and the formation of plants suitable for transplant, acclimatization and greenhouse culture. The rooting of D. congestiflora shoots coincided with this response, suggesting that rooting was related to the auxin concentration of tissues.…”