Research Article
Micropropagation, Micromorphological Studies, and In Vitro
Flowering in Rungia pectinata L.
Mahipal S. Shekhawat, M. Manokari, and C. P. Ravindran
Biotechnology Laboratory, Department of Plant Science, MGGAC, Mahe, Pondicherry 673 311, India
Correspondence should be addressed to Mahipal S. Shekhawat; smahipal3@gmail.com
Received 18 December 2015; Accepted 14 April 2016
Academic Editor: Marie-Aleth Lacaille-Dubois
Copyright © 2016 Mahipal S. Shekhawat et al. Tis is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly
cited.
A tissue culture protocol was developed for an important medicinal plant Rungia pectinata L. in the present study. Nodal shoots
were used as explants and surface-sterilized with 0.1% HgCl
2
solution. Murashige and Skoog (MS) medium was used to establish the
cultures of R. pectinata. Te bud break was reported on MS medium supplemented with 1.0 mg L
−1
6-benzylaminopurine (BAP).
About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3cm length were
recorded. Te shoots were further multiplied using MS medium augmented with 0.5 mg L
−1
each of BAP and kinetin (Kin) +
0.1 mg L
−1
indole-3 acetic acid (IAA). Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted
(4.9 roots per shoot with 3.5 cm length) on half strength MS medium fortifed with 2.0 mg L
−1
indole-3 butyric acid (IBA). In vitro
fowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations
of growth regulators used for shoot multiplication under 12/12hr light/dark photoperiod. Te plantlets were hardened in the
greenhouse for two months and fnally transferred to the feld. Te foliar micromorphological studies revealed the developmental
changes in stomata, vein density, and trichomes during the culture of shoots under in vitro conditions.
1. Introduction
Rungia pectinata L. is an ethnomedicinal plant which belongs
to the family Acanthaceae. R. parvifora var. pectinata, R.
parvifora var. muralis, and Justicia pectinata are the syn-
onyms of this plant. It is an annual erect herb with profuse
branching and distributed throughout the warmer parts of
India [1]. Te herb possesses cylindrical fragile stems which
bloom during the months of November-December [2].
Phytosterols, terpenes, tannins, glycosides, favonoids,
phenolic compounds, amino acids, fxed oils, and so forth
are important phytochemicals isolated from this plant [3].
Te fowers are reported to contain isosalipurposide, luteolin,
glucoside, lutein, delphinidin-3,5-diglucoside, and pigments
[4]. Te juice prepared from the leaves is considered as cool-
ing agent and used to cure smallpox in the infants. Bruised
leaves are externally applied to relieve painful infammations
and swellings [5]. Te paste prepared from fresh leaves mixed
with castor oil is reported to cure tinea capitis, a scaly fungoid
infection on scalp. Roots are used as febrifuge and vermifuge
by the tribal population in India [6, 7]. R. pectinata have been
claimed to exhibit antipyretic, anti-infammatory, diuretic,
analgesic, antifungal, and antimicrobial activities [4, 8, 9].
Recently, due to high demand by the drug manufacturers,
this plant has been overexploited. Te natural population of
R. pectinata is decreasing in the forests. Terefore, conserva-
tion activities have been initiated by the Wildlife Institute of
India [10].
Te transfer of vegetative phase to reproductive phase
is a complicated and fascinating process in the life cycle of
the plants. Environmental factors, photoperiod, and genetic
makeup of the plant are responsible for fowering phenomena
[11]. Reproductive phase can be induced by the artifcial
manipulation of these factors under in vitro controlled envi-
ronment [12]. In vitro fowering can be induced artifcially
by altering the concentration of plant growth regulators,
diferent light qualities, photoperiods, and so forth [13–18].
Te in vitro approach enables early fowering in the plants and
could help in understanding the suitable sites of tissues for the
fower induction.
Hindawi Publishing Corporation
Scientifica
Volume 2016, Article ID 5813851, 7 pages
http://dx.doi.org/10.1155/2016/5813851