Use of Response Surface Methodology (RSM) to Optimize Culture Media for in vitro Callogenesis in Ipomoea obscura (L.) Ker-Gawl, a Little Known Medicinal Plant | Chapter 4 | Current Strategies in Biotechnology and Bioresource Technology Vol. 1
Aims: The present study was undertaken to explore the
potentiality of in vitro micropropagation (by callus induction) of Ipomoea
obscura(L.) Ker-Gawl (Convolvulaceae), a perennial herb. Method: Axenically
grown cotyledonary leaves regenerated into profuse calli, in various
combinations of naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA),
indole-3-butyric acid (IBA) and 2,4dichlorophenoxyacetic acid (2,4 D) with
benzylaminopurine (BAP). Microroots emerged from these calli when subjected to
various combinations of NAA and BAP. Maximum number of root meristems and
microroots were formed in 1.25 mg/L NAA with the combination of 0.5 mg/L BAP
(1:4 v/v). Leaf, nodal and internodal segments as explants were cultured on MS
media containing 2,4 D at 0.5, 1.25, 2.50, 5.0 and 10.0 mg/L, for callus
induction. Only leaf tips initiated callogenesis with highest response at 10
mg/l 2,4 D and lowest with 0.5 mg/l 2.4 D. This is the first report of in vitro
response of I. obscura. Results: All the
results were analyzed on the context of response surface methodology (RSM) and
simultaneously by contour plot. Two variables lay out in X and Y axes and a
third variable Z is plotted for contour level as concentric values. An
efficient and effective micropopagation protocol was developed from the leaf of
Ipomoea obscura (L.) Ker-gawl, an important medicinal plant. Transplantation of
regenerated plants under in vivo condition has been done successfully. RSM has
been used for modeling, analyzing and optimizing in vitro responses of Ipomoea
obscura. The specific target response surface plot and contour plot are focused
to determine optimum shoot and root length against optimum hormone
concentration, number of shoot and root length
in best concentration of MS medium and finally mean number of root and
shoot in optimum NAA concentration. Depending upon variances of dry weight of
callus, rooting response and root length, response surface plots (RSM) and
contour plots were drawn where four types
of hormone combinations (IAA- BAP, IBABAP,
NAA-BAP, 2,4D-BAP) were used in different combinations.
Author(s)
Details
Dr. Tanmoy Sinha
Cytogenetics and Molecular Biology Unit, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India
A. Bandyopadhyay
Cytogenetics and Molecular Biology Unit, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India.
Dr. Tanmoy Sinha
Cytogenetics and Molecular Biology Unit, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India
A. Bandyopadhyay
Cytogenetics and Molecular Biology Unit, Department of Botany, The University of Burdwan, Burdwan-713104, West Bengal, India.
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