Medicinal and Aromatic Plant Science and Biotechnology

µM) and the same medium was used for raising cell suspension cultures. An acidic pH (i.e. 5.0-6.50) of the medium helped cell
growth, while that having alkaline pH (7.25-9.50) favoured pigment production. Almost double cell biomass (198.67g, FW.L -1 )
was recorded in medium at physiological pH (5.70), when compared to neutral (138.55 g FW.L -1 ) or alkaline levels (130.56 g
FW.L -1 ). Acetylshikonin (I) was found to be the major shikonin derivative in these cultures. The maximum content i.e., 2.049%
was recorded at alkaline pH (9.5) in comparison to 0.803% at acidic pH level (5.75) on a fresh weight basis, as revealed by
HPLC analysis of the cell suspension cultures. Microscopic examination of the cell suspensions grown at different pH revealed
that higher pH adversely affected growth and morphology of the cells. Importantly, the induction of pigment in the alkaline range
of pH was independent of the growth phase and pH showed a regulatory role in its biosynthesis.
K. Padmalatha, K. Jayaram, M.N.V. Prasad (India) A Rapid Protocol for the Isolation of Polysaccharide- and
Polyphenolic-Free Genomic DNA for RAPD Analysis of Threatened Medicinal Plants (pp 50-53)
ABSTRACT
Techniques Paper: Many procedures in molecular biology require the isolation of high quality genomic DNA which is difficult in
most plant species because of the presence of secondary metabolites that interfere with DNA isolation which effects
downstream applications such as DNA restriction, amplification and cloning. Here we describe a modified procedure based on
the hexadecyltrimethylammoniumbromide (CTAB) method to isolate DNA from leaf tissues containing high levels of
polysaccharides, polyphenols, gums, tannins and secondary metabolites. This modified CTAB (2%) protocol includes the use of
a combination of polyvinypyrrolidone (PVP) and polyvinylpolypyrrolidone (PVPP), 0.2% β mercaptoethanol for Pterocarpus
marsupium and use of 2 M NaCl, 1% PVPP, and 1% β-mercaptoethanol for other selected medicinal plants with an initial
prolonged chloroform:isoamylalcohol (24:1) step (30 min shaking) repeated twice, RNAse treatment for 1 hr and avoiding the
incubation and precipitation steps in isopropanol. The yield was approximately 20 µg DNA/200 mg of initial fresh leaf tissue.
This method solved the problem of DNA degradation and co-precipitation of the secondary metabolites. The isolated DNA
proved amenable to restriction digestion and PCR amplification. The technique is fast, reproducible and can be applied for
further molecular approaches.
Jin-ichi Sasaki, Chihiro Sutoh, Ayako Norigami, Tomomi Satoh, Nana Kumaki, Hidemitsu Uchisawa, Shinya Yamaguchi
(Japan), Vachik Lazarian (USA) Emission of Odor in Plants: A Strategy to Fight Infection? (pp 54-56)
ABSTRACT
Techniques Paper: The bacteriocidal potency of odor was studied by the Petri dish method using eight types of plant odors
from Hiba tree/Japanese cypress (Thujopsis dolabrata var. hondae), garlic (Allium sativum), onion (Allium cepa), horseradish
(Armoracia rusticana), cinnamon (Cinnamomum zeylanicum), dokudami (Houttuynia cordata), clove (Syzygium aromaticum)
and sage (Salvia officinalis). Target bacteria in the test were Pseudomonas aeruginosa, Bacillus natto, enterohemorragic
Escherichia coli O157:H7, methicillin-resistant Staphylococcus aureus (MRSA), and Candida albicans. Seven out of eight plant
odors showed antibacterial activity with a different spectrum of activity against the above described bacteria. Odor released
from Hiba tree, garlic, onion, horseradish and dried whole clove demonstrated the strongest antibacterial activity, followed by
dried cinnamon and dokudami, but the odor of dried sage did not show any activity against these bacteria. Gas chromatography
analysis clarified the presence of 15 composites in fresh garlic odor, but two of them were unidentified in an analysis 24 hours
later. When viewed by scanning electron microscopy, the bacteria surface became fluffy due to exposure to Hiba odor
accompanied with bleb formation on the surface, which may have been caused by protein degeneration, while formaldehyde (a
representative disinfectant)-exposed bacteria maintained a smooth surface without any formation of blebs. These results
suggest that anti-bacteria mechanisms of odor vary depending on the odor (i.e. chemical(s) included) emitted from plants. One
of the significances of plant odor might be the protection from attacks by harmful agents such as bacteria and insects in
environmental surroundings.
Klára Szentmihályi, Mária Hajdú, Mária Then (Hungary) Inorganic Biochemistry of Medicinal Plants (pp 57-62)
ABSTRACT
Original Research Paper: Most medicinal plants are flowering plants and they also serve as ornamental plants. Measurement
of trace element content in medicinal plants, drugs and extracts may be relevant in view of e.g. human health, animal health and
environmental relations. This fact has a great significance since about half of the plant drugs available in the trade originate
from natural habitats and the element content of herbs may originate from soil or air pollution, or relate to the soil type on which