ISSN 10214437, Russian Journal of Plant Physiology, 2010, Vol. 57, No. 1, pp. 110–117. © Pleiades Publishing, Ltd., 2010.
Published in Russian in Fiziologiya Rastenii, 2010, Vol. 57, No. 1, pp. 117–124.
Pall, a herb species of Genti
anaceae, is widely used in Chinese herb medicine
because of being rich in gentiopicroside, a secoiridoid
glucoside (Fig. 1), in its roots [1, 2], which has been
confirmed to have many medicinal functions, includ
ing abirritation, defervescence, lowering blood pres
sure, antibacterium, antiinflammation, and antirheu
matism effects [3, 4].
has been success
fully used in curing virusinduced respiration tube and
cardiovascular diseases . Among four related spe
cies with similar functions,
onstrated to be the best in their effects . However,
because the traditional Chinese medicine therapy and
modern pharmaceutical industry still mainly rely on
the roots of wild
as a major source, a
very limited wild resource can hardly meet the increas
ing demands of the market. On the other hand, chron
ically unreasonable excavation is leading to serious soil
erosion and exhaustion of the wild
This text was submitted by the authors in English.
These authors contributed equally to this research.
China. Therefore, it has become a very important and
urgent task to protect this interesting plant resource.
Hairy root (HR) cultures produced by
mediated transformation have been widely
accepted as not only an important experimental sys
tem for plant physiological and molecular research but
also a promising biotechnology for producing useful
secondary plant metabolites due to its many well
known advantages [5–7], e.g. fast growth rate, high
genetic stability, and some others. To date, the HR cul
tures from many plants, e.g.
, and others have been found being
able to produce pharmaceutical chemicals. In several
medicinally important plants, HRs have been success
fully utilized in the production of pharmaceutical
chemicals . For exploring a possibility to produce
gentiopicroside using HR cultures of
several HR lines were established by
mediated transformation over three previous years.
Recently these lines were subjected to PCR and
HPLC analyses in attempt to evaluate their genetic
stability and capacity of gentiopicroside synthesis.
Establishment of Hairy Root Lines and Analysis of Gentiopicroside
in the Medicinal Plant
H. L. Zhang
, S. H. Xue
, F. Pu, R. K. Tiwari, and X. Y. Wang
Institute of Cell Biology, School of Life Sciences, Lanzhou University, Lanzhou, 730000 P.R. China;
fax: +869318912562; email: email@example.com
Received July 4, 2008
—The leaf explants from the eightweekold
Pall sterile seedlings were precul
tured for three days on Murashige and Skoog (MS) agar medium with 1 mg/l of 6benzyladenine, and then
incubated in MS liquid medium containing
R1000 for 2 h with shaking. After
10 days of culture on hormonefree MS agar medium with 500 mg/l cefotaxime under illumination of
s) with a 16h photoperiod at 24
C, up to 18.3% of the infected explants produced typical hairy
roots (HRs). Finally, three stable HR lines with no morphologically visible differences were obtained. The
HR lines could vigorously grow and propagate on/in hormonefree halfstrength MS solid/liquid medium.
Moreover, they changed from white to green when moved from darkness to the light and spontaneously pro
, which were able later to produce adventitious buds. PCR analysis revealed that three HR lines,
which were undergone to continuous subculturing over two years, still contained the
gene from the
Ri plasmid. HPLC revealed the HR line possessing a certain capacity of synthesizing gentiopicro
side (0.11 mg/g dry wt).
Key words: Agrobacterium rhizogenes Gentiana macrophylla genetic transformation gentiopicroside hairy
HR—hairy root; MS—Murashige and Skoog nutrient medium.