ISSN 1021-4437, Russian Journal of Plant Physiology, 2009, Vol. 56, No. 2, pp. 224–231. © Pleiades Publishing, Ltd., 2009.
Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 2, pp. 247–255.
(Gaertn.) Libosch is a peren-
nial medicinal plant that originates from China and
belongs to the family Scrophulariaceae. Although there
are seven species in the genus
, only three
are used for medicinal purposes:
Makino in Japan,
(Chao and Schih)
Hsiao in China .
is far more frequently
prescribed in China than in other countries . It is gen-
erally cultivated in the provinces of China, such as
Henan, Hebei, and Zhejiang, and is used in three ways:
fresh, dried, and steamed. Each is known to have differ-
ent medicinal effects [1, 3].
is an important medicinal crop in Huai zone, Henan
province, China, to which is endemic.
is commonly used to replenish vitality, strengthen the
This text was submitted by the authors in English.
liver, kidney, and heart, and for treatment of a variety of
ailments like diabetes, anemia, and urinary tract prob-
lems according to the Chinese Pharmacopoeia .
Many components have been isolated from both the
radix and from the fresh plant, including
stigmasterol, campesterol, catalpol, aucubin, rehman-
nin, rehmannioside (A, B, C, and D), stachyose, verbas-
cose, leonuride, monomelittoside, vitamin A, and so on
[1, 4–6], of which catalpol is the main active ingredient
performing pharmacological action in rehmannia, so
that rehmannia is often identiﬁed using catalpol as stan-
dard preparation . Because of some negative effects
of continuous vegetable propagation (virus diseases
and so on) on its yield and quality,
urgently requires improve-
ment by plant biotechnology.
can cause hairy root induction , and hairy
roots are characterized by such advantages as fast
growth, unique genetic and biosynthetic stability, and
easy regeneration of intact plants . Plants infected
with the Ri plasmid usually have a phenotype consist-
ing of wrinkled leaves, shortened internodes, increased
Hairy Root Induction and Plant Regeneration of
Y. Q. Zhou
, H. Y. Duan
, C. E. Zhou
, J. J. Li
, F. P. Gu
, Z. Y. Zhang
, and Z. M. Gao
College of Life Sciences, Henan Normal University, Jianshe Road, Xinxiang, Henan, 453007China;
College of Agriculture Sciences, Henan Agriculture University, Zhengzhou, Henan, 450002 China
Received August 17, 2007
—The objective of this research was to establish an efﬁcient system of genetic transformation and
plant regeneration from hairy roots by infecting the leaf sections and stem segments of in vitro
Hsiao plantlets. Hairy roots were induced from them after co-culturing with
strain 15834 at a frequency of 32 and 29.4%, respectively. The calluses were induced from
hairy roots on half-strength Murashige and Skoog medium containing 0.2 mg/l kinetin and 3.0 mg/l benzyladenine
at a frequency of 100%, from which transgenic shoots and plantlets were developed. Transgenic plantlets did
not have differences in morphology except the shortened internodes and an increase in adventitious root formation
compared to wild-type plants. PCR and Southern-blot analyses conﬁrmed that
gene of TL-DNA was inserted
in the genome of transformed hairy roots and plantlets. RT-PCR analysis and opine paper electrophoresis
gene was expressed in the transformed hairy roots and plantlets. Conclusively, transgenic
hairy roots and transgenic plants of
Hsiao were developed for
the ﬁrst time.
Key words: Rehmannia glutinosa f. hueichingensis - Agrobacterium rhizogenes - genetic transformation - hairy
root - RT-PCR - Southern blot
: BA—6-benzyladenine; KT—kinetin; MS—Murash-
ige and Skoog nutrient medium; PGR—plant growth regulator.