09-02-2013, 10:59 AM
In vitro clonal propagation of Achyranthes aspera L. and Achyranthes
bidentata Blume using nodal explants
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ABSTRACT
Objective: To develop the reproducible in vitro propagation protocols for the medicinally plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blu immep ourstianngt annodd awl assehgemde nwtist ha sr uenxnpilnang ttsa. pM weattheor dasn:d Y toreuantge dsh woiotths 0o.f1 A%. baaspveisrtai na nadn dA .r ibnisdeedn ttawtiac ew ewriet hh adrivsetisltleedd rwinatseinr.g T whietnh tshteer ielex pdlaisnttisll ewde rwea tseurr ffaocr e3 -st4e rtiilmizeesd, nwoidtha l0 s.1e%gm (wen/vt)s HwgeCrel2 csuotl uitnitoon ss mfoarl l1e rm sieng.m Aefntetsr (s1o lcidm )b aasnadl used as the explants. The explants were placed horizontally as well as vertically on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi- Mkiendeitain, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration. Results: Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments firneoqcuuelantceyd. oMnS MmSe dbiausmal amuegdmiuenmt ewdi twhi t3h% 3 .s0u cmrogs/Le oafn dB AaPu gsmhoewnteedd twheit hh iBgAhePs ta npde rKceinn tawgieth ( 9v3a.6ri0e暲d
0n.u7m1) boef rs ohfo sohtloeotsts /feoxrmplaatniotsn for A. aspera and (94.70暲0.53) percentages for A. bidentata. Maximum in (10.60暲0.36) for A. aspera and (9.50暲0.56) for A. bidentata was observed shoMoStl emtse dwiausm o bfotratiinfieedd iwni th 5.0 mg/L of BAP. For A.
Introduction
Achyranthes aspera (A. aspera) L. and Achyranthes
bidentata (A. bidentata) Blume belong to the family
Amaranthaceae and are found to possess lots of medicinal
properties. A. aspera is a small herb found all over India
possessing valuable medicinal properties. It is useful in
cough, bronchitis, rheumatism, malarial fever, dysentery,
asthma, renal and cardiac dropsy, hypertension and
diabetes mellitus[1]. A. aspera can stimulate the immunity,
enhance the antigen clearance, potentiate antibody
production, elevate thyroid hormone levels, decrease
hepatic lipid peroxidation and also possesses spermicidal,
chemopreventive, anti-inflammatory, anti-arthritic
and hypoglycaemic activities[2-4]. In Chinese traditional
medicine, the hot water extract of the plant has been used
as an antiarthritic to alleviate arthritic pain.
Results
The frequency of explants survival and regeneration of
explants varied with concentration of HgCl2 and sterilization
time. 0.1% HgCl2 for 3 min showed the highest percentage
(95%) of survival rate of explants. 0.1% HgCl2 for 2 min showed
the highest percentage (90%) of microbial contamination. It
was observed that when the explants were sterilized with
0.1% HgCl2 for 5 min, 100% of microbes are free from explants
but the mortality rate of explants was high up to 95% to 100%.
Proliferation of multiple shoots was observed with high
frequency from nodal segments and shoot tips within five
days of inoculation.
Materials and methods
Plants of A. aspera L. and A. bidentata Blume were
collected from Kollimalai, Salem, Tamil Nadu, India and
were established in the green house and herbal garden.
Young shoots were harvested and washed with running tap
water and treated with 0.1% bavistin and rinsed twice with
distilled water. Then the explants were surface sterilized
with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with
sterile distilled water for 3-4 times, nodal segments were
cut into smaller segments (1 cm) and used as the explants.
The explants were placed horizontally as well as vertically
on solid basal Murashige and Skoog (MS)[22] medium
supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media,
Mumbai) and different concentration and combination of
6-benzyl amino purine (BAP), kinetin (Kin), naphthalene
acetic acid (NAA) and indole acetic acid (IAA) for direct
regeneration.