EFFICIENT in vitro PROPAGATION OF Amaranthus viridis L. USING NODE EXPLANTS
Tour JanPlant Tissue Culture Laboratory Department of Botany, University of Malakand, Dir Lower, KP, Pakistan
Ikram UllahPlant Tissue Culture Laboratory Department of Botany, University of Malakand, Dir Lower, KP, Pakistan
Bilal MuhammadPlant Tissue Culture Laboratory Department of Botany, University of Malakand, Dir Lower, KP, Pakistan
_ TariqPlant Tissue Culture Laboratory Department of Botany, University of Malakand, Dir Lower, KP, Pakistan
Ali MansoorPlant Tissue Culture Laboratory Department of Botany, University of Malakand, Dir Lower, KP, Pakistan
Zaheer UllahPlant Tissue Culture Laboratory Department of Botany, University of Malakand, Dir Lower, KP, Pakistan
Muhammad Asif NawazDepartment of Biotechnology, SBBU, Sheringal, Dir Upper, KP, Pakistan
Hyperhydricity is a frequently problem in plants during in vitro culture and affected micropropagation of
plants. To develop an efficient in vitro regenerated system without hyperdydricity, we demonstrated the effect
of different disinfected agents (mercuric chlorite and hypochlorite), growth regulators, their concentrations
and combinations, Agar, pH, ammonium nitrate (NH4NO3) and number of subcultures. Mercuric chlorite at
0.07% and exposing time (9–10 min) was appropriate for hygienic culture. The shoots induced by Benzyladnine
(BA) alone or in combination with α-Naphthaleneacetic acid (NAA) exhibited maximum multiplication
with symptoms of hyperhydricity than those induced by Kinetin alone or in combination with NAA. Hyperhydricity
was also reduced by increasing the concentration of agar, pH and elimination of NH4NO3 from the
macroelements of Murashig and Skoog (MS) medium. Repeated subcultures affected both multiplication and
hyperhydricity. The multiplication of shoots increased from parental culture up to 5th subculture and thereafter
declined in 6th subculture. Although shoot hyperhydricity were observed from 1st subculture (19%) and
then increased up to 85% in 6th subculture. This increased in hyperhydricity could be due to the remaining
influence of hormones. In shoots of 5th subculture the content of chlorophyll (dark green) were higher than
shoots of 6th subculture.
Keywords:agar, Amarinthus viridis, HgCl2, hyperhydricity, pH, subcultures
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