Agrobacterium mediated transformation in Phlogacanthus thyrsiflorus Nees for quality parameters
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Date
2011
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Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
on P. thyrsiflorus which has not been reported earlier. The total
carbohydrate content in hairy roots were also quantified and compared from leaves from
mother plant and bacteria infected plant.
Aseptic explants for transformation were obtained by inoculating shoot tip of
P.thyrsiflorus on MS media supplemented with BAP 5.0 mg/l and AdSO4 50.0 mg/l.
Leaves , shoot tips and nodes of in vitro grown plants were used as explants while ATCC
15834 strain of A. rhizogenes Conn was used for transformation. Full loop inocula of
A.rhizogenes were grown in YEB (Yeast Extract Beef Extract) medium supplemented
with 50 mg/l rifampicin. Optimum growth was observed after 48 h in dark at 28°C.
Acclimatization of bacteria culture in MS medium for 4 h was found effective. The
explants were treated for different time periods after which those were inoculated in solid
MS basal medium and kept at 25 ± 2 °C and 16 h light/ 8 h dark. Successful hairy root
induction was observed after 7 days on all 3 explants. Amongst different treatment
durations, 225 min was found to be the best for all the three explants. A clear variation in
response was observed within different types of explants. Out of three explants, shoot tips
were found to be the best with maximum mean percentage of root induction (93.33%) as
well as maximum mean number of roots (4.80) per explant.
Total carbohydrate content was quantified with anthrone method which revealed
that the carbohydrate content was highest (2.9 mg percent) in hairy roots followed by the
leaves from bacteria infected plant (2.0 mg percent). However, leaves of mother plant
showed comparatively minimum (1.6 mg percent) carbohydrate content.
Description
Agrobacterium mediated transformation in Phlogacanthus thyrsiflorus Nees for quality parameters
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