Putrescine and Silver Nitrate Influences Shoot Multiplication, In Vitro Flowering and Endogenous Titers of Polyamines in Cichorium intybus L. cv. Lucknow Local.

Abstract

The influence of putrescine (Put) and AgNO(3) on shoot multiplication, in vitro flowering and endogenous titers of polyamines in Cichorium intybus L. cv. Lucknow local was investigated. Exogenous administration of Put at a concentration of 40 mM resulted in maximum tissue response in terms of shoot numbers (34.6 +/- 2.61) and shoot lengths (7.6 +/- 0.57 cm) on MS media supplemented with 2-iP (2.0 mg L(-1)) and GA(3) (0.5 mg L(-1)) as observed on the 35(th) day. Exogenous application of 40 µM AgNO(3) resulted in maximum shoot number (36.8 +/- 2.63) and shoot lengths (7.9 +/- 0.76 cm) on day 35 on the same media. Endogenous titers of conjugated spermidine decreased sharply from day 7-21, whereas endogenous conjugated spermine levels peaked on day 28 (1265 +/- 94.9 nmoles g(-1) FW), after treatment with 40 mM Put. Whereas, AgNO(3) (40 µM) fed samples resulted in higher titers of endogenous conjugated spermine (1405 +/- 105.6 nmoles g(-1) FW, 3.62 fold over control) on day 14. All other treatments showed decreasing endogenous levels during the whole culture period. Both Put (40 mM) and AgNO(3) (40 µM) resulted in floral initiation and floral development on day 28 and 14 (3.76 +/- 0.16, 4.2 +/- 0.21 flowers per shoot apices), respectively. To investigate the role of Put (40 mM) and AgNO(3) (40 µM) on morphogenetic response and endogenous conjugated polyamine titers in shoots of chicory, polyamine inhibitors (DFMA and DFMO) were used. The morphogenetic response and the endogenous conjugated pool of polyamines were diminished in DFMA and DFMO treatments, but could be restored by addition of Put (40 mM) and AgNO(3) (40 µM). Under exogenous Put feeding, ethylene production was reduced in shoot cultures of chicory. This study shows for the first time the influence of polyamines on multiple shoot initiation from axillary buds of C. intybus L. cv. Lucknow local and also indicates the promotive effect of Put and AgNO(3) on autoregulation of polyamine biosynthesis, thereby regulating in vitro flowering, the endogenous pool of polyamines and shoot multiplication.