Development of a protocol for in-vitro propagation of black pepper (Piper nigrum L.) local selections
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Date
2015
Journal Title
Journal ISSN
Volume Title
Publisher
Uva Wellassa University of Sri Lanka
Abstract
Black pepper (Piper nigrum L.) belongs to family Piperaceae and it is one of the most
economically important spice crops in the world (Srinivasan, 2007; Mathew et al., 2001).
Unavailability of sufficient mother plant stock in the field, obtaining basal runners for
propagation and less success and multiplication rate of the high yielding local pepper cultivars
are the major problems faced by the farmers who cultivate. Being in vitro propagation a
promising option, this study was focused to develop a suitable protocol for in vitro propagation
of black pepper local selections.
Methodology
This research was carried out at Central Research Station, Department of Export Agriculture,
Matale. Four experiments were conducted during the research period. Experiment one was
conducted to find out the suitable surface sterilization method for the sterilization of black
pepper shoot tips. Selecting of appropriate media for the culture establishment of black pepper
shoot tips were carried out in second experiment using 1/3Murashige and Skoog (MS) medium
and 1/2 Woody Plant Medium (WPM).Experiment three was conducted to find out suitable
combination of auxin and cytokinin for the shoot multiplication of black pepper local selections.
In fourth experiment, priority was given for the selection of best media and hormonal
combination for the callus initiation of TG7 black pepper local selection. Full and half strength
MS media were used as the culture media and two different concentration levels of kinetin and
NAA were used as the growth regulators. Complete Randomized Design (CRD) was used as the
experimental design. ANOVA was used to analyze the statistical difference of parametric data
and non-parametric data were subjected for logarithmic transformation. SAS statistical software
was used to analyze the data and mean separation was performed using Least Significant
Difference (LSD).
Results and Discussion
As the results summarized in Table 1, sterilization using 10%- 20% Clorox for five to ten
minutes (T1 to T5) showed higher percentages of bacterial contamination (40 to 80 %). Lower
percentages of fungal contamination was observed in T4 to T8 within the period of three to five
days (3% to 7%).The highest survival percentage (66.6 %) was reported in T8, 0.04 % HgCl2
for five minutes. Similarly, the lowest percentages of bacterial and fungal contamination were
observed in T8. The highest percentage of phenolic browning (80%) was shown in T6 and
lowest percentage of phenolic browning (10.0%) was observed in T2, i.e. 10% Clorox for 10
minutes within four to seven days.
Description
Keywords
Export Agriculture, Agriculture, Pepper, Spices