Development of a protocol for in-vitro propagation of black pepper (Piper nigrum L.) local selections
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Date
2015
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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