Figure 1: Culture Showing in DM-DPS initiation (a), culture after 1 weak (b), 2 weak (c), 4 weak (d), 6 weak (e) and 8 weak (f). Culture in CMS shown shoot multiplication after 4 weak (g), culture in semi solid medium shown rooting after 4 weak of inoculation in rooting medium (h). Acclimatization of dual- phasic culture plants after 45 days (i).

Stages of micropropagation
Culture media used
Tissue culture responses
CMS
DPS

Bud proliferation

MS +3% sucrose
00
00
MS + 0.10mg/l BAP + 3% sucrose
67 ± 0.6b
68 ± 0.5b
MS + 0.25mg/l BAP + 3% sucrose
76 ± 0.5c
79 ± 0.7c
MS + 0.50mg/l BAP + 3% sucrose
96 ± 0.6e
97 ± 0.5e
MS + 1.00mg/l BAP + 3% sucrose
56 ± 0.7a*
63 ± 0.4a*
MS + 0.50mg/l Kn + 3% sucrose
86 ± 1.0d
89 ± 0.8d
MS + 0.5mg/l BAP + 0.50mg/l Kn + 3% sucrose
90 ± 0.8e*
91 ± 0.7e*
Shoot multiplication and growth
MS + 0.5mg/l BAP + 3% sucrose
32.6 ± 0.8a
36.3 ± 0.8a
MS + 1.5mg/l BAP + 3% sucrose
51.2 ± 0.7b
55.2 ± 0.5b
MS + 2.0mg/l BAP + 3% sucrose
76.5 ± 0.8d
81.4 ± 0.6e
MS + 2.5mg/l BAP + 3% sucrose
60.3 ± 1.0b*
63.3 ± 0.8c*
MS + 2.0mg/l Kn + 3% sucrose
66.3 ± 0.5c
71.3 ± 0.5d
MS + 2.0mg/l BAP + 0.1mg/l NAA + 3% sucrose
79.2 ± 0.8d
85.2 ± 0.8e
MS + 2.0mg/l BAP + 0.25mg/l NAA+ 3% sucrose
96.4 ± 0.8e
98.7 ± 0.4f
MS + 2.0mg/l Kn + 0.25mg/l NAA + 3% sucrose
70.2 ± 0.6d
76.2 ± 0.7d
Root induction and growth
 ½ MS +0.25mg/l IAA + 2% sucrose
44.2 ± 0.8 b
48.2 ± 0.8 b
½ MS + 0.50mg/l IAA + 2% sucrose
61.4 ± 1.0d
69.4 ± 0.7d
½ MS + 0.25mg/l IBA + 2% sucrose
53.4 ± 0.8c
57.6 ± 0.8c
½ MS + 0.50mg/l IBA + 2% sucrose
73.3 ± 1.0e
79.1 ± 0.7e
½ MS + 1.00mg/l IBA + 2% sucrose
48.3 ± 0.6b*
51.6 ± 0.7b*
½ MS + 0.50mg/l NAA + 2% sucrose
36.5 ± 0.6a
39.8 ± 0.8a
½ MS + 0.50mg/l IBA + 0.25mg/l NAA + 2% sucrose
60.0 ± 0.8c
70.6 ± 1.0d
½ MS + 0.50mg/l IBA + 0.25mg/l IAA + 2% sucrose
59.2 ± 0.6c*
63.5 ± 0.8d*
Values are mean ± SE; 20 replicates per treatment, repeated three times; *callusing at the basal end; means having a same letter within columns were not significantly different at P < 0.05.
Table 1: List of selected media used in the different stages of micropropagation in Rauwolfia serpentine and their responses
Parameters studied
Culture system
DPS †
CMS †
Percentage of shoot multiplication in the culture medium MS + 2.0mg/l BAP + 0.25mg/l NAA + 3% sucrose (mean ± SE)
98.7 ± 0.4 b
96.4 ± 0.8 a
Av. No. of multiple shoot/explants in the culture medium MS + 2.0mg/l BAP + 0.25mg/l NAA + 3% sucrose (mean ± SE)
10.4 ± 0.8 a
13.2 ± 0.7 b
Av. Shoot length (cm.) in the culture medium MS + 2.0mg/l BAP + 0.25mg/l NAA + 3% sucrose (mean ± SE)
9.8 ± 0.3 b
8.2 ± 0.5 a
Av. Shoot thickness (mm.) in the culture medium MS + 2.0mg/l BAP + 0.25mg/l NAA + 3% sucrose (mean ± SE)
5.2 ± 0.6 b
1.8 ± 0.2 a
Av. No. of roots/shoot in the culture medium ½ MS + 0.50mg/l IBA + 2% sucrose (mean ± SE)
8.7 ± 0.5 b
6.4 ± 0.7 a
Av. Root length (cm.) in the culture medium ½ MS + 0.50mg/l IBA + 2% sucrose (mean ± SE)
5.3 ± 0.8 b
4.9 ± 0.8 a
Av. Root thickness (mm.) in the culture medium ½ MS + 0.50mg/l IBA + 2% sucrose (mean ± SE)
6.1 ± 0.8 b
1.9 ± 0.5 a
Percentage of acclimatization efficiency(mean ± SE)
98.5 ± 0.7 b
78.3 ± 0.4 a

† 20 replicates per treatment, repeated three times
Values are mean ± SE, * callusing at the basal end, means having same letter within rows were not significantly different at P < 0.05
Table 2: A comparative study on micropropagation of Rauwolfia serpentine in two different culture systems after 6 weeks of culture

Name of the system
   Name of different stages
Amount of different media used/1set of experiment#
Total cost of the media used in the experiment in different system
MS
1/2MS
CMS
Initiation
500 ml
---------
Rs156.56/-
Multiplication*
1000ml
---------
Rooting
---------
500ml
Total amount of media used
1500ml
500ml
DPS
Initiation
500ml
---------
Rs100.64/-
Multiplication + Rooting
500ml†
250ml
Total amount of media used
1000ml
250ml

* Two multiplication phase in each phase 500ml media was used
† 250ml used as liquid phase along with 250ml solid ½ MS and 250ml rest of the liquid media was used to maintain the culture
# In each experiment 20 replicates were used
Table 3: Details amount of media used in different methods and their cost

Factors affecting micropropagation
Types of culture system
% of reduction in cost
CMS
DPS
Cost of Nutrient media/plant let (Total cost of media/ total no. of plants produced)
Rs 7.82/-
Rs 5.00/-
35.36 %
Energy cost /plant let
Rs 0.28/-
Rs 0.17/-
39.28 %
Labour cost/ complete experiment (initial to rooting)
Rs 15960/-
Rs 10640/-
33.33 %

Table 4: A comparison of overall production cost between CMS and DM-DPS