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Bharti Gautam, Et Al
Bharti Gautam, Et Al
*Corresponding author
ABSTRACT
The present investigation entitled, “Studies on the effect of various sterilization procedures
Keywords for in vitro propagation Of Carnation (Dianthus caryophyllus L.) was carried out at the
Plant Tissue Culture Laboratory of Department of Floriculture and Landscape
In vitro, Surface Architecture, Dr Y S Parmar University of Horticulture and Forestry, Nauni, Solan (HP) as
sterilization, Heavy
metals, Hazardous,
a refinement in already existing protocol to find suitable and less hazardous surface
Uncontaminated sterilization chemicals than Mercuric Chloride which is one of the most widely used
growing cultures surface sterilant in micropropagation of carnation. The use of this chemical is being
prohibited because of the presence of heavy metal ions in it, causing environment hazards.
Article Info The necessity to consider an alternative surface sterilization agent is therefore obvious.
The experiments were laid out in a Completely Randomized Design (factorial) consisting
Accepted:
07 January 2019
of two cultivars i.e. ‘Parendillo’ and ‘Yellow Star’. Treatment of explants with 5 %
Available Online: Calcium Hypochlorite is suggested as a potential substitute for Mercuric Chloride. 100 %
10 February 2019 uncontaminated growing cultures in cv. ‘Parendillo’ and ‘Yellow Star’ were obtained with
5 % Calcium Hypochlorite treatment of explants for 10 and 15 minutes, respectively.
effects of Mercuric Chloride have also been an autoclave at 121°C and 1.05 Kg/cm²
reported by Counter and Buchanan, 2004. The pressure (15 psi) for 20 minutes (Dodds and
necessity to consider an alternative surface Roberts, 1982). The medium was then kept at
sterilization agent is therefore obvious. A room temperature and used for culturing after
refinement in already existing protocol is, 2 days waiting period to check for any
therefore, required to find out a potential contamination in the medium.
surface sterilization chemical which could be
used as an alternative to Mercuric Chloride. Surface sterilization of explants
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Int.J.Curr.Microbiol.App.Sci (2019) 8(2): 481-485
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Int.J.Curr.Microbiol.App.Sci (2019) 8(2): 481-485
Table.2 Interaction effect of cultivars, surface sterilization treatments and explant source on
percent uncontaminated growing cultures (4 weeks after inoculation)
Surface sterilization Cultivars of carnation
Treatments (min.) Parendillo Yellow Star
Shoot tip Nodal Shoot tip Nodal
HgCl2 3 98.33 93.33 93.33 90.00
(0.1 %) (85.69)* (75.24) (75.24) (71.57)
NaOCl (3%) 5 20.00 25.33 26.67 23.33
(26.45) (30.12) (31.07) (28.67)
10 60.00 48.00 53.33 43.33
(50.79) (43.85) (46.91) (41.15)
15 83.33 86.67 83.33 85.00
(65.95) (68.62) (65.95) (67.40)
20 76.67 78.67 66.67 76.67
(61.15) (62.50) (54.75) (61.21)
25 73.33 65.33 66.00 68.33
(58.93) (53.93) (54.37) (55.85)
30 66.67 62.67 53.33 50.00
(54.75) (52.34) (46.92) (45.00)
35 63.33 54.67 46.67 45.00
(52.74) (47.68) (43.09) (42.12)
CaOCl (5%) 5 93.33 91.67 66.67 90.00
(77.71) (76.26) (54.78) (75.00)
10 100.00 100.00 80.00 90.00
(90.00) (90.00) (63.55) (75.00)
15 100.00 100.00 100.00 100.00
(90.00) (90.00) (90.00) (90.00)
20 100.00 100.00 100.00 100.00
(90.00) (90.00) (90.00) (90.00)
25 100.00 100.00 100.00 100.00
(90.00) (90.00) (90.00) (90.00)
30 93.33 85.00 90.00 100.00
(78.09) (67.41) (75.00) (90.00)
35 86.67 81.67 90.00 96.67
(68.86) (64.69) (75.00) (83.86)
*values in parenthesis are arc sine transformed values
CD0.05 for: Cultivars x Treatments x Explants: 8.35
Among the different chemicals used for Hypochlorite (CaOCl) for 10, 15, 20 and 25
surface sterilization, Calcium Hypochlorite minutes irrespective of explants source (Table
was found to be the most superior resulting in 1 and 2). On the other hand, similar results in
maximum uncontamination as compared to ‘Yellow Star’ were recorded with 5 %
Mercuric Chloride and Sodium Hypochlorite Calcium Hypochlorite (CaOCl) for 15, 20, 25
doses. and 30 minutes.
100 % uncontaminated growing cultures were Use of Calcium Hypochlorite (CaOCl) for
obtained when explants of cv. ‘Parendillo’ surface sterilization of explants has been
were surface sterilized with 5 % Calcium reported by many workers. Sangwan et al.,
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Int.J.Curr.Microbiol.App.Sci (2019) 8(2): 481-485
Bharti Gautam, Puja Sharma, Y.C. Gupta, Anil Handa, Manisha Thakur and Priyanka Sharma.
2019. Studies on the Effect of Various Sterilization Procedures for in vitro Propagation of
Carnation (Dianthus caryophyllus L.). Int.J.Curr.Microbiol.App.Sci. 8(02): 481-485.
doi: https://doi.org/10.20546/ijcmas.2019.802.054
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