Contrast analysis recommends flame sterilization for surface depuration in coconut (Cocos nucifera) meristem culture
DOI:
https://doi.org/10.48165/Keywords:
Surface sterilization, Meristematic shoot, Contamination, Flame sterilizationAbstract
Five different surface sterilization treatments were carried out along with the control treatment for obtaining sterile explants in coconut (Cocos nucifera L.) at ICAR-CPCRI, Kasaragod, Kerala, during 2019-20. The treatments included different combinations of mercuric chloride, sodium hypochoride, tween-20 and flame sterilization. While 100% of the meristem got contaminated in the control treatment which was surface sterilization with 0.1% mercuric chloride (HgCl2) treatment for 3 minutes, followed by 20% sodium hypochlorite (NaOCl ) treatment for 10 minutes, only 26.67% contamination was found in the treatment five which included 20% sodium hypochlorite with 250 µL tween 20 treatment for 10 minutes followed by tap water wash for 30 minutes, 0.1% mercuric chloride treatment for 3 minutes, 20% sodium hypochlorite treatment for 5 minutes and finally flame sterilization before chopping and inoculating the meristem. Contrast analysis was carried out for flame sterilization and difference between the means of two groups was statistically significant in both bacteria and fungi contamination revealing that flame sterilization has resulted in reduction of contamination.
Downloads
References
Cassells AC. 2012. Pathogen and biological contamination management in plant tissue culture: phytopathogens, vitro pathogens, and vitro pests. Methods in Molecular Biology 877:57-80.
Jan A, Bhat K M, Bhat S J A, Mir M A, Bhat M A, Imtiyaz A and Rather J A. 2013. Surface sterilization method for reducing microbial contamination of field grown strawberry explants intended for in vitro culture. African Journal of Biotechnology 12(39): 5749-53.
Leifert C, Morris C and Waites W M. 1994. Ecology of microbial saprophytes and pathogens in field grown and tissue cultured plants: reasons for contamination problems in vitro. CRC Critical reviews plant science 13(2): 139-83.
Lopez Del Egido L, Navarro-Miró D, Martinez-Heredia V, Toorop PE and Iannetta PPM. 2017. A spectrophotometric assay for robust viability testing of seed batches using 2,3,5- triphenyl tetrazolium chloride: using Hordeum vulgare L. as a model. Frontiers in Plant Science 8:747.
Manorama, K., Mathur, R.K., Prasad, M.V., Suresh, K., Ramachandrudu, K. and Rao, B.N., 2019. Doubling oil palm yield through technological interventions—a review. Current Horticulture 7(2), pp.28-31.
Murkute A A. 2020. In-vitro scape and bulb formation in garlic (Allium sativum). Current Horticulture 8(2): 44-46.
Nguyen Q T, Bandupriya H D, López-Villalobos A, Sisunandar S, Foale M and Adkins S W. 2015. Tissue culture and associated biotechnological interventions for the improvement of coconut (Cocos nucifera L.): a review. Planta 242(5): 1059-
Nurul H D, Shashita J and Rozi M. 2012. An improved surface sterilization technique for introducing leaf, nodal and seed explants of Aquilaria malaccensis from field sources into tissue culture. Asia Pacific Journal of Molecular Biology and Biotechnology 20(2):55-58.
Omamor I B, Asemota A O, Eke C R and Ezia E I. 2007. Fungal contaminants of the oil palm tissue culture in Nigerian Institute for Oil Palm Research (NIFOR). African Journal of Agricultural Research 2(10): 534-37.
Orlikowska T, Nowak K and Reed B. 2017. Bacteria in the plant tissue culture environment. Plant Cell, Tissue and Organ Culture 128(3): 487-08.
Sharuti V, Kuldeep Y and Narender S. 2011. Optimization of the protocols for surface sterilization, regeneration and acclimatization of Stevia rebaudiana Bertoni. American Eurasian Journal of Agricultural and Environmental Sciences 11(2):221-27.
Tiwari S, Arya A and Kumar S. 2012. Standardizing sterilization protocol and establishment of callus culture of sugarcane for enhanced plant regeneration in vitro. Research Journal of Botany 7(1): 1-7.
Tsao C W, Postman J D and Reed B M. 2000. Virus infections reduce in vitro multiplication of “Malling Landmark” raspberry. In Vitro Cellular and Developmental Biology 36(1): 65-68.
Tyagi V S A, Chauhan P, Kumari P and Kaushal S. 2011. Identification and prevention of bacterial contamination on explant used in plant tissue culture labs. Alcohol 3(1): 0-722.