Standardization of Protocol for the Surface Sterilization and Callus Induction of Saussurea lappa: An Endangered Medicinal plant

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Research Paper 01/07/2016
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Standardization of Protocol for the Surface Sterilization and Callus Induction of Saussurea lappa: An Endangered Medicinal plant

Zaib Un Nisa, Samin Jan, Safdar Hussain Shah
Int. J. Biosci. 9(1), 324-330, July 2016.
Copyright Statement: Copyright 2016; The Author(s).
License: CC BY-NC 4.0

Abstract

Saussurea lappa is an endangered medicinal plant, used against lot of diseases like fever, headache, pain, arthritis, ulcers, cough, flatulence, asthma, colic, diarrhea, dyspepsia, vomiting, tenesmus, and dysentery. The plant is endangered due to over exploitation for commercial and medicinal purposes. The study was carried out to develop a protocol for the surface sterilization and callus induction of the seeds of S. lappa. Seeds of the plant were used as explants for the sterilization and callus induction. The explants were surface sterilized with 5% solution of tween-20 for 20 minutes followed by three different concentrations (0.05%, 0.1% and 0.2%) of mercuric chloride for 8minutes in three different experiments and rinsed 5 times thoroughly with autoclaved distilled water. Results were shown in percentage. The concentration of 0.2% solution of mercuric chloride showed best sterilization (96%) compared to 0.1% (48%) and 0.05% (0.2%) solution of mercuric chloride. Solid Murashige and Skoog (MS) medium fortified with 1mgL-1Dichlorophenoxyacetic acid (2-4 D) alone and in combination with 0.25 mgL-1 kinetin (Kn) were used for the callus induction. Relative Growth Rate (RGR) of the callus was determined after 15 days of inoculation and found that the combination of 2-4 D and Kn produced maximum (0.141week-1) callus compared to 2-4D (0.083 week-1) used alone. The results showed that 0.2% HgCl2 solution is the best seed sterilizing agent while 2-4 D (1mgL-1) and Kn (0.25mgL-1) are the best combination for the callus induction and fresh weight bio mass.

Abdel-Rahim EAO, Abdel-Fatah M, Shemy HAI, AbdSamei MB. 1998. Growth of date palm callus as affected by amino acids as organic nitrogen source. First International Conference on Date Palms AlAin, UAE 8-10.

Ahmad M, Nangyal H, Imran M, Ullah F. 2016. Optimization of Protocol for Surface Sterilization and Callus Induction for Three Rice Varieties Am-Euras. Journal of Agriculture and Environmental Sciences 16, 357-361.

Amiri S, Kazemitabar SK, Ranjbar GA, Azadbakht M. 2011. In vitro propagation and whole plant regeneration from callus in Datura (Daturastramonium. L). African Journal of Biotechnology 10, 442-448.

Anolles CG, Favelukes G, Bauer WD. 1990.Optimization of surface sterilization for legume seed. Crop Science 30, 708-712. http://dx.doi.org/10.2135/cropsci1990.0011183X003000030047x

Arora R, Bhojwani SS. 1989. In vitro propagation and low temperature storage Sassurealappa C. B. Clarke – an endangered medicinal plant. Plant Cell Reports 8, 44-47. http://dx.doi.org/10.1007/BF00735776

Backer WS, Aljefri AM, Sawi NA. 2012. Possible Relief for Migraine Patients:Saussurea Lappa. Journal of US-China Medical Science 9, 51-5.

Bano R, Khan MH, Khan RS, Rashid H, Khan RS, Munir I, Swati ZA, Chudhry Z. 2010. Callogenesis and organogenesis in three genotypes of brassica juncea affected by explants source. Pakistan Journal of Botany 42, 3925-3932.

Central Potato Research Institute, Shimla, 1992.Tissue Culture technique for potato health, conservation, micro propagation and improvement. CPRI, Shimla,  1-23 P.

Daud HN, Jayaraman S, Mohamed R. 2012. An improved surface sterilization technique for introducing leaf, nodal and seed explants of Aquilariamalaccensisfrom field sources into tissue culture. Asia Pacific Journal of Molecular Biology and Biotechnology 20, 55-58.

Gupta P, Sharma S, Saxena S. 2010. Callusing in Stevia rebaudiana (Natural Sweetener) for Steviol Glycoside Production. International Journal of Agriculture and Biological Sciences 1, 30-34.

Gupta OP, Ghatak RBJ. 1967. Pharmacological investigations on Saussurealappa (Clarke). Indian Journal of Medical Research 55, 10.

Hidayatullah, Iltafullah, Jadoon SA, Rashid H. 2007. Tissue culture techniques for callus induction in rice. Sarhad Journal of Agriculture 1, 81-86.

Kirtikar KR, Basu BD. 2001. Oriental enterprises. Indian Medicinal Plants 6, 1961–1965.

Kuniyal CP, Rawat YS, Oinam SS, Kuniyal JC, Vishvakarma SCR. 2005.“S. costus (Saussurealappa) cultivation in the cold desert environment of the Lahaul valley, northwestern Himalaya, India: arising threats and need to revive socio-economic values”. Biodiversity and Conservation 14, 1035-1045.

Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. PhysiologiaPlantarum 15, 473-479. http://dx.doi.org/10.1111/j.13993054.1962.tb08052.x

Murch SJ, Krishna RS, Saxena PK. 2000. Phyto-pharmaceuticals: mass production, standardization and conservation. Herbal Medicine 4, 39–43.

Negi JS, Bisht VK, Bhandari AK. 2013. Antidiarrheal activity of methanol extract andmajor essential oil contents of Saussurealappa Clarke. African journal of pharmacy andpharmacology. 7, 474-7. http://dx.doi.org/10.5897/AJPP2012.1532

Oyebanji OB, Nweke O, Odebunmi O, Galadima NB, Idris MS, Nnodi UN, Afolabi AS, Ogbadu GH. 2009. Simple, effective and economical explant-surface sterilization protocol for cowpea, rice and sorghum seeds. African Journal of Biotechnology 8, 5395-5399.

Pandey MM, Rastogi S, Rawat AKS. 2007. Saussurea costus: botanical, chemical and pharmacological review of anayurvedic medicinal plant. Journal of Ethnopharmacology 110, 379-90. http://dx.doi.org/10.1016/j.jep.2006.12.033

Ramakrishna N, Lacey J, Smith JE. 1991. Effect of surface sterilization, fumigation and gamma irradiation on the microflora and germination of barley seeds. International Journal of Food Microbiology 13, 47-54. http://dx.doi.org/10.1016/0168-1605(91)90135-C

Shah SH, Wainwright SJ, Merrett MJ. 1990. The interaction of sodium and calcium chloride and light on growth potassium nutrition and proline accumulation in callus cultures of Medicago sativa L.  New Phytologist 116, 37-45. http://dx.doi.org/10.1111/j.14698137.1990.tb00508.x

Swedlund B,Locy RD. 1993. Sorbitol as the primary carbon source for the growth of embryogenic callus of maize. Plant Physiology 103, 1339-1346. http://dx.doi.org/10.​1104/​pp.​103.​4.​1339

Smith RH. 2005. Plant Tissue Culture: Techniques and Experiments 2 edn. Elsevier Publishers, New Delhi.

Singh A, Kandasamy T, Odhav B. 2009. In vitro propagation of Alternantherasessilis (sessile joyweed), a famine food plant. African Journal of Biotechnology 8, 5691-5695.

Tamil C, Sundram M, Suffian M, Annuar M, Khalid N. 2012. Optimization of culture condition for callus induction from shoot buds for establishment ofrapid growing cell suspension cultures of Mangoginger (Curcuma mangga). Australian Journal of Crop Sciences 6, 1139-1146.

Pierik RLM. 1990. Cultivoin vitro de lasplantas superiors. Multiprensa publisher, Madrid, España 326.

Perez-Tornero O, Burgos L, Egea J. 1999. Introduction and establishment of apricot in vitrothrough regeneration of shoots from meristem tips. InvitroCellular and Developmental Biology-  Plant 35, 249-253. http://dx.doi.org/10.1007/s11627-999-0087-9

Visarada KBR, Sailaja SM, Sarma SM. 2002. Effect of callus induction media onmorphology of embryogeniccalliin rice genotypes. Biolgia Plantarum 45, 495-502. http://dx.doi.org/10.1023/A:1022323221513

Xiao PG. 2006. Modern chinese material medica.  Beijing: Chemical Industry Press

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