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Appl Biochem Biotechnol (2014) 174:21952208
DOI 10.1007/s12010-014-1175-9
Richa Upadhyay & Sarvesh Pratap Kashyap & Chandra Shekhar Singh & Kavindra Nath Tiwari & Karuna Singh & Major Singh
Received: 8 June 2014 /Accepted: 15 August 2014 /
Published online: 30 August 2014# Springer Science+Business Media New York 2014
Abstract Germplasm storage of Phyllanthus fraternus by using synseed technology has been optimized. Synseeds were prepared from nodal segments taken from in vitro-grown plantlets. An encapsulation matrix of 3 % sodium alginate and 100 mM calcium chloride with polymerization duration up to 15 min was found most suitable for synseed formation. Maximum plantlet conversion (92.52.5 %) was obtained on a growth regulator-free -strength solid Murashige and Skoog (MS) medium. Multiple shoot proliferation was optimum on a MS medium containing 0.5 mg/l 6-benzylaminopurine (BAP). Shoots were subjected to rooting on MS media containing 1 mg/l -naphthaleneacetic acid (NAA) and acclimatized successfully. Encapsulated nodal segments can be stored for up to 90 days with a survival frequency of 47.33 %. The clonal fidelity of synseed-derived plantlets was also assessed and compared with that of the mother plant using rapid amplified polymorphic DNA and inter-simple sequence repeat analysis. No changes in molecular profiles were observed among the
R. Upadhyay : C. S. Singh : K. N. Tiwari (*)
Department of Botany, MMV, Banaras Hindu University, Varanasi, Uttar Pradesh 221005, India e-mail: [email protected]
R. Upadhyaye-mail: [email protected]
C. S. Singhe-mail: [email protected]
S. P. Kashyap : M. Singh
Crop Improvement Division, Indian Institute of Vegetable Research, Shanahshahpur, Varanasi, Uttar Pradesh 221305, India
S. P. Kashyape-mail: [email protected]
M. Singhe-mail: [email protected]
K. Singh
Department of Zoology, MMV, Banaras Hindu University, Varanasi, Uttar Pradesh 221005, India e-mail: [email protected]
Ex Situ Conservation of Phyllanthus fraternus Webster and Evaluation of Genetic Fidelity in Regenerates Using DNA-Based Molecular Marker
2196 Appl Biochem Biotechnol (2014) 174:21952208
synseed-derived plantlets and mother plant, which confirms the genetic stability of regenerates. This synseed production protocol could be useful for in vitro multiplication, short-term storage, and exchange of germplasm of this important antiviral and hepatoprotective plant.
Keywords Encapsulation . Plantlet conversion . Synseed . Phyllanthus fraternus . Genetic stability. Germplasm storage
Abbreviations
BAP 6-BenzylaminopurineNAA -Naphthaleneacetic acidMS Murashige and SkoogRAPD Random amplified polymorphic DNA ISSR Inter-simple sequence repeat analysis
Introduction
Phyllanthus fraternus is an important medicinal...