title: Ex situ conservation of Phyllanthus fraternus Webster and evaluation of genetic fidelity in regenerates using DNA-based molecular marker.
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 (92.5 ± 2.5 %) was obtained on a growth regulator-free ½-strength solid Murashige and Skoog (MS) medium. Multiple shoot proliferation was optimum on a ½ MS medium containing 0.5 mg/l 6-benzylaminopurine (BAP). Shoots were subjected to rooting on MS media containing 1 mg/l α-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 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.
authors.affiliation: Department of Botany, MMV, Banaras Hindu University, Varanasi, Uttar Pradesh, 221005, India, ru4004@gmail.com.
authors.forename: Richa
authors.initials: R
authors.lastname: Upadhyay
authors.forename: Sarvesh Pratap
authors.initials: SP
authors.lastname: Kashyap
authors.forename: Chandra Shekhar
authors.initials: CS
authors.lastname: Singh
authors.forename: Kavindra Nath
authors.initials: KN
authors.lastname: Tiwari
authors.forename: Karuna
authors.initials: K
authors.lastname: Singh
authors.forename: Major
authors.initials: M
authors.lastname: Singh
mesh_terms: D000064:Acclimatization / Q000187:drug effects / Q000235:genetics; D001593:Benzyl Compounds; D001709:Biotechnology / Q000379:methods*; D003080:Cold Temperature; D018744:DNA, Plant / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D005819:Genetic Markers / Q000235:genetics*; D007701:Kinetin; D009280:Naphthaleneacetic Acids / Q000494:pharmacology; D029803:Phyllanthus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D018517:Plant Roots / Q000187:drug effects / Q000254:growth & development; D018520:Plant Shoots / Q000187:drug effects / Q000254:growth & development; D011687:Purines; D012038:Regeneration / Q000187:drug effects / Q000235:genetics*
chemical_list: D001593:Benzyl Compounds; D018744:DNA, Plant; D005819:Genetic Markers; D009280:Naphthaleneacetic Acids; D011687:Purines; C480551:benzylaminopurine; D007701:Kinetin