ISSR and RAPD based evaluation of genetic stability of encapsulated micro shoots of Glycyrrhiza glabra following 6 months of storage.
In vitro grown axillary micro shoots of Glycyrrhiza glabra were encapsulated in alginate beads. Following 6 months of normal storage at 25 ± 2°C the re growth of encapsulated G. glabra micro shoots, reached 98% within 30 days of incubation on MS medium supplemented with 0.1 mg/l IAA. Re growth was characterized by the development of both shoot and root from single encapsulated micro shoot. Healthy plants were established to glass house with 95% survival. The genetic fidelity of plants obtained after conversion of alginate beads was ascertained through 10 RAPD and 13 ISSR primers. Of the 10 RAPD primers tested, 6 of them produced 14 clear and reproducible amplicons with an average of 2.3 bands per primer out of which 28.57% were polymorphic generated by only two primers. Eight ISSR primers produced total 37 bands ranging between 300 and 3,500 bp length. Number of scorable bands for each primer varied from 3 to 8 with an average of 4.6 bands per primer. Cluster analysis from ISSR and RAPD showed that all the tested plants including the mother plant distributed in two major groups with similarity coefficient ranging from 0.91 to 0.96 for RAPD and 0.89 to 0.97 for ISSR.
Plant Biotechnology Division, Central Institute of Medicinal and Aromatic Plants, Picnic Spot Road, P.O. CIMAP, Lucknow 226015, India. shaktimehrotra22@gmail.com
Shakti
S
Mehrotra
O
O
Khwaja
A K
AK
Kukreja
L
L
Rahman
D000464:Alginates / Q000737:chemistry; D018914:Cells, Immobilized; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D020723:Glucuronic Acid / Q000737:chemistry; D006035:Glycyrrhiza / Q000235:genetics*; D006603:Hexuronic Acids / Q000737:chemistry; D018895:Microsatellite Repeats; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA
D000464:Alginates; D017931:DNA Primers; D018744:DNA, Plant; D006603:Hexuronic Acids; D020723:Glucuronic Acid