Molecular Characterization

With the advent of molecular biology techniques, DNA-based markers very efficiently augment morphological, cytological, and biochemical characters in germplasm characterization, varietal identification, clonal fidelity testing, assessment of genetic diversity, validation of genetic relationship, phylogenetic and evolutionary studies, marker-assisted selection, and gene tagging. Owing to plasticity, ubiquity, and stability, DNA markers are easier, efficient, and less time consuming, especially in perennials where morphological markers are few. The relatively easy to use, low-cost, and highly accurate nature of the polymerase chain reaction (PCR)—based technologies such as RAPD, AFLP, and microsatellites are widely appreciated.

RAPD Profiling of Ginger Cultivars

Genomic DNA was successfully isolated in the authors' laboratory from young and fresh leaves of ginger plants using the modified cetyl amino butane (CTAB) method of Ausubel et al. (1995). Development of RAPD profiles for various ginger cultivars (see Figure 4.4a) and related species is in progress at the Indian Institute of Spices Research to study the interrelationships and to identify the core collections in the germplasm.

Figure 4.4 Molecular characterization of ginger: (a) RAPD profiles as expressed by OPERON primer OPD 11 among different cultivars of ginger collected from different geographical regions. lanes: 1 1 kb ladder, 2 Maran, 3 Australia, 4 Jamaica, 5 Gurubathani, 6 Bhainse, 7 RRE, 8 Hawaiian, 9 Sabarimala, 10 Mananthody, 11 Anamica, 12 Bisonvally, 13 China, 14 Jugijan, 15 IISR Rajitha, 16 IISR Varada, 17 Wyanad local, 18 Nadan, 19 PGS 43, 20 V2-E5-3. (b) RAPD profiles as expressed by OPERON primer OPE 01 in micropropagated plants of ginger: lanes: 1. 1 Kb Ladder, 2 RRE 1, 3 M Control, 4 MP 49-7, 5 MP 50-3, 6 MP 54-2, 7 MP 54-4, 8 MP 61-4, 9 MP 61-9, 10 MP 61-10, 11 MP 64-3, 12 MP 68-4, 13 MP 70-3, 14 MP 70-5, 15 MP 74-4, 16 MP 76-3. (c) RAPD profiles as expressed by OPERON primer OPD 10 in callus regenerated plants of ginger: lanes: 1 1 Kb Ladder, 2 RRE 1, 3 M Control, 4 CR 3, 5 CR 4, 6 CR 7, 7 CR 9, 8 CR 10-1, 9 CR 13, 10 CR 20, 11 CR 42, 12 CR 64, 13 CR 67, 14 CR 816, 15 CR 817, 16 CR 1222, 17 CR 1441, 18 AC-1. (d) RAPD profiles of microrhizome derived plants of ginger, as expressed by OPERON primer OPB 10: L—1kb ladder, lanes 1, 2, 3, 4, 5, 6, 7, 8: microrhizome derived plants, C—control.

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