Abstract
Molecular characterization of eleven clones of C. winterianus collected from eleven districts of West Bengal was carried out with 10 RAPD and 9 ISSR primers. The RAPD primers developed 81 robust loci, which revealed 74.07% polymorphism and 61 ISSR markers generated 47.50% polymorphism. Out of the different anchored primer combinations, ISSR primers with AG and GA motifs produced clear and maximum scorable markers, thus revealing a better coverage of the genome. Genetic diversity parameters (average and effective number of alleles, percent polymorphism, average heterozygosity, intralocus gene diversity, polymorphic information content (PIC)) for RAPD, ISSR, and RAPD+ISSR along with UPGMA clustering based on Jaccard's coefficient were estimated with a view to assess efficiency of the marker system in Cymbopogon. RAPD markers were more efficient than ISSR markers with regards to detection of polymorphism, number of loci scored and PIC values. However resolving power (Rp), Shannon's index, mean coefficient of gene differentiation (Gst), and gene flow estimates were better ISSR markers. Genetic variations detected among the geographically different populations of C. winterianus could be of much use for the introgression of new characters from wild counterparts to the cultivars, isolation of stable segregating markers, and selection of improved varieties and conservation of germplasm resources.
Highlights
The genus Cymbopogon possesses a large number of odoriferous species of the grass family (Poaceae), and is characterized by plants bearing aromatic essential oils in all parts
Considering the potentials of the DNA marker based genetic diversity analysis, the present study aimed to evaluate the congruency of molecular markers system viz. RAPD and Inter-simple sequence repeat (ISSR), in assessing and analyzing the nature and the extent of genetic diversity among the different elite germplasm of Cymbopogon winterianus collected from a narrow geographical region primarily from eleven different districts of West Bengal, India
Eleven germplasms of Cymbopogon winterianus collected from eleven districts of West Bengal, India were amplified with 10 RAPD and 9 ISSR (22 anchored & unanchored) markers to ascertain the level of genetic diversity within different germplasms different geographical regions (Table 2)
Summary
The genus Cymbopogon possesses a large number of odoriferous species of the grass family (Poaceae), and is characterized by plants bearing aromatic essential oils in all parts. These peculiarities of RAPDs impede direct estimations of allele frequency and can bias calculations of population differentiation (Neale and Harry, 1994) Another molecular system, Inter-simple sequence repeat (ISSR) markers, developed by Zietkiewicz et al (1994) based on the amplification of a single primer containing a microsatellite ‘core’ sequence anchored at the 5′or 3′ end by a set of 2–4 purine or pyrimidine residues and offers a high degree of reproducibility with the detection of rich level of polymorphism in a relatively simple procedure. Inter-simple sequence repeat (ISSR) markers, developed by Zietkiewicz et al (1994) based on the amplification of a single primer containing a microsatellite ‘core’ sequence anchored at the 5′or 3′ end by a set of 2–4 purine or pyrimidine residues and offers a high degree of reproducibility with the detection of rich level of polymorphism in a relatively simple procedure It has been widely used in assessments of genetic diversity (Bornet and Branchard, 2001) and cultivar identification (Prevost and Wilkinson, 1999)
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