3. Results
S.K. Sharma et al. / Biochemical Systematics and Ecology 39 (2011) 9–13 11
Eight ISSR and five DAMD primers (Table 2) were selected for final amplification programme as others produced sub- optimal, indistinct or monomorphic amplification products. The amplified DNA polymorphic fragments were scored as binary matrix for presence (1) and absence (0), and the data matrix of ISSR and DAMD phenotypes was assembled for further analysis. A total of 83 bands were generated from eight ISSR and five DAMD primers of which 79 were found to be poly- morphic (95.18%) (Table 3). Present investigation revealed that observed and effective number of alleles ranged between 1.470–1.734 and 1.322–1.536, respectively. Similarly, Nei’s gene diversity (h) and Shannon’s information index (I) ranged between 0.185 and 0.301 with overall diversity of 0.316 and 0.253–0.438 with an average value of 0.243, respectively. These values revealed that the population of district Barmer showed the highest variability whereas district Nagaur showed the lowest diversification. Percentage of polymorphic loci (Pp) was estimated in the range of 49.99–73.49 with a relatively high mean value of 95.18. The present study showed that P. cineraria is more diverse at intra-populational level with high percentage of variation (88.2%) than at inter-populational level with comparatively low variation (11.8%). The gene flow value (1.713) and the diversity among population (0.226) demonstrated elevate genetic variation within the population thereby all population can be predicted to be in Hardy–Weinberg proportions (Table 3).
AMOVA was performed at three hierarchical levels to examine differences among and within populations of five districts of Rajasthan. The significance of this analog was evaluated by 1000 random permutations of sequences among populations. The result of AMOVA with a high significant value (P < 0.001) showed that most of the genetic variation (88.2%) observed within the populations, whereas the variance among populations was 11.8% only. Relationships of populations were esti- mated from ISSR and DAMD data using UPGMA clustering methods on the basis of Nei’s (1978) unbiased genetic diversity. Dendrogram (Fig. 1) revealed that populations from Barmer, Nagaur and Bikaner are closely related, while population from Jaisalmer and Jodhpur showed the most diverse populations.