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20 Hairul Azman ROSLAN et al. Figure 7. Clade 6 Penicillium P6 isolate. (a) Colony surface on CYA, (b) Colony reverse on CYA, (c) Colony surface on MEA, (d) Colony reverse on MEA, (e) Conidiophore structure at 40X magnification (Terverticillata), (f) Conidia elliptical shape. Molecular groupings of Penicillium isolates Single, simple repetitive PCR primers have been designed to amplify the microsatelite regions of chromosomal DNA. In most applications these primers have provided similar levels of specificity to those seen with RAPD, and the results have been used to group fungi at species level (Meyer et al., 1992; Schlick et al., 1994; Bridge et al., 1997). Two sets of primers were used, M13 and OPD-10 primers to analyse the variations between the 20 isolates of Penicillium spp. Six isolates were excluded from the molecular study because either the DNA could not be isolated or amplification was not reproducible. Each sample was repeated at least two times to determine its reproducibility and consistency. Bands were scored for each primer based on the presence (1) or absence (0) of amplicon migration in the gel. Figure 8 and Figure 9 represent the RAPD profile of M13 and OPD-10 respectively. Figure 10 is a dendogram generated from M13 data.
Genetic diversity of Penicillium species in Sawarak, Malaysia 21 Figure 8. RAPD band profile generated using M13 primer visualized on 1.3% (v/v) agarose. The lane markings correspond to the isolate number. Lane M: 1kbp DNA ladder (Fermentas) and Lane N: 100bp DNA ladder (Seegene) Figure 9. RAPD band profile generated using OPD10 primer visualized on 1.3% (v/v) agarose gel. The lane markings correspond to the isolate number. Lane M: 1kbp DNA ladder (Fermentas) and Lane N: 100bp DNA ladder (Seegene)
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