RAPD-PCR in two species of Catopsilia (Pieridae, Lepidoptera)

Abstract

RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) technique was used to analyze the polymorphism among the individuals of the two species of butterflies, namely Catopsilia pyranthe and Catopsilia crocale, which were captured from their natural habitat. Three decanucleotide primers viz., primer A(AAGAGCCCGT), primer B(AACGCGCAAC) and primer C(CCCGTCAGCA) were used to amplify the gDNA from the adult individuals of both the species. A series of bands ranging from below 125 bp to 1500 bp were produced by these primers. Standard curves were prepared and used to calculate the number of base pairs present in all the amplified fragments. The results obtained with these primers revealed sufficient amount of relatedness and variations in these two species. Primer A amplified some common DNA fragments of the same size in both the species, revealing the presence of conserved regions and thereby indicating their ancestral relationship. Some of the fragments were unique to the individuals of both the species which could be used to generate their diagnostic profiles. Primer B produced single bands but of different sizes in the invididuals of both the species revealing the presence of intraspecific genetic variation. Primer C did not amplify the gDNA of the individuals of C.pyranthe but produced 4 bands ranging from 480 to 1160 bp length in C.crocale. These results indicate that the RAPD-PCR technique is useful in the studies on molecular taxonomy of Lepidoptera.

Key words:

• Catopsilia
• RAPD
• PCR
• Primers