ABSTRACT. In this study, we report a modified CTAB-PVP method combined with silicon dioxide (silica) treatment for the extraction of high quality genomic DNA from a single larva or pupa. This method efficiently obtains DNA from small specimens, which is difficult and challenging because of the small amount of starting tissue. Maceration with liquid nitrogen, phenol treatment, and the ethanol precipitation step are eliminated using this methodology. The A 260 /A 280 absorbance ratios of the isolated DNA were approximately 1.8, suggesting that the DNA is pure and can be used for further molecular analysis. The quality of the isolated DNA permits molecular applications and represents a fast, cheap, and effective alternative method for laboratories with low budgets.

Macaria mirthae Vargas et al (Lepidoptera: Geometridae) is a geometrid moth native to the northern Atacama Desert of Chile. Its oligophagous larvae are associated with native hosts of the plant family Fabaceae, the most important of which is Acacia macracantha. The invasive tree Leucaena leucocephala (Fabaceae) was recently recorded as a host plant for M. mirthae based on morphology. The taxonomic status of larvae collected on A. macracantha and L. leucocephala was assessed using sequences of the DNA barcode fragment of the cytochrome c oxidase subunit I (COI) gene. Genetic divergence between samples from the host plants was found to be 0%-0.8% (Kimura 2-parameter model). Neighbor-joining and maximum likelihood analyses were also performed, including additional barcode sequences of Neotropical geometrid moths from GenBank and BOLD databases. Sequences of the larvae from both host plants clustered in a single clade with high statistical support in both analyses. Based on these results, it is concluded that M. mirthae has effectively expanded its host range and its larvae are currently feeding on the exotic tree L. leucocephala. Additionally, the importance of this new host association in a highly disturbed habitat is briefly discussed in terms of the field biology of this native geometrid moth.