Molecular differentiation of Dichelops melacanthus (Hemiptera: Pentatomidae) and D. furcatus (Hemiptera: Pentatomidae) from the Southern Neotropical Region

M. GABRIELA MURÚA; DE OLIVEIRA A. G.; GÓMEZ, DANIEL R. SOSA

Jornada; XVI JORNADAS FITOSANITARIAS ARGENTINAS; 2018

The identification of Dichelops species is the initial step in biology and behavioral studies, as well as, in the practical point of view to adopt control measures, since species may differ in their susceptibility to insecticides and biological control agents. Morphological studies of Dichelops eggs features with scanning electron microscopy did not provide clear differentiation between both species. Therefore, a molecular diagnostic method using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis was developed to distinguish Dichelops species commonly found in the Southern Neotropical region. A 658-bp region of the mtDNA COI gene was amplified using PCR and then sequenced. The pentatomid species were distinguished by PCR-RFLPs analysis using restriction enzymes BspMI and Vsp I. Bsp MI did not digest the amplicons from D. furcatus, but digested D. melacanthus amplicons producing to bands of 465 and 193 bps. RFLP patterns produced by the restriction enzyme Vsp I digested the PCR products of both species with two, clear patterns, two bands of 288 and 370 bps when the template DNA was from D. melacanthus and two bands of 197 and 461 bps, when the DNA was from D. furcatus. PCR-RFLP can be performed in a short period and it is useful to distinguish one of the most important pentatomids of corn and wheat pests.