Matrilineage differentiation of the genus Tetragonisca using mitochondrial DNA markers and the polymerase chain reaction-restriction fragment length polymorphism technique

Abstract

ABSTRACT. The Meliponinae are important pollinators of plant species, and one of the most managed species is Tetragonisca angustula. Initially, two subspecies were identified in T. angustula: T. angustula angustula and T. angustula fiebrigi. Subsequently, T. a. fiebrigi was considered a species, based on the coloration of its mesepisternum. The objective of the present study was to obtain genetic markers that could differentiate the two species by amplifying regions of mitochondrial DNA and conducting polymerase chain reaction-restriction fragment length polymorphism analysis. Worker bees were collected in three Brazilian states: Paraná (Maringá, Altônia, and Foz do Iguaçu), São Paulo (Dracena, São Carlos, and Santa Cruz do Rio Pardo), and Rondônia (Ariquemes). Ten pairs of insect heterologous primers were tested and four were used (primer pair 1, ND2 and COI; primer pair 2, COI; primer pair 8, 16S and 12S; and primer pair 9, COII). For the restriction analysis, 13 enzymes were tested: EcoRI, EcoRV, HindIII, HinfI, RsaI, PstI, XbaI, HaeIII, ClaI, XhoI, BglII, PvuII, and ScaI. Markers were obtained (primer pair 8 cleaved with EcoRV and XbaI and primer pair 9 cleaved with HaeIII, RsaI, and XbaI) that enabled matrilineage identification in the nests studied, which confirmed that hybridization could occur between both Tetragonisca species. The beginning of speciation was probably recent, and secondary contact has resulted in crosses between T. angustula females and T. fiebrigi males. Because of this hybridization, it would be appropriate to consider them as two subspecies of T. angustula.

Research Article Mitochondrial polymorphism in <i>Tetragonisca</i> <i>angustula</i> and <i>Tetragonisca</i> weyrauchi</i> (Apidae) in northern Brazil

“…The results of our study were consistent with those reported by Santos et al (2015), as the Tetragonisca samples amplified for the ND2, COI (primer 1) region, approximately 2,400 bp in size, and cleavage occurred using the same restriction enzymes (EcoRI and EcoRV) in T. angustula from the state of Rondônia, except for the cleavage using the RsaI enzyme, which occurred only in T. weyrauchi.…”

“…The 1,850-bp fragment resulted from the amplification of (COI) primer 2 region observed in T. angustula and T. weyrauchi was also observed by Santos et al (2015) in T. angustula and T. fiebrigi. However, these authors observed the cleaving of this region using two restriction enzymes (EcoRV and HinfI).…”

“…Using ND2, COI (primer 1) in T. angustula and T. fiebrigi collected in three Brazilian states, Santos et al (2015) observed polymorphism in samples of T. angustula from Porto Velho city (Rondônia State), where some individuals amplified a fragment 900 bp in size.…”

“…According to Santos et al (2015), only one queen originates all the workers present in the Tetragonisca nest. Therefore, no variation in worker mtDNA was detected.…”

Supernumerary B Chromosomes of <i>Tetragonisca fiebrigi</i> Share Repeat Content with Standard Chromosome Set of both <i>T. fiebrigi</i> and <i>Tetragonisca angustula</i> (Apidae: Meliponini)

“…While Tetragonisca angustula (Latreille, 1811) occurs in practically all Brazilian regions, Tetragonisca fiebrigi [Schwarz, 1938] has a more central-to-south distribution [Camargo and Pedro, 1836]. The taxonomic status of these two species is still discussed, and several authors have called attention to the possibility of hybridization between them Contel, 1995, 2005;Koling and Moretto, 2010;Francisco et al, 2014;Santos et al, 2015;Kulza et al, 2022].…”