Then the gonocoxites is laterally compressed, additionally the gonobase was retracted ( Figure 3I), increasing the angulation involving the set of manhood valves and you can ensuing regarding perforation towards membranous pockets, beneath the gonopore, and you can subsequent connection ( Figure 3J).
Immediately after affixed because of the pair of penis regulators, this new genital product was irreversibly decoupled away from male’s muscles, lodged when you look at the people vaginal chamber, and you may leaving precisely the gonostyli, obvious away from girls human anatomy ( Profile cuatro and you will Secondary Contour S2 ). This new mating plug is actually in the end affixed, a man forever will lose their genital pill, keeping unchanged within his human anatomy the latest sterna 
S-VIII and S-IX.
From inside the recently mated queens, gonostyli will always be establish and you can noticeable exterior their body and the gonocoxites harbor a large part of your seminal vesicles ( Supplementary Shape S2 ). In the long run, gonostyli try shed, such as for instance found in elderly M. fasciculata queens ( Shape step one), having mating plugs becoming visible to the women just lower than review out-of this new vaginal chamber ( Second Shape S1 ).
Repair away from mating pair interactions
Whenever a male in-flight finds out a lady worldwide ( Profile 4A), the guy mounts this new female’s system and you may begins moving their wings uninterruptedly since copulation begins ( Figure 4B). From the positioning himself totally along side female, a man securely changes per set of base at particular urban centers of queen’s muscles: (1) their forelegs anchors within the femurs of forelegs of your female; (2) their midlegs, within the trochanter of hindlegs of one’s girls; and you can (3) his hindlegs anchor beneath the idea off woman’s abdomen, thereby training it ( Shape 4C). A man cities the end out-of their gut on the woman’s gut, building a good “J,” moving it backward and up ( Profile 4C). Shortly after vaginal protrusion, and you may next mating plug creating, an effectively mated male departs their genital capsule into the female’s genital chamber, which is conveyed of the presence out-of an apparent set of gonostyli ( Profile 4D and Secondary Shape S2 ).
Discussion
The analysis shows that mating when you look at the eight Neotropical stingless bees are a harmful male–female correspondence, a traumatic mating system. According to (1) observations regarding mated and you will unmated lady vaginal morphology and muscle integrity, (2) the male genital procedure breakdown, and you will (3) the latest choices of your own mating partners, we understood you to mating plugs hop out attribute marks to the mated queens, that development and you will condition ones marks satisfy the mating connect morphology, and amount of scratching corresponds to new almost certainly level of queen mating people sensed in almost any variety confirmed by molecular investigation on books. Especially, our research implies that brand new molecular proof of multiple patrilines one of brood off M. seminigra ( Francini 2013) happens to be supported by facts with the quantity of mating sets portrayed by the amount of copulatory marks used in mated queens. Our results suggest that harrowing mating plugs may be pervading inside the stingless bees, but one to for example a process might not totally stop king remating in some species of this group. A button applying of all of our studies are that copulatory lesion activities is a reputable and you can reasonable-pricing diagnostic device to understand mating standards (age.grams., unmated, once mated, otherwise remated) off Meliponini queens.
In all seven studied species, mated queens shown melanized patches, whether or not virgins failed to ( Contour step one). This is actually the earliest proof of traumatic mating contained in this class, recommending male–lady harmful connections during the mating. Copulatory lesions due to penile spines regarding the ladies genital system was indeed before thought inside the ants (Atta sp. for the Baer and you can Boomsma 2006; Formica japonica in the Kamimura 2008). The latest pattern are also known as harrowing penetration, thus definition harmful penetration, but innocuous insemination ( Lange ainsi que al. 2013), that also pertains to this new stingless bees learnt here. This means that that injury to female can get simply result from an excellent solid real anchorage from male vaginal structures to make certain copulation. From inside the stingless bees, thicker mating swarms all are ( Imperatriz-Fonseca mais aussi al. 1998; Van Veen and you may Sommeijer 2000; Sommeijer et al. 2003; Koffler mais aussi al. 2016; Veiga mais aussi al. 2018), in which intercourse proportion is extremely skewed so you’re able to men, in particular regarding the low-Melipona species ( Boomsma ainsi que al. 2005; Koffler mais aussi al. 2016), hence implying one to opponent males you will definitely dislodge brand new mating couple. In this circumstances off extreme male–male competition, both men and women will benefit out-of piercing formations, therefore harrowing mating may also have an adaptive really worth, such improved paternity or fecundity pleasure ( Lange mais aussi al. 2013). Indeed, it’s been shown your technical arousal out-of mating plugs triggers ovarian activation into the about 1 types of stingless bee (Meters. quadrifasciata; Melo et al. 2001). However, whether natural or adaptive, negative effects of copulatory lesions considering traumatic mating plugs toward females reproductive exercise are entitled to after that studies.