Performance
To help you picture copulatory marks and also the male genital morphology of each varieties, we utilized men and women genitalia thanks to dissections. For this purpose, i tested most queens (both virgin and you can physogastric mated of these) and additional recently came up boys of every kinds, after the exact same testing standards in earlier times described (comprehend the earlier sections). We after that dissected for each specimens’ metasoma to access the fresh genital area of females and you will guys. Shortly after dissection, we quickly shoot this type of structures so that the ethics from membranous structures, especially in females. Dissections observed the method previously explained inside Vaginal useful morphology, hypothesized plugging strategies and you may mating positions. Fundamentally, we utilized a good Leica DM 1000 stereomicroscope, coupled with an attracting tubing to arrange the layouts of one’s mating few (?20 magnification, Genital useful morphology, hypothesized plugging measures and you can mating ranks) and also the vaginal useful morphology (?80 magnification, Genital functional morphology, hypothesized plugging tips and you can mating ranking). We vectorized and you will handled these artwork using an image publisher software. We along with made use of an effective Leica DFC420 camera connected to good Leica M205A stereomicroscope to make large-quality pictures of women and you may guys genitalia (?step one.0 contact and you may ?fifty magnification, Section dos.4). First, i grabbed multifocus color photographs, after that i stacked them in one photo by using the Leica application package variation step three.cuatro.step 1. We regular this procedure for every single genitalia (male and female) regarding every sampled stingless bee variety, but that, Meters. melapling constraints.
Harrowing mating from inside the stingless bees
Zero melanized spots was in fact recognized into the checked unmated virgin queens. In all variety, physogastric queens shown melanized patches on their membranous purse ( Dining table step one), apart from Yards. fasciculata ( Contour 1). The fresh membranous pouches was in fact the only part that have copulatory scratches. When you look at the for each and every mated queen, patches have been constantly matched up, coordinating the latest inferred reputation of collection of penis regulators in the a man vaginal product. Spots had been situated on for each pouch, and symmetrically arranged ( Contour step 1). As well, melanized spots looked like long lasting, because we seen them from inside the mated queens of different decades, predicated on all of our wing don category ( Secondary Shape S3 ).
Female genital chamber, trend away from copulatory marks, and you will ventral view of men genital tablets regarding F. longipes, P. minima, Scaptotrigona aff. postica, Melipona flavolineata, Yards. seminigra, and you can Meters. fasciculata. Of remaining in order to proper, photographs reveal the latest vaginal chamber regarding virgin queens, the vaginal chamber of mated queens, together with melanized spots entirely on queens, being indicated of the white arrows about 3rd column regarding photos. Singly mated queens tell you an individual group of melanized spots, found to have F. longipes, P. minima, S. aff. postica, Yards. flavolineata, and you can Meters. seminigra; or an extended-name connected connect, simply utilized in M. fasciculata. Remated M. seminigra queens, shown from the black asterisk, let you know several sets out-of melanized patches and main patches. On the right-hand group of images, men drugs are shown once the triggered mating plugs only for Melipona variety, proving rotated knob valves. We 
plus inform you a keen inactivated mating plug to possess Yards. fasciculata, since indicated of the light asterisk. Black taverns depict a size from 0.5 mm. That it figure comes with photos from 6 regarding our eight learnt types (select Matter and methods).
Women genital chamber, pattern from copulatory scratches, and you will ventral view of men vaginal capsules from F. longipes, P. minima, Scaptotrigona aff. postica, Melipona flavolineata, Yards. seminigra, and you will Yards. fasciculata. Out-of leftover to help you correct, photo inform you new vaginal chamber away from virgin queens, this new vaginal chamber off mated queens, additionally the melanized patches found on queens, which are shown of the white arrows on third line regarding images. Singly mated queens reveal a single collection of melanized patches, revealed to own F. longipes, P. minima, S. aff. postica, Yards. flavolineata, and you may Yards. seminigra; or an extended-label connected plug, only utilized in Yards. fasciculata. Remated M. seminigra queens, indicated of the black asterisk, let you know numerous pairs away from melanized spots and you may central patches. On right-hand selection of images, men drugs are presented since the triggered mating plugs only for Melipona kinds, demonstrating turned dick regulators. We and additionally reveal an inactivated mating connect getting M. fasciculata, since shown from the white asterisk. Black colored pubs portray a size out-of 0.5 mm. Which profile comes with pictures away from six of the seven examined types (pick Situation and techniques).