Kevin Maebe

16S rRNA Amplicon Sequencing Demonstrates that Indoor-Reared Bumblebees (Bombus terrestris) Harbor a Core Subset of Bacteria Normally Associated with the Wild Host

A MiSeq multiplexed 16S rRNA amplicon sequencing of the gut microbiota of wild and indoor-reared Bombus terrestris (bumblebees) confirmed the presence of a core set of bacteria, which consisted of Neisseriaceae (Snodgrassella), Orbaceae (Gilliamella), Lactobacillaceae (Lactobacillus), and Bifidobacteriaceae (Bifidobacterium). In wild B. terrestris we detected several non-core bacteria having a more variable prevalence. Although Enterobacteriaceae are unreported by non next-generation sequencing studies, it can become a dominant gut resident. Furthermore the presence of some non-core lactobacilli were associated with the relative abundance of bifidobacteria. This association was not observed in indoor-reared bumblebees lacking the non-core bacteria, but having a more standardized microbiota compared to their wild counterparts. The impact of the bottleneck microbiota of indoor-reared bumblebees when they are used in the field for pollination purpose is discussed.

Discrimination of haploid and diploid males of Bombus terrestris (Hymenoptera; Apidae) based on wing shape

Presence of diploid males in wild bees reflects inbreeding and provides information about the health status of a colony or population. Detection of diploid males, and discrimination from haploid males and workers, has, however, been limited to molecular diagnostics. Here we present a novel method based on differences in wing shape, e.g., venation patterns in wings. The method is easy to apply and results, for Bombus terrestris, in very high discrimination success. Possible applications of the method are discussed.

Microsatellite Analysis of Museum Specimens Reveals Historical Differences in Genetic Diversity between Declining and More Stable Bombus Species

Worldwide most pollinators, e.g. bumblebees, are undergoing global declines. Loss of genetic diversity can play an essential role in these observed declines. In this paper, we investigated the level of genetic diversity of seven declining Bombus species and four more stable species with the use of microsatellite loci. Hereto we genotyped a unique collection of museum specimens. Specimens were collected between 1918 and 1926, in 6 provinces of the Netherlands which allowed us to make interspecific comparisons of genetic diversity. For the stable species B. pascuorum, we also selected populations from two additional time periods: 1949–1955 and 1975–1990. The genetic diversity and population structure in B. pascuorum remained constant over the three time periods. However, populations of declining bumblebee species showed a significantly lower genetic diversity than co-occurring stable species before their major declines. This historical difference indicates that the repeatedly observed reduced genetic diversity in recent populations of declining bumblebee species is not caused solely by the decline itself. The historically low genetic diversity in the declined species may be due to the fact that these species were already rare, making them more vulnerable to the major drivers of bumblebee decline.

Molecular characterization of Vietnamese cocoa genotypes (Theobroma cacao L.) using microsatellite markers

Vietnam has the appropriate climate, soil, and humidity for cocoa cultivation and is growing as a cocoa-producing country. To supply the international cocoa market, trees have been planted in Southern Vietnam. Cocoa quality depends on various factors, such as the genotype/cultivar, environment, and post-harvest processing. Until now, little research has been done on the genetic background of Vietnamese cocoa. Therefore, this study focused on the genetic relationships of 75 cocoa cultivars, sampled in Vietnam. Fourteen microsatellite markers were used to assay the genetic diversity and to genotype and differentiate the accessions. Descriptive statistics showed that most of the used microsatellite markers were sufficient to be used in further analysis, of which mTcCIR 15, 33, and 37 were the most polymorphic. The principal coordinate analysis (PCoA) and Bayesian clustering approach divided the samples in two groups, which were linked to the Trinitario and Forastero varieties. The Vietnamese Can Tho (CT) cultivars showed little variation, while the Thu Duc (TD) cultivars showed a continuous variation between the different reference cultivars. This indicates that the CT cultivars were Trinitario and that the TD cultivars were hybrids of Forastero or Forastero and Trinitario. The molecular characterization is an important step towards developing a strong genetic basis for the Vietnamese cocoa industry. It can be used to conserve valuable genetic material and to select promising cocoa cultivars which are disease resistant, high yielding, and fine flavored. In this way, the high-quality Vietnamese cocoa production can be improved and maintained.

Chemical reproductive traits of diploid Bombus terrestris males: Consequences on bumblebee conservation

The current bumblebee decline leads to inbreeding in populations that fosters a loss of allelic diversity and diploid male production. As diploid males are viable and their offspring are sterile, bumblebee populations can quickly fall in a vortex of extinction. In this article, we investigate for the first time a potential premating mechanism through a major chemical reproductive trait (male cephalic labial gland secretions) that could prevent monandrous virgin queens from mating with diploid males. We focus our study on the cephalic labial gland secretions of diploid and haploid males of Bombus terrestris (L.). Contrary to initial expectations, our results do not show any significant differentiation of cephalic labial gland secretions between diploid and haploid specimens. Queens seem therefore to be unable to avoid mating with diploid males based on their compositions of cephalic labial gland secretions. This suggests that the vortex of extinction of diploid males could not be stopped through premating avoidance based on the cephalic labial gland secretions but other mechanisms could avoid mating between diploid males and queens.

A century of temporal stability of genetic diversity in wild bumblebees.

Since the 1950s, bumblebee (Bombus) species are showing a clear decline worldwide. Although many plausible drivers have been hypothesized, the cause(s) of this phenomenon remain debated. Here, genetic diversity in recent versus historical populations of bumblebee species was investigated by selecting four currently restricted and four currently widespread species. Specimens from five locations in Belgium were genotyped at 16 microsatellite loci, comparing historical specimens (1913–1915) with recent ones (2013–2015). Surprisingly, our results showed temporal stability of genetic diversity in the restricted species. Furthermore, both historical and recent populations of restricted species showed a significantly lower genetic diversity than found in populations of co-occurring widespread species. The difference in genetic diversity between species was thus already present before the alleged recent drivers of bumblebee decline could have acted (from the 1950’s). These results suggest that the alleged drivers are not directly linked with the genetic variation of currently declining bumblebee populations. A future sampling in the entire distribution range of these species will infer if the observed link between low genetic diversity and population distribution on the Belgium scale correlates with species decline on a global scale.

Molecular Variability and Genetic Structure of Chrysodeixis includens (Lepidoptera: Noctuidae), an Important Soybean Defoliator in Brazil

This study provides the first genetic characterization of the soybean looper, Chrysodeixis includens (Walker, 1857), an important defoliating pest species of soybean crops in Brazil. Population genetic variability and the genetic structure of C. includens populations were evaluated by using ISSR markers with samples from the major soybean producing regions in Brazil in the growing seasons 2011/2012. Seven different primers were applied for population characterization of the molecular variability and genetic structure of 8 soybean looper populations from 8 states of Brazil. The seven ISSR loci generated 247 bands in 246 individuals of C. includens sampled. The expected heterozygosity (H E) in the populations varied between 0.093 and 0.106, while the overall H E was 0.099, indicating low genetic diversity. The analysis of molecular variance indicated that 98% of the variability was expressed among individuals within populations (F ST = 0.021, p = 0.001). The low level of polymorphism over all populations, the high levels of gene flow, and the low genetic structure are indicatives of the exchange of genetic information between the different sampled regions. Population structuring suggests the presence of two major groups which do not correlate with their geographic sampling location in Brazil. These results may indicate recent recolonization of C. includens in Brazil or migration patterns following source-sink dynamics. Furthermore, the presence of two groups within C. includens suggests that a study on development of resistance or any other genetic-based trait needs to be evaluated on both groups, and pest management in soybean fields should be aware that differences may come to the control strategies they use.

Temporal changes in genetic variability in three bumblebee species from Rio Grande do Sul, South Brazil

In order to protect biodiversity, conservation genetics are of great importance. Until now, a few population genetic studies of Neotropical bumblebees are available but studies of temporal stability in genetic diversity are lacking. Here, genetic variability of three South Brazilian species Bombus bellicosus, B. pauloensis, and B. morio was investigated over time. Hereto, museum collection specimens of 1946 until 2012, from eight locations in Rio Grande do Sul, were genotyped at 16 microsatellite loci. During an additional sampling in the foraging season of 2015, no bumblebees could be collected possibly due to the super El Niño of 2015–2016. Our results on the collection specimens demonstrated a significantly higher genetic diversity in B. morio than in B. pauloensis. Genetic variability in B. pauloensis gradually and significantly decreased over time from 1946 until 2012; while in B. morio, genetic variability remained stable until the last time period (2010–2012). For B. bellicosus, not enough data was available. Although the studied populations became more vulnerable over time, for the conservation of Neotropical bumblebees, still more information is needed and could include more frequent monitoring of bumblebees. Adding B. bellicosus to the Brazilian list of threatened species is suggested.

Quantitative trait loci for light sensitivity, body weight, body size, and morphological eye parameters in the bumblebee, Bombus terrestris

Bumblebees such as Bombus terrestris are essential pollinators in natural and managed ecosystems. In addition, this species is intensively used in agriculture for its pollination services, for instance in tomato and pepper greenhouses. Here we performed a quantitative trait loci (QTL) analysis on B. terrestris using 136 microsatellite DNA markers to identify genes linked with 20 traits including light sensitivity, body size and mass, and eye and hind leg measures. By composite interval mapping (IM), we found 83 and 34 suggestive QTLs for 19 of the 20 traits at the linkage group wide significance levels of p = 0.05 and 0.01, respectively. Furthermore, we also found five significant QTLs at the genome wide significant level of p = 0.05. Individual QTLs accounted for 7.5-53.3% of the phenotypic variation. For 15 traits, at least one QTL was confirmed with multiple QTL model mapping. Multivariate principal components analysis confirmed 11 univariate suggestive QTLs but revealed three suggestive QTLs not identified by the individual traits. We also identified several candidate genes linked with light sensitivity, in particular the Phosrestin-1-like gene is a primary candidate for its phototransduction function. In conclusion, we believe that the suggestive and significant QTLs, and markers identified here, can be of use in marker-assisted breeding to improve selection towards light sensitive bumblebees, and thus also the pollination service of bumblebees.