1. Introduction
Tephritidae family includes fruit flies of major economic importance worldwide [
1,
2,
3]. They are among the main insect pests of horticulture due to their life cycle being dependent on host plants, which provide breeding grounds and sources of nutrients for the development of the larvae [
2,
4,
5]. They are responsible for huge economic losses in the countries affected, due to crop losses [
1,
6]. The most damaging fruit fly species belong to the following genera:
Anastrepha,
Bactrocera,
Ceratitis,
Dacus,
Rhagoletis and
Zeugodacus [
3]. The genus
Bactrocera comprises around 460 species organized into subgroups, including the
Bactrocera dorsalis complex [
7,
8].
The
Bactrocera dorsalis s.l. complex comprises around a hundred morphologically similar species. Most are economically less important, with the exception of
B. dorsalis (Hendel), commonly known as the oriental fruit fly.
Bactrocera dorsalis (Hendel) and its synonymous species such as
B. papayae Drew & Hancock,
B. philippinensis Drew & Hancock and
B. invadens Drew, Tsuruta & White, are notorious insect pests, highly polyphagous, with over 250 host plant species, and extremely invasive. They are particularly fecund and highly resilient to environmental stresses [
3,
5,
9,
10].
Bactrocera dorsalis (Hendel) has a worldwide distribution. Its presence in Africa was detected in Kenya in 2003 and it was first described as
B. invadens [
11], a species closely related to
B. dorsalis, as were
B. philippinensis, described in the Philippines, and
B. papayae, described in Thailand, Malaysia and Indonesia [
12,
13,
14].
B. dorsalis has now spread throughout sub-Saharan Africa, where its capacity for invasion and destruction has made it the main insect pest of the horticultural sector, with damage estimated at nearly US
$2 billion per year [
15,
16,
17,
18].
In Burkna Faso,
B. dorsalis was detected for the first time in 2005 [
1,
15,
19]. It is now the main problem in the fruit sector, affecting a dozen cultivated and/or wild host plants, the main ones being mango (
Mangifera indica) and shea (
Vitellaria paradoxa). The damage to mangoes is much more significant, with crop losses of up to 100% in some orchards, and quarantine measures affecting the export of fruit from infested countries, contributing to significant economic losses [
1,
19].
The control of
B. dorsalis is essentially based on the use of chemical pesticides, which unfortunately pollute the environment, affecting the food chain and the well-being of humans. Furthermore, these pesticides are nowadays inefficient due to resistance problems developed by insect pests [
16,
18]. Eco-friendly techniques such as sterile insect techniques (SIT), entomopathogenic fungi, parasitoids and predatory ants have been adopted in control programs [
16,
20,
21]. Unfortunately, these resources are limited by their cost, accessibility and/or the complexity of their implementation.
Recent advances in genetic engineering have led to the emergence of innovative tools for controlling insect pests. Genetic control appears to be a promising technology, based on the introduction of genetic traits aimed to interfere with the ability of insects to develop or reproduce thereby suppressing their populations [
22]. These new technologies are potentially more species-specific, accessible, non-polluting, easy to deploy and applicable on a larger scale compared to existing methods [
23,
24]. However, effective control of insect pests, especially through genetic control strategies, would require an understanding of genetic diversity, genetic structure and gene flow within target populations [
23,
25,
26].
Microsatellites are genetic markers of nuclear origin that are non-coding, codominant and highly polymorphic. They are composed of short nucleotide sequences repeated in tandem, distributed throughout the eukaryotic genome, and are powerful tools for the genetic analysis of natural populations [
20,
27]. In this study, we exploited the potential of microsatellite markers to elucidate the genetic diversity, gene flow and genetic structure of
B. dorsalis populations in Burkina Faso.
4. Discussion
Bactrocera dorsalis is currently a serious threat for horticulture in sub-Saharan Africa. Clearly, effective control of this insect pest would require a good understanding of its ecology, biology, population genetics and other relevant parameters. In sub-Saharan Africa, despite efforts, much remains to be done in the investigation of this insect pest. Several studies were carried out to inventory the main and intermediate host plants of
B. dorsalis among cultivated and wild plants, both commercial and non-commercial or to elucidate the seasonal abundance and population dynamics of this insect pest [
14,
39]. However, data are lacking on a number of parameters, including the distribution of insecticide resistance and genetic diversity and structuring. An initial study by Khamis et al. (2009) looked at the diversity and genetic structuring of
B. dorsalis populations in West Africa, Central Africa, East Africa and Sri Lanka [
30]. Another study by Qin et al. (2018) gave a global overview of the different structures of
B. dorsalis across locations in Africa, Asia and Hawaii. In Africa, 11 countries including Burkina Faso were involved [
5]. Two other studies by Faye et al. (2020) and Diallo et al. (2021) pointed out the genetic structuring of
B. dorsalis in certain regions of Senegal [
40,
41]. This study is the first to provide a specific overview of the genetic structuring of
B. dorsalis in Burkina Faso.
The results of transversal collections of
B. dorsalis scales during July 2021 across three types of plant formations in seven sampling sites in Burkina Faso showed that the highest proportion of flies was observed in Kénédougou while the lowest proportion was observed in Sissili. The month of July was indicated as the outbreak period for
B. dorsalis in the west part of Burkina Faso, and this is thought to be mainly influenced by the availability of host fruits and the suitability of climatic conditions with a favorable atmospheric humidity and temperature [
1,
42]. Moreover, a study in Benin showed that the abundance of
B. dorsalis was strongly correlated with relative humidity and rainfall [
43]. Therefore, the uneven distribution of rainfall would be a factor influencing the distribution of
B. dorsalis in Burkina Faso. The results showed that the most watered areas, such as Comoé, Bougouriba, Kénédougou, Houet and Poni in the Sudanian climate, had higher proportions of flies, while less irrigated areas such as Boulkiemdé-Sanguié and Sissili in the Sudano-Sahelian climate, had lower proportions (
Figure 2). Of the three types of plant formations surveyed, the results showed that the high proportion of flies was observed in mango orchards, while the low proportion was observed in natural formations. In Burkina Faso, the natural formations are composed of plants species such as
Annona senegalensis,
Landolphia heudelotii,
Opilia celtidifolia,
Sarcocephalus latifolius,
Uvaria chamae, etc., which are secondary hosts for
B. dorsalis, whereas mangoes (
Mangifera indica) are its preferred host fruits [
43]. Furthermore, this knowledge of differential distribution of
B. dorsalis abundance in Burkina Faso would make it possible to better guide the implementation of the various control interventions for greater impact at a national scale.
Analysis of the genetic diversity of
Bactrocera dorsalis in Burkina Faso through polymorphisms of the 10 microsatellite loci showed a high genetic diversity in all subpopulations with an average Shannon Information Index (I) of 0.722. The highest genetic diversity was observed in Comoé (I=0.771) while the lowest was observed in Sissili (I=0.657). The same finding was reported by Khamis et al. (2009), who observed similarly high genetic diversity in West Africa (0.59), and this diversity was also slightly higher than that observed in East Africa (0.54) [
30]. Qin et al. (2018) also observed high genetic diversity (0.55) in
B. dorsalis populations in certain African localities, including Burkina Faso [
5]. These results show that
B. dorsalis has successfully adapted in West Africa, and populations are expanding after more than fifteen years of evolution since its invasion around 2005 [
1,
15,
19,
30,
44]. Indeed, the West African region is a major fruit production and export area, more specifically for mango, which is a key host for
B. dorsalis. In addition,
B. dorsalis is also known to have a very high capacity for invasion and adaptation [
10,
30]. Fixation indices (F) in the different subpopulations ranged from 0.062 (Boulkiemdé-Sanguié) to 0.269 (Comoé) with an average of 0.166, showing that all seven
B. dorsalis subpopulations deviated from the Hardy-Weinberg equilibrium with heterozygote deficiencies. The Hardy-Weinberg equilibrium is based on a population model with an infinite size where reproduction is sexual and panmictic, there is no overlapping of generations, there is no mutation, no selection and no migration [
45].
B. dorsalis is a highly fertile sexual species in which males and females mate with several partners. Females reproduce several times during their lifetime, lasting around three months. The flies reach sexual maturity in around 30 days after undergoing successive embryonic, larval, pupal and imaginal development, giving rise to overlapping generations of flies. The species is also known for its great dispersal capacity, often facilitated by commercial trade in fruit [
18]. Populations of
B. dorsalis therefore deviate from the Hardy-Weinberg model because of the overlapping generations and migrations that characterize them. The indices of genetic differentiation (Fst) varying from 0.009 (Comoé-Poni) to 0.023 (Kénédougou-Sissili) observed between the sub-populations were low according to Wright's scale, which considers that an Fst between 0 and 0.05 indicates low genetic differentiation [
46]. These low genetic differentiations led to high gene flows, varying from 10.62 between Kénédougou and Sissili, to 27.53 between Comoé and Poni. In fact, gene flow is a factor in the adaptation and persistence of the species in new environments through the dispersal of selected resilience traits such as heat or cold resistance alleles and insecticide resistance alleles [
23,
46]. In addition, local gene flow is a crucial advantage for the success of a genetic control intervention by facilitating the dispersal of the gene of interest [
23]. However, it should also be noted that the observed gene flow could constitute an additional complication in the choice of trial sites for a potential evaluation of a genetically modified strain in the natural environment. Indeed, trial sites with a good level of containment, exchanging fewer or no individuals with the surrounding sites, are ideal candidates. These sites offer settings that minimize the risk of invasion of genetically modified insects into non-target sites and maximize the chances of success by preventing bias in the experiment through potential invasions of surrounding wild strains into the trial sites [
48]. The genetic differentiations observed were correlated with the Nei's genetic distance observed between the sub-populations, ranging from 0.04 between Kénédougou and Sissili to 0.009 between Comoé and Poni. The high flight capacity and variety of host fruits of
B. dorsalis would facilitate its dispersal from one site to another [
30]. These migrations help to reduce the genetic difference and/or the genetic distance between the different populations.
Bayesian admixture analysis in STRUCTURE showed that the
B. dorsalis populations in Burkina Faso were organized into three genetic groups derived from three ancestral origins. A proportion of 30.8% of the total population belonged to the first ancestral group, 49.5% to the second group and 19.7% to the third group. This is consistent with the three different invasion events of
B. dorsalis in Burkina Faso as suggested by Khamis et al. (2009) for the invasion of Africa [
30]. Within the sites, the second ancestral group was more represented in Bougouriba, Boulkiemdé-Sanguié, Comoé, Kénédougou and Houet, the first ancestral group was more represented in Poni and Sissili, and the third ancestral group was not dominant in any of the sites but had a significant proportion in Comoé, Kénédougou and Poni. These results show that the population of
B. dorsalis fruit flies is not geographically structured in Burkina Faso, but it would appear that the second ancestral group is much more dominant in the western part of the country, with Houet as the epicenter, while the first group is more dominant in the southern part.
DAPC is a multivariate analysis used to infer a model of genetic variation providing information on genetic differentiation, both between subgroups and between individuals, while minimizing information within subgroups [
34]. The DAPC of
B. dorsalis in Burkina Faso, using microsatellite analysis, showed that the populations were structured into three main genetic groups that were not very well separated. The first group consisted of individuals from the localities Bougouriba, Comoé, Houet, Poni and Sissili, the second group consisted of individuals from Kénédougou and the third group consisted of individuals from Boulkiemdé-Sanguié. Kénédougou, which accounts for almost half of the country's mango production, is a major production area, exporting fruit to other regions of Burkina Faso and abroad. The area imports relatively few fruits from other regions. Interregional trade in fruit is the main means by which fruit flies, including
B. dorsalis, are exchanged [
49], so the site of Kénédougou constitutes a sort of enclave that could evolve into a special structure. Boulkiemdé-Sanguié is also a site that could evolve into a particular structure due to the geographical distance separating it from the other sites, unlike the case of Kénédougou. Furthermore, the low level of separation between the genetic groups observed demonstrates an absence of geographical structure, which can be explained by the fact that the introduction of
B. dorsalis in Burkina Faso is recent and also the genetic structure is a variable strongly influenced by the gene flow factor [
23,
46].
Figure 1.
Map showing the sampling sites of Bactrocera dorsalis. The green markers represent mango orchards, the yellow markers represent shea agroforestry parks, and the pink markers represent wild plant formations.
Figure 1.
Map showing the sampling sites of Bactrocera dorsalis. The green markers represent mango orchards, the yellow markers represent shea agroforestry parks, and the pink markers represent wild plant formations.
Figure 2.
Distribution of Bactrocera dorsalis abundance in Burkina Faso during July 2021. The names of the different localities and plant formations are abbreviated as follows: Bougouriba (BGRB), Boulkiemdé-Sanguié (BK-SG), Comoé (CMOE), Houet (HOUET), Kénédougou (KNDG), Poni (PONI), Sissili (SSLI), Mango orchard (MO), Sea Agroforestry Park (SAP) and Wild (WLD). Each locality is represented with the same three colors pink, green and blue, corresponding respectively to MO, SAP and WLD. The y-axis represents the number of flies collected per trap per day, while the x-axis represents the sampling sites.
Figure 2.
Distribution of Bactrocera dorsalis abundance in Burkina Faso during July 2021. The names of the different localities and plant formations are abbreviated as follows: Bougouriba (BGRB), Boulkiemdé-Sanguié (BK-SG), Comoé (CMOE), Houet (HOUET), Kénédougou (KNDG), Poni (PONI), Sissili (SSLI), Mango orchard (MO), Sea Agroforestry Park (SAP) and Wild (WLD). Each locality is represented with the same three colors pink, green and blue, corresponding respectively to MO, SAP and WLD. The y-axis represents the number of flies collected per trap per day, while the x-axis represents the sampling sites.
Figure 3.
Analysis of Molecular variance. The pie chart shows the percentages of molecular variance among populations (Among Pops), among individuals (Among Indiv) and within individuals (Within Indiv). The blue color within the pie chart represents the genetic variation observed between populations (2%), the orange color represents the genetic variation observed between individuals (57%) and the grey color represents the genetic variation observed within individuals (41%).
Figure 3.
Analysis of Molecular variance. The pie chart shows the percentages of molecular variance among populations (Among Pops), among individuals (Among Indiv) and within individuals (Within Indiv). The blue color within the pie chart represents the genetic variation observed between populations (2%), the orange color represents the genetic variation observed between individuals (57%) and the grey color represents the genetic variation observed within individuals (41%).
Figure 4.
Heatmap of Nei’s Genetic distance (NGD). The intensity of the colors observed between the populations on the heatmap, varying from pink to red, is proportional to the genetic distance separating them. The lowest genetic distance was observed between Comoé (CMOE) and Kénédougou (KNDG) while the highest was observed between Kénédougou (KNDG) and Sissili (SSLI).
Figure 4.
Heatmap of Nei’s Genetic distance (NGD). The intensity of the colors observed between the populations on the heatmap, varying from pink to red, is proportional to the genetic distance separating them. The lowest genetic distance was observed between Comoé (CMOE) and Kénédougou (KNDG) while the highest was observed between Kénédougou (KNDG) and Sissili (SSLI).
Figure 5.
Genetic structuring of B. dorsalis populations based on STRUCTURE analysis at K=3. (A) Sort by Q plot, (B) Group by pop Id plot. Each vertical bar represents an individual bearing the genotypic proportions of belonging to the different ancestral groups. The first ancestral group (Red), the second ancestral group (Green) and the third ancestral group (Blue). Bougouriba (BGRB), Boulkiemdé-Sanguié (BK-SG), Comoé (CMOE), Houet (HOUET), Kénédougou (KNDG), Poni (PONI), Sissili (SSLI).
Figure 5.
Genetic structuring of B. dorsalis populations based on STRUCTURE analysis at K=3. (A) Sort by Q plot, (B) Group by pop Id plot. Each vertical bar represents an individual bearing the genotypic proportions of belonging to the different ancestral groups. The first ancestral group (Red), the second ancestral group (Green) and the third ancestral group (Blue). Bougouriba (BGRB), Boulkiemdé-Sanguié (BK-SG), Comoé (CMOE), Houet (HOUET), Kénédougou (KNDG), Poni (PONI), Sissili (SSLI).
Figure 6.
DAPC of Bactrocera dorsalis population in Burkina Faso. Bougouriba (BGRB), Boulkiemdé-Sanguié (BK-SG), Comoé (CMOE), Houet (HOUET), Kénédougou (KNDG), Poni (PONI) and Sissili (SSLI). Three weakly differentiated clusters stand out, with individuals from Comoé, Houet, Bougouriba, Poni and Sissili in the first cluster, individuals from Kénédougou in the second and from Boulkiemdé in the third.
Figure 6.
DAPC of Bactrocera dorsalis population in Burkina Faso. Bougouriba (BGRB), Boulkiemdé-Sanguié (BK-SG), Comoé (CMOE), Houet (HOUET), Kénédougou (KNDG), Poni (PONI) and Sissili (SSLI). Three weakly differentiated clusters stand out, with individuals from Comoé, Houet, Bougouriba, Poni and Sissili in the first cluster, individuals from Kénédougou in the second and from Boulkiemdé in the third.
Table 1.
Summary of genetic statistics and Wright’s F-statistic for each microsatellite locus.
Table 1.
Summary of genetic statistics and Wright’s F-statistic for each microsatellite locus.
Locus |
N |
Na |
Ne |
I |
Ho |
He |
Ht |
Fis |
Fit |
Fst |
Nm |
Bd15 |
44,286 |
1,286 |
1,007 |
0,018 |
0,003 |
0,006 |
0,006 |
0,491 |
0,499 |
0,015 |
16,819 |
Bd19 |
44,286 |
4 |
3,131 |
1,239 |
0,622 |
0,673 |
0,7 |
0,075 |
0,111 |
0,039 |
6,139 |
Bi1 |
44,143 |
3 |
1,527 |
0,611 |
0,356 |
0,34 |
0,345 |
-0,044 |
-0,03 |
0,014 |
17,553 |
Bi5 |
44,429 |
3,571 |
1,454 |
0,594 |
0,276 |
0,304 |
0,31 |
0,092 |
0,109 |
0,019 |
12,915 |
Bi8 |
44,714 |
2 |
1,147 |
0,223 |
0,035 |
0,118 |
0,125 |
0,704 |
0,719 |
0,052 |
4,561 |
Bi10 |
44,143 |
3 |
1,834 |
0,773 |
0,292 |
0,447 |
0,462 |
0,347 |
0,368 |
0,032 |
7,577 |
MS12A |
43,857 |
3 |
1,894 |
0,787 |
0,589 |
0,47 |
0,477 |
-0,253 |
-0,233 |
0,016 |
15,846 |
Bd85b |
45 |
3,714 |
2,613 |
1,055 |
0,438 |
0,616 |
0,644 |
0,288 |
0,32 |
0,044 |
5,368 |
MS4 |
44,286 |
4,429 |
3,013 |
1,235 |
0,791 |
0,664 |
0,68 |
-0,191 |
-0,164 |
0,023 |
10,732 |
MS3 |
44 |
3,143 |
1,623 |
0,688 |
0,162 |
0,378 |
0,384 |
0,571 |
0,577 |
0,015 |
16,091 |
Mean |
44,314 |
3,114 |
1,924 |
0,722 |
0,356 |
0,402 |
0,413 |
0,208 |
0,228 |
0,027 |
11,36 |
SE |
0,111 |
0,130 |
0,089 |
0,047 |
0,031 |
0,026 |
0,048 |
0,103 |
0,101 |
0,004 |
1,623 |
Table 2.
Genetic diversity parameters among the seven populations of B. dorsalis.
Table 2.
Genetic diversity parameters among the seven populations of B. dorsalis.
Pop |
N |
Na |
Ne |
I |
Ho |
He |
uHe |
F |
PPL |
HOUET |
44,6 |
3,3 |
1,895 |
0,718 |
0,347 |
0,390 |
0,394 |
0,128 |
80% |
KNDG |
44,8 |
3,5 |
1,963 |
0,741 |
0,328 |
0,397 |
0,402 |
0,202 |
80% |
CMOE |
43,2 |
3,2 |
2,032 |
0,771 |
0,348 |
0,431 |
0,436 |
0,269 |
100% |
BGRB |
44,6 |
3,1 |
1,916 |
0,740 |
0,384 |
0,423 |
0,428 |
0,167 |
90% |
PONI |
44,5 |
2,9 |
1,926 |
0,716 |
0,365 |
0,398 |
0,402 |
0,159 |
90% |
SSLI |
44,6 |
2,8 |
1,819 |
0,657 |
0,354 |
0,370 |
0,374 |
0,164 |
90% |
BK-SG |
43,9 |
3,0 |
1,919 |
0,712 |
0,369 |
0,403 |
0,407 |
0,062 |
90% |
Mean |
44,3 |
3,1 |
1,924 |
0,722 |
0,356 |
0,402 |
0,406 |
0,166 |
88.6 % |
Table 3.
Pairwize Nm (above diagonal) and Fst (below diagonal) values among the seven populations of Bactrocera dorsalis.
Table 3.
Pairwize Nm (above diagonal) and Fst (below diagonal) values among the seven populations of Bactrocera dorsalis.
Pop |
BGRB |
BK-SG |
CMOE |
HOUET |
KNDG |
PONI |
SSLI |
BGRB |
- |
16.416 |
20.583 |
11.114 |
12.250 |
22.477 |
12.908 |
BK-SG |
0.015 |
- |
17.607 |
11.655 |
11.114 |
13.639 |
12.250 |
CMOE |
0.012 |
0.014 |
- |
14.456 |
22.478 |
27.528 |
13.639 |
HOUET |
0.022 |
0.021 |
0.017 |
- |
14.456 |
13.639 |
14.456 |
KNDG |
0.020 |
0.022 |
0.011 |
0.017 |
- |
17.607 |
10.620 |
PONI |
0.011 |
0.018 |
0.009 |
0.018 |
0.014 |
- |
24.75 |
SSLI |
0.019 |
0.020 |
0.018 |
0.017 |
0.023 |
0.010 |
- |