Parasite and virus dynamics in the honeybee Apis mellifera unicolor on a tropical island recently invaded by Varroa destructor.

In La Réunion, the established honeybee subspecies Apis mellifera unicolor, an endemic subspecies of African lineage, is facing considerable challenges. Since the introduction of the Varroa destructor mite in 2017 high colony losses have been recorded. We investigated the dynamics of V. destructor and two viruses, the Deformed Wing Virus (DWV), known to be transmitted by the mite, and the Chronic Bee Paralysis Virus (CBPV), in A. m. unicolor. Colonies from two apiaries located at 300 and 900 m a.s.l were monitored twice for one year without any acaricide treatment. The brood area, V. destructor infestation rates, DWV and CBPV prevalence and load were recorded monthly. A. m. unicolor maintained brood rearing throughout the year. Varroa destructor infestation resulted in high colony mortality (up to 85 %) and high phoretic mite rates (up to 52 mites per hundred bees). The establishment of DWV in colonies occurred after that of V. destructor and the mite infestation rate had a significant effect on the virus prevalence and load. CBPV appeared only transiently throughout the surveys. The data showed that, in tropical colonies with permanent brood rearing, V. destructor and DWV can reach high levels, but are still subject to seasonal variations that appear to be influenced by environmental conditions. This suggests that beekeeping practices could be adapted by favouring sites and periods for transhumance or acaricide treatment.

A comprehensive framework for the delimitation of species within the Bemisia tabaci cryptic complex, a global pest-species group.

Identifying cryptic species poses a substantial challenge to both biologists and naturalists due to morphological similarities. Bemisia tabaci is a cryptic species complex containing more than 44 putative species; several of which are currently among the world's most destructive crop pests. Interpreting and delimiting the evolution of this species complex has proved problematic. To develop a comprehensive framework for species delimitation and identification, we evaluated the performance of distinct data sources both individually and in combination among numerous samples of the B. tabaci species complex acquired worldwide. Distinct datasets include full mitogenomes, single-copy nuclear genes, restriction site-associated DNA sequencing, geographic range, host speciation, and reproductive compatibility datasets. Phylogenetically, our well-supported topologies generated from three dense molecular markers highlighted the evolutionary divergence of species of the B. tabaci complex and suggested that the nuclear markers serve as a more accurate representation of B. tabaci species diversity. Reproductive compatibility datasets facilitated the identification of at least 17 different cryptic species within our samples. Native geographic range information provides a complementary assessment of species recognition, while the host range datasets provide low rate of delimiting resolution. We further summarized different data performances in species classification when compared with reproductive compatibility, indicating that combination of mtCOI divergence, nuclear markers, geographic range provide a complementary assessment of species recognition. Finally, we represent a model for understanding and untangling the cryptic species complexes based on the evidence from this study and previously published articles.

Bacterial plant pathogens affect the locomotor behavior of the insect vector: a case study of Citrus volkameriana-Triozae erytreae-Candidatus Liberibacter asiaticus system.

Plant pathogens can alter the behavior of their insect vectors as well as their survival and reproduction. The African psyllid, Trioza erytreae, is one of the vectors of Huanglongbing, a citrus disease caused mainly by "Candidatus Liberibacter asiaticus" (CLas). The purpose of this study was to characterize the effects of CLas on the psyllid, T. erytreae using Citrus volkamerina plants as the study system. The study focused more specifically on the CLas effects prior to and after its acquisition by the psyllid T. erytreae. Our results did not support the hypothesis that CLas effects psyllid probing behavior prior to acquisition; few differences were observed between uninfected T. erytrea feeding on CLas-infected versus control plants. On the other hand, compared to psyllids that had completed their development on control plants, the ones that had completed their development on a CLas-infected plant exhibited changes in their behavior (greater velocity), physiology (smaller mass) and biochemistry (lower water and lipid content). Altogether, our results confirm the existence of a marked postacquisition effect on the vector locomotor behavior and a minor preacquisition effect of CLas on the vector behavior, which can be partially explained by physiological and biochemical changes.

Geographic population structure of the honeybee microsporidian parasite Vairimorpha (Nosema) ceranae in the South West Indian Ocean.

The microsporidian Vairimorpha (Nosema) ceranae is one of the most common parasites of the honeybee. A single honeybee carries many parasites and therefore multiple alleles of V. ceranae genes that seem to be ubiquitous. As a consequence, nucleotide diversity analyses have not allowed discriminating genetic structure of parasite populations. We performed deep loci-targeted sequencing to monitor the haplotype frequencies of genome markers in isolates from discontinuous territories, namely the tropical islands of the South West Indian Ocean. The haplotype frequency distribution corroborated the suspected tetraploidy of the parasite. Most major haplotypes were ubiquitous in the area but with variable frequency. While oceanic isolates differed from European and Asian outgroups, parasite populations from distinct archipelagoes also differed in their haplotype distribution. Interestingly an original and very divergent Malagasy isolate was detected. The observed population structure allowed formulating hypotheses upon the natural history of V. ceranae in this oceanic area. We also discussed the usefulness of allelic distribution assessment, using multiple informative loci or genome-wide analyses, when parasite population is not clonal within a single host.

Evolutionarily diverse origins of deformed wing viruses in western honey bees.

Novel transmission routes can allow infectious diseases to spread, often with devastating consequences. Ectoparasitic varroa mites vector a diversity of RNA viruses, having switched hosts from the eastern to western honey bees (Apis cerana to Apis mellifera). They provide an opportunity to explore how novel transmission routes shape disease epidemiology. As the principal driver of the spread of deformed wing viruses (mainly DWV-A and DWV-B), varroa infestation has also driven global honey bee health declines. The more virulent DWV-B strain has been replacing the original DWV-A strain in many regions over the past two decades. Yet, how these viruses originated and spread remains poorly understood. Here, we use a phylogeographic analysis based on whole-genome data to reconstruct the origins and demography of DWV spread. We found that, rather than reemerging in western honey bees after varroa switched hosts, as suggested by previous work, DWV-A most likely originated in East Asia and spread in the mid-20th century. It also showed a massive population size expansion following the varroa host switch. By contrast, DWV-B was most likely acquired more recently from a source outside East Asia and appears absent from the original varroa host. These results highlight the dynamic nature of viral adaptation, whereby a vector's host switch can give rise to competing and increasingly virulent disease pandemics. The evolutionary novelty and rapid global spread of these host-virus interactions, together with observed spillover into other species, illustrate how increasing globalization poses urgent threats to biodiversity and food security.

Tethered-flight performance of thermally-acclimated pest fruit flies (Diptera: Tephritidae) suggests that heat waves may promote the spread of Bactrocera species.

BACKGROUND: Thermal history may induce phenotypic plasticity in traits that affect performance and fitness. One type of plastic response triggered by thermal history is acclimation. Because flight is linked to movement in the landscape, trapping and detection rates, and underpins the success of pest management tactics, it is particularly important to understand how thermal history may affect pest insect flight performance. We investigated the tethered-flight performance of Ceratitis capitata, Bactrocera dorsalis and Bactrocera zonata (Diptera: Tephritidae), acclimated for 48 h at 20, 25 or 30 °C and tested at 25 °C. We recorded the total distance, average speed, number of flight events and time spent flying during 2-h tests. We also characterized morphometric traits (body mass, wing shape and wing loading) that can affect flight performance. RESULTS: The main factor affecting most flight traits was body mass. The heaviest species, B. dorsalis, flew further, was faster and stopped less often in comparison with the two other species. Bactrocera species exhibited faster and longer flight when compared with C. capitata, which may be associated with the shape of their wings. Moreover, thermal acclimation had sex- and species-specific effects on flight performance. Flies acclimated at 20 °C stopped more often, spent less time flying and, ultimately, covered shorter distances. CONCLUSION: Flight performance of B. dorsalis is greater than that of B. zonata and C. capitata. The effects of thermal acclimation are species-specific. Warmer acclimation temperatures may allow pest fruit flies to disperse further and faster. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Genetic diversity, distribution, and structure of Bemisia tabaci whitefly species in potential invasion and hybridization regions of East Africa.

Outbreaks of whitefly, Bemisia tabaci species in East and Central Africa, have become increasingly prevalent during the previous 25 years and are responsible for driving the spread of plant-virus diseases, such as cassava mosaic disease and cassava brown steak disease. Epidemics of these diseases have expanded their ranges over the same period, spreading from Uganda into other sub-Saharan African countries. It was hypothesised that a highly abundant 'invader' population of B. tabaci was responsible for spreading these diseases from Uganda to neighbouring countries and potentially hybridising with the resident cassava B. tabaci populations. Here, we test this hypothesis by investigating the molecular identities of the highly abundant cassava B. tabaci populations from their supposed origin in Uganda, to the northern, central, eastern and coastal regions of Tanzania. Partial mitochondrial cytochrome oxidase I (mtCOI) barcoding sequences and nuclear microsatellite markers were used to analyse the population genetic diversity and structure of 2734 B. tabaci collected from both countries and in different agroecological zones. The results revealed that: (i) the putative SSA1 species is structured according to countries, so differ between them. (ii) Restricted gene flow occurred between SSA1-SG3 and both other SSA1 subgroups (SG1 and SG2), even in sympatry, demonstrating strong barriers to hybridization between those genotypes. (iii) Not only B. tabaci SSA1-(SG1 and SG2) was found in highly abundant (outbreak) numbers, but B. tabaci SSA1-SG3 and the Indian Ocean (IO) species were also recorded in high numbers in several sites in Tanzania. (iv) The SSA1-(SG1 and SG2) species was distributed in both countries, but in Tanzania, the B. tabaci IO and SSA1-SG3 species predominated. These data confirm that multiple, local Tanzanian B. tabaci species produce highly abundant populations, independent of the spread of the putative invasive B. tabaci SSA1-(SG1 and SG2) populations.

Bactrocera dorsalis in the Indian Ocean: A tale of two invasions.

An increasing number of invasive fruit fly pests are colonizing new grounds. With this study, we aimed to uncover the invasion pathways of the oriental fruit fly, Bactrocera dorsalis into the islands of the Indian Ocean. By using genome-wide SNP data and a multipronged approach consisting of PCA, ancestry analysis, phylogenetic inference, and kinship networks, we were able to resolve two independent invasion pathways. A western invasion pathway involved the stepping-stone migration of B. dorsalis from the east African coast into the Comoros, along Mayotte and into Madagascar with a decreasing genetic diversity. The Mascarene islands (Reunion and Mauritius), on the contrary, were colonized directly from Asia and formed a distinct cluster. The low nucleotide diversity suggests that only a few genotypes invaded the Mascarenes. The presence of many long runs of homozygosity (ROH) in the introduced populations is indicative of population bottlenecks, with evidence of a more severe bottleneck for populations along the western migration pathway than on the Mascarene islands. More strict phytosanitary regulations are recommended in order to prevent the further spread of B. dorsalis.

Tri-trophic interactions among Fopius arisanus, Tephritid species and host plants suggest apparent competition.

When several polyphagous herbivore species share a parasitoid, the tri-trophic interaction networks can be difficult to predict. In addition to direct effects, the parasitoid may influence the herbivore community by mediating indirect interactions among hosts. The plant species can also modulate the parasitoid preference for a specific host. One of the indirect effects is apparent competition, a negative interaction between individuals as a result of the action of shared natural enemies. Here, we focus on the interactions between the parasitoid Fopius arisanus (Braconidae) and two generalist fruit fly pests: Bactrocera dorsalis and Bactrocera zonata (Tephritidae). This parasitoid was introduced into La Réunion in 2003 to control populations of B. zonata and can also interact with B. dorsalis since its invasion in 2017. Our main objective is to characterize the tri-trophic interactions between F. arisanus, fruit fly and host plant species. We developed a long-term field database of fruit collected before and after the parasitoid introduction and after the B. dorsalis invasion in order to compare parasitism rate and fruit fly infestation for the different periods. In laboratory assays, we investigated how the combination of fruit fly species and fruit can influence the preference of F. arisanus. In the field, before the invasion of B. dorsalis, the parasitism rate of F. arisanus was low and had a little impact on the fruit fly infestation rate. After the B. dorsalis invasion, we observed an increase in parasitism rate from 5% to 17%. A bioassay showed that females of F. arisanus could discriminate between eggs of different fruit fly and host plant species. The host plant species preference changed in relation to the fruit fly species inoculated. Field observations and laboratory experiments suggest the possible existence of apparent competition between B. dorsalis and B. zonata via F. arisanus.

Genomes of the cosmopolitan fruit pest Bactrocera dorsalis (Diptera: Tephritidae) reveal its global invasion history and thermal adaptation.

INTRODUCTION: The oriental fruit fly Bactrocera dorsalis is one of the most destructive agricultural pests worldwide, with highly debated species delimitation, origin, and global spread routes. OBJECTIVES: Our study intended to (i) resolve the taxonomic uncertainties between B. dorsalis and B. carambolae, (ii) reveal the population structure and global invasion routes of B. dorsalis across Asia, Africa, and Oceania, and (iii) identify genomic regions that are responsible for the thermal adaptation of B. dorsalis. METHODS: Based on a high-quality chromosome-level reference genome assembly, we explored the population relationship using a genome-scale single nucleotide polymorphism dataset generated from the resequencing data of 487 B. dorsalis genomes and 25 B. carambolae genomes. Genome-wide association studies and silencing using RNA interference were used to identify and verify the candidate genes associated with extreme thermal stress. RESULTS: We showed that B. dorsalis originates from the Southern India region with three independent invasion and spread routes worldwide: (i) from Northern India to Northern Southeast Asia, then to Southern Southeast Asia; (ii) from Northern India to Northern Southeast Asian, then to China and Hawaii; and (iii) from Southern India toward the African mainland, then to Madagascar, which is mainly facilitated by human activities including trade and immigration. Twenty-seven genes were identified by a genome-wide association study to be associated with 11 temperature bioclimatic variables. The Cyp6a9 gene may enhance the thermal adaptation of B. dorsalis and thus boost its invasion, which tended to be upregulated at a hardening temperature of 38 °C. Functional verification using RNA interference silencing against Cyp6a9, led to the specific decrease in Cyp6a9 expression, reducing the survival rate of dsRNA-feeding larvae exposed to extreme thermal stress of 45 °C after heat hardening treatments in B. dorsalis. CONCLUSION: This study provides insights into the evolutionary history and genetic basis of temperature adaptation in B. dorsalis.

Spatio-temporal changes in endosymbiont diversity and composition in the African cassava whitefly, Bemisia tabaci SSA1.

Sap-sucking insects, including whiteflies, are amongst the most devastating and widely distributed organisms on the planet. They are often highly invasive and endosymbiont communities within these insects help them adapt to new or changing environments. Bemisia tabaci (Gennadius; Hemiptera: Aleyrodidae) whitefly species are vectors of more than 500 known plant-viruses and harbour highly diverse endosymbionts communities. To date, however, whitefly-endosymbiont interactions, community structure and their spatio-temporal changes are still poorly understood. In this study, we investigated the spatio-temporal changes in the composition and diversity of bacterial endosymbionts in the agricultural crop pest whitefly species, Bemisia tabaci sub-Saharan Africa 1-subgroup 1 and 2 (SSA1-SG1 and SSA1-SG2). 16S rRNA amplicon sequencing analysis was carried out to characterise endosymbiont compositionsin field-collected SSA1 (SSA1-SG1 and SSA1-SG2) populations infesting cassava in Uganda in 1997 and 2017. We detected Portiera, Arsenophonus, Wolbachia, Hamiltonella and Hemipteriphilus, with Arsenophonus and Wolbachia infections being predominant. Hemipteriphilus and Hamiltonella frequencies were very low and were detected in seven and two samples, respectively. Bacterial diversity based on three independent parameters including Simpson index, number of haplotypes and Bray-Curtis dissimilarity matrix was significantly higher in 1997 than in 2017. This period also coincided with the advent of super-abundant cassava-whitefly populations on cassava crops in Uganda. We discuss how endosymbionts may influence the biology and behaviour of whiteflies leading to population explosions.

Virome and nrEVEome diversity of Aedes albopictus mosquitoes from La Reunion Island and China.

BACKGROUND: Aedes albopictus is a public health threat for its worldwide spread and ability to transmit arboviruses. Understanding mechanisms of mosquito immunity can provide new tools to control arbovirus spread. The genomes of Aedes mosquitoes contain hundreds of nonretroviral endogenous viral elements (nrEVEs), which are enriched in piRNA clusters and produce piRNAs, with the potential to target cognate viruses. Recently, one nrEVE was shown to limit cognate viral infection through nrEVE-derived piRNAs. These findings suggest that nrEVEs constitute an archive of past viral infection and that the landscape of viral integrations may be variable across populations depending on their viral exposure. METHODS: We used bioinformatics and molecular approaches to identify known and novel (i.e. absent in the reference genome) viral integrations in the genome of wild collected Aedes albopictus mosquitoes and characterize their virome. RESULTS: We showed that the landscape of viral integrations is dynamic with seven novel viral integrations being characterized, but does not correlate with the virome, which includes both viral species known and unknown to infect mosquitoes. However, the small RNA coverage profile of nrEVEs and the viral genomic contigs we identified confirmed an interaction among these elements and the piRNA and siRNA pathways in mosquitoes. CONCLUSIONS: Mosquitoes nrEVEs have been recently described as a new form of heritable, sequence-specific mechanism of antiviral immunity. Our results contribute to understanding the dynamic distribution of nrEVEs in the genomes of wild Ae. albopictus and their interaction with mosquito viruses.

Genetic diversity of whitefly species of the Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) species complex, associated with vegetable crops in Côte d'Ivoire.

Since several years, whiteflies of the species complex of Bemisia tabaci (Gennadius) are causing several damages on vegetable crops in Côte d'Ivoire. These sap-sucking insects are the main vector of many viruses on tomato and several species of this complex have developed resistances against insecticides. Nevertheless, there is very little information about whitefly species on vegetable crops in Côte d'Ivoire. Here, we investigated the species diversity and their genetic diversity and structuring on samples from vegetable crops in the major tomato production areas of Côte d'Ivoire. To assess this diversity, 535 whitefly samples from different localities and plant species were collected and analysed with nuclear (microsatellite) and mitochondrial (mtCOI) markers. In each site, and ecological data were recorded, including whiteflies abundance and plant species colonised by B. tabaci. The analysis of mtCOI sequences of whiteflies indicated the presence of four cryptic species on tomato and associated crops in Côte d'Ivoire. These were MED ASL, MED Q1, SSA 1 and SSA3. The MED ASL species dominated over all samples in the different regions and plant species. One haplotype of MED ASL out of the 15 identified predominated on most plant species and most sites. These results suggested that MED ASL is probably the main phytovirus vector in the Ivorian vegetable cropping areas. In contrast, only five haplotypes of MED Q1 were identified on vegetables but in the cotton-growing areas of the country. Its low prevalence, low nuclear and mitochondrial diversity might indicate a recent invasion of this species on vegetable crops in Côte d'Ivoire. The Bayesian nuclear analysis indicated the presence of hybrid genotypes between the two main species MED ASL and MED Q1, however in low prevalence (10%). All these results highlight the need to maintain whitefly populations monitoring for a more effective management in Côte d'Ivoire.

Eating eggplants as a cucurbit feeder: Dietary shifts affect the gut microbiome of the melon fly Zeugodacus cucurbitae (Diptera, Tephritidae).

While contemporary changes in feeding preferences have been documented in phytophagous insects, the mechanisms behind these processes remain to be fully clarified. In this context, the insect gut microbiome plays a central role in adaptation to novel host plants. The cucurbit frugivorous fruit fly Zeugodacus cucurbitae (Diptera, Tephritidae) has occasionally been reported on "unconventional" host plants from different families, including Solanaceae. In this study, we focus on wild parental (F0 ) adults and semiwild first filial (F1 ) larvae of Z. cucurbitae from multiple sites in La Réunion and explore how the gut microbiome composition changes when this fly is feeding on a noncucurbit host (Solanum melongena). Our analyses show nonobvious gut microbiome responses following the F0 -F1 host shift and the importance of not just diet but also local effects, which heavily affected the diversity and composition of microbiomes. We identified the main bacterial genera responsible for differences between treatments. These data further stress the importance of a careful approach when drawing general conclusions based on laboratory populations or inadequately replicated field samples.

Distribution of invasive versus native whitefly species and their pyrethroid knock-down resistance allele in a context of interspecific hybridization.

The invasion success of a species in an agrosystem is greatly influenced by environmental factors such as the use of insecticides, by the intrinsic evolutionary capabilities of the species, and also by interactions with resident species. On the island of La Réunion, the successive invasions of MEAM1 and MED whitefly species over the last 20 years have not only led an increased use of insecticides, but have also challenged the resident IO species. To trace the evolution of the 3 species, and the distribution of the kdr mutation (resistance to pyrethroid) in the para-type voltage-gated sodium channel, we genotyped 41 populations (using neutral nuclear markers) and look at the prevalence of the kdr allele. MEAM1 was predominantly present in agrosystems showing quasi fixation of the resistant kdr allele whereas IO was mainly in natural environments and did not have any resistant allele. Hybridization between the two former species was detected in low frequency but has not led to introgression of resistant alleles in the resident species so far. MED showed a limited distribution in agrosystems but all individuals displayed a resistant allele. These highly contrasting patterns of distribution and resistant mutations between invasive and resident whitefly species are further discussed.

Life history parameters and predation capacities of Nesidiocoris volucer: a new biological control agent for tomato crop.

Whiteflies are one of the major pests of tomato under greenhouses, and their control partly relies on biocontrol strategies. Among those biocontrol agents, parasitoids or predators are widely used. However, the introduction of a biocontrol agent in a new area is not trivial. For that reason, we investigated the use of a tropical native mirid, Nesidiocoris volucer (Hemiptera: Miridae), for the biological control of whiteflies among other insect pests on tomato crops under greenhouses in the subtropical island of La Réunion, France. Nesidiocoris volucer life history traits and plant injury were examined. Nymphs developed and survived between 15 and 30°C and required on average 49.41 days at 15°C and on average 10.50 days at 30°C to develop (nymph survival >94%). At 25°C, each female produced on average 65 eggs. Nesidiocoris volucer was able to feed on several prey species, but performed better on whiteflies than on spider mites or thrips. No N. volucer feeding injury was observed on tomato. Nesidiocoris volucer has also been found in tropical countries of Africa, and we believe that the data presented on this natural enemy could be of great importance for the biocontrol of whiteflies in tropical areas.

The African citrus psyllid Trioza erytreae: An efficient vector of Candidatus Liberibacter asiaticus.

Introduction: Huanglonbing (HLB) is the most serious disease of citrus in the world, associated with three non-cultivable phloem-restricted bacteria Candidatus Liberibacter asiaticus (CLas), Ca L. africanus (CLaf) and Ca L. americanus (CLam). CLas is transmitted by the Asian citrus psyllid Diaphorina citri, and has spread to several countries. The African psyllid Trioza erytreae, the vector of CLaf occurs in Africa and neighbouring islands. Only two major citrus-growing regions - Australia/New Zealand and the Mediterranean Basin - are still HLB-free in the world. However, T. erytreae has recently been introduced into continental Europe (Portugal and Spain) and has become a potential threat to citrus production. The transmission of CLas by T. erytreae had been postulated but never tested. To evaluate the risk of T. erytreae transmitting CLas, comparative transmissions of CLas by T. erytreae and D. citri were assessed. Methods: Transmission tests were performed on excised leaves and seedlings of Citrus volkameriana with different inoculation access periods (in series) for both insect species. Quantifications of bacterial titers were made in excised leaves, seedlings three and six months after inoculation and on individual insects. Results: Our results showed that T. erytreae was able to efficiently acquire CLas. Furthermore, T. erytreae carried significantly higher bacterial titers than D. citri, and was able to efficiently transmit the bacteria to seedlings at a similar rate that D. citri highlighting the high risk of spread of the most aggressive variant of HLB (CLas) by T. erytreae in Europe. Discussion: Thus, extreme precautions to prevent any entry of CLas into Europe should be adopted.

Microbiome diversity and reproductive incompatibility induced by the prevalent endosymbiont Arsenophonus in two species of African cassava Bemisia tabaci whiteflies.

A minimum of 13 diverse whitefly species belonging to the Bemisia tabaci (B. tabaci) species complex are known to infest cassava crops in sub-Saharan Africa (SSA), designated as SSA1-13. Of these, the SSA1 and SSA2 are the predominant species colonizing cassava crops in East Africa. The SSA species of B. tabaci harbor diverse bacterial endosymbionts, many of which are known to manipulate insect reproduction. One such symbiont, Arsenophonus, is known to drive its spread by inducing reproductive incompatibility in its insect host and are abundant in SSA species of B. tabaci. However, whether Arsenophonus affects the reproduction of SSA species is unknown. In this study, we investigated both the reproductive compatibility between Arsenophonus infected and uninfected whiteflies by inter-/intraspecific crossing experiments involving the sub-group three haplotypes of the SSA1 (SSA1-SG3), SSA2 species, and their microbial diversity. The number of eggs, nymphs, progenies produced, hatching rate, and survival rate were recorded for each cross. In intra-specific crossing trials, both male and female progenies were produced and thus demonstrated no reproductive incompatibility. However, the total number of eggs laid, nymphs hatched, and the emerged females were low in the intra-species crosses of SSA1-SG3A+, indicating the negative effect of Arsenophonus on whitefly fitness. In contrast, the inter-species crosses between the SSA1-SG3 and SSA2 produced no female progeny and thus demonstrated reproductive incompatibility. The relative frequency of other bacteria colonizing the whiteflies was also investigated using Illumina sequencing of 16S rDNA and diversity indices were recorded. Overall, SSA1-SG3 and SSA2 harbored high microbial diversity with more than 137 bacteria discovered. These results described for the first time the microbiome diversity and the reproductive behaviors of intra-/inter-species of Arsenophonus in whitefly reproduction, which is crucial for understanding the invasion abilities of cassava whiteflies.

Genome Sequence Resource of 'Candidatus Liberibacter asiaticus' from Diaphorina citri Kuwayama (Hemiptera: Liviidae) from La Réunion.

'Candidatus Liberibacter asiaticus' is an insect-transmitted, phloem-restricted α-proteobacterium associated with huanglongbing. Here, we provide the whole genome sequence of 'Ca. L. asiaticus' strain ReuSP1 from its insect vector Diaphorina citri (Hemiptera: Liviidae) collected in La Réunion. The genome is composed of 1,230,064 bp and has a 36.5% G+C content. This study reports the first 'Ca. L. asiaticus' genome sequence from La Réunion, which will add to 'Ca. L. asiaticus' genome resources and help elucidate our understanding of the introduction pathway into La Réunion.