mtCOI Gene Sequences Research Articles

Population genetic diversity and structure of Tephritis angustipennis and Campiglossa loewiana (Diptera: Tephritidae) based on COI DNA barcodes in the three-river source region, China.

Tephritis angustipennis (Diptera: Tephritidae)and Campiglossa loewiana (Diptera: Tephritidae) are phytophagous pests in China. Their damage has significantly impacted the collection and cultivation of germplasm resources of native Asteraceae plants. However, the genetic characteristics and structure of their population are unclear. This study focused on the highly damaging species of T. angustipennis and C. loewiana collected from the three-river source region (TRSR). We amplified the mitochondrial cytochrome C oxidase subunit I (mtCOI) gene sequences of these pests collected from this area and compared them with COI sequences from GenBank. We also analyzed their genetic diversity and structure. In T. angustipennis, 5 haplotypes were identified from 5 geographic locations; the genetic differentiation between France population FRPY (from Nylandia, Uusimaa) and China populations GLJZ (from Dehe Longwa Village, Maqin County), GLDR (from Zhique Village, Dari County), and GLMQ (from Rijin Village, Maqin County) was the strongest. GLJZ exhibited strong genetic differentiation from GLDR and GLMQ, with relatively low gene flow. For C. loewiana, 11 haplotypes were identified from 5 geographic locations; the genetic differentiation between the Chinese population GLMQ-YY (from Yangyu Forest Farm, Maqin County) and Finnish population FDNL (from Nylandia, Uusimaa) was the strongest, with relatively low gene flow, possibly due to geographical barriers in the Qinghai-Tibet plateau. Only 1 haplotype was identified across GLDR, GLMQ, and GLBM. High gene flow between distant locations indicates that human activities or wind dispersal may facilitate the dispersal of fruit flies and across different geographic. Geostatistical analysis suggested a recent population expansion of these 2 species in TRSR. Our findings provide technical references for identifying pests in the TRSR region and theoretical support for managing resistance, monitoring pest occurrences, analyzing environmental adaptability, and formulating biological control strategies for Tephritidae pests on Asteraceae plants.

Species Delimitation of Some Melanargia Species (Lepidoptera, Nymphalidae, Satyrinae) in The Southeastern Anatolia Region Based On The mtCOI Gene

Among the Palearctic species, butterflies of the genus Melanargia are known for their black and white wing patterns. The morphological character polarization of this genus is full of varying combinations of subgenus, species complex, and subspecies status. Its taxonomy is open to debate, especially in species and subspecies categories, with definitions mostly based on wing color. In recent years, cryptic species, phenotypically masked species, and species with intense intraspecific variation have been identified through the determination of lineages under the leadership of molecular systematics. The mtCOI gene, which is especially described as a species signature, is an important DNA barcode used for Lepidoptera. In the presented study, the mtCOI gene sequence of the populations of Melanargia larissa, M.grumi, M.hylata, M.syriaca, and M.russiae species in the southeastern Anatolia region was determined for the first time. To determine the boundaries of these species, gene characterization and genetic distances were carried out according to the Kimura-2 Parameter, and putative species analyses were carried out by the ABGD method. Trees were constructed with Maximum likelihood and Bayesian inference algorithms to determine the phylogenetic relationships between species of the genus. In light of these analyses, it has been shown that the genetic distance of morphological species M. larissa, M. grumi, M.hylata, and M. syriaca is not at the species level and that M.larissa maintains its species status according to the principle of priority. In addition, the M.russiae population presented in this study forms a monophyletic clade with other populations of the same species in the phylogenetic tree, proving that this taxon is a stable species.

Molecular identification and genetic diversity of ten Pierid butterflies based on mitochondrial COI gene

The IUCN Bangladesh has documented a total of 31 species belonging to the Pieridae family in Bangladesh. There is currently limited knowledge regarding the genetic diversity and molecular relationships among these butterflies. The present investigation involved the generation of mitochondrial COI (mtCOI) gene sequences for ten species of butterflies belonging to the Pierid family. After that, BLAST analysis showed 96% to 100% similarity, and then the sequences were submitted to NCBI’s GenBank. MEGA10 and BioEdit were used to identify a 584 bp COI gene fragment with 179 variable sites and 128 parsimony-informative sites. The COI sequences' average base compositions were 38.56% T, 17.12% C, 30.18% A, and 14.12% G. While a significant AT bias (68.74%) existed among the Pierid species. The genetic distance between ten Pierid species was assessed using the Kimura 2-Parameter (K2P) algorithm, and the nucleotide differences ranged from 0.00172% to 0.24261%. Afterwards, the Maximum Likelihood (ML) method was used to construct a phylogenetic tree using ten sequences of Pierid species. These species belongs to two subfamilies, Pierinae and Coliadinae in Bangladesh. On the tree, the Pierinae subfamily of Pieridae formed a paraphyletic arrangement whereas the Coliadinae subfamily was shown to be monophyletic. The data analyses support the following relationships between the two subfamilies as follows: [(Appias + Leptosia) + (Pieris + Appias) + (Delias) + (Eurema + Catopsilia) + (Pareronia)]. This supported the theory that Pierinae and Coliadinae are sister taxa. Nonetheless, Pareronia hippia remains outside the main Pierinae group, requiring further study to resolve this issue. Finally, these studies generated ten mtCOI gene sequences that have the potential to serve as valuable references for the accurate identification of Pierid species. In addition, this result could be used in the future to reveal the subfamily relationships within the Pieridae taxonomic classification. Bangladesh J. Zool. 51(3): 289-299, 2023

A new species of the genus Scirtothrips (Thysanoptera, Thripidae) from India.

A new species of Scirtothrips is described from India, S. donumdei, collected on leaves of Senegalia pennata (Fabaceae). Partial mitochondrial cytochrome c oxidase subunit 1 (mtCOI) gene sequence of the species was sequenced and the annotated sequence was submitted to NCBI GenBank.

The use of mtCOI gene sequences in identifying Butis species in the Southwest of Vietnam

Butis genus is characterised by their small body size and morphological variability, allowing them to adapt to different habitats. This paper analyses the mitochondrial cytochrome C oxidase subunit I gene sequences and morphology of Butis for identification purposes and to understand genetic relationships. The morphological characteristics of Butis koilomatodon differed obviously from Butis humeralis and Butis butis. After classification based on morphology, the total deoxyribonucleic acid of fish samples was isolated, and the mitochondrial cytochrome C oxidase subunit I genes were successfully amplified using the polymerase chain reaction method. At approximately 617bp, the obtained mitochondrial cytochrome C oxidase subunit I gene sequences were highly similar to the reference sequences on Genbank (85.90–100%). The phylogenetic graphic was divided into five distinct groups, where B. koilomatodon was grouped in one group; and B. humeralis and B. butis were grouped together. The results suggest that B. humeralis was an entirely different species from B. butis, with a mean genetic divergence of up to 14%. However, further studies using a combination of other types of deoxyribonucleic acid barcoding together with morphological features should be undertaken to confirm these findings.

The first DNA barcode of medically important cockroaches in Bangladesh

Cockroaches can spread various pathogenic agents which are accountable for food adulteration and the dispersing of foodborne pathogens. Pest management depends on proper identification. Nowadays, the COI gene of mitochondrial DNA has been anticipated as a recent systematic method functional in taxonomy and evolutionary study on species identification. The present research work is an initiative to identify the medically important cockroaches based on mitochondrial COI gene sequences. Eight (8) cockroach species (Periplaneta americana, Periplaneta brunnea, Periplaneta australasiae, Blatella germanica, Hebardina concinna, Pycnoscleus surinamensis, Blaberidae sp. and Balta notulata) were identified. Among them, four cockroach species (Balta notulata, Blaberidae sp., Hebardina Concinna and Pycnoscelus surinamensis) were the new record from Bangladesh. AT base content in DNA fragments of cockroaches was higher than GC base content. The highest AT content was 70% and the lowest GC content was 30%. The interspecific genetic divergence range of medically important cockroach species was 0.01-0.25. According to mutational steps, Pycnoscelus surinamensis was mostly diverged from its common ancestor by 88 mutational steps. Phylogenetic analysis revealed that species belonging to the same family were in the same major clade. This research is the first molecular approach to identify the medically important cockroach species based on MT-COI gene sequences in Bangladesh.

Mitochondrial genetic diversity of Thrips tabaci (Thysanoptera: Thripidae) in onion growing regions of the United States.

Onion thrips (Thrips tabaci Lindeman, Thysanoptera: Thripidae) causes severe damage to many horticultural and agronomic crops worldwide. It also acts as a vector of several plant viruses. T. tabaci is a key pest of Allium cepa in the United States. However, there is limited information available on the genetic variation within and between T. tabaci populations in the United States and its key evolutionary parameters. In the current study, 83 T. tabaci specimens were collected from A. cepa from 15 different locations comprising four states of the United States. A total of 92 mtCOI gene sequences of T. tabaci from A. cepa were analyzed to understand the genetic diversity and structure of T. tabaci collected from onion host. Seven distinct haplotypes of T. tabaci infesting A. cepa were identified from the current collection, while nine T. tabaci sequences retrieved from GenBank comprised 5 haplotypes. Overall, 15 haplotypes of T. tabaci infesting A. cepa were identified in the world that includes the ten haplotypes in the United States. In the phylogenetic analysis, all the populations collected during the study clustered with thelytokous lineage, while T. tabaci sequences retrieved from GenBank corresponded to leek-associated arrhenotokous lineage. The highest genetic variation was found in Elba and Malheur populations with 3 haplotypes identified in each. The results suggest that haplotypes 1 and 7 are more frequently prevailing haplotypes in the north-western United States, with haplotype 1 being the predominant all over the country. The eastern United States appears to have a more diverse group of haplotypes. The populations from Hungary constituted distinct haplotypes and a haplotype from Kingston linked it with the predominant haplotype.

Insecticide Resistance, Resistance Mechanisms, and Phylogeny of Three <em>Myzus persicae</em> Populations in Cabbage from Three Agroclimatic Zones of Sri Lanka

The aphid, Myzus persicae is an important sucking pest and a virus vector in many crops. The management of M. persicae is mainly through synthetic pesticides. The development of resistance against common pesticides has resulted in control failures of M. persicae. The objective of this study was to examine the status of pesticide resistance and the genetic structure of M. persicae populations. Samples were collected from cabbage cultivations in the dry (Anuradhapura), wet (Kandy), and intermediate (Badulla) zones of Sri Lanka during 2017-2019. Bioassays were conducted against five insecticides using the leaf dip method. Biochemical assays examined the activity of insecticide detoxifying enzymes and the insensitivity of the organophosphate/carbamate target site, acetylcholinesterases (AChEs) in the Badulla and Kandy populations. A 636 bp mtCOI gene sequence was used to infer the phylogeny. The resistance spectrum of the populations was low to moderate; 17-25% resistance to thiamethoxam, 11-15% to acetamiprid, 20-28 % to carbosulfan, 20-41 % to etofenprox, and 4-9 % to profenofos. Elevated activity of esterases (specific activities 0.91-1.11 μmol/min/mg protein) and glutathione S-transferases (1.27-1.82 μmol/min/mg protein) were observed for increased metabolism of pesticides. Monooxygenase-based detoxification was noticeable in neonicotinoid resistance. Insensitive AChEs contribute to organophosphate and carbamate resistance. Phylogenetic analysis revealed that 2 haplotypes of the studied mtCOI gene sequence are present in the three populations tested, indicating that environmental and biological pressure, including pesticide selection pressure, may be contributing to the emergence of novel genotypes of M. persicae.

Ecological Factors Associated with the Distribution of Bemisia tabaci Cryptic Species and Their Facultative Endosymbionts

The sweetpotato whitefly, Bemisia tabaci species complex, comprises at least 44 morphologically indistinguishable cryptic species, whose endosymbiont infection patterns often varied at the spatial and temporal dimension. However, the effects of ecological factors (e.g., climatic or geographical factors) on the distribution of whitefly and the infection frequencies of their endosymbionts have not been fully elucidated. We, here, analyzed the associations between ecological factors and the distribution of whitefly and their three facultative endosymbionts (Candidatus Cardinium hertigii, Candidatus Hamiltonella defensa, and Rickettsia sp.) by screening 665 individuals collected from 29 geographical localities across China. The study identified eight B. tabaci species via mitochondrial cytochrome oxidase I (mtCOI) gene sequence alignment: two invasive species, MED (66.9%) and MEAM1 (12.2%), and six native cryptic species (20.9%), which differed in distribution patterns, ecological niches, and high suitability areas. The infection frequencies of the three endosymbionts in different cryptic species were distinct and multiple infections were relatively common in B. tabaci MED populations. Furthermore, the annual mean temperature positively affected Cardinium sp. and Rickettsia sp. infection frequencies in B. tabaci MED but negatively affected the quantitative distribution of B. tabaci MED, which indicates that Cardinium sp. and Rickettsia sp. maybe play a crucial role in the thermotolerance of B. tabaci MED, although the host whitefly per se exhibits no resistance to high temperature. Our findings revealed the complex effects of ecological factors on the expansion of the invasive whitefly.

A remarkable new genus of Thripinae (Thysanoptera, Thripidae) without anteocellar setae from India.

Nandithripspouzolziaegen. et sp. nov. (Thripidae, Thripinae) is described from the flowers of Pouzolziapetandrasubsp.wightii (Urticaceae) found in Nandi hills, Karnataka, India. This new genus is characterised by an apomorphy, ocellar setae pairs I and II are both absent, and also has a unique discontinuous pore plate distribution, with a single circular or oval pore plate medially on abdominal sternites II and V-VII of males. Partial mitochondrial cytochrome c oxidase subunit 1 (mtCOI) gene sequence of N.pouzolziae was sequenced and the annotated sequence was submitted to NCBI GenBank.

A method for DNA extraction and molecular identification of Aphids

Aphid species (Insecta, Hemiptera) are economically important invasive pest throughout the world, though their identification is intricate due to tiny size and inconspicuous nature of morphology. Mitochondrial cytochrome c oxidase I (mtCOI) region has been proven to be a standard barcode to identify the diverse array of insect groups. Isolation of good quality DNA is a fundamental first step in insect DNA barcoding which is obtained by standardizing the DNA isolation method. In this study, we demonstrate a modified CTAB method for the isolation of DNA to maximize the quality and yield from small aphids. This method will help the researchers to efficiently isolate DNA from small aphid and the method can be utilized for other small insects as well. We evaluated the quality of the isolated DNA and the mtCOI gene region were subjected to PCR amplification. Further, the gene segment was sequenced and gene annotation was done by NCBI BLAST program through which the insect was found to be Aphis gossypii. This study provides a set of molecular tools that can be used for identification of insect at species level through DNA barcoding and biodiversity analysis.•Detailed method to maximize quality and quantity of genomic DNA isolated from aphids.•Molecular identification of aphids using mtCOI gene amplification and sequence validation.•First report on Aphis gossypii infecting Solanum trilobatum provides insights of pest identification and management.

First Report of a Paucibranchia (Polychaeta, Eunicidae) Species without Lateral Palps in Korean Subtidal Waters, with Genetic Evidence for Its Taxonomic Position

An undescribed species belonging to the family Eunicidae was detected in a sublittoral habitat of the southern coast of Korea. This Korean eunicid species was initially thought to belong to the genus Lysidice based on its general appearance, including the absence of prostomial lateral palps and peristomial cirri. However, a more detailed characterization of the morphological features of the maxillae and mandible coupled with mtCOI and 18S rRNA gene sequence analyses confirmed that this species is a member of the genus Paucibranchia. The absence of lateral palps found in the intact adult specimens with 153 segments is a unique feature not previously reported in species belonging to Paucibranchia. Thus, the new species, Paucibranchia triantennata sp. nov., can be easily distinguished from other known congeneric species. Except for the absence of lateral palps, P. triantennata sp. nov. resembled P. conferta, P. gathofi, and P. patriciae in the shape of the prostomium, brief location and shape of branchiae, and absence of compound spinigers. However, P. triantennata could be clearly distinguished from these species by the shorter prostomial antennae, a large number of subacicular hooks, and the morphological features of the maxillae and compound falcigers.

Landscape genetics across the Andes mountains: Environmental variation drives genetic divergence in the leaf-cutting ant Atta cephalotes.

Distinguishing among the mechanisms underlying the spatial distribution of genetic variation resulting from the environmental or physical barriers from those arising due to simple geographic distance is challenging in complex landscapes. The Andean uplift represents one of the most heterogeneous habitats where multiple mechanisms may interact, confounding their relative roles. We explore this broad question in the leaf-cutting ant Atta cephalotes, a species that is distributed across the Andes mountains, using nuclear microsatellite markers and mtCOI gene sequences. We investigate spatial genetic divergence across the western range of the northern Andes in Colombia by testing the relative role of alternative scenarios of population divergence, including isolation by geographic distance (IBD), climatic conditions (IBE), and the physical barriers presented by the Andes mountains (IBB). Our results reveal substantial genetic differentiation among A. cephalotes populations for both types of markers, but only nuclear divergence followed a hierarchical pattern with multiple models of genetic divergence imposed by the western range. Model selection showed that the IBD, IBE (temperature and precipitation), and IBB (Andes mountains) models, often proposed as individual drivers of genetic divergence, interact, and explain up to 33% of the genetic divergence in A. cephalotes. The IBE model remained significant after accounting for IBD, suggesting that environmental factors play a more prominent role than IBB. These factors, in combination with the idiosyncratic dispersal patterns of ants, appear to determine the hierarchical patterns of gene flow. This study enriches our understanding of the forces shaping population divergence in complex habitat landscapes.

Distribution and Mitotype Diversity of <i>Bemisia tabaci</i>

Comparative assessment of the mitochondrial cytochrome oxidase I (mtCOI) gene sequence uncovers the Bemisia tabaci mitotypes. The current study aims to define B. tabaci mitotypes found in various agro ecological zones across India. Phylogenetic analysis of 13 samples from various agroclimatic regions revealed presence of four mitotypes: Asia 1, Asia II 1, Asia II 5 and Asia II 7. Among, Asia II 1 was found to be abundant and widespread distribution in the thirteen locations studied. From the 13 sequences examined, 7 haplotypes viz., three of Asia II 1, two of Asia II 5, each one of Asia 1 and Asia II 7 were identified. Nucleotide diversity was observed to be greater in Asia II 5 (0.00388) than in Asia II 1 (0.00258), and haplotype diversity was observed to be greater in Asia II 5 (1.000) than in Asia II 1 (0.600).

Characterization of the Asian Citrus Psyllid-'Candidatus Liberibacter Asiaticus' Pathosystem in Saudi Arabia Reveals Two Predominant CLas Lineages and One Asian Citrus Psyllid Vector Haplotype.

In Saudi Arabia (SA), the citrus greening disease is caused by ‘Candidatus Liberibacter asiaticus’ (CLas) transmitted by the Asian citrus psyllid (ACP) Diaphorina citri. The origin and route(s) of the ACP-CLas pathosystem invasion in SA have not been studied. Adult ACP were collected from citrus trees in SA and differentiated by analysis of the mitochondrial cytochrome oxidase I (mtCOI) and nuclear copper transporting protein (atox1) genes. A phylogenetic analysis of the Wolbachia spp. surface protein (wsp) gene was used to identify the ACP-associated Wolbachia spp. A phylogenetic analysis of the atox1 and mtCOI gene sequences revealed one predominant ACP haplotype most closely related to the Indian subcontinent founder populations. The detection and identification of CLas in citrus trees were carried out by polymerase chain reaction (PCR) amplification and sequencing of the 16S rDNA gene. The CLas-integrated prophage genomes were sequenced, annotated, and used to differentiate CLas populations. The ML and ASTRAL trees reconstructed with prophages type 1 and 2 genome sequences, separately and concatenated, resolved two major lineages, CLas-1 and -2. The CLas-1 clade, reported here for the first time, consisted of isolates from SA isolates and Pakistan. The CLas-2 sequences formed two groups, CLas-2-1 and -2-2, previously the ‘Asiatic’ and ‘Floridian’ strains, respectively. Members of CLas-2-1 originated from Southeast Asia, the USA, and other worldwide locations, while CLas-2-2 was identified only in Florida. This study provides the first snapshot into the status of the ACP-CLas pathosystem in SA. In addition, the results provide new insights into the pathosystem coevolution and global invasion histories of two ACP-CLas lineages with a predicted center of origin in South and Southeast Asia, respectively.

Xylotrechus smei (Cerambycidae: Cerambycinae: Clytini): a potential threat to red sanders cultivation

Red sanders (Pterocarpus santalinus L.f.) is an economically important, endangered timber species endemic to the restricted forest tracts of the Eastern Ghats, India. Here, we report the occurrence of long horned beetle Xylotrechus smei (Laporte de Castelnau & Gory, 1841) infestation on red sanders for the first time in two plantations located in Telangana, India. Xylotrechus smei is an economically important polyphagous pest with over forty recorded hosts. In red sanders, the infestation percentage ranged from 0.63 to 2.5% across plantations. Infestation was characterized by gradual chlorosis starting from the lower crown followed by complete dying of standing trees, presence of galleries and bore holes in cut stems. In addition, supplementary description of X. smei, including the terminal abdominal segments and genitalia of the sexes, immatures and its illustrations are provided for more authentic identification of this important pest. Further, the identity of the insect as X. smei was confirmed through mtCOI based DNA barcode. Phylogenetic and sequence divergence analyses based on partial mtCOI gene sequence showed that X. smei isolate obtained in the current study was closely related to X. smei isolate WA13 10.01r that was intercepted in wood packaging material in one of the ports of entry of the United States (U.S.). Further studies on devising management strategies for X. smei in red sanders are needed for improving the quality and quantity of the economic produce.

Population Genetic Structure of the Bean Leaf Beetle Ootheca mutabilis (Coleoptera: Chrysomelidae) in Uganda.

Bean leaf beetle (BLB) (Ootheca mutabilis) has emerged as an important bean pest in Uganda, leading to devastating crop losses. There is limited information on the population genetic structure of BLB despite its importance. In this study, novel microsatellite DNA markers and the partial mitochondrial cytochrome oxidase subunit I (mtCOI) gene sequences were used to analyze the spatial population genetic structure, genetic differentiation and haplotype diversity of 86 O. mutabilis samples from 16 (districts) populations. We identified 19,356 simple sequence repeats (SSRs) (mono, di-, tri-, tetra-, penta-, and hexa-nucleotides) of which 81 di, tri and tetra-nucleotides were selected for primer synthesis. Five highly polymorphic SSR markers (4-21 alleles, heterozygosity 0.59-0.84, polymorphic information content (PIC) 50.13-83.14%) were used for this study. Analyses of the 16 O. mutabilis populations with these five novel SSRs found nearly all the genetic variation occurring within populations and there was no evidence of genetic differentiation detected for both types of markers. Also, there was no evidence of isolation by distance between geographical and genetic distances for SSR data and mtCOI data except in one agro-ecological zone for mtCOI data. Bayesian clustering identified a signature of admixture that suggests genetic contributions from two hypothetical ancestral genetic lineages for both types of markers, and the minimum-spanning haplotype network showed low differentiation in minor haplotypes from the most common haplotype with the most common haplotype occurring in all the 16 districts. A lack of genetic differentiation indicates unrestricted migrations between populations. This information will contribute to the design of BLB control strategies.

Rapid Identification of Western Flower Thrips, Frankliniella occidentalis (Thysanoptera: Thripidae) in Shade Houses in Western Cape of South Africa

The western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thripidae: Thysanoptera), is one of the most economically important pests in shade houses. However, their identification is usually difficult, with them being easily confused with other thrips species. Accurate, rapid and reliable identification of insect pests is a crucial first step in implementing an efficacious management program. In this study the identity of F. occidentalis was verified, using molecular methods using the mtCOI gene sequences. Molecular analysis of thrips specimens collected from chrysanthemums and blueberries in the Western Cape Province, South Africa revealed the presence of Frankliniella. occidentalis and four other thrips species. Two of the five the specimens were F. occidentalis and one of the specimens was identified as Gynaikothrips ficorum. The remaining specimens were identified only to a generic level as Pseudophilothrips sp and to an ordinal level as Thrips sp. Failure to identify the two specimens to specific levels might have resulted from lack of sequences of identified thrips species in the molecular data bases. Considering the speed, specificity as well as sensitivity of the method, mtCOI sequences can be used as a swift and efficient method for western flower thrips identification and detection of new thrips pest invasion.

A new species of freshwater crab of the genus Barytelphusa (Decapoda, Brachyura, Gecarcinucidae) from the Eastern Ghats of India

Abstract A new species of gecarcinucid crab, Barytelphusa choprai sp. nov., is described here from the Eastern Ghats in Andhra Pradesh state of India. The new species differs from all three congeners by its convex and trapezoidal carapace, pale white colouration, and by a distinct structure of the male gonopod G1. It differs from its congeners by genetic distance (p-distance) in the range of 5.7-13.6% in partial mtCOI gene sequence. The habitat at the type locality is a slow flowing shallow stream with rocky substratum, covered with silt and debris, and devoid of aquatic vegetation.

Integrative Techniques Confirms the Presence of Bemisia tabaci Parasitoids: Encarsia formosa, Encarsia porteri and Eretmocerus mundus (Hymenoptera: Aphelinidae) on Soybean and Tomatoes in South Brazil.

Parasitoid wasps from the Aphelinidae family (Hymenoptera) are important control agents of Bemisia tabaci (Gennadius, 1889) cryptic species, both through reproduction and feeding processes. Identifying native parasitoid species within agricultural systems affected by Bemisia whitefly species is the first step to developing guidelines for the creation and release of biological control agents aiming at this highly damaging pest species complex. Taxonomic and phylogenetic analyses based on morphological and molecular characters, respectively, confirmed the occurrence of Encarsia formosa (Gahan, 1924) in greenhouse tomatoes from Santa Maria, Encarsia porteri (Mercet, 1928) in open-field soybean from Santa Maria, and Eretmocerus mundus Mercet, 1931 in greenhouse tomatoes from São José do Hortêncio, all within Rio Grande do Sul state (South Brazil). This is the first report of En. formosa, En. porteri and Er. mundus parasitising B. tabaci in South Brazil, and the first En. porteri partial mtCOI gene sequence being reported and characterised. The high temperature inside the tomato greenhouses can be a possible cause for the predominance of Er. mundus in São José do Hortêncio, and sex ratios in the surveyed populations point to female and male prevalence within Encarsia and Eretmocerus genera, respectively. The combined use of taxonomic and molecular characterisation highlights the importance of combining both morphological and molecular approaches in the assessment of previously unidentified whitefly parasitoids.