Research Article
Print
Research Article
First record of Tamarixia dahlsteni Zuparko (Hymenoptera, Eulophidae), a parasitoid of Trioza eugeniae Froggatt (Hemiptera, Triozidae) and current status of the Tamarixia species in Mexico
expand article infoKenzy I. Peña-Carrillo, Antonio Rodríguez-Rivas§, Sara G. Díaz-Ramos§, Mayra A. Gómez-Govea|, Patricia Zambrano-Robledo|, Iram P. Rodríguez-Sánchez|, María de Lourdes Ramírez-Ahuja|
‡ INIFAP, Campo Experimental General Terán, Terán, Mexico
§ Universidad de Guadalajara, Guadalajara, Mexico
| Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Mexico
Open Access

Abstract

Parasitic wasps of the genus Tamarixia represent important biological control agents of members of the true bug group, Psylloidea, and are host specific; therefore, they can be used to control insect pests. In this study we report, for the first time, the presence of the parasitoid Tamarixia dahlsteni in Mexico and its mitochondrial barcode region of the cytochrome oxidase I gene (COI). We also review the species diversity of the genus Tamarixia in Mexico.

Keywords

Biological control, COI, eugenia psyllid, parasitic wasps

Introduction

Biological control agents represent a sustainable pest management option that help to maintain pest populations under accepted levels (Wang et al. 2019; Al-Ani et al. 2020). Parasitic wasps from the family Eulophidae represent a very important group of biological control agents as they have a wide range of insect hosts with different degrees of specialization. The genus Tamarixia Mercet represents one example of parasitoids with a high degree of host specificity (Urbaneja-Bernat et al. 2019). Most species act as ectoparasitoids, but in some cases endoparasitism has been reported (Noyes 2022). Species of the genus Tamarixia parasitize free-living and gall-forming species of true bugs, Psylloidea. It was proposed that they reached this host restriction through a specialization on their host (LaSalle 1994), although the species has also been reported parasitizing aphids (Zuparko et al. 2011). According to the Universal Chalcidoidea database, the genus Tamarixia comprises 54 species to date, which are distributed worldwide (Noyes 2022).

In Mexico, five species (both native and exotic) of Tamarixia are found: Tamarixia aguacatensis Yefremova (Yefremova et al. 2014), Tamarixia leucaenae Boucek (McClay 1990), Tamarixia radiata (Waterston) (González-Hernández et al. 2009), Tamarixia schina Zuparko (Zuparko et al. 2011), and Tamarixia triozae (Burks) (Lomelí-Flores and Bueno 2002). Some of these species have already been part of integrated pest management strategies with remarkable results. For instance, T. triozae, the parasitoid of the potato psyllid Bactericera cockerelli, was introduced to New Zealand for the biological control of the psyllids that vector the bacterium Candidatus Liberibacter solanacearum (CLso) (Workman and Whiteman 2009). This bacterium has been linked to different diseases in plants of the nightshade family Solanaceae (Munyaneza et al. 2007). In Mexico, T. triozae was found naturally in tomato crops and, according to parasitism evaluation, the percentage of parasitism reached by T. triozae on B. cockerelli has been up to eighty percent when insecticides are not used to control the psyllid populations (Lomelí-Flores and Bueno 2002). Tamarixia triozae is commercially available in Mexico through Koppert Mexico, and several studies have been carried out on its biological cycle (Rojas et al. 2015), its release into the environment either individually or in combination with other natural enemies for the control of B. cockerelli (Cerón-González et al. 2014; Ramírez-Ahuja et al. 2017). Other examples of Tamarixia species used for biological control are T. schina, which was introduced in California for the control of Calophya schini Tuthill (Psyllidae: Calophyidae), T. dahlsteni Zuparko, which was introduced for the control of Trioza eugeniae Froggatt (Hemiptera: Triozidae) (Zuparko et al. 2011) and T. radiata. The latter is native to Pakistan (Chen and Stansly 2014), but has been introduced into countries such as Taiwan, the United States and France (Guadeloupe), to control populations of the psyllid Diaphorina citri Kuwayama (Hemiptera: Liviidae), a vector of the bacterium Candidatus Liberibacter asiaticus (Chien et al. 1989; Michaud 2002; De León and Sétamou 2010). In Texas, a reduction of more than ninety percent of D. citri populations has been observed in regions where T. radiata was released (Flores and Ciomperlik 2017). In Mexico, the parasitoid was reported as an accidentally introduced species (De León and Sétamou 2010).

Currently, DNA barcodes are important tools for species identification with potential for bio-surveillance programs in agriculture (Ashfaq and Hebert 2016). DNA barcodes have been useful to identify important arthropod pests even at immature stages (Ashfaq and Hebert 2016). The common barcoding method used for animal identification is based on the sequencing of a part of the mitochondrial gene cytochrome oxidase subunit I (COI). COI has been used to create universal and public databases of sequences, such as the Barcode of Life Data System (BOLD), which includes agriculturally important insect sequences (Hebert et al. 2003). In this regard, here we report the occurrence of T. dahlsteni in Mexico for the first time, and the first mitochondrial cytochrome oxidase subunit one (COI) sequences for this species. We also discuss the potential of additional species of Tamarixia in Mexico.

Materials and methods

Biological samples

We obtained parasitoids emerged from nymphs of Trioza eugeniae feeding on Syzygium paniculatum, collected in an urban area from Zapopan, Jalisco, Mexico [Colonia Las Palomas, Tesistán (20.7890, -103.4831) and Club Deportivo UdeG (20.7793, -103.6075)]. The nymphs were taken to the laboratory (HR 70%, T 25 ± 2 °C) and were placed into Petri dishes until the parasitoids emerged. The parasitoids were placed in 96% ethanol for morphological and molecular determination.

Morphological determination

According to Zuparko et al. (2011), the psyllid Trioza eugeniae is parasitized by Tamarixia dahlsteni; therefore, we employed Zuparko’s morphological description to identify the newly emerged parasitoids. Voucher specimens of the recovered parasitoids were deposited in the Beneficial Insects Collection of the Universidad Autónoma de Nuevo León (CIBE–UANL). All individuals followed the same diagnosis and, given the low number of specimens recovered, only one female was photographed with a scanning electronic microscope (JEOL JSM–6510LV) in order to illustrate its diagnostic characteristics.

Barcoding determination

Genomic DNA was non-destructively isolated according to the protocol described by Giantsis et al. (2016). We extracted three individual specimens that corresponded to T. dahlsteni, two specimens of T. triozae and two of T. schina. Polymerase chain reaction (PCR) was carried out to amplify the DNA barcode region of the cytochrome oxidase subunit I (COI) using the LCO1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HCO2198 (5‘-TAAACTTCAGGGTGACCAAAAAATCA-3’) primers (Folmer et al. 1994). PCRs were performed in a 20 µl reaction volume: 2 µl of DNA, 2 µl of 10× Qiagen PCR buffer containing 15 mM MgCl2, 0.9 µl of each primer (10 um), 0.6 µl of dNTPs (25 mM each), 0.2 µl of (5 U/µl) Taq DNA Polymerase (Qiagen, Hilden, Germany), and 13.4 µl of H2O. PCR conditions were as follows: 94 °C for 3 min, followed by 40 cycles of 94 °C for 30 s, 52 °C for 1 min, 72 °C for 1 min with a final extension at 72 °C for 10 min. All PCR products were electrophoresed through an agarose gel (1%) and sequenced in both directions in an Applied Biosystems model 3500 automated sequencer in Lanbama–Ipicyt (San Luis Potosí, Mexico).

Phylogenetic analysis

We employed the resulting COI sequences to reconstruct the phylogenetic relations of the emerged parasitoids. For this objective, we included COI sequences of all available species of the genus Tamarixia, which were downloaded from the BOLD Systems database and GenBank. All sequences were aligned in Mesquite ver. 3.70 (Maddison and Maddison 2021) with the program MUSCLE ver. 3 (Edgar 2004). Later, the alignment was used for the phylogenetic analysis inferred with the Maximum Likelihood method in the online server IQ-tree ver. 1.6.12 (Trifinopoulos et al. 2016), and the model GTR+F+I+G4 which was inferred with the function Model Finder (Kalyaanamoorthy et al. 2017). Branch support was obtained with the ultrafast bootstrap approximation (Hoang et al. 2018) with 10 000 replicates. Sequences generated in this study were deposited in GenBank.

Results

Morphological and barcoding determination

From the collected material for this study, seven females and seven males of the genus Tamarixia emerged. They agreed with the diagnosis of T. dahlsteni: the entire ventral surface of the gaster was yellow, and in the dorsal part the yellow color extending to, or slightly beyond, the apex of the second tergite (Fig. 1). The specimens had a paraspicular carina posteriorly bifurcated and located medial to the propodeal spiracle (Figs 2, 3). Regarding the barcoding determination, the sequences generated in this study represent the first barcoding evidence for the species T. dahlsteni (GenBank accession ON491415, ON491416, ON491417) and T. schina (GenBank accession ON548243, ON684328).

Figures 1–3. 

Tamarixia dahlsteni 1 gaster (dorsal view), arrow pointing the yellow color extending to the apex of the second tergite 2 mesosoma (dorsal view) 3 arrow pointing paraspicular carina. Scale bars: 100 μm (2); 50 μm (3).

Phylogenetic analysis

Our sequence alignment contained 893 bp, and included sequences of the species Tamarixia drukyulensis Yefremova and Yegorenkova, Tamarixia dryi Waterston, Tamarixia pronomus Walker, Tamarixia pubescens Nees, T. radiata, T. triozae and Tamarixia upis Walker, obtained from public databases and those of T. dahlsteni and T. schina generated in this study. In the phylogenetic reconstruction, sequences of each species were clustered in individual subclades with high support (ultrabootstrap values >95) (Fig. 4). Therefore, the barcoding region appears to be useful for the molecular identification of the Tamarixia species included in this study. Our analysis did not resolve interspecific relations due to the low bootstrap support for interior branches. On the other hand, the presence of highly supported (ultrabootstrap values >95) intraspecific subclades within T. drukyulensis and T. dryi suggests intraspecific genetic structure.

Figure 4. 

Phylogenetic tree showing the genetic relations between species of the genus Tamarixia. Aprostocetus cerricola was included as an outgroup and Tamarixia dahlsteni is highlighted in gray color. Bootstrap branch support ≥ 95% is shown above branches; samples in bold represent sequences generated in this study, while samples with an asterisk (*) were downloaded from the BOLD Systems database. The rest of the samples were downloaded from GenBank. BOLD Systems specimen records and GenBank accession numbers are shown before the species name.

Discussion

According to the literature, seven of the 54 existing species of Tamarixia are reported from the Nearctic and Neotropical regions (Noyes 2022). Mexico spans both regions and, following this study, the number of Tamarixia species was updated to six. In addition, our phylogenetic analysis suggested the COI barcoding region to be a useful molecular marker for the distinction of Tamarixia species. The phylogeny obtained suggested intraspecific genetic structure for some of the species, which unveils the necessity of robust and wider phylogenetic analysis at the genus level.

The host for T. dahlsteni is the eugenia psyllid Trioza eugeniae. Both species were found in Australia associated with the ornamental tree Syzygium paniculatum, and in 1988 the psyllid was found for the first time in California, USA. The damage caused by the psyllid on S. paniculatum trees prompted a search for its natural enemies in Australia. As a result, the wasp T. dahlsteni was identified as the primary parasitoid of T. eugeniae and was later imported to the USA to control eugenia psyllid populations (Dahlsten et al. 1993). A similar case is the one of Tamarixia schina. The wasp was reported as a natural enemy of the exotic psyllid Calophya rubra (Blanchard) which feeds on Schinus molle trees (Álvarez-Zagoya and Cibrián-Tovar 1999). According to Zuparko et al. (2011), new undescribed Tamarixia species were found in both California and Florida, USA, parasitizing psyllid species which have potential distribution in Mexico.

In biological research, names of species are essential to ensure comparable results when working with model organisms (Pante et al. 2015), and in agriculture they are also required for biosecurity and quarantine concerns (Lyal et al. 2008). Notwithstanding, in some cases species identification is not an easy task and deep taxonomic studies are needed. For instance, in 2019 the misidentification of the eugenia psyllid T. eugeniae was uncovered by Taylor and Martoni (2020), who indicated that the true name of the species should be Trioza adventicia Tuthill. Taylor and Martoni mentioned that the two exotic species resemble each other, and only a detailed study based on a series of morphological characters and DNA barcoding supported the separation and validity of both species.

As already mentioned, psyllids are main hosts for the Tamarixia species and because of their possible broad dietary tolerance some species might migrate and disperse to new geographic regions (Percy et al. 2012). This might also promote the introduction and dispersion of exotic or new species of parasitoids in countries like Mexico. For example, T. schina apparently migrated to Mexico from California (Yefremova et al. 2014). According to the study by Percy et al. (2012), in the same region (California, USA), different species of psyllids attacked by some unidentified Tamarixia species exist, hosted by plants with potential distribution in Mexico. Moreover, in this country more than 114 species of psyllids (Méndez-Tobar 2015) exist, which may also represent possible hosts for exotic Tamarixia species. Therefore, the diversity of this genus might be currently underestimated in Mexico.

In agriculture, species identification protocols based on DNA represent powerful tools for the success of early detection programs, or monitoring of species (Lyal et al. 2008; Boykin et al. 2012; Poland and Rassati 2019). However, for some groups of insects, the lack of reference barcodes, errors in databases, scarcity of voucher specimens and presence of cryptic species represent strong limitations. As an example, recently a new Tamarixia species (T. aguacatensis) was described based on morphological characters, but because their sampling seems to be seasonally restricted (Yefremova et al. 2014), the generation of barcodes for further studies on their biology and phylogeny represents a challenge.

Conclusion

Besides reporting the presence of Tamarixia dahlsteni in Mexico, we also provided barcodes that may be employed as a reference for further monitoring programs or studies about this economically important group of wasps. Moreover, our phylogenetic analysis suggests the need for a deeper and wider taxonomic revision of the genus.

Acknowledgements

The authors are grateful to Dave Burns and Laurie Jo Elliot for providing English editing of the manuscript.

References

  • Al-Ani LKT, Aguilar Marcelino L, Fiorotti J, Sharma V, Sarker MS, Furtado EL, Wijayawardene NN, Herrera-Estrella A (2020) Biological control agents and their importance for plant health. In: Singh JS, Vimal SR (Eds) Microbial Services in Restoration Ecology, Elsevier, Netherlands, 13–36. https://doi.org/10.1016/B978-0-12-819978-7.00002-6
  • Ashfaq M, Hebert PD (2016) DNA barcodes for bio-surveillance: Regulated and economically important arthropod plant pests. Genome 59(11): 933–945. https://doi.org/10.1139/gen-2016-0024
  • Boykin LM, Armstrong KF, Kubatko L, De Barro P (2012) Species delimitation and global biosecurity. Evolutionary Bioinformatics Online 8: 1–37. https://doi.org/10.4137/EBO.S8532
  • Cerón-González C, Lomelí-Flores JR, Rodríguez-Leyva E, Torres-Ruíz A (2014) Fertility and feeding of Tamarixia triozae (Hymenoptera: Eulophidae) on potato psyllid Bactericera cockerelli. Revista Mexicana de Ciencias Agrícolas 5: 893–899. https://doi.org/10.29312/remexca.v5i5.912
  • Chen X, Stansly PA (2014) Biology of Tamarixia radiata (Hymenoptera: Eulophidae), Parasitoid of the Citrus Greening Disease Vector Diaphorina citri (Hemiptera: Psylloidea): A Mini Review. The Florida Entomologist 97(4): 1404–1413. https://doi.org/10.1653/024.097.0415
  • Dahlsten DL, Kent DM, Rowney DL, Cooper WA, Young TE, Tassan RL (1993) Parasitoid shows potential for biocontrol of eugenia psyllid. California Agriculture 49(4): 36–40. https://doi.org/10.3733/ca.v049n04p36
  • De León JH, Sétamou M (2010) Molecular Evidence suggests that populations of the Asian Citrus Psyllid parasitoid Tamarixia radiata (Hymenoptera: Eulophidae) from Texas, Florida, and Mexico represent a single species. Annals of the Entomological Society of America 103(1): 100–110. https://doi.org/10.1603/008.103.0113
  • Flores D, Ciomperlik M (2017) Biological control using the ectoparasitoid, Tamarixia radiata, against the Asian citrus psyllid, Diaphorina citri, in the Lower Rio Grande Valley of Texas. Southwestern Entomologist 42(1): 49–59. https://doi.org/10.3958/059.042.0105
  • Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3: 294–299. https://pubmed.ncbi.nlm.nih.gov/7881515/
  • Giantsis IA, Chaskopoulou A, Bon MC (2016) Mild-Vectolysis: Nondestructive DNA extraction method for vouchering sand flies and mosquitoes. Journal of Medical Entomology 53(3): 692–695. https://doi.org/10.1093/jme/tjv236
  • González-Hernández A, Arredondo-Bernal HC, Robles-González M, Martínez-Carrillo JL, Pérez J, López-Arroyo JI (2009) Determinación de especies de parasitoides del psílido asiático de los cítricos Diaphorina citri (Hemiptera: Psyllidae) en México. Entomología Mexicana 8: 373–377.
  • Hebert PDN, Ratnasingham S, de Waard JR (2003) Barcoding of animal life: Cytochrome c oxidase subunit 1 divergence among closely related species. Proceedings. Biological Sciences 270: S96–S99. https://doi.org/10.1098/rsbl.2003.0025
  • Hoang DT, Chernomor O, von Haeseler A, Minh BQ, Vinh LS (2018) UFBoot2: Improving the ultrafast bootstrap approximation. Molecular Biology and Evolution 35(2): 518–522. https://doi.org/10.1093/molbev/msx281
  • Kalyaanamoorthy S, Minh B, Wong TKF, von Haeseler A, Jermiin LS (2017) ModelFinder: Fast model selection for accurate phylogenetic estimates. Nature Methods 14(6): 587–589. https://doi.org/10.1038/nmeth.4285
  • Lomelí-Flores JR, Bueno R (2002) Nuevo registro de Tamarixia triozae (Burks) parasitoide del psílido del tomate Paratrioza cockerelli (Sulc) (Homoptera: Psyllidae) en México. Folia Entomologica Mexicana 41: 375–376.
  • McClay AS (1990) Distribution of Leucaena psyllid and its natural enemies in México: Implications for biological control. In: Napompeth B, MacDicken KG (Eds) Leucaena psyllid: problems and management. Proceedings of an international workshop held in Bogor (Indonesia), January 16–21 1989. Winrock International Institute for Agricultural Development, Bangkok, 139–143.
  • Munyaneza JE, Crosslin JM, Upton JE (2007) Association of Bactericera cockerelli (Homoptera: Psyllidae) with ‘Zebra chip’, a new potato disease in Southwestern United States and Mexico. Journal of Economic Entomology 100(3): 656–663. https://doi.org/10.1093/jee/100.3.656
  • Pante E, Schoelinck C, Puillandre N (2015) From integrative taxonomy to species description: One step beyond. Systematic Biology 64(1): 152–160. https://doi.org/10.1093/sysbio/syu083
  • Ramírez-Ahuja ML, Rodríguez-Leyva E, Lomelí-Flores JR, Torres-Ruíz A, Guzmán-Franco AW (2017) Evaluating combined use of a parasitoid and a zoophytophagous bug for biological control of the potato psyllid, Bactericera cockerelli. Biological Control 106: 9–15. https://doi.org/10.1016/j.biocontrol.2016.12.003
  • Rojas P, Rodríguez-Leyva E, Lomelí-Flores JR, Liu TX (2015) Biology and life history of Tamarixia triozae, a parasitoid of the potato psyllid Bactericera cockerelli. BioControl 60(1): 27–35. https://doi.org/10.1007/s10526-014-9625-4
  • Taylor GS, Martoni F (2020) Case of mistaken identity: resolving the taxonomy between Trioza eugeniae Froggatt and T. adventicia Tuthill (Psylloidea: Triozidae). Bulletin of Entomological Research 110(3): 340–351. https://doi.org/10.1017/S0007485319000695
  • Trifinopoulos J, Nguyen LT, von Haeseler A, Minh BQ (2016) W-IQ-TREE: A fast online phylogenetic tool for maximum likelihood analysis. Nucleic Acids Research 44(W1): W232–W235. https://doi.org/10.1093/nar/gkw256
  • Urbaneja-Bernat P, Pérez-Rodríguez J, Krüger K, Catalán J, Rizza R, Hernández-Suárez E, Tena A (2019) Host range testing of Tamarixia dryi (Hymenoptera: Eulophidae) sourced from South Africa for classical biological control of Trioza erytreae (Hemiptera: Psyllidae) in Europe. Biological Control 135: 110–116. https://doi.org/10.1016/j.biocontrol.2019.04.018
  • Yefremova Z, González-Santarosa G, Lomeli-Flores JR, Bautista-Martínez N (2014) A new species of Tamarixia Mercet (Hymenoptera, Eulophidae), parasitoid of Trioza aguacate Hollis and Martin (Hemiptera, Triozidae) in Mexico. ZooKeys 368: 23–35. https://doi.org/10.3897/zookeys.368.6468
  • Zuparko LR, De Queiroz DL, LaSalle J (2011) Two new species of Tamarixia (Hymenoptera: Eulophidae) from Chile and Australia, established as biological control agents of invasive psyllids (Hemiptera: Calophyidae, Triozidae) in California. Zootaxa 2921(1): 13–27. https://doi.org/10.11646/zootaxa.2921.1.2
login to comment