Diagnostic molecular markers to detect and identify primary parasitoids (Hymenoptera: Braconidae) of Ericaphis fimbriata on highbush blueberry

Snehlata Mathur, David A. Raworth, Keith S. Pike, Sheila M. Fitzpatrick

Abstract


This research provides mitochondrial COI sequences and species-specific reverse primers for Aphidius ericaphidis Pike and Starý, Ephedrus incompletus Provencher, and Praon unicum Smith (Hymenoptera: Braconidae), which are primary parasitoids of Ericaphis fimbriata (Richards) (Hemiptera: Aphididae), an important vector of Blueberry scorch virus (BlScV) on highbush blueberry Vaccinium corymbosum L. (Ericales: Ericaceae) in the Pacific Northwest. The COI sequences are deposited under accession numbers EU574902, EU574904, GU237131, KC211020-211024, KC211027-211032.  The combination of three primers in a multiplex PCR assay detects and differentiates DNA from adults of all three parasitoid species.  When individual field-collected aphids were challenged with the multiplex PCR assay, immatures of the two numerically dominant species, A. ericaphidis and P. unicum, were readily detected as was multiparasitism by these two species.  The uncommon species, E. incompletus, was detected less frequently by multiplex PCR assay than by rearing from the aphid hosts.  The diagnostic molecular markers are useful tools for estimating of rates of parasitism and for identifying immatures of parasitoid species within aphid hosts, particularly if used in combination with rearing and dissection assays of field-collected aphids.

Keywords


Aphids; Parasitoids; mtDNA; COI; Multiplex PCR; Multiparasitism

Full Text:

PDF

References


Acheampong S., D. R. Gillespie, and D. J. M. Quiring. 2012. Survey of parasitoids (Hymenoptera) of the green peach aphids, Myzus persicae and foxglove aphid, Aulacorthum solani (Hemiptera: Aphididae) in British Columbia. Journal of Entomological Society of British Columbia 109: 12-23.

Bristow, P. R., R. R. Martin, and G. E. Windom. 2000. Transmission, field spread, cultivar response, and impact on yield in high bush blueberry infected with Blueberry scorch virus. Phytopathology 90: 474-479.

Carroll, D. P., and S. C. Hoyt. 1986. Hosts and habitats of parasitoids (Hymenoptera: Aphidiidae) implicated in biological control of apple aphid (Homoptera: Aphididae). Environmental Entomology 15: 1171-1178.

Chow, F. J., and M. Mackauer. 1984. Inter- and intraspecific larval competition in Aphidius smithi and Praon pequodorum (Hymenoptera: Aphidiidae). The Canadian Entomologist 116: 1097-1107.

Chow, F. J., and M. Mackauer. 1985. Multiple parasitism of the pea aphid: stage of development of parasite determines survival of Aphidius smithi and Praon pequodorum (Hymenoptera: Aphidiidae). The Canadian Entomologist 117: 133-134.

Ehret, D. L., B. Frey, T. Forge, T. Helmer, and D. R. Bryla. 2012. Effects of drip irrigation configuration and rate on yield and fruit quality of young highbush blueberry plants. Hortscience 47: 414-421.

Folmer, O., M. Black, W. Hoeh, R. Lutz, and R. Vrijenhoek. 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3: 294-299.

Gariepy, T. D., U. Kuhlmann, C. Gillott, and M. Erlandson. 2007. Parasitoids, predators and PCR: the use of diagnostic molecular markers in biological control of arthropods. Journal of Applied Entomology 131: 225-240.

Hebert, P. D. N., S. Ratnasingham, and J. R. deWaard. 2003. Barcoding animal life: cytochome oxydase subunit 1 divergences among closely related species. Proceedings of the Royal Society of London, Series B 270 (supplement 1): 96-99.

Johnson, J. W. 1987. A revision of the species of Praon Haliday in North America north of Mexico (Hymenoptera: Aphidiidae). The Canadian Entomologist 119: 999-1025.

Pike, K. S., P. Starý, T. Miller, D. Allison, L. Boydston, G. Graf, and R. Gillespie. 1997. Small-grain aphid parasitoids (Hymenoptera: Aphelinidae and Aphidiidae) of Washington: distribution, relative abundance, seasonal occurrence, and key to known North American species. Environmental Entomology 26: 1299-1311.

Pike, K. S., P. Starý, T. Miller, G. Graf, D. Allison, L. Boydston, and R. Miller. 2000. Aphid parasitoids (Hymenoptera, Braconidae, Aphidiinae) of Northwest USA. Proceedings of the Entomological Society of Washington 102: 688-740.

Pike, K. S., P. Starý, G. Graf, D. A. Raworth, S. Mathur, L. K. Tanigoshi, and T. Murray. 2011. A new Aphidius Nees (Hymenoptera, Braconidae, Aphidiinae) of Ericaphis fimbriata (Richards) (Hemiptera, Aphidiidae) and key to parasitoids of blueberry aphid in the Pacific Northwest. Zootaxa 2802: 58-62.

Raworth, D. A. 2004. Ecology and management of Ericaphis fimbriata (Hemiptera: Aphidiidae) in relation to the potential for spread of Blueberry scorch virus. The Canadian Entomologist 136: 711-718.

Raworth, D. A., and D. Schade. 2006. Life history parameters and population dynamics of Ericaphis fimbriata (Hemiptera: Aphididae) on blueberry, Vaccinium corymbosum. The Canadian Entomologist 138: 205-217.

Raworth, D. A., K. S. Pike, L. K. Tanigoshi, S. Mathur, and G. Graf. 2008. Primary and secondary parasitoids (Hymenoptera) of aphids (Hemiptera: Aphididae) on blueberry and other Vaccinium in the Pacific Northwest. Environmental Entomology 37: 472-477.

Rozen, S., and H. J. Skaletsky. 2000. Primer3 on the WWW for general users and for biologist programmers pp. 365-386 In S. Krawetz and S. Misener, Bioinformatics Methods and Protocols: Methods in Molecular Biology. Humana Press. New Jersey.

Smith, C. F. 1944. The Aphidiinae of North America (Hymenoptera: Braconidae). Ohio State University Contributions in Zoology and Entomology 6: 1-154.

SYSTAT Software Inc. 2007. SYSTAT ® 13. 1735 Technology Dr., Ste. 430, San Jose, CA 95110 USA.

Vafaie, E. K., S. M. Fitzpatrick, and J. S. Cory. Under revision. Does rearing an aphid parasitoid on one host affect its ability to parasitize another species? Agricultural and Forest Entomology.

Weathersbee A. A., K. A. Shufran, T. D. Panchal, P. M. Dang, and G.A. Evans. 2004. Detection and differentiation of parasitoids (Hymenoptera: Aphidiidae and Aphelinidae) of the brown citrus aphid (Homoptera: Aphididae): species-specific polymerase chain reaction amplification of 18S rDNA. Annals of the Entomological Society of America 97: 286-292.


Refbacks

  • There are currently no refbacks.