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Genetic structure of varroa mite populations in A. mellifera syriaca.

Introduction

Beekeeping and hunting of wild honey have been important in the Middle East generally and in Jordan particularly for thousands of years[6]. The honeybee of the eastern Mediterranean, A. mellifera syriaca, is found in the regions of Syria, Lebanon, Israel, Gaza Strip and Jordan. First detailed descriptions were given by Buttel-Reepen[2], and its racial status was later confirmed by Ruttner based on samples collected by Brother Adam in[1].

Due to the high defensive behavior of the A. mellifera syriaca, many beekeepers started in the late seventies a world wide import of honey bee queens and bee colonies (MoA Jordan), but resent research confirmed that it is possible to find A. mellifera syriaca in some areas of Jordan kept in traditional bee hives[6] . In 1986 the beekeeping sector faced major losses, with over 50% of the beehives dying. This happened one year after the first detection of the ectoparasitic varroa mite[9] , but until now little information about honeybees in general and Varroa mites in particular are available from the Middle East.

The aim of this study was to gain insight into the genetic traits and invasion process of V. destructor at both local and worldwide scale and on the interaction between the Korea and Japan Varroa mites types in particular.

Material and methods

a. Populations

The studied populations consisted of a total of 60 female mites belonging to 10 A.m. ligustica from the north of Jordan, where the bees are kept in modern bee hives and 20 mites from A. mellifera syriaca populations in 2 different locations, where the 1st location in the south where the bees are kept in traditional wooden hives while the 2nd location in the northern part of Jordan where the bees are kept in clay mode hives, these two traditional apiaries where conformed as the closest to the historical samples collected by Brother Adam in the 1952, using morphometric[6] and mitochondrial DNA methods[7].

b. DNA extraction and sequencing of Varroa mites.

Individual mites were dried in sterile microcentrifuge tubes, and then ground using disposable pestles. DNA was extracted by suspending these samples in 100 ul 10% Chelex-100 (Bio-Rad), followed by incubation at 95[degrees]C for 10 minutes. A 1:10 dilution of this suspension was used for PCR. using specific oligonucleotide primers CO1F.F and CO1N.R[3], sequences 5'- CAGATCGAAATTTTAATA C-3 ' and 5'- TACAGCTCCTATAGATAAAA-3',respectively). Reaction mixes consisted of 2 ul mite DNA extract, 2 U Taq DNA polymerase with recommended buffer 2 (Boehringer), 1mM DNTP mix, 2mM Mg[Cl.sub.2] , and 0.2 [micro]M of each primer. PCR was carried out on an MJ-Research PTC-100 thermal cycler using 30 cycles of 93[degrees]C 1 min, 55[degrees]C 1 min, 72[degrees]C 1 min. Bands of an appropriate size were confirmed by agarose gel electrophoresis, then PCR products were purified directly (Gene Pure). Products were sequenced using Big Dye 2.0 end-terminal cycle sequencing (Applied Biosystems), followed by separation and analysis on an Applied Biosystems 3130 DNA Analysis machine. Sequencing was carried out in one direction from the 5' end using primer CO1F.F.

C. Sequence analyses

Sequences were checked and trimmed using the software program Sequencher (Gene Codes). Sequences were then aligned with each other and with known COI sequences for Varroa mites 7 samples from the Genbank database, one haplotype from Algeria[10], and nine samples from Bolivia, South America (Evans, J.D. and Hopkins, D., unpublished). Of the Bolivian samples, three (QB1, QB5, and QB9) fell into the 'Japan' haplotype, while the rest were of the more widespread 'Korean' haplotype.

Results and discussions

A total of 16 Varroa mites collected from several sites in Jordan in addition to those from Algeria, Bolivia, South Korea, Vietnam, China, Japan, Nepal and Sri lanka were sequenced successfully. The Jordanian samples were grouped in one cluster and were matches with the 'Korean' haplotype of V. destructor, which has historically been the more virulent of the two widespread V. destructor haplotypes. The sequenced mites were collected equally from the two bee subspecies. This is the first analysis of V destructor types that are parasitizing A. mellifera syriaca. We took advantage of the sequence of the entire mitochondrial genome of V. destructor[3,8] to provide a more precise picture of the genetic structure of Varroa populations in Jordan in comparing with worldwide picture because of the unique geographical position of Jordan as a transcendent area between the African and the Asian continents. Further collections within A. m. syriaca can help confirm that the mites in this subspecies are, as with ligustica, predominantly the Korean haplotype. If so, any differences in mite resistance between A. m. syriaca and A. m. ligustica will likely reflect resistance traits of these two bee subsepecies, rather than those of their mite parasites.

References

[1.] Brother Adam., 1954. In Search of the Best Strains of Bee: Second Journey. Bee World, 35: 193-203.

[2.] Buttel-Reepen, H., 1906. Apistica. Beitrage zur Systematik, Biologie, sowie zur geschichtlichen und geographischen Verbreitung der Honigbiene (Apis mellifica L), ihrer Varietaten und der ubrigen Apis-Arten. Veroff. Zool. Mus. Berlin., pp: 118-120.

[3.] Evans, J.D. and D.L. Lopez, 2002. Complete mitochondrial DNA sequence of the important honey bee pest, Varroa destructor (Acari: Varroidae). Exp. Appl. Acarol., 27: 69-79.

[4.] Evans, J.E., 2000. Microsatellite loci in the honey bee parasitic mite Varroa jacobsoni. Mol. Ecol., 9: 1436-1438.

[5.] Haddad, N. and S. Fuchs, 2004. Honeybee agrobiodiversity: a project in conservation of Apis mellifer syriaca in Jordan. Uludag bee Journal., 3: 116-120.

[6.] Haddad, N., 2005. Bees for Development Journal, 75: 6-7.

[7.] Meixner M., N. Haddad and S. Fuchs, 2006. Variation of mitochondrial DNA in honey bees of Jordan. Proceedings of The Second European Conference of Apidology, pp: 66.

[8.] Navajas, M., M. Solignac, Le Y. Conte, S. Cros-Arteil and J.M. Cornuet, 2002. The complete sequence of the mitochondrial genome of the honey-bee ectoparasite Varroa destructor (Acari: Mesostigmata). Mol. Biol. Evol., 19: 2313-2317.

[9.] Nazer, I.K., J.M. Rateb, 1990. Status of honey bee diseases and pests in Jordan. pp: 210-212. In: WRitter (Ed.) Proceedings of the international symposium on recent research on bee pathology, 5-7 September, 1990, Gent, Belgium. Rijksstation voor Nematologe en Entomology on behalf of Apimondia. Merelbeke.

[10.] Solignac, M, J.M. Cornuet, D. Vautrin, Le Y. Conte, D. Anderson, J.D. Evans, S. Cros-Arteil and M. Navajas, 2005. The invasive Korea and Japan types of Varroa destructor, ectoparasitic mites of the Western honey bee (Apis mellifera), are two partially isolated clones, Proc. Royal Society, Series B, 272: 411-420.

[11.] Solignac, M., D. Vautrin, A. Pizzo, M. Navajas, Y. Le Conte and J.M. Cornuet, 2003. Characterization of microsatellite markers for the apicultural pest Varroa destructor (Acari: Varroidae) and its relatives. Mol. Ecol. Notes, 3: 556-559.

Corresponding Author: Nizar Haddad N, National Center for Agricultural Research and Technology Transfer, Baqa' 19381 Jordan, E-mail: drnizarh@ncartt.gov.jo.

(1) National Center for Agricultural Research and Technology Transfer, Baqa' 19381 Jordan,

(2) United States Department of Agriculture, Agricultural Research Service, Bee Research Laboratory, Beltsville, Maryland 20705.

Nizar Haddad N, Hussein Migdadi, Jay Evans J, Jeff Pettis J.,: Genetic Structure of Varroa Mite Populations in A. Mellifera syriaca, Adv. Environ. Biol., 1(1): 1-3, 2007
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Title Annotation:Original Article
Author:N., Nizar Haddad; Migdadi, Hussein; J., Jay Evans; J., Jeff Pettis
Publication:Advances in Environmental Biology
Article Type:Report
Date:Sep 1, 2007
Words:1213
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