15-20 September, Apimondia 2009, Montpellier, France. Mohamed Alburaki

Transcription

1 Centre National de la Recherche Scientifique Université Paris VI (Pierre et Marie Curie) September, Apimondia 2009, Montpellier, France Mohamed Alburaki Ph.D student, Laboratory LEGS/CNRS, Genetic, biodiversity and honeybee behavior University of Paris VI (Pierre et Marie Curie), France. Collaboration: CNRS/LEGS & Damascus University Supervised by: Dr. Lionel Garnery Univ. de Versailles, CNRS / LEGS, France Prof. Ali Alburaki, Univ. of Damascus, Syria Funding: French scholarship (AUF): Agence Universitaire de la Francophonie & CNRS/ LEGS ( Laboratoire d Evolution, Genomes et Speciation)

2 Genetic analyses of Syrian honeybee diversity Apis mellifera syriaca 1. Local subspecies found in Syria, Lebanon & Jordan 2. Well productive subspecies 3. Defensive honeybee 4. Very well adapted to Syrian environment conditions 12/7/2009 Alburaki.M 2

3 The purposes of this study 1. What is the genetic relationship between A.m.syriaca & other honeybee subspecies? 2. To which evolutionary lineage belongs the Syrian honeybee populations? 3. what kind of haplotypes is present in this honeybee subspecies? 4. Are we assisting to genetic changes, evolutions or even degradations of our local honeybee due to the uncontrolled and random inportation of other honeybee subspecies? 5. Trying to determine the geographical areas of introgression/ importations? With which other subspecies? 6. Leading to set up genetic conservatories in the most adapted Syrian areas for our local honeybee Apis mellifera syriaca. 3

4 Honeybee evolutionary lineages Repartition of the 26 subspecies of honeybee Apis mellifera (Ruttner, 1988; Franck et al., 2000b). M: West Mediterranean C: North Mediterranean O: Oriental A: African 4

5 Sampling Genetic analyses of Syrian honeybee diversity Apis mellifera syriaca 5

6 The molecular markers 1. Mitochondrial marker: mtdna, COI-COII region. Powerful marker (honeybee queen passes this DNA on without modification to all her descendents). Non coding region bp CAPS/PCR-RFLP technique Sequencing in ND6 & CytB regions 6

7 1. Mitochondrial marker: mtdna, COI-COII region. A- Identification of the evolutionary lineages: 1. Extraction of the DNA from honeybee thorax, (Chelex method). 2. PCR amplification of the intergenic region COI-COII, (H2, E2 primers). 3. Electrophoresis migration on agarose gel, (1.4% concentration). 7

8 1. Mitochondrial marker: mtdna, COI-COII region. A- Identification of the evolutionary lineages: Q: C lineage, P0Q: A lineage, P0QQ: A lineage 8

9 1. Mitochondrial marker: mtdna, COI-COII region. A- Identification of the evolutionary lineages: A C An overview of the two evolutionary lineages distribution on Syrian map. Z & A belong to the African lineage A C belongs to the North Mediterranean lineage C 9

10 1. Mitochondrial marker: mtdna, COI-COII region. 1. CAPS technique. B- Haplotypes identification : 2. Digestion of PCR products with DraI, (Incubation 37 C, 48h). 3. Electrophoresis migration on poly-acrylamide gel, (7.5% concentration). 10

11 1. Mitochondrial marker: mtdna, COI-COII region. B- Haplotypes identification : Local honeybee A.m. syriaca Haplotype Z Haplotype C2 : A.m. ligustica 11

12 1. Mitochondrial marker: mtdna, COI-COII region. B- Haplotypes identification : A.m. syriaca : Haplotype Z Haplotype C2 : A.m. ligustica Haplotype C3 : Buckfast bee 12

13 1. Mitochondrial marker: mtdna, COI-COII region. C- Sequencing of the haplotypes Z characterizing the local Syrian honeybee : mtdna / intergenic region COI-COII / sequenced with 4 capillary ABI genetic analyser 3130 Alignment/ BioEdit software 13

14 1. Mitochondrial marker: mtdna, COI-COII region. Restriction map/ Profile of the Syrian haplotypes Z Damascus: C1, C2, Z1, Z2, Z3, Z4, Z5, Z7, Z15, Z16, A2, A3. Souwaida: C1, C2, Z1, Z2, Z3, Z4, Z5, Z7. Idlib: C1, C2, Z3, Z4, Z5, Z7. Latakia: C1, C2, C3, Z1, Z2, Z3, Z4, Z5, Z7, Z14. Kounitera: C1, Z1, Z2, Z3, Z7, Z14, Z15. Alburaki et al., (in prep.) 14

15 The molecular markers 2. Microsatellite marker: 14 loci on the genomic DNA. Genomic DNA 14 loci SSR technique DNA fragments were identified using ABI genetic analyser Neighbor-Joining trees were performed using specific software. 15

16 2. Microsatellite markers: 14 loci on the genomic DNA. TreeMaker\Syrian Populations, Neighbor Joining, Bootstaping, Cavalli-Sforza O C A 16 M

17 Conclusion Syrian honeybee A.m.syriaca is present in almost Syrian regions (according to our sampling) with less introgressed area in the South of the country. Nearly all Syrian honeybee populations contain importation of foreign honeybee subspecies but in a low percentages. We have recorded 26 specific haplotypes (named Z haplotypes, Alburaki et al., in prep.) related to this honeybee subspecies. Some regions are promising to establish in them genetic conservatories as the level of introgression is very low and other encouraged factors. Focusing more on this area in further studies as it is an area of interaction and contact between different evolutionary lineages. 12/7/2009 M. Alburaki 17

18 Many thanks to: Syrian beekeepers to have contributed in the sampling Syrian beekeeper association Hélène Legout, the technician of our team for her help during 2 years of work. All member of our laboratory LEGS/CNRS. mohamed.alburaki@legs.cnrs-gif.fr 12/7/2009 M. Alburaki 18