Διάλεξη 3 Φαινόμενα ειςβολήσ και προςαρμογήσ φυςικών πληθυςμών (Κ. Ματθιόπουλοσ)
Η εισβολή της Αφρικανικής μέλισσας στην Αμερική Η φυσική έκταση της Apis mellifera Εκτείνεται από το βόρεια Ευρώπη μέχρι τη νότια Αφρική και από τα Βρετανικά νησιά μέχρι τα Ουράλια όρη, το δυτικό Ιράν και την Αραβική χερσόνησο. Η μελέτη των μιτοχονδριακών απλοτύπων αποκάλυψε 4 χαρακτηριστικές γραμμές: Δυτικής Ευρώπης Ανατολικής Ευρώπης Αφρικής Μέσης Ανατολής Before humans began large-scale transportation and mixing of A. mellifera populations, the four lineages were probably allopatric in distribution. At least three, and probably all four, mitochondrial lineages have been introduced into the New World: The west European dominated d 16th through h 18th century introductions into N and S America The east European lineage dominated subsequent introductions N African lineages were also introduced and this was present al low frequency in feral N American population prior to the arrival of the African bee from Latin America. Η αρχή της εφόδου From 1954 to 1955, beekeeping agencies, government departments and private beekeeping cooperatives in Brazil initiated projects to increase the low honey production of European colonies kept by commercial beekeepers. In 1956, WE Kerr traveled to Africa to select queens of the best stocks The nucleus formed by: One of six queens from Tanganyika 46 of 132 queens from Pretoria, S. Africa mtdna haplotype frequencies PCR RFLP diagnostic sites A: African, M: W&N European, C: S-E European 1
A. lsrna cut with EcoRI B. CO-I with HincII C. COI/COII length polymorphism D. COI/COII with XbaI 1: Italy, 2: Russia, 3-4: Spain, 5-6: France, 7-10: S. Africa A. lsrna cut with EcoRI B. CO-I with HincII C. COI/COII length polymorphism D. COI/COII with XbaI 1-5: Arizona, USA; 6-10: feral swarms, Honduras mtdna maternal ancestry Paternal contribution to the history and pattern of gene flow? Nuclear markers: allozymes RAPDs nuclear RFLPs microsatellites Microsatellites 1985, compare sample from Yucatan (21) plus reference samples from Venezuela (28) with regard to microsatellite allele frequencies Yucatan: East European Venezuela: African / w European 1998, new set of samples (312 colonies) Yucatan: most were African / w European only four were east European Summary of molecular data Allele frequencies vs diagnostic alleles Mitochondrial i haplotypes vs nuclear markers When African bees first expand into a new location, both E and W European nuclear and mt markers are found in the feral population. Within 5-10 years, African markers predominate. East European markers decrease to <10%. West European markers remain constant at 16-30%. This may suggest that: West European markers have entered the African population during the initial introduction and then carried along during the invasion process Despite the 50 years of contact, the African bees have mainly retained their genetic identity. 2
Factors contributing in the preservation of African characteristics Colony growth and swarming rates Negative heterosis in hybrid bees Mating advantages for African drones African-patriline advantages during queen replacement Dominance of African alleles Nest usurpation Adaptation of natural olive fly populations Olive cultivation around the world Περιοχές καλλιέργειας της ελιάς 1. Analysis of olive fly populations adaptation in nature and in laboratory conditions 2. Genome organization 3. Spread of insecticide resistance Molecular mechanisms of resistance 98% of olive trees are grown in the Mediterranean basin A complex microsatellite on an acrylamide gel 1. Analysis of olive fly populations with the use of microsatellites G A T C Primer F A 10 (GA) 9 Primer R 3
Polymorphism analysis of microsatellite markers 8 repeats 9 repeats Electrophoresis of PCR products in denaturing acrylamide gels Primer F Primer R Primer F Primer R Allele β (GA) Allele α (GA) 8 9 (GA) 9 Electrophoresis in acrylamide αα ββ αβ 86 90 75 67 Gimmaraes-PO Arrhenys-SP Murcia-SP Lisbon-PO Madrid-SP 82 71 39 35 53 Aidin-TU Vasto-IT Kos-GR Ithaka-GR Lefkas-GR 38 100 29 20 14 Isolation of 64 microsatellite markers. Use 12 markers in order to analyze 671 flies from 19 locations in the European side of the Mediterranean. 30 53 46 Limassol-CY Nicosia-CY D2 1,5 1,0 0,5 D0,0 Li-PO Ma-SP Gi-PO Ar-SP Mu-SP It-GR Cr-GR Pa-GR Al-GR Man-GR Mal-GR Ky-GR Ni-CY Li-CY 0,2 0,18 0,16 0,14 0,12 0,1 0,08 0,06 y = 2E-05x + 0,0233 R 2 = 0,4664-0,5-1,0 Lef-GR Fa-IT Kos-GR Va-IT Aid-TU 0,04 0,02 0 0 500 1000 1500 2000 2500 3000 3500 4000-1,5-3 -2-1 0 1 2 3 D1 4
0,7 0,6 0,5 0,4 y = 3E-05x + 0,5084 R 2 = 0,8383 0,3 0,2 0,1 0 0 500 1000 1500 2000 2500 3000 3500 4000 Olive transport in the Americas Olive fly invasion in California Today, table olive production in California is a ~ $80 mil business yearly UPGMA dendrogram after 100 boostrap resamples Structure analysis of natural olive fly populations West med Group Central med Group East med Group 5
Principal Component Analysis of molecular variance. The first two axes explain 71.56% of the total variance Conclusions I Higher genetic variability in California than in Iberia Private alleles in California that are not present in Iberia Most likely origin of California population is the East Mediterranean Remaining questions Adaptation of invading population in the new environment Diachronic samples from California Single or multiple invasions?? San Luis Obispo 2004 & 2008 San Diego 2002, 2005 & 2009 Diachronic sample analysis UPGMA dendrogram Adaptation in the laboratory 6
SIT attempts against the olive fly ('70): No sex separation Low competitiveness of released flies COMPETITIVENESS OF LABORATORY STRAIN Department of Entomology, Institute of Plant Protection, Agricultural Research Organization, Beit-Dagan Colonization of wild Israeli population Hybrid strain International Atomic Energy Agency (IAEA, FAO, Vienna) Aim Concerns / Questions Adaptation in laboratory diet and oviposition conditions Determination of genetic differentiation during colonization Determination of kind and frequency of enrichments in order to maintain mating competitiveness Changes in genetic structure Reduction of competitiveness Genetic analysis of consecutive generations of the colonized Israeli strain with microsatellite markers Augustinos, PhD thesis 7
Genetic differentiation-1 Principal Coordinates Generation * na (mean) Coo ord. 2 Pop26 Pop27 Coord. 1 WILD F 1 F 2 F 5 F 11 4.5714 3.8571 3.5714 3.4286 2.4286 n a (mean) 5 4 3 2 1 0 Genetic differentiation Wild F1 F2 F5 F11 * na = Observed Number of Alleles Augustinos, Cosmides et al., last week Genetic differentiation-2 Generation WILD F 1 F 2 F 5 F 11 * ne (mean) 3.3031 2.7790 2.7287 2.7235 1.5926 * ne = Effective Number of Alleles n) n e (mean 4 3 2 1 0 Genetic differentiation Wild F1 F2 F5 F11 Generation WILD F 1 F 2 F 5 F 11 Genetic differentiation-3 * Ho (mean) 0.6204 0.6038 0.6921 0.5718 0.3488 * Ho = Observed Heterozygosity Ho (mean) 1.0 08 0.8 0.6 0.4 0.2 0.0 Genetic differentiation Wild F1 F2 F5 F11 D84 allele frequencies D64 allele frequencies 8
Conclusions-2 What does disappearance or fixation of an allele mean? Allele Frequency fluctuation Allele disappearance Allele fixation microsatellite (neutral marker) allele Α selection "oviposition" gene allele Β * adaptive mutation in artificial oviposition material Allele Β hitch-hikes with an adaptive mutation of a gene that favors oviposition in artificial material 9