title: Cytogenetic characterization and AFLP-based genetic linkage mapping for the butterfly Bicyclus anynana, covering all 28 karyotyped chromosomes.
abstract: BACKGROUND
  The chromosome characteristics of the butterfly Bicyclus anynana, have received little attention, despite the scientific importance of this species. This study presents the characterization of chromosomes in this species by means of cytogenetic analysis and linkage mapping.
  
  
  RESULTS
  Physical genomic features in the butterfly B. anynana were examined by karyotype analysis and construction of a linkage map. Lepidoptera possess a female heterogametic W-Z sex chromosome system. The WZ-bivalent in pachytene oocytes of B. anynana consists of an abnormally small, heterochromatic W-chromosome with the Z-chromosome wrapped around it. Accordingly, the W-body in interphase nuclei is much smaller than usual in Lepidoptera. This suggests an intermediate stage in the process of secondary loss of the W-chromosome to a ZZ/Z sex determination system. Two nucleoli are present in the pachytene stage associated with an autosome and the WZ-bivalent respectively. Chromosome counts confirmed a haploid number of n = 28. Linkage mapping had to take account of absence of crossing-over in females, and of our use of a full-sib crossing design. We developed a new method to determine and exclude the non-recombinant uninformative female inherited component in offspring. The linkage map was constructed using a novel approach that uses exclusively JOINMAP-software for Lepidoptera linkage mapping. This approach simplifies the mapping procedure, avoids over-estimation of mapping distance and increases the reliability of relative marker positions. A total of 347 AFLP markers, 9 microsatellites and one single-copy nuclear gene covered all 28 chromosomes, with a mapping distance of 1354 cM. Conserved synteny of Tpi on the Z-chromosome in Lepidoptera was confirmed for B. anynana. The results are discussed in relation to other mapping studies in Lepidoptera.
  
  
  CONCLUSIONS
  This study adds to the knowledge of chromosome structure and evolution of an intensively studied organism. On a broader scale it provides an insight in Lepidoptera sex chromosome evolution and it proposes a simpler and more reliable method of linkage mapping than used for Lepidoptera to date.
authors.affiliation: Department of Evolutionary Biology, Institute of Biology, Leiden University, Leiden, The Netherlands.
authors.forename: Arjen E
authors.initials: AE
authors.lastname: Van't Hof
authors.forename: Frantisek
authors.initials: F
authors.lastname: Marec
authors.forename: Ilik J
authors.initials: IJ
authors.lastname: Saccheri
authors.forename: Paul M
authors.initials: PM
authors.lastname: Brakefield
authors.forename: Bas J
authors.initials: BJ
authors.lastname: Zwaan
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis; D000818:Animals; D002080:Butterflies / Q000166:cytology / Q000235:genetics*; D002874:Chromosome Mapping; D002875:Chromosomes / Q000235:genetics*; D005260:Female; D005819:Genetic Markers; D007399:Interphase; D007621:Karyotyping; D008297:Male; D008540:Meiosis; D009697:Nucleolus Organizer Region / Q000235:genetics; D012730:Sex Chromosomes / Q000235:genetics
chemical_list: D005819:Genetic Markers