Skip to main content

Genetic and physical maps around the sex-determining M-locus of the dioecious plant asparagus

Molecular Genetics and Genomics volume 278, pages 221–234 (2007)Cite this article

Abstract

Asparagus officinalis L. is a dioecious plant. A region called the M-locus located on a pair of homomorphic sex chromosomes controls the sexual dimorphism in asparagus. The aim of this work was to clone the region determining sex in asparagus from its position in the genome. The structure of the region encompassing M should be investigated and compared to the sex-determining regions in other dioecious model species. To establish an improved basis for physical mapping, a high-resolution genetic map was enriched with AFLP markers closely linked to the target locus by carrying out a bulked segregant analysis. By screening a BAC library with AFLP- and STS-markers followed by chromosome walking, a physical map with eight contigs could be established. However, the gaps between the contigs could not be closed due to a plethora of repetitive elements. Surprisingly, two of the contigs on one side of the M-locus did not overlap although they have been established with two markers, which mapped in a distance as low as 0.25 cM flanking the sex locus. Thus, the clustering of the markers indicates a reduced recombination frequency within the M-region. On the opposite side of the M-locus, a contig was mapped in a distance of 0.38 cM. Four closely linked BAC clones were partially sequenced and 64 putative ORFs were identified. Interestingly, only 25% of the ORFs showed sequence similarity to known proteins and ESTs. In addition, an accumulation of repetitive sequences and a low gene density was revealed in the sex-determining region of asparagus. Molecular cytogenetic and sequence analysis of BACs flanking the M-locus indicate that the BACs contain highly repetitive sequences that localize to centromeric and pericentromeric locations on all asparagus chromosomes, which hindered the localization of the M-locus to the single pair of sex chromosomes. We speculate that dioecious Silene, papaya and Asparagus species may represent three stages in the evolution of XX, XY sex determination systems. Given that asparagus still rarely produces hermaphroditic flowers and has homomorphic sex chromosomes, this species may be an ideal system to further investigates early sex chromosome evolution and the origins of dioecy.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2

References

  • Arumuganathan K, Earle ED (1991) Nuclear DNA content of some important plant species. Plant Mol Biol Rep 9:208–218

    CAS  Google Scholar 

  • Bendahmane A, Kanyuka K, Baulcombe DC (1997) High-resolution genetical and physical mapping of the Rx gene for extreme resistance to potato virus X in tetraploid potato. Theor Appl Genet 95:153–162

    Article  CAS  Google Scholar 

  • Boyko E, Kalendar R, Korzun V, Fellers J, Korol A, Schulman AH, Gill BS (2002) A high-density cytogenetic map of the Aegilops tauschii genome incorporating retrotransposons and defense-related genes: insights into cereal chromosome structure and function. Plant Mol Biol 48:767–790

    PubMed  Article  CAS  Google Scholar 

  • Copenhaver GP, Nickel K, Kuromori T, Benito MI, Kaul S, Lin X, Bevan M, Murphy G, Harris B, Parnell LD, McCombie WR, Martienssen RA, Marra M, Preuss D (1999) Genetic definition and sequence analysis of Arabidopsis centromeres. Science 286:2468–2474

    PubMed  Article  CAS  Google Scholar 

  • Dellaporta SL, Calderon-Urrea A (1993) Sex determination in flowering plants. Plant Cell 5:1241–1251

    PubMed  Article  CAS  Google Scholar 

  • Ewing B, Green P (1998) Base-calling of automated sequencer traces using phred. II. Error probabilities. Genome Res 8:186–194

    PubMed  CAS  Google Scholar 

  • Ewing B, Hillier L, Wendl MC, Green P (1998) Base-calling of automated sequencer traces using phred. I. Accuracy assessment. Genome Res 8:175–185

    PubMed  CAS  Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13

    PubMed  Article  CAS  Google Scholar 

  • Fleischer B, Gindullis F, Dechyeva D, Schmidt T (2002) Beetle1, a Ty3-gypsy retrotransposon highly amplified at centromeres of Beta procumbens chromosomes. Plant and Animal Genome X Conference, San Diego, USA

  • Flory WS (1932) Genetic and Cytological Investigations on Asparagus officinalis L. Genet Princeton 17:432–467

    CAS  Google Scholar 

  • Fransz PF, Alonso-Blanco C, Liharska TB, Peeters AJ, Zabel P, de Jong JH (1996) High-resolution physical mapping in Arabidopsis thaliana and tomato by fluorescence in situ hybridization to extended DNA fibres. Plant J 9:421–430

    PubMed  Article  CAS  Google Scholar 

  • Galli MG, Bracale M, Falavigna A, Soave C (1988) Sexual differentiation in Asparagus officinalis L.: I. DNA characterization and mRNA activities in male and female flowers. Sex Plant Reprod 1:202–207

    Article  Google Scholar 

  • Gindullis F, Desel C, Galasso I, Schmidt T (2001) The large-scale organization of the centromeric region in Beta species. Genome Res 11:253–265

    PubMed  Article  CAS  Google Scholar 

  • Gorman SW, Banasiak D, Fairley C, McCormick S (1996) A 610 kb YAC clone harbors 7 cM of tomato (Lycopersicon esculentum) DNA that includes the male sterile 14 gene and a hotspot for recombination. Mol Gen Genet 251:52–59

    PubMed  CAS  Google Scholar 

  • Hosouchi T, Kumekawa N, Tsuruoka H, Kotani H (2002) Physical map-based sizes of the centromeric regions of Arabidopsis thaliana chromosomes 1, 2, and 3. DNA Res 9:117–121

    PubMed  Article  CAS  Google Scholar 

  • Jakse J, Telgmann A, Jung C, Khar A, Melgar S, Cheung F, Town CD, Havey MJ (2006) Comparative sequence and genetic analysis of asparagus BACs reveal no microsynteny with onion or rice. Theor Appl Genet 114:31–39

    PubMed  Article  CAS  Google Scholar 

  • Jamsari A, Nitz I, Reamon-Büttner SM, Jung C (2004) BAC-derived diagnostic markers for sex determination in asparagus. Theor Appl Genet 108:1140–1146

    PubMed  Article  CAS  Google Scholar 

  • Jiang C, Lewis ME, Sink KC (1997) Combined RAPD and RFLP molecular linkage map of asparagus. Genome 40:69–76

    PubMed  CAS  Google Scholar 

  • Jiang J, Birchler JA, Parrott WA, Dawe RK (2003) A molecular view of plant centromeres. Trends Plant Sci 8:570–575

    PubMed  Article  CAS  Google Scholar 

  • Jiao Y, Jia P, Wang X, Su N, Yu S, Zhang D, Ma L, Feng Q, Jin Z, Li L, Xue Y, Cheng Z, Zhao H, Han B, Deng XW (2005) A tiling microarray expression analysis of rice chromosome 4 suggests a chromosome-level regulation of transcription. Plant Cell 17:1641–1657

    PubMed  Article  CAS  Google Scholar 

  • Jin W, Lamb JC, Vega JM, Dawe RK, Birchler JA, Jiang J (2005) Molecular and functional dissection of the maize B chromosome centromere. Plant Cell 17:1412–1423

    PubMed  Article  CAS  Google Scholar 

  • Jin W, Melo JR, Nagaki K, Talbert PB, Henikoff S, Dawe RK, Jiang J (2004) Maize centromeres: organization and functional adaptation in the genetic background of oat. Plant Cell 16:571–581

    PubMed  Article  CAS  Google Scholar 

  • Kato A, Lamb JC, Birchler JA (2004) Chromosome painting using repetitive DNA sequences as probes for somatic chromosome identification in maize. Proc Natl Acad Sci USA 101:13554–13559

    PubMed  Article  CAS  Google Scholar 

  • Kato A, Albert PS, Vega JM, Birchler JA (2006) Sensitive fluorescence in situ hybridization signal detection in maize using directly labeled probes produced by high concentration DNA polymerase nick translation. Biotech Histochem 81: 71–78

    PubMed  Article  CAS  Google Scholar 

  • Kejnovsky E, Kubat Z, Macas J, Hobza R, Mracek J, Vyskot B (2006) Retand: a novel family of gypsy-like retrotransposons harboring an amplified tandem repeat. Mol Gen Genet 276:254–263

    CAS  Google Scholar 

  • Kosambi DD (1944) The estimation of map distances from recombination values. Ann Eugen 12:172–175

    Google Scholar 

  • Kumekawa N, Hosouchi T, Tsuruoka H, Kotani H (2000) The size and sequence organization of the centromeric region of Arabidopsis thaliana chromosome 5. DNA Res 7:315–321

    PubMed  Article  CAS  Google Scholar 

  • Kumekawa N, Hosouchi T, Tsuruoka H, Kotani H (2001) The size and sequence organization of the centromeric region of Arabidopsis thaliana chromosome 4. DNA Res 8:285–290

    PubMed  Article  CAS  Google Scholar 

  • Lahaye T, Hartmann S, Töpsch S, Freialdenhoven A, Yano M, Schulze-Lefert P (1998) High-resolution genetic and physical mapping of the Rar1 locus in barley. Theor Appl Genet 97:526–534

    Article  CAS  Google Scholar 

  • Lai CW, Yu Q, Hou S, Skelton RL, Jones MR, Lewis KL, Murray J, Eustice M, Guan P, Agbayani R, Moore PH, Ming R, Presting GG (2006) Analysis of papaya BAC end sequences reveals first insights into the organization of a fruit tree genome. Mol Gen Genet 276:1–12

    CAS  Google Scholar 

  • Lamb JC, Birchler JA (2006) Retroelement genome painting: cytological visualization of retroelement expansions in the genera Zea and Tripsacum. Genetics 173:1007–1021

    PubMed  Article  CAS  Google Scholar 

  • Lengerova M, Kejnovsky E, Hobza R, Macas J, Grant SR, Vyskot B (2004) Multicolor FISH mapping of the dioecious model plant, Silene latifolia. Theor Appl Genet 108:1193–1199

    PubMed  Article  CAS  Google Scholar 

  • Liu Z, Moore PH, Ma H, Ackerman CM, Ragiba M, Yu Q, Pearl HM, Kim MS, Charlton JW, Stiles JI, Zee FT, Paterson AH, Ming R (2004) A primitive Y chromosome in papaya marks incipient sex chromosome evolution. Nature 427:348–352

    PubMed  Article  CAS  Google Scholar 

  • Löptien H (1979) Identification of the sex chromosome pair in asparagus (Asparagus officinalis L.). Z Pflanzenzüchtg 82:162–173

    Google Scholar 

  • Ma H, Moore PH, Liu Z, Kim MS, Yu Q, Fitch MM, Sekioka T, Paterson AH, Ming R (2004) High-density linkage mapping revealed suppression of recombination at the sex determination locus in papaya. Genetics 166:419–436

    PubMed  Article  CAS  Google Scholar 

  • Ma J, Bennetzen JL (2006) Recombination, rearrangement, reshuffling, and divergence in a centromeric region of rice. Proc Natl Acad Sci USA 103:383–388

    PubMed  Article  CAS  Google Scholar 

  • Mariotti B, Navajas-Perez R, Lozano R, Parker JS, de la HR, Rejon CR, Rejon MR, Garrido-Ramos M, Jamilena M (2006) Cloning and characterization of dispersed repetitive DNA derived from microdissected sex chromosomes of Rumex acetosa. Genome 49:114–121

    PubMed  CAS  Google Scholar 

  • Marra MA, Kucaba TA, Dietrich NL, Green ED, Brownstein B, Wilson RK, McDonald KM, Hillier LW, McPherson JD, Waterston RH (1997) High throughput fingerprint analysis of large-insert clones. Genome Res 7:1072–1084

    PubMed  CAS  Google Scholar 

  • Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88:9828–9832

    PubMed  Article  CAS  Google Scholar 

  • Nicolas A (1998) Relationship between transcription and initiation of meiotic recombination: toward chromatin accessibility. Proc Natl Acad Sci U S A 95:87–89

    PubMed  Article  CAS  Google Scholar 

  • Nicolas M, Marais G, Hykelova V, Janousek B, Laporte V, Vyskot B, Mouchiroud D, Negrutiu I, Charlesworth D, Moneger F (2005) A gradual process of recombination restriction in the evolutionary history of the sex chromosomes in dioecious plants. PLoS Biol 3:e4 Epub

  • Pelissier T, Tutois S, Deragon JM, Tourmente S, Genestier S, Picard G (1995) Athila, a new retroelement from Arabidopsis thaliana. Plant Mol Biol 29:441–452

    PubMed  Article  CAS  Google Scholar 

  • Pereira V (2004) Insertion bias and purifying selection of retrotransposons in the Arabidopsis thaliana genome. Genome Biol 5:R79

    PubMed  Article  Google Scholar 

  • Reamon-Büttner SM, Jung C (2000) AFLP-derived STS markers for the identification of sex in Asparagus officinalis L. Theor Appl Genet 100:432–438

    Article  Google Scholar 

  • Reamon-Büttner SM, Schmidt T, Jung C (1999) AFLPs represent highly repetitive sequences in Asparagus officinalis L. Chromosome Res 7:297–304

    PubMed  Article  Google Scholar 

  • Reamon-Büttner SM, Schondelmaier J, Jung C (1998) AFLP markers tightly linked to the sex locus in Asparagus officinalis L. Mol Breed 4:91–98

    Article  Google Scholar 

  • Roberts PA (1965) Difference in the behavior of eu- and hetero-chromatin: cross-over. Nature 205:725–726

    PubMed  Article  CAS  Google Scholar 

  • Sakamoto K, Abe T, Matsuyama T, Yoshida S, Ohmido N, Fukui K, Satoh S (2005) RAPD markers encoding retrotransposable elements are linked to the male sex in Cannabis sativa L. Genome 48:931–936

    PubMed  CAS  Google Scholar 

  • Sakamoto K, Ohmido N, Fukui K, Kamada H, Satoh S (2000) Site-specific accumulation of a LINE-like retrotransposon in a sex chromosome of the dioecious plant Cannabis sativa. Plant Mol Biol 44:723–732

    PubMed  Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York, USA

    Google Scholar 

  • Shibata F, Hizume M, Kuroki Y (1999) Chromosome painting of Y chromosomes and isolation of a Y chromosome-specific repetitive sequence in the dioecious plant Rumex acetosa. Chromosoma 108:266–270

    PubMed  Article  CAS  Google Scholar 

  • Soderlund C, Humphray S, Dunham A, French L (2000) Contigs build with Fingerprints, Markers, and FPC V4.7. Genome Res 10:1772–1787

    PubMed  Article  CAS  Google Scholar 

  • Spada A, Caporali E, Marziani G, Portaluppi P, Restivo FM, Tassi F, Falavigna A (1998) A genetic map of Asparagus officinalis based on integrated RFLP, RAPD and AFLP molecular markers. Theor Appl Genet 97:1083–1089

    Article  CAS  Google Scholar 

  • Staden R (1996) The Staden sequence analysis package. Mol Biotechnol 5:233–241

    PubMed  CAS  Article  Google Scholar 

  • Storey WB (1938) Segregation of sex types in Solo papaya and their application to the selection of seed. Am Soc Hortic Sci Proc 35:83–85

    Google Scholar 

  • Tai T, Dahlbeck D, Stall RE, Peleman J, Staskawicz BJ (1999) High-resolution genetic and physical mapping of the region containing the Bs2 resistance gene of pepper. Theor Appl Genet 99:1201–1206

    Article  CAS  Google Scholar 

  • Tanksley SD, Ganal MW, Prince JP, Devicente MC, Bonierbale MW, Broun P, Fulton TM, Giovannoni JJ, Grandillo S, Martin GB, Messeguer R, Miller JC, Miller L, Paterson AH, Pineda O, Röder MS, Wing RA, Wu W, Young ND (1992) High density molecular linkage maps of the tomato and potato genomes. Genetics 132:1141–1160

    PubMed  CAS  Google Scholar 

  • The Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 408:796–815

    Google Scholar 

  • Uno Y (2002) Haploid Production from polyembryonic seeds of Asparagus officinalis L. Acta Hort (ISHS) 589:217–221

    Google Scholar 

  • Van Ooijen JW, Voorrips RE (2001) JoinMap® 3.0, software for the calculation genetic linkage maps. Plant Research International, Wageningen, The Netherlands

  • Vyskot B, Hobza R (2004) Gender in plants: Sex chromosomes are emerging from the fog. Trends Genet 20:432–438

    PubMed  Article  CAS  Google Scholar 

  • Wang Y, Tang X, Cheng Z, Mueller L, Giovannoni J, Tanksley SD (2006) Euchromatin and pericentromeric heterochromatin: comparative composition in the tomato genome. Genetics 172:2529–2540

    PubMed  Article  CAS  Google Scholar 

  • Waugh R, McLean K, Flavell AJ, Pearce SR, Kumar A, Thomas BB, Powell W (1997) Genetic distribution of Bare-1-like retrotransposable elements in the barley genome revealed by sequence-specific amplification polymorphisms (S-SAP). Mol Gen Genet 253:687–694

    PubMed  Article  CAS  Google Scholar 

  • Weiblen GD, Oyama RK, Donoghue MJ (2000) Phylogenetic analysis of dioecy in monocotyledons. Am Nat 155:46–58

    PubMed  Article  Google Scholar 

  • Wiegant J, Verwoerd N, Mascheretti S, Bolk M, Tanke HJ, Raap AK (1996) An evaluation of a new series of fluorescent dUTPs for fluorescence in situ hybridization. J Histochem Cytochem 44:525–529

    PubMed  CAS  Google Scholar 

  • Wu J, Yamagata H, Hayashi-Tsugane M, Hijishita S, Fujisawa M, Shibata M, Ito Y, Nakamura M, Sakaguchi M, Yoshihara R, Kobayashi H, Ito K, Karasawa W, Yamamoto M, Saji S, Katagiri S, Kanamori H, Namiki N, Katayose Y, Matsumoto T, Sasaki T (2004) Composition and structure of the centromeric region of rice chromosome 8. Plant Cell 16:967–976

    PubMed  Article  CAS  Google Scholar 

  • Yu A, Zhao C, Fan Y, Jang W, Mungall AJ, Deloukas P, Olsen A, Doggett NA, Ghebranious N, Broman KW, Weber JL (2001) Comparison of human genetic and sequence-based physical maps. Nature 409:951–953

    PubMed  Article  CAS  Google Scholar 

  • Yu Q, Hou S, Feltus FA, Moore RC, Moore PH, Alam M, Jiang J, Paterson AH, Ming R (2006) The papaya Y chromosome evolved recently and shows gene paucity and DNA sequence expansion. Plant and Animal Genome XV Conference 2006, San Diego, USA

Download references

Acknowledgments

We thank C. Kruse, B. Defant, M. Bruisch and E. Danklefsen for their technical assistance. This work was financially supported by the German research foundation DFG (grant No. JU205/2-5 and 205/11-1). J. Chris Pires acknowledges funding from the US National Science Foundation.

Author information

Affiliations

  1. Plant Breeding Institute, Christian-Albrechts-University Kiel, Olshausenstr. 40, Kiel, 24098, Germany

    Alexa Telgmann-Rauber & Christian Jung

  2. Plant Breeding Section, Faculty of Agriculture, Andalas University, 25163, Padang-West Sumatra, Indonesia

    Ari Jamsari

  3. Division of Biological Sciences, University of Missouri-Columbia, 371 B Life Sciences Center, Columbia, MO, 65211-7400, USA

    Michael S. Kinney & J. Chris Pires

Authors
  1. Alexa Telgmann-Rauber
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ari Jamsari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael S. Kinney
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. Chris Pires
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Christian Jung
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Christian Jung.

Additional information

Communicated by S. Hohmann.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Telgmann-Rauber, A., Jamsari, A., Kinney, M.S. et al. Genetic and physical maps around the sex-determining M-locus of the dioecious plant asparagus. Mol Genet Genomics 278, 221–234 (2007). https://doi.org/10.1007/s00438-007-0235-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00438-007-0235-z

Keywords

  • Asparagus officinalis
  • FISH
  • Centromere
  • Sex chromosome
  • Dioecy