Theoretical and Applied Genetics

, Volume 129, Issue 3, pp 535–545 | Cite as

High-resolution tyramide-FISH mapping of markers tightly linked to the male-fertility restoration (Ms) locus of onion

  • Ludmila KhrustalevaEmail author
  • Jiming Jiang
  • Michael J. Havey
Original Article


Key message

Tyramide FISH was used to locate relatively small genomic amplicons from molecular markers linked to Ms locus onto onion chromosome 2 near the centromere, a region of relatively low recombination.


Fluorescence in situ hybridization (FISH) has not been readily exploited for physical mapping of molecular markers in plants due to the technical challenge of visualizing small single-copy probes. Signal amplification using tyramide (tyr) FISH can increase sensitivity up to 100-fold. We used tyr-FISH to physically locate molecular markers tightly linked to the nuclear male-fertility (Ms) restoration locus of onion onto mitotic metaphase, pachytene, and super-stretched pachytene chromosomes. Relatively short genomic amplicons (846–2251 bp) and a cDNA clone (666 bp) were visualized in 9–42 % of observed cells. The markers were assigned to proximal locations close to the centromere on the long arm of chromosome 2, a region of lower recombination, revealing that tightly linked markers may be physically distant from Ms. This result explains why several labs have identified molecular markers tightly linked to the Ms locus after screening relatively few DNA clones or primers and segregating progenies. Although these markers are still useful for marker-aided selection, our results indicate that map-based cloning of Ms will likely be difficult due to reduced recombination near this gene.


Cytoplasmic Male Sterility Restriction Fragment Length Polymorphism Mitotic Metaphase Pachytene Chromosome Mitotic Metaphase Chromosome 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We gratefully acknowledge the support of a research fellowship to LK (IIE Grantee ID 68130146) from the Fulbright-Hayes Program.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


Names are necessary to report factually on available data; however, the U.S. Department of Agriculture (USDA) neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Ludmila Khrustaleva
    • 1
    Email author
  • Jiming Jiang
    • 2
  • Michael J. Havey
    • 3
  1. 1.Center of Molecular Biotechnology, Department of Genetics and BiotechnologyRussian State Agrarian University-Timiryazev Agricultural AcademyMoscowRussia
  2. 2.Department of HorticultureUniversity of WisconsinMadisonUSA
  3. 3.USDA-ARS and Department of HorticultureUniversity of WisconsinMadisonUSA

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