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Dioecy is relatively rare among plant species, and distinguishable sex chromosomes have been reported in few dioecious species. The multiple sex chromosome system (XX/XY1Y2) of    Humulus   japonicus    Siebold et Zuccarini, 1846 differs from that of other members of the family   Cannabaceae  , in which the XX/XY chromosome system is present. Sex chromosomes of    H.   japonicus    were isolated from meiotic chromosome spreads of males by laser microdissection with the P.A.L.M. MicroLaser system. The chromosomal DNA was directly amplified by degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR). Fast fluorescence  in situ  hybridization (FAST-FISH) using a labeled, chromosome-specific DOP-PCR product as a probe showed preferential hybridization to sex chromosomes. In addition, the DOP-PCR product was used to construct a short-insert,    H.   japonicus    sex chromosomes-specific DNA library. The randomly sequenced clones showed that about 12% of them have significant homology to    H.   lupulus    and 88% to    Cannabis   sativa    Linnaeus, 1753 sequences from GenBank database. Forty-four percent of the sequences show homology to plant retroelements. It was concluded that laser microdissection is a useful tool for isolating the DNA of sex chromosomes of    H.   japonicus    and for the construction of chromosome-specific DNA libraries for the study of the structure and evolution of sex chromosomes. The results provide the potential for identifying unique or sex chromosome-specific sequence elements in    H.   japonicus    and could aid in the identification of sex chromosome-specific repeat and coding regions through chromosome isolation and genome complexity reduction.

comparative cytogenetics

Use of laser microdissection for the construction of    Humulus   japonicus    Siebold et Zuccarini, 1846 (  Cannabaceae  ) sex chromosome-specific DNA library and cytogenetics analysis