title: Microdissection and painting of the Y chromosome in spinach (Spinacia oleracea).
abstract: Spinach has long been used as a model for genetic and physiological studies of sex determination and expression. Although trisomic analysis from a cross between diploid and triploid plants identified the XY chromosome as the largest chromosome, no direct evidence has been provided to support this at the molecular level. In this study, the largest chromosomes of spinach from mitotic metaphase spreads were microdissected using glass needles. Degenerate oligonucleotide primed polymerase chain reaction was used to amplify the dissected chromosomes. The amplified products from the Y chromosome were identified using the male-specific marker T11A. For the first time, the largest spinach chromosome was confirmed to be a sex chromosome at the molecular level. PCR products from the isolated chromosomes were used in an in situ probe mixture for painting the Y chromosome. The fluorescence signals were mainly distributed on all chromosomes and four pair of weaker punctate fluorescence signal sites were observed on the terminal region of two pair of autosomes. These findings provide a foundation for the study of sex chromosome evolution in spinach.
authors: College of Life Science, Henan Normal University, Xinxiang, 453007, People's Republic of China. dcl75@yahoo.cn
authors: Chuan-Liang
authors: CL
authors: Deng
authors: Rui-Yun
authors: RY
authors: Qin
authors: Ying
authors: Y
authors: Cao
authors: Jun
authors: J
authors: Gao
authors: Shu-Fen
authors: SF
authors: Li
authors: Wu-Jun
authors: WJ
authors: Gao
authors: Long-Dou
authors: LD
authors: Lu
mesh_terms: D001483:Base Sequence; D020223:Chromosome Painting / Q000379:methods*; D032461:Chromosomes, Plant / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D004171:Diploidy; D015723:Gene Library; D005819:Genetic Markers / Q000235:genetics*; D018745:Genome, Plant / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008677:Metaphase; D042282:Microdissection / Q000379:methods*; D008938:Mitosis; D008969:Molecular Sequence Data; D018517:Plant Roots / Q000166:cytology / Q000235:genetics; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D018724:Spinacia oleracea / Q000166:cytology / Q000235:genetics*
chemical_list: D017931:DNA Primers; D018744:DNA, Plant; D005819:Genetic Markers
references: Chromosoma. 1999 Aug;108(4):266-70
references: 10460415
references: EMBO J. 1999 Aug 2;18(15):4169-79
references: 10428956
references: Plant J. 1996 Oct;10(4):679-89
references: 8893544
references: Chromosome Res. 2000;8(3):229-36
references: 10841050
references: Chromosome Res. 2004;12(8):767-76
references: 15702415
references: Theor Appl Genet. 2005 Sep;111(5):923-31
references: 16044269
references: Curr Top Dev Biol. 1998;38:167-223
references: 9399079
references: Plant Cell Rep. 2011 Jun;30(6):965-71
references: 21301852
references: Cytogenet Genome Res. 2006;114(2):175-7
references: 16825771
references: Curr Genomics. 2007 Mar;8(1):67-72
references: 18645627
references: Genome. 1995 Aug;38(4):706-14
references: 18470198
references: Hum Genet. 1988 Nov;80(3):224-34
references: 3192212
references: Dev Genes Evol. 2005 Mar;215(3):132-42
references: 15660251
references: Cytogenet Cell Genet. 1995;68(1-2):17-8
references: 7956351
references: Annu Rev Plant Biol. 2011;62:485-514
references: 21526970
references: J Genet. 2012 Aug;91(2):209-12
references: 22942092
references: Genetics. 1962 Sep;47(9):1225-41
references: 17248129
references: Genome. 2009 Jan;52(1):68-76
references: 19132073
references: Nature. 1998 May 21;393(6682):263-6
references: 9607762
references: Genetics. 1955 Jul;40(4):429-37
references: 17247563
references: Chromosome Res. 2012 Jan;20(1):111-25
references: 22198613
references: Genome. 1995 Dec;38(6):1082-90
references: 8654909
references: Hum Genet. 1988 Nov;80(3):235-46
references: 3192213
references: Genomics. 1992 Nov;14(3):769-74
references: 1427905
references: Mol Genet Genomics. 2009 Mar;281(3):249-59
references: 19085011
references: Plant J. 1993 Nov;4(5):899-903
references: 8275107
references: Planta. 2009 Feb;229(3):507-21
references: 19005675
references: Plant Cell Physiol. 1999 Jan;40(1):60-8
references: 10189703
references: BMC Plant Biol. 2010 Mar 12;10:46
references: 20226063
references: Chromosoma. 2009 Feb;118(1):43-51
references: 18685858
references: Mol Cell Biol. 1986 Nov;6(11):3826-30
references: 3796596
references: Proc Natl Acad Sci U S A. 1988 Dec;85(23):9138-42
references: 2973607
references: Genetica. 2006 Sep-Nov;128(1-3):167-75
references: 17028949
references: Chromosoma. 1981;82(2):205-16
references: 6452995
references: Plant Mol Biol. 1992 Nov;20(3):503-11
references: 1421153
references: Chromosome Res. 2004;12(3):245-50
references: 15125638
references: Hum Mol Genet. 1998;7(10):1619-26
references: 9735383
references: Am J Bot. 2007 Feb;94(2):141-50
references: 21642216
references: Planta. 2005 Oct;222(2):284-92
references: 15940462
references: Chromosoma. 1996;104(5):315-20
references: 8575242
references: Nature. 1989 Mar 23;338(6213):348-50
references: 2784197
references: Cell. 1984 Mar;36(3):783-8
references: 6697397
references: Genetica. 2011 Aug;139(8):1065-72
references: 21927842
references: BMC Plant Biol. 2008 Mar 18;8:28
references: 18366673
references: Genome. 1997 Oct;40(5):705-15
references: 9352647
references: Chromosoma. 2011 Jun;120(3):287-96
references: 21327830
references: Chromosome Res. 1997 Feb;5(1):57-65
references: 9088644