Chromosome observation method at metaphase and pro-metaphase stages in diploid and octoploid strawberries

Nathewet Preeda, Tomohiro Yanagi, Kazumi Sone, Shin Taketa, Nobuyuki Okuda

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Chromosome observation is necessary to elucidate the structure, function, organization, and evolution of octoploid strawberry plants' genes and genomes. However, distinguishing strawberries' chromosomes from one another using light microscopy is extremely difficult, not only because of their small size and large number, but also because current chromosome observation methods are insufficient. Chromosome preparation and staining using maceration enzymes, acetic acid, and DAPI (4′,6-diamidino-2-phenylindole) were improved for this study to obtain clear images of somatic chromosomes in Fragaria vesca (2n = 14) and Fragaria×ananassa (2n = 56). Collected root tips of octoploid plants were placed in 0.002 M 8-hydroxyquinoline solution for 1 h and stored at 4 °C for 16 h. Subsequently, they were fixed using 3:1 absolute alcohol:glacial acetic acid for 40 min, hydrolyzed in the 1N HCl solution at room temperature for 2 h, macerated using an enzyme solution for 25 min at 42 °C, and stained in 1.5% lacto-propionic orcein solution. On the other hand, in case of DAPI staining, the macerated root tips of octoploid plants were soaked in 60% acetic acid for 5 min before staining. Clear digital images of F. vesca and F.×ananassa were obtained using light and fluorescent microscopy. Their respective 14 and 56 chromosomes were counted. Fluorescent microscopy yielded clear chromosome images at the pro-metaphase in F. vesca and F.×ananassa. This chromosome observation method alleviates the difficulties that have heretofore hindered chromosome analyses of strawberry plants.

Original languageEnglish
Pages (from-to)133-137
Number of pages5
JournalScientia Horticulturae
Volume114
Issue number2
DOIs
Publication statusPublished - Oct 2 2007
Externally publishedYes

Fingerprint

metaphase
strawberries
diploidy
chromosomes
Fragaria vesca
Ananas
acetic acid
methodology
root tips
microscopy
8-hydroxyquinoline
maceration
digital images
enzymes
light microscopy
ambient temperature
alcohols
genome

Keywords

  • Chromosome
  • DAPI
  • Fragaria vesca
  • Fragaria×ananassa
  • Strawberry

ASJC Scopus subject areas

  • Horticulture
  • Plant Science

Cite this

Chromosome observation method at metaphase and pro-metaphase stages in diploid and octoploid strawberries. / Preeda, Nathewet; Yanagi, Tomohiro; Sone, Kazumi; Taketa, Shin; Okuda, Nobuyuki.

In: Scientia Horticulturae, Vol. 114, No. 2, 02.10.2007, p. 133-137.

Research output: Contribution to journalArticle

Preeda, Nathewet ; Yanagi, Tomohiro ; Sone, Kazumi ; Taketa, Shin ; Okuda, Nobuyuki. / Chromosome observation method at metaphase and pro-metaphase stages in diploid and octoploid strawberries. In: Scientia Horticulturae. 2007 ; Vol. 114, No. 2. pp. 133-137.
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