ArticleMolecular cytogenetics2015
A modified method for preparing meiotic chromosomes based on digesting pollen mother cells in suspension.
Article in Molecular cytogenetics, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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3 citing papers in PubMed.
- The mechanism of tetraploidization in tree peony, and its implications for speciation and evolution of genusFrontiers in plant science · 2025Article
- Fluorescence in situ hybridization in plants: recent developments and future applications.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2019Review
- Application of a modified drop method for high-resolution pachytene chromosome spreads in two Phalaenopsis species.Molecular cytogenetics · 2016Article
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6 authors.
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Abstract
backgroundMeiotic chromosome preparation is a key step in plant meiotic research. Pollen mother cell (PMC) wall elimination is beneficial to cytogenetic experimental procedures. Without wall interference, these procedures are easier and more successful. In existing methods it is difficult to eliminate PMC walls completely and uniformly. In this paper, we present an improved method for digesting PMC walls, and one for providing massive chromosomal spreads on a slide for other cytogenetic experimental procedures.
resultsThree plants were selected to exhibit the modified meiotic chromosome preparation method. PMCs were dispersed as single cells and incubated in a mixed enzyme solution (3 % cellulose + 0.3 % pectinase + 1 % snailase) for 1.5-2.5 h. In total, 28.28 % cells were lost during this process. There were 800-1900 spreads on every slide and no PMC wall interference was found on any of the slides. The spreads were also evenly distributed on the slides. More spreads were obtained when PMC and protoplast densities in the suspension were increased. All three plants' spreads were successfully used to locate a 5 s rDNA conserved sequence. The Nicotiana hybrid's spreads were successfully used to identify the hybrid's parental genome.
conclusionThis is an alternative method for meiotic chromosome preparation. Through this method, PMC walls can be completely and uniformly eliminated, and hundreds of spreads on every slide can be obtained. These spreads can be successfully used for DNA in situ hybridization.
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