Development of a genomic in situ hybridization method using Technovit 7100 sections of early wheat embryo.
It is difficult to observe the behavior of chromosomes in early wheat embryos because they are wrapped in several cell layers of the ovary. Here we conducted genomic in situ hybridization on sections of ovary embedded in Technovit 7100, a resinous compound suitable for in situ hybridization of mRNA in sectioned tissues. With this resin it is possible to make thin sections with high resolution, no autofluorescence, and good water permeability. These features enable histochemical study using fluorescence microscopy. We established the most suitable conditions for the denaturation of target DNA embedded in Technovit resin, and performed GISH on them. Using this method, we identified Leymus mollis chromosomes in the young ovary of F1 hybrids between wheat and L. mollis. Furthermore, we observed the behavior of maize chromosomes in early wheat x maize hybrid embryos.
Mochida
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K
Kihara Institute for Biological Research and Graduate School of Integrated Science, Yokohama City University, Japan.
Tsujimoto
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H
D000180:Acrylic Resins / Q000737:chemistry*; D002467:Cell Nucleus / Q000235:genetics; D002678:Chimera; D002875:Chromosomes; D018744:DNA, Plant / Q000032:analysis; D017403:In Situ Hybridization / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence; D012116:Resins, Plant / Q000737:chemistry; D012639:Seeds / Q000235:genetics*; D012680:Sensitivity and Specificity; D014908:Triticum / Q000235:genetics*; D003313:Zea mays / Q000235:genetics
D000180:Acrylic Resins; D018744:DNA, Plant; D012116:Resins, Plant; C053012:Technovit 7100