Abstract
A high-yield method for preparation of suspensions of intact Norway spruce [Picea abies (L.) Karst.] chromosomes was developed for the first time. To accumulate meristem root tip cells at metaphase, actively growing roots were subjected to subsequent treatments with 0.625 mM hydroxyurea for 18 h and after 8 h recovery in distilled water with 0.05 % (m/v) colchicine for 8 h. These treatments resulted in 50 % metaphase indices. Synchronized root tips were fixed in 2 % formaldehyde for 10 min and chromosomes were released into a lysis buffer by mechanical homogenisation, producing 5 × 105 chromosomes from 50 root tips, at average. The isolated chromosomes were morphologically intact and suitable for flow cytometric analysis. Flow karyotypes obtained after the analysis of DAPI-stained chromosomes indicated a possibility to sort at least three different chromosome types.
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Űberall, I., Vrána, J., Bartoš, J. et al. Isolation of Chromosomes from Picea abies and their Analysis by Flow Cytometry. Biologia Plantarum 48, 199–203 (2004). https://doi.org/10.1023/B:BIOP.0000033445.15336.04
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DOI: https://doi.org/10.1023/B:BIOP.0000033445.15336.04