Abstract
BAC FISH (fluorescence in situ hybridization using bacterial artificial chromosome probes) is a useful cytogenetic technique for physical mapping, chromosome marker screening, and comparative genomics. As a large genomic fragment with repetitive sequences is inserted in each BAC clone, random BAC FISH without adding competitive DNA can unveil complex chromosome organization of the repetitive elements in plants. Here we performed the comparative analysis of the random BAC FISH in monocot plants including species having small chromosomes (rice and asparagus) and those having large chromosomes (hexaploid wheat, onion, and spider lily) in order to understand a whole view of the repetitive element organization in Poales and Asparagales monocots. More unique and less dense dispersed signals of BAC FISH were observed in species with smaller chromosomes in both the Poales and Asparagales species. In the case of large-chromosome species, 75–85% of the BAC clones were detected as dispersed repetitive FISH signals along entire chromosomes. The BAC FISH of Lycoris did not even show localized repetitive patterns (e.g., centromeric localization) of signals.
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Abbreviations
- BAC:
-
Bacterial artificial chromosome
- FISH:
-
Fluorescence in situ hybridization
- FITC:
-
Fluorescein isothiocyanate
- PFGE:
-
Pulsed field gel electrophoresis
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Acknowledgments
This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT) and the Japan Society for the Promotion of Science (JSPS), KAKENHI (18075003, 20780240, 23113001 and 23113006 to G.S., 20380014 to A.K., 22580004 and 19380194 to Y.M.).
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Communicated by K. Toriyama.
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Representative results of PFGE analysis of NotI-digested BAC clones in rice, asparagus, and spider lily (Lycoris). BAC-vector (pBeloBAC11) fragments are indicated by asterisks. Other fragments represent the BAC inserts.
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Suzuki, G., Ogaki, Y., Hokimoto, N. et al. Random BAC FISH of monocot plants reveals differential distribution of repetitive DNA elements in small and large chromosome species. Plant Cell Rep 31, 621–628 (2012). https://doi.org/10.1007/s00299-011-1178-8
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DOI: https://doi.org/10.1007/s00299-011-1178-8