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Plant and Cell Physiology, 1985, Vol. 26, No. 7 1401-1409
© 1985

Article

Extrachromosomal Circular DNA from Nuclear Fraction of Higher Plants

Yasuhiro Kinoshita1, Noboru Ohnishi1, Yasuyuki Yamada1, Takahiro Kunisada2 and Hideo Yamagishi2

1Research Center for Cell and Tissue Culture, Faculty of Agriculture, Kyoto University Kyoto 606, Japan
2Department of Biophysics, Faculty of Science, Kyoto University Kyoto 606, Japan

To determine the cellular location of extrachromosomal circular DNA in higher plants, wheat (Triticum aestivum) and tobacco (Nicotiana tabacum) nuclei were isolated and purified, and their circular DNAs were examined by electron microscopy. Covalently closed circular (ccc) DNAs were found in nuclear fractions from both species. They showed a heterogeneous size distribution ranging from 0.1 µm to more than 5 µm in contour length, with a mean of 1.7 µm (5.3 kbp) for T. aestivum and 1.5 µm (4.7 kbp) for N. tabacum, respectively. This distribution is significantly different from that for small circular DNAs in mitochondria. Small polydisperse circular (spc) DNA/protein complexes were also observed by a rapid microscale method of mica-press-adsorption for electron microscopy. Complexes of spcDNA/protein showed a similar size distribution to cccDNA. The average number of spcDNA/protein complexes per nucleus was estimated at more than hundred. The origin and biological functions of the nuclear circular DNAs are discussed.

(Received February 13, 1985; Accepted August 8, 1985)
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