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Association of particles that contain double-stranded RNAs with algal chloroplasts and mitochondria

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Abstract

Linear dsRNAs (double-stranded RNAs) belonging to several distinct size classes were found to be localized in chloroplasts and mitochondria of Bryopsis spp., raising the possibility that these dsRNAs are prokaryotic in nature. The algal cytosol and nuclei did not contain dsRNAs. The amount of the dsRNAs in the organelles appeared constant, and there were about 500 copies per chloroplast. The four major dsRNAs from Bryopsis chloroplasts were about 2 kbp (kilobase pairs) in length and originated from discrete isometric particles of about 25 nm diameter. These virus-like particles were purified by CsCl density gradient centrifugation after extraction from isolated chloroplasts with chloroformbutanol and subsequent precipitation with polyethylene glycol. They had a buoyant density of about 1.40 g · cm−3 and contained four major and three minor proteins. Mitochondrial dsRNAs were about 4.5 kbp in length and formed less-stable particles of about 40 nm in diameter with a buoyant density of 1.47 g · cm−3. Some observations support the hypothesis that vertical transmission of the protein-coated, non-infectious dsRNAs occurs within cell organelles. Double-stranded RNAs of various sizes were found in most green, red, and brown algae. The characteristics of the algal dsRNAs are compared with those of dsRNAs from higher plants and the biological significance of the dsRNAs in cell organelles is discussed.

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Abbreviations

dsRNA:

double-stranded RNA

kbp:

kilobase pairs

SDS:

sodium dodecyl sulfate

SSC 0.15 M NaCl:

0.015 M sodium citrate

PAGE:

polyacrylamide gel electrophoresis

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Author notes

  1. J. Ishihara

    Present address: Shimadzu-Seisakusyo Co. Ltd., 604, Kyoto, Japan

  2. J. Y. Pak

    Present address: Laboratory of Biology, Korea University, Kodaira, 187, Tokyo, Japan

Authors and Affiliations

  1. Laboratory of Biology, Tokyo University of Agriculture and Technology, Fuchu, 183, Tokyo, Japan

    J. Ishihara, J. Y. Pak, T. Fukuhara & T. Nitta

Authors
  1. J. Ishihara
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  2. J. Y. Pak
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  3. T. Fukuhara
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  4. T. Nitta
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Additional information

The authors would like to express their gratitude to Dr. T. Natsuaki, Utsunomiya University, and Dr. D. Hosokawa, Tokyo University of Agriculture and Technology, for their helpful suggestions throughout this research. They are also much indebted to Dr. B. Wang, Institute of Genetics, Academia Sinica, Beijing, PRC, for his suggestions on rice dsRNA, and to Dr. T. Kohbara, Senshu University, on Bryopsis cells. Sincere thanks are also due to Dr. T. Misonou, Yamanashi University, and Dr. K. Masuda, Akita Prefectural College of Agriculture, for supplying plant materials; to Dr. N. Sonoki, Azabu University, for nucleotide analysis of dsRNAs; and to Y. Koshino for technical assistance. This research was supported in part by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan.

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Ishihara, J., Pak, J.Y., Fukuhara, T. et al. Association of particles that contain double-stranded RNAs with algal chloroplasts and mitochondria. Planta 187, 475–482 (1992). https://doi.org/10.1007/BF00199965

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