1989 Fiscal Year Final Research Report Summary
ANALYSES OF GENE STRUCTURE AND FUNCTION BY ELECTRONMICROSCOPY
| Project/Area Number |
61440011
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物保護
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
SHIKATA Eishiro HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (00001389)
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| Co-Investigator(Kenkyū-buntansha) |
SANO Teruo HOKKAIDO UNIV., FACULTY OF AGR., ASSISTANT, 農学部, 助手 (30142699)
UEDA Ichiro HOKKAIDO UNIV., FACULTY OF AGR., ASSOC. PROF., 農学部, 助教授 (10113523)
IIZUKA Toshihiko HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (50001441)
OGOSHI Akira HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (80109503)
KINOSHITA Toshiro HOKKAIDO UNIV., FACULTY OF AGR., PROFESSOR, 農学部, 教授 (10001421)
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| Project Period (FY) |
1986 – 1989
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| Keywords | Electron microscopy / Nucleic acid / Gene |
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| Research Abstract |
1) Cytochrome C method for visualizing double-standed DNA, double-stranded RNA, and single-stranded RNA was developed. The nucleic acids were rotary shadowed with Pt-Pd. BAC method was also applied. The method was simple and rapid when compared with the Cytochrome C method.
2) All the double-stranded genome segments of rice dwarf, rice black-streaked, and rice ragged stunt viruses were examined under the electron microscope for sizing. The sizes were in good agreement with those determined by nucleotide sequencing.
3) Non-chromosomal genetic elements of Rhizoctonia, a plant pathogen, were examined under the microscope. Linear dsRNA molecules were detected.
4) Hop stunt viroid and viroid from plum were seen as a rigid rod under the microscope. However, they were seen as a circular molecule 90-100 nm in length under the denaturing condition.
5) Mitochondrial DNAs from normal(N) and male sterile(S) cytoplasms of sugar beet were investigated by electron microscopy. The DNA was composed of a heterogeneous population of circular molecules. Their contour lengths varied from 0.28-51 mum. N and S cytoplasms were shown to contain their own characteristic combinations of small circular DNA species with lengths between 0.28-0.6 mum. Chloroplast DNA was a circular molecule of 45 mum in length.
6) The native plasmids bearing the crystal protein gene of Bacillus thuringiensis were observed by electron microscopy. The size of several large plasmids from subsop. sotto, kurstaki HD-1, and aizawai were determined by computer analysis of micrographs. The plasmids were a circular molecule and its estimated size was about 7OKb.
7) Viroplasm was isolated from rice dwarf virus infected rice plants by differential centrifugation. Viroplasm was seen in 5,000g fraction.
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