KINOSHITA Toshiro Faculty of Agriculture, Hokkaido University, 農学部, 教授 (10001421)
OGIHARA Yasunari Kihara Institute for Biological Research, Yokohama City University, 木原生物学研究所, 助手 (40185533)
YABUNO Tomosaburo Faculty of Agriculture, University of Osaka Prefecture, 農学部, 教授 (50081485)
HIRAI Atsushi Faculty of Agriculture, Nagoya University, 農学部, 助教授 (60023470)
YAMAGUCHI Hikoichi Faculty of Agriculture, University of Tokyo, 農学部, 教授 (40011829)
藤井 太朗 国立遺伝学研究所, 遺伝実験生物保存研究センター, 助教授 (90000249)
|Budget Amount *help
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1987: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1986: ¥5,000,000 (Direct Cost: ¥5,000,000)
The present cooperative research aimed to clarify cytoplasmic variation in rice, wheat and sugar beet, including their wild relatives, by various molecular techniques, and to utilize the variation in breeding of these crops. The research was conducted on the following seven items:
1) Restriction endonuclease analysis and construction of the physical map of organellar DNAs: Detailed physical map of chloroplast (ct) DNA was conctructed in Japanese rice, common wheat and sugar beet. Restriction endonuclease analysis was carried out using a large number of accessions of diploid Oryza species, all Triticum and Aegilops species, and 11 Beta species. From the results, a phylogenetic tree of the chloroplast genome was constructed for each genus.
2) Construction of the organellar DNA library: CtDNA library was constructed in rice, common wheat and sugar beet. About 70% of the HindIII fragments of wheat mitochondrial (mt) DNA were cloned.
3) Studies on small circle mtDNAs: Two small circle DNAs, B-
1 and B-2, in mitochondria of male sterile rice were cloned, and partially or completely sequenced, respectively. These DNA species disappeared from mitochondria of the male fertile revertants. Eight different molecular species of small circle DNA were present in different combinations in ten male fertile or sterile accessions of sugar beet.
4) Analysis on chloroplast genes: Ten to 18 chloroplast genes were located inthe physical map of each rice, common wheat and sugar beet. Complete nucleotide sequence of the genes, rbcL, atpB, aptE and psbA of rice, and rbcL genes of common wheat and three Aegilops species was determined.
5) Cloning and sequencing the ars regions of mtDNA: Two sequences having the ars activity were isolated from rice mtDNA,and sequenced. They had the characteristic structures of the ars region.
6) Genetic characterization of alien cytoplasms introduced into crop species: O. sativa cytoplasm was transferred into O. glaberrima, and the male sterile line was produced. Four new male sterile cytoplasms were identified among those introduced from O. rufipogon. Ae. searsii was found to be the cytoplasm donor to two tetraploid species, Ae. kotschyi and Ae. variabilis. Some alloplasmic wheat lines showed different photosynthetic and/or electron transport activities from the normal wheat line.
7) Use of alien cytoplasms: Triple cross whear hybrids produced using Ae. kotschyi cytoplasm were more productive than the F_1 hybrids produced using t. timopheevi. cytoplasm. A common wheat line yielding twin seedlings at a high frequency was also produced using Ae. kotschyi cytoplasm. Less