Hiroshi Ikehashi

Repetitive sequences: cause for variation in genome size and chromosome morphology in the genus Oryza.

Large variation in genome size as determined by the nuclear DNA content and the mitotic chromosome size among diploid rice species is revealed using flow cytometry and image analyses. Both the total chromosomal length (r_0.939) and the total chromosomal area (r_0.927) correlated well with the nuclear DNA content. Among all the species examined, Oryza australiensis (E genome) and O. brachyantha (F genome), respectively, were the largest and smallest in genome size. O. sativa (A genome) involving all the cultivated species showed the intermediate genome size between them. The distribution patterns of genome-specific repetitive DNA sequences were physically determined using fluorescence in situ hybridization (FISH). O. brachyantha had limited sites of the repetitive DNA sequences specific to the F genome. O. australiensis showed overall amplification of genome-specific DNA sequences throughout the chromosomes. The amplification of the repetitive DNA sequences causes the variation in the chromosome morphology and thus the genome size among diploid species in the genus Oryza.

Linkage analysis among loci for RAPDs, isozymes and some agronomic traits in Brassica campestris L.

F2 populations from a hybrid between an inbred line of Chinese cabbage (Brassica campestris L. var. pekinensis, 2n = 20) and an inbred line of Mizu-na (B. campestris L. var. japonica 2n = 20) were grown in two seasons, and were analyzed with RAPD and isozyme markers to construct a linkage map following our previous linkage analyses for 19 isozyme loci. A total of 52 RAPD markers were integrated into ten linkage groups, which is the same as the haploid chromosome number of B. campestris. Genes for morphological traits, such as leaf shape, period to bolting, leaf hairiness, the self-incompatibility (the S-glycoprotein) and the NS-glycoprotein which has a high degree of structural homology with the S-glycoprotein, were found to link to the RAPD markers by QTL or linkage analyses. Leaf shape was found to be controlled by multiple genes in two linkage groups. Leaf hairiness seemed to be controlled by a single gene and to be linked to a group of RAPD markers. The period to bolting was found to be controlled by at least two independent loci in the genome. A RAPD marker showing linkage to the self-incompatibility locus showed a recombination value of 20.2%. The NS-glycoprotein locus was linked to the ACP-1 isozyme locus. Linkages between markers and traits are useful for developing marker-assisted selection of important traits.

Identification of two types of differentiation in cultivated rice (Oryza sativa L.) detected by polymorphism of isozymes and hybrid sterility

Magnitudes of genetic variation within each of major varietal groups of cultivated rice were surveyed in terms of isozyme polymorphism and allelic differentiation of hybrid sterility loci, both of which are considered to have litt le selective value. Allelic differences for 20 isozyme loci were examined in a total of 337 accessions, including Indica and Japonica rices. Aus cultivars from India, Javanica cultivars and both landraces and cultivars from China. Eleven out of the 20 isozyme loci were polymorphic. The Aus cultivars contained more alleles per locus than others. The hybrid sterilities in the crosses among Chinese cultivars, Indica and Japonica cultivars were mainly controlled by locus S-5, whereas the hybrid sterilities of Aus cultivars, when they were crossed to Indica, Japonica or Javanica cultivars, were found to be controlled by allelic interaction at hybrid sterility loci S-5, S-7, S-9 and S-15. Also in terms of the number of alleles at S-5 and S-7, Aus cultivars contained more alleles than others. While the Aus group showed an extreme diversity for both hybrid sterility alleles and isozymes, modern cultivars from the south of YangZi River in China were classified into Indica type and those from the north were into Japonica, which were almost the same as those in Japan. On the basis of the measured polymorphism the Indica-Japonica differentiation was explained by founder effects, i.e., through selection and distribution of two original sources each with a unique set of markers. Contrastingly, the continuous and pronounced diversity in the Aus group was attributed to the absence of such a process. The intermediate groups in Yunnan province and Tai-hu Lake region of China are considered to be isolated from such founder effects, retaining an intermediate diversity in terms of isozymes and hybrid sterility genes. Since hybrid sterility hampers the exchange of genes between cultivars of different groups, the understanding of its genetic basis will be important in rice breeding, particularly in hybrid rice breeding.

Identification of segregation-distortion-neutral alleles to improve pollen fertility of indica-japonica hybrids in rice (Oryza sativa L.)

Utilization of the pronounced heterosis of indica-japonica hybrids in rice had been difficult due to panicle sterility caused by male and female gamete abortion in such hybrids. But the female gamete abortion in the most indica-japonicahybrids has been solved by using an abortion-neutral gene S-5n called as wide compatibility gene. On the other hand, the problem of pollen sterility remains to be solved for such hybrids. This study was conducted to identify abortion-neutral genes for pollen that may be utilized to alleviate pollen sterility in distant crosses. Some cultivars like Ketan Nangka (KN) and Dular, which are known as wide compatibility variety (WCV), were tested to find whether they possess neutral alleles for segregation distortion of marker genes. The distorted segregation of markers as well as normal segregation were confirmed among progeny lines of three-way crosses, WCV/indica//japonica and japonica//indica/WCV. Neutral alleles for segregation distortion were identified at two gamete gene loci (ga), i.e., ga-14(t) on chromosome 3 and ga-11 on chromosome 7. A new locus ga-14(t) was identified in between Pgi-1 and bc-1 and independent of ga-3 on chromosome 3. The neutral alleles at the two loci may be used to solve pollen sterility in indica-japonica hybrids.