Rieko Shishido

Ribosomal DNAs: an exception to the conservation of gene order in rice genomes

Abstract rDNA (18S-5.8S-25S rDNA) and 5S rDNA loci were visualized on the chromosomes of six species of the genus Oryza by fluorescence in situ hybridization (FISH) and the labeled rice chromosomes were identified based on their condensation patterns. As a result, the chromosomes harboring rDNA and/or 5S rDNA loci were determined in the complement for all the known rice genomes. Variation in the location of the rDNA loci indicated the transpositional nature of the rDNAs in the genus Oryza, as also suggested in Triticeae and Allium. Comparative analysis of the locations of rDNA loci among rice, maize and wheat revealed that variability in the physical location of the rDNA loci was characteristic of the genus Oryza and also of the genera of Gramineae. This variability in the location of the rDNA loci between evolutionarily related species is in sharp contrast to the conservation of the general order of genes in their genomes.

Evaluation of Genetic Diversity of Wild Rice (Oryza rufipogon Griff.) in Myanmar using Simple Sequence Repeats (SSRs)

As a part of an in situ survey of wild rice (Oryza rufipogon Griff.) in Myanmar (Burma), 16 strains of wild rice were collected, and analyzed for allelic diversity over 74 loci with simple sequence repeat (SSR) markers to obtain a basic information for their conservation. Three each of indica and japonica cultivars were added for a comparison. In the six cultivars and 16 strains of wild rice, three to 15 alleles were detected per locus with an average of 7.9. The wild rice revealed a large number of unique alleles throughout their chromosomes with much wider ranges of variation than those detected in the six cultivars of O. sativa L.. The alleles found in the wild rice were classified into those specific to wild rice, common to wild rice and cultivars, and those similar to indica or japonica cultivars. According to the classification, the genotype of each of the 16 strains of wild rice was schematically depicted. The genetic variation among individual strains within a collection site was larger than the variation among the collection sites.

Evaluation of Genetic Variation Among Wild Populations and Local Varieties of Rice

Cultivated rice (Oryza sativa L.) is derived from Asian wild rice (Oryza rufipogon Griff). Vietnamese local varieties and wild natural populations in Vietnam and Myanmar were examined to evaluate the levels of genetic variation in cultivated and wild rice. In total, 222 Vietnamese local varieties were analyzed with ten microsatellite markers. Using marker genotype and gene diversity data, the local varieties were differentiated based on geographical distribution, cropping season, and human preference. A total of 976 wild plants were collected at six natural sites of wild populations (three each in Myanmar and Vietnam), and the degrees of variation among populations were analyzed with five microsatellite markers. Phylogenetic analyses revealed wide genetic differentiation among wild populations. The diversity values detected in a single wild population in Vietnam were higher than those in whole Vietnamese local varieties. These results indicate that wild rice has much greater genetic variation than cultivated rice.

Chromosome painting as a tool for rice genetics and breeding.

Chromosome painting and genomic in situ hybridization (GISH) are both effective methods for basic genetic research and practical breeding. These methods were applied even in the typically small chromosomes of rice. This manuscript describes in detail, highly reproducible, complete protocols for chromosome painting and GISH in rice chromosomes. Examples of useful applications of these methods are also presented.

Quantitative chromosome map of a representative indica rice

The somatic chromosomes of standard indica diploid rice, IR 36, were squashed on glass slides and stained with Giemsa. The condensation patterns (CP) of prometaphase chromosomes were quantitatively analysed using CHIAS III software. The relative length and centromeric index (CI) were converted from CHIAS III to numerical data and calculated by EXCEL program. The ideogram based on CP of indica rice was established. There were 2 pairs of satellite chromosomes and the result was confirmed by fluorescence in situ hybridization using 45S rDNA as a probe.

Evaluation of genetic variation among wild rice populations in Cambodia

A total of 448 samples in five natural populations of wild rice (Oryza rufipogon) were collected in Cambodia. They were examined using 12 SSR and two chloroplast markers to evaluate the degree of variation among populations and the genetic structure within populations. In the two annual populations, the number of plants with homozygous alleles at all 12 SSR loci were high (66.3% and 79.5%), suggesting that these plants propagate mainly through self-pollination. In the three perennial populations, no individuals had all homozygous genotypes, but redundant genotypes resulted from clonal propagation were observed. Percentages of the redundant genotypes were highly varied (3.6%, 29.2% and 86.0%). This may be due to the different stable levels of environmental conditions. As for chloroplast genome, most of the wild plants showed the same chloroplast types as most Indica-type cultivars have. However, plants with different chloroplast types were maintained, even in the same population. In tropical Asian countries, many wild rice populations were observed under similar ecological conditions examined in this study. Therefore, the present results concerning population structure will be important to further elucidate genetic features of wild rice, and will also give strong clues to utilize and conserve wild natural genetic resources.