Masahiro Akimoto

Physical strength and its relation to leaf anatomical characteristics of nine forage grasses

Nine species of forage grasses (five C3 species and four C4 species) were planted in a controlled-environment glasshouse. The C3 plants were Festuca arundinacea Schreb, Dactylis glomerata L., Phleum pratense L., Lolium perennel L. and Poa pratensis L.; the C4 plants were Chloris gayana Kunch., Cynodon dactylon (L.) Pers., Paspalum dilatatum Poir. and Sorghum halenpense (L.) Pers. The number of major vascular bundles and minor vascular bundles, cross-sectional area, the area and proportion of sclerenchyma in a cross-section, thickness of leaf blade, and tensile and shear strength were investigated in order to determine the relationship between physical strength and anatomical characteristics. Physical strength and anatomical characteristics of leaf blades showed significant (P < 0.01) variation between species. Significant correlations were detected between tensile strength and cross-sectional area in forage grasses except Festuca arundinacea. Festuca arundinacea, Dactylis glomerata, Phleum pratense, Chloris gayana and Sorghum halenpense showed significant correlations of tensile strength with the number of major vascular bundles. Festuca arundinacea, Dactylis glomerata and Lolium perennel showed significant correlations of shear strength with cross-sectional area. Festuca arundinacea, Dactylis glomerata and Paspalum dilatatum showed significant correlations of shear strength with the number of major vascular bundles. The proportion of sclerenchyma in a cross-section showed poor correlations with tensile and shear strength. Thickness of leaf blade showed poor correlations with tensile and shear strength except in Dactylis glomerata. Physical strength and anatomical characteristics of leaf blades of the C3 group differed significantly (P < 0.01) when compared with the C4 group except for cross-sectional area. Tensile and shear strength showed significant correlations with cross-sectional area, sclerenchyma area and the number of vascular bundles when all nine species were treated as one group.

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.

Growth promotion and inhibition of the Amazonian wild rice species Oryza grandiglumis to survive flooding.

Abstract In Asian cultivated rice (Oryza sativa), distinct mechanisms to survive flooding are activated in two groups of varieties. Submergence-tolerant rice varieties possessing the SUBMERGENCE1A (SUB1A) gene display reduced growth during flash floods at the seedling stage and resume growth after the flood recedes, whereas deepwater rice varieties possessing the SNORKEL1 (SK1) and SNORKEL2 (SK2) genes display enhanced growth based on internodal elongation during prolonged submergence at the mature stage. In this study, we investigated the occurrence of these growth responses to submergence in the wild rice species Oryza grandiglumis, which is native to the Amazon floodplains. When subjected to gradual submergence, adult plants of O. grandiglumis accessions showed enhanced internodal elongation with rising water level and their growth response closely resembled that of deepwater varieties of O. sativa with high floating capacity. On the other hand, when subjected to complete submergence, seedlings of O. grandiglumis accessions displayed reduced shoot growth and resumed normal growth after desubmergence, similar to the response of submergence-tolerant varieties of O. sativa. Neither SUB1A nor the SK genes were detected in the O. grandiglumis accessions. These results indicate that the O. grandiglumis accessions are capable of adapting successfully to flooding by activating two contrasting mechanisms as the situation demands and that each mechanism of adaptation to flooding is not mediated by SUB1A or the SK genes.

Ethylene is not involved in adaptive responses to flooding in the Amazonian wild rice species Oryza grandiglumis.

The Amazonian wild rice Oryza grandiglumis has two contrasting adaptation mechanisms to flooding submergence: a quiescence response to complete submergence at the seedling stage and an escape response based on internodal elongation to partial submergence at the mature stage. We investigated possible factors that trigger these responses. In stem segments excised from mature O. grandiglumis plants, complete submergence only slightly promoted internodal elongation with increased ethylene levels in the internodes, while partial submergence substantially promoted internodal elongation without increased ethylene levels in the internodes. Incubation of non-submerged stem segments under a continuous flow of humidified ethylene-free air promoted internodal elongation to the same extent as that observed for partially submerged segments. Applied ethylene had little effect on the internodal elongation of non-submerged segments irrespective of humidity conditions. These results indicate that the enhanced internodal elongation of submerged O. grandiglumis plants is not triggered by ethylene accumulated during submergence but by the moist surroundings provided by submergence. The growth of shoots in O. grandiglumis seedlings was not promoted by ethylene or complete submergence, as is the case in O. sativa cultivars possessing the submergence-tolerant gene SUB1A. However, because the genome of O. grandiglumis lacks the SUB1A gene, the quiescence response of O. grandiglumis seedlings to complete submergence may be regulated by a mechanism distinct from that involved in the response of submergence-tolerant O. sativa cultivars.

Inconsistent diversities between nuclear and plastid genomes of AA genome species in the genus Oryza

AA genome species in the genus Oryza are valuable resources for improvement of cultivated rice. Oryza rufipogon and O. barthii were progenitors of two domesticated rice species, O. sativa and O. glaberrima, respectively. We used chloroplast single-nucleotide repeats (RCt1-10) to evaluate genetic diversity among AA genome species. Higher diversity was detected in the American species O. glumaepatula and the Asian species O. rufipogon. Other chloroplast sequences indicated that O. glumaepatula shares high similarity with O. longistaminata. Insertions of retrotransposable elements, however, showed a close relation between O. barthii and O. glumaepatula. To clarify phylogenetic relationships among AA genomes, whole-genome sequences obtained from different species were used to develop chloroplast INDEL markers. The INDEL patterns clearly showed multiple maternal origins of O. glumaepatula. The complicated origins have resulted in high genetic diversity in this species. In contrast, the Australian endemic species O. meridionalis tended to show narrower diversity than the other species. High variation in O. rufipogon, reconfirmed using the chloroplast INDELs, covered the variation in O. meridionalis and part of the variation in O. glumaepatula. Maternal lineages including O. barthii, O. longistaminata and the remainder of O. glumaepatula were phylogenetically close to each other and carried low genetic diversity. They were separated from independent lineages, suggesting that they had diverged from a single ancestral maternal lineage, but diverged later to keep gene flow within respective species, as SSR compositions suggested. Genetic relationships among AA genome species indicate how these species have evolved and become distributed across four continents.

Internodal elongation under submergence in the Amazonian wild rice species Oryza glumaepatula: the growth response is induced by hypoxia but not by ethylene

Whole plants of Oryza glumaepatula (accession W1246) displayed rapid internodal elongation under partial submergence, comparable with those of deepwater or floating Oryza sativa cultivars. In excised stem segments of O. glumaepatula, submergence and hypoxia equally induced internodal elongation, whereas ethylene, which plays an important role in the rapid internodal elongation of deepwater rice, induced internodal elongation of only one-third relative to submergence or hypoxia. Pretreatments of stem segments with 1-methylcyclopropene (an ethylene action inhibitor) almost completely blocked ethylene-induced internodal elongation but had limited effects on submergence-induced internodal elongation. These results indicate that hypoxia, but not ethylene, triggers rapid internodal elongation during submergence in O. glumaepatula. O. glumaepatula W1246 possessed a homologue of SNORKEL1 (SK1) and two homologues of SNORKEL2 (SK2). SK1 and SK2 encode ethylene response factors and evoke rapid internodal elongation in response to ethylene in O. sativa. However, none of these homologues showed an increased expression level in internodes of stem segments treated with hypoxia.

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.