Lesion-based analysis of leaf blast suppression in mixture of rice cultivar and a resistant near-isogenic line

Leaf blast suppression in multilines was evaluated based on the number of susceptible lesions observed in a pure stand of susceptible rice cultivar Sasanishiki, and in 1 : 1 and 1 : 3 mixtures of Sasanishiki and a resistant near-isogenic line, Sasanishiki BL4 or BL7, from 1998 to 2001. The number of lesions first observed in fields in the 1 : 1 and 1 : 3 mixtures were close to theoretical numbers calculated using the number of lesions observed in the pure stands and the ratios of the susceptible Sasanishiki in the mixtures. The ratio of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number in the pure stand was 0.29 and 0.09, respectively. The relationship between these ratios and the ratios of susceptible Sasanishiki in mixtures was defined in an equation to estimate the degree of leaf blast suppression. Validation studies for the ratios of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number in the pure stand were conducted in two different locations and showed that the ratios are almost acceptable. The calculated autoinfection to alloinfection ratio was 1.3 and 1.4 in the 1 : 1 and 1 : 3 mixtures, respectively, suggesting that the calculated ratio will affect the degree of leaf blast suppression. Thus, predictors were obtained to estimate leaf blast suppression for effective blast control in multilines.     

Simple diagnosis using ethanol immersion of strawberry plants with latent infection by Colletotrichum acutatum, Dendrophoma obscurans, and Fusarium oxysporum f. sp. fragariae

Simple diagnosis by ethanol immersion (SDEI) to detect Glomerella cingulata was used to detect three other fungi that also cause latent infection of strawberry plants. Signs on strawberry leaves with asymptomatic latent infection by Colletotrichum acutatum became visible using SDEI. Salmon-pink conidial masses were produced in the acervuli on the treated leaves 5 days after incubation at 28°C. In the case of Dendrophoma obscurans, pycnidia with amber conidial masses formed 5 days after incubation at 28°C. The pycnidia were observed mainly on the ribs, and conidial masses exuded from the ostiole. These macroscopic conidial masses were similar to those of G. cingulata and C. acutatum. When water was dripped onto a lesion caused by D. obscurans, the pycnidia exuded white filamentous conidial masses, making the distinction of D. obscurans from G. cingulata or C. acutatum. On petioles with latent infection by Fusarium oxysporum f. sp. fragariae, white aerial hyphae grew out from the vascular tissues on the cut surface 3 days after incubation at 28°C and were easily observed by eye or with a loupe. Thus, SDEI was also useful for diagnosing latent infection of strawberry plants by C. acutatum, D. obscurans, and F. oxysporum f. sp. fragariae.     

Characteristics of Monomeric Al Concentration in An Acidified River

The Kusurisambetsu River is a tributary of the Noboribetsu River. These rivers flow through Hokkaido Prefecture, Japan. Because the upper reaches of the river are acidified by a caldera lake and basin, the river water contains aluminum in high concentration, together with iron and sulfate. The drainage area at the lower reaches is not acidified. Therefore, the acidified water of the river is neutralized by the alkalinity of water from the drainage area. The water at the mouth is neutral in pH, although about 1 mg/L of aluminum is detected. Heavy rainfall occurred in Autumn 1993, and many adult salmon were killed at the lower reaches of the river. This prompted study of the neutralization process of river water, the Al concentration change, and the influence of rainfall. Determination of Al found total Al concentration to be roughly constant at the mouth of the Noboribetsu River. However, the concentration of monomeric Al varied with pH, which varied according to rainfall and flow volume. Monomeric Al showed a minimum concentration of roughly pH 6.0. At pH of greater than 6.0, the presence of some kinds of polymeric Al is determined by filtration fraction. This suggests that the death of salmon is caused by monomeric Al when conditions are acidic. Liming test of the river water confirmed that the monomeric Al that was toxic under acidic conditions became non-toxic under alkaline conditions.     

Irreversible unfolding of myoglobin in an aqueous solution by supercritical carbon dioxide

The conformational changes in myoglobin, treated by microbubbling of supercritical carbon dioxide (SC-CO(2)), were investigated by measuring the circular dichroism spectra in the ultraviolet range and compared with those in other proteins (ovoalbumin, bovine serum albumin, and beta-lactoglobulin). Irreversible unfoldings were observed after the microbubbling of SC-CO(2) at 35 degrees C and 30 MPa for 30 min. The degree of unfolding depended on the number of intramolecular S-S bonds. alpha-Helix contents of myoglobin decreased with increasing density of SC-CO(2). Unfoldings of myoglobin induced by heating, pH-lowering, and the addition of a denaturant were reversible. The irreversible unfolding of myoglobin was also observed by the bubbling of gaseous CO(2) under atmospheric pressure, but heating was required.     

Construction of a linkage map and identification of DNA markers linked to Fom-1, a gene conferring resistance to Fusarium oxysporum f.sp. melonis race 2 in melon

Resistance to Fusarium oxysporum f.sp. melonis race 2 is conferred by a single dominant gene, Fom-1 in melon. Here, we identified DNA markers tightly linked to Fom-1 that could be used for marker assisted selection in breeding programs. First, we developed 125 F₂ plants derived from the cross between melon lines P11 (fom-1fom-1) and MR-1 (Fom-1Fom-1). Using the F₂ population, we constructed a linkage map including 14 SSR markers which had not been mapped previously. Fom-1 was confirmed to be allocated to linkage group 7. Then, we identified four AFLP markers using bulked segregant analysis. The AFLP marker TAG/GCC-470 was completely linked to Fom-1 and other three markers were mapped near Fom-1. TAG/GCC-470 and TCG/GGT-400 were respectively converted to STS and CAPS markers. Usefulness of DNA markers was confirmed in the analysis with several melon cultivars and lines.     

A Novel Pair of HMW Glutenin Subunits from Aegilops searsii Improves Quality of Hexaploid Wheat

This study involved screening of wild species of wheat in search of functionally useful seed storage proteins for improvement of breadmaking quality of wheat (Triticum aestivum). After screening of 177 disomic addition lines (DALs) of wheat belonging to different wild species, Aegilops searsii DALs were selected and studied in detail. These DALs of Ae. searsii were from chromosome 1S^s to 7S^s in the background of cultivated wheat cv. Chinese Spring (CS). By analyzing these addition lines, genetic loci of actively expressed genes for the high molecular weight glutenin subunits (HMW‐GS) and gliadin were found on the chromosome 1S^s for the first time and have been designated as Glu‐S^s1 and Gli‐S^s1, respectively. Disomic addition line of chromosome 1S^s (DAL1S^s) showed improved dough strength in different generations compared with CS. SDS sedimentation value and specific sedimentation of DAL1S^s were higher than CS. Mixograph peak height and band width were higher, with no difference in mixing peak time from CS. All these factors indicate a positive effect of quantity as well as quality of gluten proteins of Ae. searsii. This was further supported by increased polymer formation in DAL1S^s because the ratio of unextractable polymeric protein to total polymeric protein (UPP/TPP%) of DAL1S^s was significantly higher than CS. Genes for HMW‐GS (major determinant of end‐product quality in wheat) of Ae. searsii were cloned and sequenced from the DAL1S^s. Phylogenetic analysis of deduced amino acid sequences showed that both x and y HMW‐GS were more similar to that of D genome rather than B genome of wheat. Although S genome is structurally more similar to B genome of wheat, functionally it is more similar to the D genome of wheat and possesses good quality HMW‐GS required for improvement of breadmaking quality of wheat.     

Genetic polymorphisms in Japanese fragrant landraces and novel fragrant allele domesticated in northern Japan

Rice fragrance is an important characteristic for Southeast Asian consumers, and fragrant landraces from Japan were first recorded in the 17th century. Principal component analysis clearly showed that Japanese fragrant landraces were genetically different from non-Japanese fragrant landraces. Japanese fragrant landraces were composed of six clades, none of which carried the most common fragrance mutation, an 8-bp deletion in exon 7 of Badh2. Fragrant landraces comprised two major groups carrying different Badh2 mutations. One group carried a known SNP at exon13 and the other a SNP at the exon1-intron1 junction as splicing donor site. The latter was considered to be a potential splicing mutant group as a novel allele at Badh2. Heterozygosity (He) scores in the two fragrant groups were not significantly different from non-fragrant landraces and modern cultivars. However, lower He scores were found around the Badh2 locus in the two groups. The potential splicing mutant group showed a more extended haplotype than the E13 SNP group. A likely causal factor responsible for loss of function is a novel splicing mutation allele that may have been generated quite recently. The fragrance allele has dispersed as a result of out-crossing under local environmental conditions.     

Estimation of the outcrossing rate for annual Asian wild rice under field conditions

The Asian wild rice, Oryza rufipogon, has partial outcrossing behavior and shows high levels of genetic variation. To estimate an accurate outcrossing rate of annual form of O. rufipogon, two backcross lines (Lines R1 and R2) between Oryza sativa Nipponbare and O. rufipogon W630 were examined under field conditions. A chromosome survey confirmed that these lines had wild chromosomal segments at more than 92% of marker loci. As for the traits of glume, stamen and pistil, Line R1 showed similar floral morphology as that of O. rufipogon W630, whereas Line R2 had larger glumes. In 2005, 2006 and 2008, a total of 22 backcross plants were planted in the middle of wild rice plots. The successive progenies of each plant were examined using microsatellite markers that could clearly detect self-pollination and outcrossing. The outcrossing rates of Line R1 plants ranged from 4.04% to 25.50% with an average of 10.20%. This indicates that cross-pollination of wild rice is a chance event affected by many environmental factors. The outcrossing rates of Line R2 plants also varied, however, no significant difference was observed between the averages of Lines R1 and R2, suggesting that the glume sizes are not critical for outcrossing ability.     

Evaluation of genetic characteristics of wild and cultured populations of the Japanese pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis> by using AFLP markers

Genetic characteristics of four wild (Mie, Fukui, Shimane and Nagasaki) and five cultured populations (selectively bred for 12 years with an origin of Ehime population) of the Japanese pearl oyster, Pinctada fucata martensii, were evaluated by using AFLP markers. Six primer pairs generated 1019 loci in total, among which 45.2–55.1 % was polymorphic among populations. Although there was no significant difference in gene diversity between wild (0.170–0.174) and cultured (0.158–0.173) populations, genetic relatedness in cultured populations (0.316–0.450) was about three times higher than that in wild populations (0.110–0.165). In addition, genetic differentiation was about twenty times larger in cultured populations (Nei’s distance: 0.0111) than in wild populations (Nei’s distance: 0.0005). These results mean that selective breeding can cause marked inbreeding as well as large genetic differentiation among cultured populations in a short period. On the other hand, it was suggested that genetic homogenization in the wild, probably due to a large-scale transport of cultured oysters, had progressed in the sea around Japan. In conclusion, it is necessary to prevent inbreeding by the reconsideration of the style of selective breeding in cultured populations, while the escape of gametes or spats of cultured strains in the wild should be avoided for the preservation of genetic characteristics in native populations of P. f. martensii.     

Stable isotope analysis suggests the existence of multiple populations of streaked spinefoot (<Emphasis Type="Italic">Siganus javus</Emphasis> L.) in Bandon Bay,Southern Thailand

The stock unit used in fisheries resource assessment and management is generally based on the morphological and genetic characteristics of a particular population or species to avert problems caused by the treatment of multiple populations as one stock, which can lead to the overestimation of population sizes and genetic pollution. Furthermore, since the linkage of microhabitats is an important factor affecting the reproduction of marine organisms in coastal areas, an understanding of the food web in each microhabitat is essential to establish sustainable fisheries management practices. We investigated spatial variations in the food sources and feeding habits of immature stage of Siganus javus using genetic population analyses and stable isotope analyses (δ¹³C and δ¹⁵N). These species are commonly harvested by small-scale fisheries, and it inhabits Bandon Bay in the Surat Thani Province of Southern Thailand. Genetic variation within sampling sites was greater than that between sites. The δ¹³C values of S. javus differed between sites, which suggest that the different ecological habitats exhibit different rates and patterns of carbon flow even among sites located in the same bay. Our results suggest that studies combining genetic population analyses and stable isotope analyses are required to confirm the delineation of fine-scale management units intended for the development of coastal fishery resources.