Potential application of simple easy-to-use insertion-deletion (InDel) markers in citrus cultivar identification

We previously developed insertion-deletion (InDel) markers that distinguish three genotypes (two homozygous and one heterozygous) of diverse citrus cultivars. These InDel markers were codominant and could be clearly detected by using simple agarose gel electrophoresis. We sought to establish a method for cultivar identification using these 28 InDel markers to genotype 31 citrus cultivars. The results revealed that a minimum of 6 markers were required to identify individuals using the three-genotype classification method. Furthermore, we found that a simple method for distinguishing between two genotypes (homozygous and heterozygous) could be used to identify individuals using a minimum of 7 markers. Our findings provide a basis for the development of simple and rapid citrus cultivar identification methods.     

The characterization of autoallotetraploid hybrids between Iris ensata Thunb. and I. laevigata Fisch.

The characteristics of autoallotetraploid hybrids obtained from the cross between Iris ensata cv. Raspberry Rimmed (4X) and amphidiploids of I. laevigata × I. ensata were examined and compared with those of their parents. The color of inner and outer perianths in the autoallotetraploids were bluish purple and similar to those of the amphidiploid parent. However, the autoallotetraploids exhibited low pollen fertility. In addition, the autoallotetraploids were characterized by 17 or 19 anthocyanins and had high resemblance to their parents in the anthocyanin expression. Among these anthocyanins, malvidin 3RGac5G and petunidin 3RGac5G were regarded as major anthocyanins in the autoallotetraploids and their parents, but the differences in the ratios of malvidin 3RGac5G:petunidin 3RGac5G between the autoallotetraploids and their parents were ca. 2:1 for the former and ca. 1:1 for the latter. No viable hybrid seeds were obtained from the reciprocal crosses between I. ensata (2X and 4X) and the autoallotetraploids. Finally, the interspecific cross-breeding of I. ensata using the autoallotetraploids is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Offshore currents explain the discontinuity of a fish community in the seagrass bed along the Japanese archipelago

Oceanographic conditions can affect spatial variability in fish community structures by influencing the temperature‐dependent latitudinal distribution of adult fishes and transport during their young stages. In order to examine latitudinal variability in the fish community structure within a single coastal ecosystem, quantitative sampling was conducted in the sub‐tidal zone of seagrass Zostera marina beds over a broad latitudinal scale (31.31–43.0°N: from subtropical to sub‐boreal zones, covering 80% of the latitudinal range of seagrass distribution in Japan) in the western North Pacific based on a uniform methodology. Cluster analysis with the similarity of fish communities showed that 13 sampling sites were divided into two clusters. The border between the two clusters corresponded with the area of mixing of two dominant currents, Oyashio and Kuroshio, which form a border between the warm temperate zone and the cool temperate zone off the Pacific coast of Japan. Oceanographic properties, such as major currents off the coast, are suggested to affect the latitudinal variability in the fish communities in the coastal ecosystem in the western North Pacific.     

Structural identification of anthocyanins and analysis of concentrations during growth and flowering in buckwheat (Fagopyrum esculentum Moench) petals

The anthocyanin profiles and variety/breeding-line differences of anthocyanin concentrations in petals of common buckwheat flowers have been studied. Four anthocyanins, cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-rhamnoside, and cyanidin 3-O-galactosyl-rhamnoside were isolated from the petals of common buckwheat (Fagopyrum esculentum Moench), separated using high performance liquid chromatography and identified using reversed-phase liquid chromatography-electrospray ionization-tandem mass spectrometry techniques. In every variety/breeding line tested, cyanidin 3-O-rutinoside was detected as the major anthocyanin and the next is cyanidin 3-O-glucoside whereas cyanidin 3-O-rhamnoside and cyanidin 3-O-galactosyl-rhamnoside were trace or not detectable in white and pink flowered buckwheat. Of all the varieties/breeding lines tested, Gan-Chao, a Chinese variety, contained the highest amount of anthocyanins. The largest part of cyanidin moiety was presented as a proanthocyanidin form (PAs-Cy). Anthocyanins and PAs-Cy in petals were increased along with increase of flower development stages. Therefore, fully developed petals of red flowered buckwheat, especially Gan-Chao, are promising as a new anthocyanin-rich material for food processing.     

Protectivity of an immunoaffinity-purified 39 kDa capsular protein of avian Pasteurella multocida in mice

A 39 kDa protein of avian Pasteurella multocida strain P-1059 (serovar A:3) was purified from a crude capsular extract by immunoaffinity chromatography by using a ligand of purified mouse monoclonal antibody to the 39 kDa capsular protein of the strain. Protective activity of the purified 39 kDa protein antigen was determined by inoculation of ddY mice twice with 25 or 125 mug of the protein and challenge-exposure with 10 or 50 LD(50) of strains P-1059 or X-73 (serovar A:1). The results showed that the antigen gave high protection (60 to 100%). These results indicated that the 39 kDa protein of avian P. multocida is a cross-protective antigen over serovars A:1 and A:3.     

Barley lines showing prominent high green shoot regeneration in cultures derived from immature embryos

Eighty‐four cultivars of barley (Hordeum vulgare) and 95 wild strains (82 of H. spontaneum and 13 of H. agriocrithon) were surveyed for the production of callus, callus growth, and shoot regeneration in cultures derived from immature embryos. All cultivars except for ‘Turkey 381′, induced calli from more than 90% of embryos. On the other hand, the wild lines showed a large variation in the percentage of callus induction from 0 to 100%. Among the cultivars, those with the brittle rachis genotype, bt Bt₂, on chromosome 3H, regenerated shoots with a significantly higher percentage than the cultivars with the Bt bt₂ genotype. Green shoots were produced in a higher ratio (0.84) in the cultivars than in the wild lines (0.52). Among the lines examined,‘Lenins’ regenerated shoots efficiently (90.4%) and produced the highest number of calli with green shoots per embryo (4.77) followed by ‘Golden Promise’ (3.15). Examination of callus growth and shoot regeneration from embryos at different developmental stages revealed that scutellum development affected the quantity and quality of callus and shoot regeneration.