A new species of tetramethylammonium-assimilating bacteria was isolated from an activated sludge which was used for the treatment of tetramethylammonium hydroxide contained in the wastewater from semiconductor manufacturing processes. Cells of the bacteria were gram-negative, nonmotile, short rods (0.5 to 0.8 pm by 0.7 to 1.1 pm). The major respiratory quinone component of the bacteria was Q-lo. The G+C content was 71 mol% . Isolates are mesophilic and assimilate methylated amines such as tetramethylammonium, trimethylamine, dimethylamine, and methylamine under neutral conditions. The isolates resemble Purucoccus species with respect to morphology but were distinguishable from the known species of the genus. We propose Paracoccus kocurii sp. nov. The type strain is strain B (= JCM 7684).

Here we established a DNA marker-based method to identify cultivars of loquat (Eriobotrya japonica Lindl.) using simple sequence repeat (SSR) markers derived from pears and apples. A total of 24 loquat cultivars commercially grown in Japan were used for genetic identification, including 15 diploid-, 6 triploid-, and 3 tetraploid cultivars. Analysis using the 88 SSR markers derived from pears and apples, which belong to the same subfamily (family Rosaceae, subfamily Maloideae) as the loquat, indicated that 26 SSR markers were applicable to the identification of loquat cultivars. A total of 82 putative SSR alleles were obtained by the 26 SSR markers. SSR analysis enabled the identification of all loquat varieties tested, except for 'Tomihusa' and '4N-Tomihusa'. In addition, the parentages of several cultivars, including 'Oohusa' and 'Mizuho', were confirmed from the SSR genotype data obtained. Triploid and tetraploid varieties could be identified from the SSR genotypes because some SSR loci generated 2 or more alleles for polyploids. The relationship between 'Kibou' and its parents was confirmed since SSR alleles of the seedless triploid offspring 'Kibou' were clearly inherited from the female parent '4N-Tanaka 1' and the male parent 'Nagasakiwase'. The phenogram obtained by cluster analysis showed no distinctive separation of Japanese commercial cultivars from Chinese cultivars. This result was consistent with the hypothesis that Japanese commercial cultivars were derived from introduced Chinese loquat.

Cucumber mosaic virus (CMV) is a tripartite, positive sense RNA virus causing infections and yield losses to many plant species. Here, we generated a construct containing inverted repeat of 1,793 bp fragment of defective CMV replicase gene derived from RNA2 of cucumber mosaic virus strain O (CMV-O). The replicase gene was modified by deleting a 9 bp region between nucleotides 1909-1918. This caused a deletion in the active centre motif of polymerases, producing defective translated product 9 nucleotides shorter than the full length protein. The RNAi construct containing inverted repeat of the defective gene was used to produce transgenic tobacco lines expressing CMV-derived double-stranded RNA via Agrobacterium-mediated transformation. Of the four transgenic lines inoculated with CMV-O or CMV-Y in vitro and ex vivo, three lines (T1, T4 and T5) showed immunity to both strains of CMV as no symptoms were detected, whereas one line (T7) exhibited high resistance with mild symptoms limited to inoculation portions. No virus could be detected in uninoculated new leaves of the transgenic lines after RT-PCR and Dot-immunobinding assay analyses. Small interfering RNAs present in transgenic lines before and after virus challenge indicates that the resistance was acquired through RNA silencing.

The effect of alternative carbohydrate sources to sucrose for plant regeneration from long-term cell cultures of creeping bentgrass (Agrostis palustris Huds.cv.Penncross) and japonica rice (Oryza sativa L.cv.Nipponbare) was studied. Both maltose and lactose supported a higher degree of regeneration compared to sucrose; in 8-and 19-month-old cultures of creeping bentgrass, the frequencies of regenerating calli remained at 76-93% and the numbers of plants regenerated were 8 to 36-fold higher. In 35-month-old cultures of japonica flee, 2-4% of the calli were capable of regeneration on maltose and lactose media. These results indicate that loss of plant regeneration in long-term cultures is caused, at least in part, by specific cultural conditions and not by genetic changes.