The aim of this study was to investigate the genetic diversity and relationships among 'Candidatus Liberibacter asiaticus' isolates from different hosts and distinct geographical areas in Southeast Asia. Genetic diversity among 'Ca. Liberibacter asiaticus' was estimated by sequencing four well-characterized DNA fragments: the 16S ribosomal DNA (rDNA) and 16S/23S intergenic spacer regions; the outer membrane protein (omp) gene region; the trmU-tufB-secE-nusG-rplKAJL-rpoB region (gene cluster region); and the bacteriophage-type DNA polymerase region. The sequences of the 16S rDNA and 16S/23S intergenic spacer regions were identical among all 'Ca. Liberibacter asiaticus' isolates. In contrast, nucleotide substitutions were observed in both the omp gene and the gene cluster regions. However, extended bacteriophage-type DNA polymerase sequences acquired by thermal asymmetric interlaced polymerase chain reaction provided the most sequence diversity among isolates. Phylogenetic analysis of the bacteriophage-type DNA polymerase sequences revealed three clusters in the Southeast Asian 'Ca. Liberibacter asiaticus' population. All Indonesian 'Ca. Liberibacter asiaticus' isolates clustered in one group. The other clusters were not correlated with geographic distribution. The differences in genetic sequences did not reflect differences in the original citrus host (mandarin or pummelo). These results suggest that the bacteriophage-type DNA polymerase region would be useful for molecular differentiation between different Southeast Asian 'Ca. Liberibacter asiaticus' isolates.

Studying subduction zone fluid at shallow seismogenic depths is important to understand the nature of fault rocks at the updip limit of the seismogenic zone because fluid-rock interactions affect heat and mass transfer, and fault strength. In this study, we conducted detailed analyses of distribution of shear veins, and estimation of pressuretemperature conditions for shear vein formation for the Yokonami mélange, Shikoku, Southwest Japan, which is tectonic mélange zone in an on-land accretionary complex. We found a seismogenic fault at the upper boundary of the Yokonami mélange, indicating that the Yokonami mélange was active at seismogenic depth. The field-transect distribution of shear veins was examined. The frequency, the total and mean thicknesses of the shear veins were about 3.7 per meter, about 10 mm per meter, and about 3 mm per shear vein, respectively. Quartz within the shear veins shows elongate-blocky textures, suggesting precipitation from advective flow. The pressure and temperature conditions for shear vein formation were examined by fluid inclusion analysis, ranging 175-225°C and 143-215 MPa, respectively. The temperature is almost consistent with the paleotemperature determined from vitrinite reflectance, suggesting that the shear veins were formed at up to the maximum depth. The depth might be consistent with that where the seismogenic fault was formed. On the basis of the pressure and temperature conditions and the distribution of shear veins, we estimated the minimum volumetric ratio of fluid to host rocks, assuming that the shear veins had precipitated from advective flow. The estimated amount of fluid is about 106 m 3 per cubic meter of host rocks. The results suggest that a large amount of fluid migrates through mélange zones at shallow seismogenic depths. This fluid possibly originates from the dehydration of clay minerals from underthrusted sediments and an altered subducting slab.

Isolates of Banana bunchy top virus (BBTV), which causes bunchy top disease in bananas, were collected in field surveys on seven islands in Okinawa Prefecture, Japan. From 44 banana samples, one isolate from each island was selected, and the DNA-1 and DNA-3 components were sequenced. Analysis of the major common region of DNA-1 showed that BBTV in Okinawa belongs to the Asian group of BBTV. DNA-1 and DNA-3 analysis revealed that Okinawan BBTV had a closer relationship with isolates from Taiwan and the Philippines than with some isolates from China and Vietnam. All the Okinawan BBTV isolates had high homology in the nucleotide sequences of DNA-1 and DNA-3 (%) because of a single, recent BBTV invasion of this area.

In Japan and Southeast Asia, 'Candidatus Liberibacter asiaticus' (Las) is the dominant causal agent of citrus greening (huanglongbing) disease. Using PCR techniques, we determined the 11168-nucleotide sequence of the wserA-trmU-tufB-secE-nusG-rplKAJL-rpoB gene cluster and the flanking regions for 51 Japanese, four Taiwanese, four Indonesian, and three Vietnamese isolates of Las. The sequence is identical in 62 isolates collected from Japan, Taiwan, Indonesia, and Vietnam, except for nucleotide substitutions at 11 positions. Some Las isolates from Sakishima Islands near Taiwan had unique nucleotide mutations, but all Las isolates around Okinawa Main Island were homologous. On the basis of the pattern of single nucleotide polymorphisms (SNPs) of the 11 nucleotide substitutions, the 62 Las isolates from Japan, Taiwan, Indonesia, and Vietnam could be divided into 12 pattern groups, and the 51 Japanese isolates consisted of six patterns. The results suggested that one unique genetic group is dominant around Okinawa Main Island, whereas several different are commonly distributed around islands near Taiwan.

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