Publications

4369

Huanglongbing also known in Indonesia as citrus vein phloem degeneration (CVPD) is a devastating disease in citrus plantation worldwide, especially in Asia, Africa, and America. In Asian countries including Indonesia, Candidatus Liberibacter asiaticus (CLas) has been confirmed as the causal agent of huanglongbing disease on citrus. Distribution of CLas in Indonesia has been reported in West Borneo, East Nusa Tenggara, Bali, Yogyakarta, Central Java and East Java.  The purpose of this study was to detect CLas in several Indonesia’s citrus plantations that has not and has been reported previously and to study its genetic diversity and their relationship. DNA of plant samples, i.e. citrus leaves, was extracted using CTAB method and CLas was amplified using PCR with the A2/J5 primer pair. DNA amplification results showed that infection of CLas was positively detected from samples from Bogor and Cibodas (West Java), Tuban and Jember (East Java), as well as Katung, Bayung Gede, Kerta, and Pancasari (Bali). Alignment of nucleotide sequences from positive samples showed that their ribosomal protein β-operon has high similiarity to that of CLas OK901 originated from Okinawa (Japan). Isolates of CLas originated from Bogor, Cibodas, Tuban, Jember, and Katung have been known to be identical to other CLas of Indonesian origins reported earlier.  Isolates of CLas originated from Bayung Gede, Pancasari, and Kerta have single nucleotide polymorphisms at 6 points of bases of the 539 total bases compared in their conservative regions, although only 3 of the 6 bases could affect their amino acid sequences. 

AbstractThe olive fruit fly,Bactrocera oleae, is the most destructive pest of olive orchards worldwide. The monophagous larva has the unique capability of feeding on olive mesocarp, coping with high levels of phenolic compounds and utilizing non-hydrolyzed proteins present, particularly in the unripe, green olives. On the molecular level, the interaction betweenB. oleaeand olives has not been investigated as yet. Nevertheless, it has been associated with the gut obligate symbiotic bacteriumCandidatus Erwinia dacicola. Here, we used aB.oleaemicroarray to analyze the gene expression of larvae during their development in artificial diet, unripe (green) and ripe (black) olives. The expression profiles ofCa. E. dacicolawere analyzed in parallel, using the Illumina platform. Several genes were found overexpressed in the olive fly larvae when feeding in green olives. Among these, a number of genes encoding detoxification and digestive enzymes, indicating a potential association with the ability ofB. oleaeto cope with green olives. In addition, a number of biological processes seem to be activated inCa. E. dacicoladuring the development of larvae in olives, with the most notable being the activation of amino-acid metabolism.

ABSTRACT SAR11 bacteria are small, heterotrophic, marine alphaproteobacteria found throughout the oceans. They thrive at the low nutrient concentrations typical of open ocean conditions, although the adaptations required for life under those conditions are not well understood. To illuminate this issue, we used cryo-electron tomography to study “ Candidatus Pelagibacter ubique” strain HTCC1062, a member of the SAR11 clade. Our results revealed its cellular dimensions and details of its intracellular organization. Frozen-hydrated cells, which were preserved in a life-like state, had an average cell volume (enclosed by the outer membrane) of 0.037 ± 0.011 μm 3 . Strikingly, the periplasmic space occupied ∼20% to 50% of the total cell volume in log-phase cells and ∼50% to 70% in stationary-phase cells. The nucleoid occupied the convex side of the crescent-shaped cells and the ribosomes predominantly occupied the concave side, at a relatively high concentration of 10,000 to 12,000 ribosomes/μm 3 . Outer membrane pore complexes, likely composed of PilQ, were frequently observed in both log-phase and stationary-phase cells. Long filaments, most likely type IV pili, were found on dividing cells. The physical dimensions, intracellular organization, and morphological changes throughout the life cycle of “ Ca. Pelagibacter ubique” provide structural insights into the functional adaptions of these oligotrophic ultramicrobacteria to their habitat. IMPORTANCE Bacterioplankton of the SAR11 clade ( Pelagibacterales ) are of interest because of their global biogeochemical significance and because they appear to have been molded by unusual evolutionary circumstances that favor simplicity and efficiency. They have adapted to an ecosystem in which nutrient concentrations are near the extreme limits at which transport systems can function adequately, and they have evolved streamlined genomes to execute only functions essential for life. However, little is known about the actual size limitations and cellular features of living oligotrophic ultramicrobacteria. In this study, we have used cryo-electron tomography to obtain accurate physical information about the cellular architecture of “ Candidatus Pelagibacter ubique,” the first cultivated member of the SAR11 clade. These results provide foundational information for answering questions about the cell architecture and functions of these ultrasmall oligotrophic bacteria.