Publications

4865

AbstractBackground:  “Candidatus Helicobacter heilmannii” induce chronic gastritis, which eventually leads to gastric B‐cell type mucosa‐associated lymphoid tissue (MALT) lymphoma. This study was performed using an animal model of infection with “Candidatus Helicobacter heilmannii” to elucidate how this chronic inflammation is induced or maintained.Materials and Methods:  BALB/c mice were infected with the “Candidatus Helicobacter heilmannii” isolate SH4. The animals were examined at 8, 26, 54, and 83 weeks after the infection. The stomach of the animals was resected and immunostained for peripheral lymph node addressin (PNAd) and mucosal addressin cell adhesion molecule 1 (MAdCAM‐1), “Candidatus Helicobacter heilmannii,” and CD45R/B220. An in vitro binding assay with L‐ and E‐selectin·IgM chimeric proteins was performed. Real‐time polymerase chain reaction was used to evaluate transcripts of N‐acetylglucosamine‐6‐O‐sulfotransferases (GlcNAc6STs), which direct the expression of the PNAd and MAdCAM‐1.Results:  Chronic gastritis developed in the infected animals, and its severity increased with the duration of the infection. B‐cell type MALT lymphoma developed in some animals at 54 and 83 weeks after infection. PNAd‐ and MAdCAM‐1‐expressing high endothelial venule (HEV)‐like vessels were induced in infected animals which developed chronic gastritis and MALT lymphoma. The number of HEV‐like vessels increased as chronic inflammation progressed. The induced HEV‐like vessels were bound by L‐ and E‐selectin·IgM chimeric protein. mRNA expressions of GlcNAc6ST‐1 and MAdCAM‐1 increased in the infected animals.Conclusions:  HEV‐like vessels expressing GlcNAc6ST‐1‐mediated L‐selectin ligand carbohydrate and MAdCAM‐1 may play a crucial role in the pathogenesis of “Candidatus Helicobacter heilmannii”‐induced chronic gastritis and MALT lymphoma.

Changes in carbohydrate metabolism in sweet orange ( Citrus sinensis ) infected with ‘ Candidatus Liberibacter asiaticus’, a purported cause of citrus Huanglongbing (HLB), were investigated. Starch levels in HLB‐infected leaves with and without symptoms increased 3·1‐ and 7·9‐fold, respectively, compared to healthy controls. In symptomless leaves, sucrose and fructose accumulated significantly ( P  <   0·05) in both midribs and lobes, and glucose only in the midribs (greater than fivefold); whereas maltose levels were reduced to 64·6% and 86·8% in the midribs and foliar lobes, respectively, of the values in healthy leaves. In leaves with symptoms, sucrose and glucose remained at high levels compared to healthy leaves, whilst no accumulation of fructose was observed; by contrast, the maltose content decreased to as low as 49·6% of that in healthy leaves. Fourfold induction of cell‐wall‐bound invertase activity was detected in both types of leaves on diseased plants. Additionally, the expression profiles of starch breakdown genes suggested that the transcription of DPE2 and MEX1 was down‐regulated. Together with the reduction of maltose, it is suggested that the impairment of starch breakdown may contribute to the starch accumulation in infected leaves. The imbalance of carbohydrate partitioning and its relation to disease physiology are discussed.

ABSTRACT Mealybugs (Coccoidea: Pseudococcidae) are sap-sucking plant parasites that harbor bacterial endosymbionts within specialized organs. Previous studies have identified two subfamilies, Pseudococcinae and Phenacoccinae, within mealybugs and determined the primary endosymbionts (P-endosymbionts) of the Pseudococcinae to be Betaproteobacteria (“ Candidatus Tremblaya princeps”) containing Gammaproteobacteria secondary symbionts. Here, the P-endosymbionts of phenacoccine mealybugs are characterized based on 16S rRNA from the bacteria of 20 species of phenacoccine mealybugs and four outgroup Puto species (Coccoidea: Putoidae) and aligned to more than 100 published 16S rRNA sequences from symbiotic and free-living bacteria. Phylogenetic analyses recovered three separate lineages of bacteria from the Phenacoccinae, and these are considered to be the P-endosymbionts of their respective mealybug hosts, with those from (i) the mealybug genus Rastrococcus belonging to the Bacteroidetes , (ii) the subterranean mealybugs, tribe Rhizoecini, also within Bacteroidetes , in a clade sister to cockroach endosymbionts ( Blattabacterium ), and (iii) the remaining Phenacoccinae within the Betaproteobacteria , forming a well-supported sister group to “ Candidatus Tremblaya princeps.” Names are proposed for two strongly supported lineages: “ Candidatus Brownia rhizoecola” for P-endosymbionts of Rhizoecini and “ Candidatus Tremblaya phenacola” for P-endosymbionts of Phenacoccinae excluding Rastrococcus and Rhizoecini. Rates of nucleotide substitution among lineages of Tremblaya were inferred to be significantly faster than those of free-living Betaproteobacteria . Analyses also recovered a clade of Gammaproteobacteria , sister to the P-endosymbiont lineage of aphids (“ Candidatus Buchnera aphidicola”), containing the endosymbionts of Putoidae, the secondary endosymbionts of pseudococcine mealybugs, and the endosymbionts of several other insect groups.

Citrus huanglongbing, putatively caused by the associated bacterium ‘Candidatus Liberibacter asiaticus’, is the greatest threat to the world citrus industry today. The bacterium is spread locally and regionally by the citrus psyllid Diaphorina citri, and also can be disseminated by propagation of contaminated scion budwood that is grafted to the appropriate rootstock. The planting of ‘Ca. Liberibacter asiaticus’-free trees is a component of a comprehensive strategy to manage huanglongbing. In contrast to the scion budwood, the rootstocks used to produce these trees are grown from seed. This research was undertaken to provide evidence as to whether or not ‘Ca. L. asiaticus’ can be transmitted through seed. Two groups of 360 or more seedlings each of various citrus species were grown from seed removed from fruit on trees that were symptomatic for huanglongbing and confirmed to be infected with ‘Ca. L. asiaticus’ by polymerase chain reaction (PCR) tests. These seedlings were tested multiple times over periods of up to 3 years. No symptoms typical of huanglongbing, such as blotchy leaf mottle, chlorotic shoots, or dieback of branches, were observed in these seedlings, and none of these 723 seedlings tested positive for the presence of ‘Ca. L. asiaticus’ even after repeated testing by sensitive quantitative PCR assays. Some sour orange seedlings did have quite pronounced and atypical growth, including stunting and mild to severe leaf malformation. These atypical growth habits were limited to seedlings that arose from zygotic embryos as determined by expressed-sequence tag simple-sequence repeat analyses. Thus, no evidence of transmission of ‘Ca. L. asiaticus’ via seed was obtained, and an earlier report of transmission of the pathogen through seed was not confirmed.