Transcriptome analysis of 'Candidatus Liberibacter solanacearum' in its psyllid vector, Bactericera cockerelli [sexed insects]
ABSTRACT: We sequenced mRNA from the bacterial pathogen 'Candidatus Liberibacter solanacearum" during its association with the psyllid vector Bactericera cockerelli. Overall design: Total RNA was purified from psyllids, insect and bacterial rRNAs were depleted. PolyA RNA was purified using Dynabeads. PolyA purified RNA and depleted RNA were sequenced.
Frontiers in Cellular and Infection Microbiology 20160609
"Candidatus Liberibacter solanacearum" (Lso) has emerged as a serious threat world-wide. Five Lso haplotypes have been identified so far. Haplotypes A and B are present in the Americas and/or New Zealand, where they are vectored to solanaceous plants by the potato psyllid, Bactericera cockerelli (Šulc) (Hemiptera: Triozidae). The fastidious nature of these pathogens has hindered the study of the interactions with their eukaryotic hosts (vector and plant). To understand the strategies used by the ...[more]
Project description:We seqeunced mRNA from the bacterial pathogen 'Candidatus Liberibacter solanacearum" during its association with the psyllid vector Bactericera cockerelli. Total RNA was purified from psyllids, insect and bacterial rRNAs were depleted. PolyA RNA was purified using Dynabeads. PolyA purified RNA and depleted RNA were sequenced.
Project description:In this study we compare the transcriptome response of two potato varieties Atlantic and NY138 to the infection by Candidatus Liberibacter solanacearum. Four weeks old potato plant grown in growth chamber were infested with potato psyllid to transmit the pathogen Candidatus Liberibacter solanacearum. Three weeks after infestation leaf samples were collected for RNA extraction and transcriptome analysis. This is the first transcriptome study on this potato disease. Overall design: Two factorial design experiment with two potato varieites Atlantic and NY138, and two treatment: no infection compared to infection by Candidatus liberibacter solanacearum
Project description:This study evaluated the transcriptional reprogramming of a susceptible genotype (Pera sweet orange) challenged with the pathogenic bacteria Candidatus Liberibacter americanus (CaLam), using a customized 385K microarray containing about 32,000 unigene transcripts. For the microarray experiment were used symptomatic leaves from two Pera sweet orange plants inoculated with either bark or bud pieces infected with Candidatus Liberibacter americanus and two non-infected control plants.
Project description:HLB is suggested to be caused by the phloem-limited fastidious prokaryotic α-proteobacterium “Candidatus Liberibacter spp.” Previous studies focused on the proteome and transcriptome analyses of citrus 5 to 35-week-after “Ca. L. spp.” inoculation. In this study, gene expression profiles was analyzed using mandarin of Citrus reticulate Blanco cv. jiaogan leaves after 2-year infection with “Ca. L. asiaticus”. The Affymetrix GeneChip® citrus genome were applied to study the molecular pathways mediated by “Ca. L. asiaticus” inoculated 3-year-old jiaogan seedlings. Each of them was graft-inoculated with one sweet orange scions with or without “Ca. L. asiaticus” in Dectember, 2009. RNA samples from three mandarin trees infected with 'Candidatus Liberibacter asiaticus' and three uninfected trees were used for affymatrix genochip
Project description:Citrus greening or huanglongbing (HLB) is a devastating disease of citrus. HLB is associated with the phloem-limited fastidious prokaryotic alpha-proteobacterium Candidatus Liberibacter spp. In this report, we used sweet orange (Citrus sinensis) leaf tissue infected with 'Ca. Liberibacter asiaticus' and compared this with healthy controls. Investigation of the host response was examined with citrus microarray hybridization based on 30,171 sets expressed sequence tag sequences from several citrus species and hybrids. The microarray analysis indicated that HLB infection significantly affected expression of 624 genes whose encoded proteins were categorized according to function. The categories included genes associated with sugar metabolism, plant defense, phytohormone, and cell wall metabolism, as well as 14 other gene categories. Young, healthy Valencia sweet orange (C. sinensis) plants were graft inoculated with budwood from Ca. L. asiaticus-infected citrus plants. Prior to the innocualtion, the plants were confirmed to be Ca. L. asiaticus-free in ordinary and quantitative PCR tests. The presence of the bacteria in the inoculated plants was confirmed in both conventional and quantitative PCR with specific primers to Ca. L. asiaticus. The stem and root samples used for RNA extraction and hybridization on Affymetrix microarrays were obtained from three symptomatic and three healthy control trees of similar size, approximately 1 year after inoculation.
Project description:Candidatus Liberibacter asiaticus infection of citrus is characterized by symptom variability within and among organs. In order to identify molecular processes involved in the regulation of organ response to Ca. Liberibacter infection, the gene expression patterns in C. sinensis leaf, stem, and root was examined in Affymetrix microarray. Our analyses showed that Ca. L. asiaticus reprograms several cellular and metabolic processes in C. sinensis, with most categories regulated in leaves, followed by stems and least in roots. Among them, we identified genes whose expression is regulated in organ-specific manner, reflecting organ specialization in the molecular response to Ca. L. asiaticus. Differences in gene expression were expected between these organs because of functional divergence among them. Overall design: Two-year old Valencia sweet orange (C. sinensis) plants were graft inoculated with budwood from Ca. L. asiaticus-infected citrus plants. Successful infection of the inoculated plants was confirmed in both conventional and quantitative PCR with specific primers to Ca. L. asiaticus. The stem and root samples used for RNA extraction and hybridization on Affymetrix microarrays were obtained from three symptomatic and three healthy control trees of similar size, 16 months after the inoculation.