Genetic and metabolic analyses of Candidatus Liberibacter solanacearum infecting carrot

Insect-vectored plant bacterial pathogens are gaining attention in recent years due to crop threatening outbreaks around the world. Candidatus Liberibacter spp. are infecting crops of different botanical families: Solanaceae, Rutaceae, and Apiaceae and are vectored by psyllids. Five genetic haplotypes (A-E) have been described thus far for the species Ca. Liberibacter solanacearum (Lso). Haplotypes A and B infecting solanaceous plants, haplotypes C-E infecting Apiaceae crops. To better understand the genetic basis that governs host specificity of Lso haplotypes, we sequenced the genome of haplotype D (LsoD). The LsoD genome size is 1.23 Mbp, with a GC content of 34.8% and 1167 predicted genes. Enzyme Commission (EC) numbers were assigned using the JGI software tool and 358 ECs were identified. ECs were mapped to metabolic pathways and compared with other sequenced Liberibacters. Phylogenetic analysis based on ECs and assigned metabolic pathways shows that LsoD groups together with Lso haplotypes (A and B) and is clearly different than Liberibacter species infecting citrus. Differences between LsoD and LsoA/B haplotypes were also found, hinting on host specific enzymes. The LsoD genome was also scanned to identify putatively secreted proteins using the SignalP tool. Thirty-one putative genes were identified, most of them with unknown function. While some genes have homologous in other Lso haplotypes, some were unique to LsoD. By quantitative-PCR we examined the expression of the putatively secreted proteins in the different hosts; the psyllid vector Bactericera trigonica, and carrot. Several genes with significantly higher expression levels in carrot compared with psyllid and vice versa were identified. These genes may have host specific functions. Overall, our analyses reveal genetic and metabolic elements differentiating the carrot-infecting Lso from Lso haplotypes infecting potato/tomato. Research is underway to identify the function of these elements

Genome Analysis of Haplotype D of Candidatus Liberibacter Solanacearum

Candidatus Liberibacter solanacearum (Lso) haplotype D (LsoD) is a suspected bacterial pathogen, spread by the phloem-feeding psyllid Bactericera trigonica Hodkinson and found to infect carrot plants throughout the Mediterranean. Haplotype D is one of six haplotypes of Lso that each have specific and overlapping host preferences, disease symptoms, and psyllid vectors. Genotyping of rRNA genes has allowed for tracking the haplotype diversity of Lso and genome sequencing of several haplotypes has been performed to advance a comprehensive understanding of Lso diseases and of the phylogenetic relationships among the haplotypes. To further pursue that aim we have sequenced the genome of LsoD from its psyllid vector and report here its draft genome. Genome-based single nucleotide polymorphism analysis indicates LsoD is most closely related to the A haplotype. Genomic features and the metabolic potential of LsoD are assessed in relation to Lso haplotypes A, B, and C, as well as the facultative strain Liberibacter crescens. We identify genes unique to haplotype D as well as putative secreted effectors that may play a role in disease characteristics specific to this haplotype of Lso

Correction:Transgenic expression of the dicotyledonous pattern recognition receptor EFR in rice leads to ligand-dependent activation of defense responses

[This corrects the article DOI: 10.1371/journal.ppat.1004809.]

Transgenic expression of the dicotyledonous pattern recognition receptor EFR in rice leads to ligand-dependent activation of defense responses

Plant plasma membrane localized pattern recognition receptors (PRRs) detect extracellular pathogen-associated molecules. PRRs such as Arabidopsis EFR and rice XA21 are taxonomically restricted and are absent from most plant genomes. Here we show that rice plants expressing EFR or the chimeric receptor EFR::XA21, containing the EFR ectodomain and the XA21 intracellular domain, sense both Escherichia coli- and Xanthomonas oryzae pv. oryzae (Xoo)-derived elf18 peptides at sub-nanomolar concentrations. Treatment of EFR and EFR::XA21 rice leaf tissue with elf18 leads to MAP kinase activation, reactive oxygen production and defense gene expression. Although expression of EFR does not lead to robust enhanced resistance to fully virulent Xoo isolates, it does lead to quantitatively enhanced resistance to weakly virulent Xoo isolates. EFR interacts with OsSERK2 and the XA21 binding protein 24 (XB24), two key components of the rice XA21-mediated immune response. Rice-EFR plants silenced for OsSERK2, or overexpressing rice XB24 are compromised in elf18-induced reactive oxygen production and defense gene expression indicating that these proteins are also important for EFR-mediated signaling in transgenic rice. Taken together, our results demonstrate the potential feasibility of enhancing disease resistance in rice and possibly other monocotyledonous crop species by expression of dicotyledonous PRRs. Our results also suggest that Arabidopsis EFR utilizes at least a subset of the known endogenous rice XA21 signaling components

Transgenic expression of the rice Xa21 pattern‐recognition receptor in banana (Musa sp.) confers resistance to Xanthomonas campestris pv. musacearum

Banana Xanthomonas wilt (BXW), caused by the bacterium Xanthomonas campestris pv. musacearum (Xcm), is the most devastating disease of banana in east and central Africa. The spread of BXW threatens the livelihood of millions of African farmers who depend on banana for food security and income. There are no commercial chemicals, biocontrol agents or resistant cultivars available to control BXW. Here, we take advantage of the robust resistance conferred by the rice pattern-recognition receptor (PRR), XA21, to the rice pathogen Xanthomonas oryzae pv. oryzae (Xoo). We identified a set of genes required for activation of Xa21-mediated immunity (rax) that were conserved in both Xoo and Xcm. Based on the conservation, we hypothesized that intergeneric transfer of Xa21 would confer resistance to Xcm. We evaluated 25 transgenic lines of the banana cultivar 'Gonja manjaya' (AAB) using a rapid bioassay and 12 transgenic lines in the glasshouse for resistance against Xcm. About 50% of the transgenic lines showed complete resistance to Xcm in both assays. In contrast, all of the nontransgenic control plants showed severe symptoms that progressed to complete wilting. These results indicate that the constitutive expression of the rice Xa21 gene in banana results in enhanced resistance against Xcm. Furthermore, this work demonstrates the feasibility of PRR gene transfer between monocotyledonous species and provides a valuable new tool for controlling the BXW pandemic of banana, a staple food for 100 million people in east Africa

Small Protein-Mediated Quorum Sensing in a Gram- Negative Bacterium

The rice XA21 pattern recognition receptor binds a type I secreted sulfated peptide, called axY S 22, derived from the Ax21 (activator of XA21-mediated immunity) protein. The conservation of Ax21 in all sequenced Xanthomonas spp. and closely related genera suggests that Ax21 serves a key biological function. Here we show that the predicted N-terminal sequence of Ax21 is cleaved prior to secretion outside the cell and that mature Ax21 serves as a quorum sensing (QS) factor in Xanthomonas oryzae pv. oryzae. Ax21-mediated QS controls motility, biofilm formation and virulence. We provide genetic evidence that the Xoo RaxH histidine kinase serves as the bacterial receptor for Ax21. This work establishes a critical role for small protein-mediated QS in a Gram-negative bacterium