Overexpression of SSBXoc, a Single-Stranded DNA-Binding Protein From Xanthomonas oryzae pv. oryzicola, Enhances Plant Growth and Disease and Salt Stress Tolerance in Transgenic Nicotiana benthamiana

We previously reported that SSBXoc, a highly conserved single-stranded DNA-binding protein from Xanthomonas spp., was secreted through the type III secretion system (T3SS) and functioned as a harpin-like protein to elicit the hypersensitive response (HR) in the non-host plant, tobacco. In this study, we cloned SsbXoc gene from X. oryzae pv. oryzicola (Xoc), the causal agent of bacterial leaf streak in rice, and transferred it into Nicotiana benthamiana via Agrobacterium-mediated transformation. The expression of SsbXoc in transgenic N. benthamiana enhanced growth of both seedling and adult plants. When inoculated with the harpin Hpa1 or the pathogen Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), the accumulation of reactive oxygen species (ROS) was increased more in SsbXoc transgenic lines than that in wild-type (WT) plants. The expression of pathogenesis-related protein genes (PR1a and SGT1), HR marker genes (HIN1 and HSR203J) and the mitogen-activated protein kinase pathway gene, MPK3, was significantly higher in transgenic lines than in WT after inoculation with Pst DC3000. In addition, SsbXoc transgenic lines showed the enhanced resistance to the pathogenic bacteria P. s. tabaci and the improved tolerance to salt stress, accompanied by the elevated transcription levels of the defense- and stress-related genes. Taken together, these results indicate that overexpression of the SsbXoc gene in N. benthamiana significantly enhanced plant growth and increased tolerance to disease and salt stress via modulating the expression of the related genes, thus providing an alternative approach for development of plants with improved tolerance against biotic and abiotic stresses

The cAMP pathway is important for controlling the morphological switch to the pathogenic yeast form of Paracoccidioides brasiliensis

Paracoccidioides brasiliensis is a human pathogenic fungus that switches from a saprobic mycelium to a pathogenic yeast. Consistent with the morphological transition being regulated by the cAMP-signalling pathway, there is an increase in cellular cAMP levels both transiently at the onset (< 24 h) and progressively in the later stages (> 120 h) of the transition to the yeast form, and this transition can be modulated by exogenous cAMP. We have cloned the cyr1 gene encoding adenylate cyclase (AC) and established that its transcript levels correlate with cAMP levels. In addition, we have cloned the genes encoding three Gα (Gpa1–3), Gβ (Gpb1) and Gγ (Gpg1) G proteins. Gpa1 and Gpb1 interact with one another and the N-terminus of AC, but neither Gpa2 nor Gpa3 interacted with Gpb1 or AC. The interaction of Gpa1 with Gpb1 was blocked by GTP, but its interaction with AC was independent of bound nucleotide. The transcript levels for gpa1, gpb1 and gpg1 were similar in mycelium, but there was a transient excess of gpb1 during the transition, and an excess of gpa1 in yeast. We have interpreted our findings in terms of a novel signalling mechanism in which the activity of AC is differentially modulated by Gpa1 and Gpb1 to maintain the signal over the 10 days needed for the morphological switch

The MinCDE Cell Division System Participates in the Regulation of Type III Secretion System (T3SS) Genes, Bacterial Virulence, and Motility in Xanthomonas oryzae pv. oryzae

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial leaf blight (BLB) in rice, which is one of the most severe bacterial diseases in rice in some Asian countries. The type III secretion system (T3SS) of Xoo encoded by the hypersensitive response and pathogenicity (hrp) genes is essential for its pathogenicity in host rice. Here, we identified the Min system (MinC, MinD, and MinE), a negative regulatory system for bacterial cell division encoded by minC, minD, and minE genes, which is involved in negative regulation of hrp genes (hrpB1 and hrpF) in Xoo. We found that the deletion of minC, minD, and minCDE resulted in enhanced hrpB1 and hrpF expression, which is dependent on two key hrp regulators HrpG and HrpX. The minC, minD, and minCDE mutants exhibited elongated cell lengths, and the classic Min system-defective cell morphology including minicells and short filamentations. Mutation of minC in Xoo resulted in significantly impaired virulence in host rice, swimming motility, and enhanced biofilm formation. Our transcriptome profiling also indicated some virulence genes were differentially expressed in the minC mutants. To our knowledge, this is the first report about the Min system participating in the regulation of T3SS expression. It sheds light on the understanding of Xoo virulence mechanisms

Image_1_Overexpression of SSBXoc, a Single-Stranded DNA-Binding Protein From Xanthomonas oryzae pv. oryzicola, Enhances Plant Growth and Disease and Salt Stress Tolerance in Transgenic Nicotiana benthamiana.PDF

<p>We previously reported that SSB_Xoc, a highly conserved single-stranded DNA-binding protein from Xanthomonas spp., was secreted through the type III secretion system (T3SS) and functioned as a harpin-like protein to elicit the hypersensitive response (HR) in the non-host plant, tobacco. In this study, we cloned Ssb_Xoc gene from X. oryzae pv. oryzicola (Xoc), the causal agent of bacterial leaf streak in rice, and transferred it into Nicotiana benthamiana via Agrobacterium-mediated transformation. The expression of Ssb_Xoc in transgenic N. benthamiana enhanced growth of both seedling and adult plants. When inoculated with the harpin Hpa1 or the pathogen Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), the accumulation of reactive oxygen species (ROS) was increased more in Ssb_Xoc transgenic lines than that in wild-type (WT) plants. The expression of pathogenesis-related protein genes (PR1a and SGT1), HR marker genes (HIN1 and HSR203J) and the mitogen-activated protein kinase pathway gene, MPK3, was significantly higher in transgenic lines than in WT after inoculation with Pst DC3000. In addition, Ssb_Xoc transgenic lines showed the enhanced resistance to the pathogenic bacteria P. s. tabaci and the improved tolerance to salt stress, accompanied by the elevated transcription levels of the defense- and stress-related genes. Taken together, these results indicate that overexpression of the Ssb_Xoc gene in N. benthamiana significantly enhanced plant growth and increased tolerance to disease and salt stress via modulating the expression of the related genes, thus providing an alternative approach for development of plants with improved tolerance against biotic and abiotic stresses.</p

Complete Genome Sequence Reveals Evolutionary and Comparative Genomic Features of Xanthomonas albilineans Causing Sugarcane Leaf Scald

International audienceLeaf scald (caused by Xanthomonas albilineans) is an important bacterial disease affecting sugarcane in most sugarcane growing countries, including China. High genetic diversity exists among strains of X. albilineans from diverse geographic regions. To highlight the genomic features associated with X. albilineans from China, we sequenced the complete genome of a representative strain (Xa-FJ1) of this pathogen using the PacBio and Illumina platforms. The complete genome of strain Xa-FJ1 consists of a circular chromosome of 3,724,581 bp and a plasmid of 31,536 bp. Average nucleotide identity analysis revealed that Xa-FJ1 was closest to five strains from the French West Indies and the USA, particularly to the strain GPE PC73 from Guadeloupe. Comparative genomic analysis between Xa-FJ1 and GPE PC73 revealed prophage integration, homologous recombination, transposable elements, and a clustered regulatory interspaced short palindromic repeats (CRISPR) system that were linked with 16 insertions/deletions (InDels). Ten and 82 specific genes were found in Xa-FJ1 and GPE PC73, respectively, and some of these genes were subjected to phage-related proteins, zona occludens toxin, and DNA methyltransferases. Our findings highlight intra-species genetic variability of the leaf scald pathogen and provide additional genomic resources to investigate its fitness and virulence

The Destructive Citrus Pathogen, ‘<em>Candidatus</em> Liberibacter asiaticus’ Encodes a Functional Flagellin Characteristic of a Pathogen-Associated Molecular Pattern

<div><p>Huanglongbing (HLB) is presently the most devastating citrus disease worldwide. As an intracellular plant pathogen and insect symbiont, the HLB bacterium, ‘<em>Candidatus</em> Liberibacter asiaticus’ (Las), retains the entire flagellum-encoding gene cluster in its significantly reduced genome. Las encodes a flagellin and hook-associated protein (Fla) of 452 amino acids that contains a conserved 22 amino acid domain (flg22) at positions 29 to 50 in the N-terminus. The phenotypic alteration in motility of a <em>Sinorhizobium meliloti</em> mutant lacking the <em>fla</em> genes was partially restored by constitutive expression of Fla<em>_Las</em>. <em>Agrobacterium</em>-mediated transient expression <em>in planta</em> revealed that Fla<em>_Las</em> induced cell death and callose deposition in <em>Nicotiana benthamiana</em>, and that the transcription of <em>BAK1</em> and <em>SGT1</em>, which are associated with plant innate immunity, was upregulated. Amino acid substitution experiments revealed that residues 38 (serine) and 39 (aspartate) of Fla<em>_Las</em> were essential for callose induction. The synthetic flg22<em>_Las</em> peptide could not induce plant cell death but retained the ability to induce callose deposition at a concentration of 20 µM or above. This demonstrated that the pathogen-associated molecular pattern (PAMP) activity of flg22 in Las was weaker than those in other well-studied plant pathogenic bacteria. These results indicate that Fla<em>_Las</em> acts as a PAMP and may play an important role in triggering host plant resistance to the HLB bacteria.</p> </div

Next Generation Sequencing and Comparative Genomic Analysis Reveal Extreme Plasticity of Two <i>Burkholderia glumae</i> Strains HN1 and HN2

Burkholderia glumae is an important rice pathogen, thus the genomic and evolutionary history may be helpful to control this notorious pathogen. Here, we present two complete genomes of the B. glumae strains HN1 and HN2, which were isolated from diseased rice seed in China. Average nucleotide identity (ANI) analysis shows greater than 99% similarity of the strains HN1 and HN2 with other published B. glumae genomes. Genomic annotation revealed that the genome of strain HN1 consists of five replicons (6,680,415 bp) with an overall G + C content of 68.06%, whereas the genome of strain HN2 comprises of three replicons (6,560,085 bp) with an overall G + C content of 68.34%. The genome of HN1 contains 5434 protein-coding genes, 351 pseudogenes, and 1 CRISPR, whereas the genome of HN2 encodes 5278 protein-coding genes, 357 pseudogenes, and 2 CRISPR. Both strains encode many pathogenic-associated genes (143 genes in HN1 vs. 141 genes in HN2). Moreover, comparative genomic analysis shows the extreme plasticity of B. glumae, which may contribute to its pathogenicity. In total, 259 single-copy genes were affected by positive selection. These genes may contribute to the adaption to different environments. Notably, six genes were characterized as virulence factors which may be an additional way to assist the pathogenicity of B. glumae