Function and Application of the CRISPR-Cas System in the Plant Pathogen<i>Erwinia amylovora</i>

Parcey, Michael; Gayder, Steven; Castle, Alan J.; Svircev, A. M.

doi:10.1128/aem.02513-21

Cited by 7 publications

(6 citation statements)

References 39 publications

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“…The type I CRISPR system in E . amylovora was recently reported to be involved in resistance against invasive plasmids and not necessarily against phages [ 55 ]. No link has been identified to c-di-GMP regulation in this regard.…”

Section: Discussionmentioning

confidence: 99%

“…CRISPR elements have been used in genotypically categorizing and tracking pathogenic evolution in E. amylovora [52][53][54]. The type I CRISPR system in E. amylovora was recently reported to be involved in resistance against invasive plasmids and not necessarily against phages [55]. No link has been identified to c-di-GMP regulation in this regard.…”

Section: Plos Pathogensmentioning

confidence: 99%

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A complete twelve-gene deletion null mutant reveals that cyclic di-GMP is a global regulator of phase-transition and host colonization in Erwinia amylovora

2022

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Cyclic-di-GMP (c-di-GMP) is an essential bacterial second messenger that regulates biofilm formation and pathogenicity. To study the global regulatory effect of individual components of the c-di-GMP metabolic system, we deleted all 12 diguanylate cyclase (dgc) and phosphodiesterase (pde)-encoding genes in E. amylovora Ea1189 (Ea1189Δ12). Ea1189Δ12 was impaired in surface attachment due to a transcriptional dysregulation of the type IV pilus and the flagellar filament. A transcriptomic analysis of surface-exposed WT Ea1189 and Ea1189Δ12 cells indicated that genes involved in metabolism, appendage generation and global transcriptional/post-transcriptional regulation were differentially regulated in Ea1189Δ12. Biofilm formation was regulated by all 5 Dgcs, whereas type III secretion and disease development were differentially regulated by specific Dgcs. A comparative transcriptomic analysis of Ea1189Δ8 (lacks all five enzymatically active dgc and 3 pde genes) against Ea1189Δ8 expressing specific dgcs, revealed the presence of a dual modality of spatial and global regulatory frameworks in the c-di-GMP signaling network.

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Section: Discussionmentioning

confidence: 99%

Section: Plos Pathogensmentioning

confidence: 99%

A complete twelve-gene deletion null mutant reveals that cyclic di-GMP is a global regulator of phase-transition and host colonization in Erwinia amylovora

2022

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“…Among the group of strains, Bacillus megaterium , Bacillus aryabhattai , Erwinia spp. and Caballeronia glathei were excluded from further study, as they were previously reported to be potential plant pathogens ( Abdel-Monaim et al., 2012 ; Narsing Rao et al., 2019 , Dobritsa and Samadpour, 2019 , Parcey et al., 2022 ). A more detailed quantitative determination of bacterial phosphorus solubilization efficiency in liquid media (NBRIP) with different phosphorus contents showed that Rhizobium spp., Pantoea agglomerans , and Paenibacillus spp.…”

Section: Resultsmentioning

confidence: 99%

Large effect of phosphate-solubilizing bacteria on the growth and gene expression of Salix spp. at low phosphorus levels

Koczorski,

Furtado,

Baum

et al. 2023

Front. Plant Sci.

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Phosphorus is one of the most important nutrients required for plant growth and development. However, owing to its low availability in the soil, phosphorus is also one of the most difficult elements for plants to acquire. Phosphorus released into the soil from bedrock quickly becomes unavailable to plants, forming poorly soluble complexes. Phosphate-solubilizing bacteria (PSB) can solubilize unavailable phosphorus-containing compounds into forms in which phosphorus is readily available, thus promoting plant growth. In this study, two willow species, Salix dasyclados cv. Loden and Salix schwerinii × Salix viminalis cv. Tora, were inoculated with two selected bacterial strains, Pantoea agglomerans and Paenibacillus spp., to evaluate the plant growth parameters and changes in gene expression in the presence of different concentrations of tricalcium phosphate: 0 mM (NP), 1 mM (LP), and 2 mM (HP). Inoculation with PSB increased root, shoot and leaf biomass, and for the HP treatment, significant changes in growth patterns were observed. However, the growth responses to plant treatments tested depended on the willow species. Analysis of the leaf transcriptomes of the phosphate-solubilizing bacterium-inoculated plants showed a large variation in gene expression between the two willow species. For the Tora willow species, upregulation of genes was observed, particularly for those involved in pathways related to photosynthesis, and this effect was strongly influenced by bacterial phosphate solubilization. The Loden willow species was characterized by a general downregulation of genes involved in pathway activity that included ion transport, transcription regulation and chromosomes. The results obtained in this study provide an improved understanding of the dynamics of Salix growth and gene expression under the influence of PSB, contributing to an increase in yield and phosphorus-use efficiency.

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Section: Discussionmentioning

confidence: 99%

“…actinidiae (Psa) [50], Pseudomonas chlororaphis [51], Agrobacterium spp. [52], and Erwinia amylovora [53]. Regarding gene knockout, the ectopic expression of the type I-C CRISPR/Cas3 system from P. aeruginosa in Pst DC3000 resulted in 67-92% efficiency of genome editing, characterized by large deletions (55-100 kb) at four target sites [25]; Application of CRISPR/SpCas9 in Psa generated very few transformants with one targeting sgRNA and 64.29% positive colonies carried 7.82-64 kb deletions.…”

Section: Plos Pathogensmentioning

confidence: 99%

CRISPR/FnCas12a-mediated efficient multiplex and iterative genome editing in bacterial plant pathogens without donor DNA templates

et al. 2023

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CRISPR-based genome editing technology is revolutionizing prokaryotic research, but it has been rarely studied in bacterial plant pathogens. Here, we have developed a targeted genome editing method with no requirement of donor templates for convenient and efficient gene knockout in Xanthomonas oryzae pv. oryzae (Xoo), one of the most important bacterial pathogens on rice, by employing the heterologous CRISPR/Cas12a from Francisella novicida and NHEJ proteins from Mycobacterium tuberculosis. FnCas12a nuclease generated both small and large DNA deletions at the target sites as well as it enabled multiplex genome editing, gene cluster deletion, and plasmid curing in the Xoo PXO99A strain. Accordingly, a non-TAL effector-free polymutant strain PXO99AD25E, which lacks all 25 xop genes involved in Xoo pathogenesis, has been engineered through iterative genome editing. Whole-genome sequencing analysis indicated that FnCas12a did not have a noticeable off-target effect. In addition, we revealed that these strategies are also suitable for targeted genome editing in another bacterial plant pathogen Pseudomonas syringae pv. tomato (Pst). We believe that our bacterial genome editing method will greatly expand the CRISPR study on microorganisms and advance our understanding of the physiology and pathogenesis of Xoo.

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Function and Application of the CRISPR-Cas System in the Plant PathogenErwinia amylovora

Abstract: Erwinia amylovora is an economically significant agricultural pathogen found throughout the world. In North America, E. amylovora has developed streptomycin resistance and therefore alternative treatments using phages have received increased attention.

Cited by 7 publications

References 39 publications

A complete twelve-gene deletion null mutant reveals that cyclic di-GMP is a global regulator of phase-transition and host colonization in Erwinia amylovora

A complete twelve-gene deletion null mutant reveals that cyclic di-GMP is a global regulator of phase-transition and host colonization in Erwinia amylovora

Large effect of phosphate-solubilizing bacteria on the growth and gene expression of Salix spp. at low phosphorus levels

CRISPR/FnCas12a-mediated efficient multiplex and iterative genome editing in bacterial plant pathogens without donor DNA templates

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