Article ; Online: First Report of Pseudomonas viridiflava Causing Fruit Rot on Sweet Cherry Trees in Chile
Plant Disease. 2023 Apr. 03, v. 107, no. 4 p.1216-
2023
Abstract: Chile is an important producer of sweet cherry (Prunus avium L.), with a total of 356,385 t exported in the 2021 to 2022 season. The production area includes most of the country’s regions (http://www.iqonsulting.com/yb/). Bacterial samples were isolated ... ...
| Abstract | Chile is an important producer of sweet cherry (Prunus avium L.), with a total of 356,385 t exported in the 2021 to 2022 season. The production area includes most of the country’s regions (http://www.iqonsulting.com/yb/). Bacterial samples were isolated in 2017 and 2018 from 18 commercial sweet cherry orchards with canker disease (Beltrán et al. 2021). From one of these samples, collected in the spring of 2018, was isolated the strain A2M176 from buds of trees that presented canker and gum exudation in Malloa locality (34°23′46″S and 71°01′39″W). The strain produced fluorescent pigment on King’s B agar medium. It was oxidase and arginine dihydrolase negative, potato soft rot positive, and showed a slight degree of tobacco hypersensitivity. It was able to grow in up to 0.8 mM (200 ppm) of CuSO₄·5H₂O (Cazorla et al. 2002). The strain A2M176 was deposited in the Colección Chilena de Recursos Genéticos Microbianos (CChRGM) under the no. RGM 3342. The DNA of this strain was extracted from a pure culture using silica spin columns (Epoch Life Science, Sugar Land). The complete DNA was sequenced using HiSeq with 150 bp paired-end at GENEWIZ (New York, U.S.A.). Raw data were checked using FASTQC (Andrews 2010) and trimmed with BBDuk (Bushnell 2014). The genome was assembled using Unicycler v0.4.9 (Wick et al. 2017) with default settings and annotated with Prokaryotic Genome Annotation Pipeline (PGAP) v4.3 (Tatusova et al. 2014). The reads and genomes were uploaded to GenBank under the BioProyect no. PRJNA750090, BioSample no. SAMN26870984, and assembly no. GCA_022936465.1. The sequenced genome was compared through the Average Nucleotide Identity algorithm (ANI) using FastANI v1.33 (Jain et al. 2018) to compare with the closest complete genome available on NCBI. The strain A2M176 was identified as P. viridiflava with an ANI value of 98.06% with the strain p22.E7 (GCF_900585495). Maximum likelihood phylogenetic estimation clustered strain A2M176 with other P. viridiflava strains with 95% bootstrap. The pathogenicity of the strain was tested by inoculating immature cherry fruits with a needle with a bacterial suspension (1 × 10⁸ CFU/ml). The inoculated fruits were placed in a humid chamber at room temperature for 10 days. Soft rot lesions were observed, which appeared at 6 days postinoculation. The control fruits treated with sterile water did not show symptoms. Further analyses in the genome of strain A2M176 identified genes related to pathogenicity, such as the effector gene avrE and the regulator gene HrpL, suggesting the pathogenicity ability of the strain. Also, genes were identified of two known Pseudomonas copper resistance mechanisms, the cus and cop operon. These genes were found as part of the copABCDns cluster, similar to what was observed in Pseudomonas from mango (Aprile et al. 2021). The presence of P. viridiflava strains causing fruit rot in P. avium is not surprising since P. viridiflava has a wide host range and causes a variety of symptoms in different plant parts, including stems, leaves, and blossoms (Lipps and Samac 2022). P. viridiflava represents one of the multiple phylogroups found within the P. syringae species complex (Bartoli et al. 2014). To our knowledge, this is the first report of a strain of copper-resistant P. viridiflava causing infection on sweet cherries in Chile. |
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| Keywords | DNA ; Prunus avium ; Pseudomonas viridiflava ; agar ; algorithms ; ambient temperature ; arginine deiminase ; cherries ; exudation ; fluorescence ; host range ; hypersensitivity ; mangoes ; operon ; oxidoreductases ; pathogenicity ; phylogeny ; plant rots ; potatoes ; regulator genes ; silica ; spring ; statistical analysis ; sugars ; tobacco ; Chile ; bacterial infection ; copper resistance ; plant pathogen ; prokaryotic genome |
| Language | English |
| Dates of publication | 2023-0403 |
| Publishing place | The American Phytopathological Society |
| Document type | Article ; Online |
| ZDB-ID | 754182-x |
| ISSN | 0191-2917 |
| ISSN | 0191-2917 |
| DOI | 10.1094/PDIS-07-22-1638-PDN |
| Database | NAL-Catalogue (AGRICOLA) |
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