The identification and characterization of endopolygalacturonases in a South African isolate of Phytophthora cinnamomi

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Authors

Miyambo, Tsakani Magdeline
Backer, Robert
Engelbrecht, Juanita
Joubert, Fourie
Van der Merwe, Nicolaas Albertus (Albie)
Van den Berg, Noelani

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MDPI

Abstract

Phytophthora cinnamomi is an economically important plant pathogen that has caused devastating losses to the avocado industry worldwide. To facilitate penetration and successful colonization of the host plant, pathogens have been reported to secrete polygalacturonases (PGs). Although a large PG gene family has been reported in P. cinnamomi, in-depth bioinformatics analyses and characterization of these genes is still lacking. In this study we used bioinformatics tools and molecular biology techniques to identify and characterize endopolygalacturonases in the genome of a South African P. cinnamomi isolate, GKB4. We identified 37 PGs, with 19 characteristics of full-length PGs. Although eight PcPGs were induced in planta during infection, only three showed significant up- and down-regulation when compared with in vitro mycelial growth, suggesting their possible roles in infection. The phylogenetic analysis of PcPGs showed both gain and loss of introns in the evolution of PGs in P. cinnamomi. Furthermore, 17 PGs were related to characterized PGs from oomycete species, providing insight on possible function. This study provides new data on endoPGs in P. cinnamomi and the evolution of introns in PcPG genes. We also provide a baseline for future functional characterization of PGs suspected to contribute to P. cinnamomi pathogenicity/virulence in avocado.

Description

SUPPLEMENTARY MATERIAL : FIGURE S1: strategy flow diagram for the identification and characterization of polygalacturonases in Phytophthora cinnamomi; FIGURE S2: pairwise alignment (zoomed out) of intron-containing PG genomic sequences with their corresponding coding sequences; FIGURE S3: pairwise alignment (zoomed in) of intron-containing PG genomic sequences with their corresponding coding sequences; FIGURE S4: a multiple sequence alignment of the polygalacturonase protein sequences from Phytophthora cinnamomi; TABLE S1: polygalacturonase sequences from different Phytophthora spp. used for the multiple sequence alignment; TABLE S2: polygalacturonase sequences from outgroup species and their ID as depicted on the phylogenetic trees; TABLE S3: polygalacturonase sequences from oomycete species and their ID as depicted on the phylogenetic tree; TABLE S4: representation of the intron exon boundaries of polygalacturonase genes from Phytophthora cinnamomi-GKB4 genome (accession: JAFJYM000000000) using Augustus for gene prediction; TABLE S5: protein sequence similarity of full-length and partial putative polygalacturonase proteins of Phytophthora cinnamomi to PG proteins from Phytophthora spp. based on BLASTP analyses; TABLE S6: full-length and partial polygalacturonase sequences from Phytophthora cinnamomic; TABLE S7: log 2-Fold Change gene expression of PG genes from P. cinnamomi at 12 hpi, 24 hpi and 120 hpi during colonization of the susceptible avocado rootstock (R. 012) normalized to mycelia. References [15,23,71,72] cited in Supplementary Materials.
DATA AVAILABILITY STATEMENT : Data generated or analyzed during this study are included in this published article and its supplementary information files. Sequences used in this study are available on Genbank (NCBI) accession numbers OL334941 to OL334977

Keywords

Phytophthora cinnamomi, Endopolygalacturonases, Bioinformatics, Evolution, Phyhylogenetics

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Citation

Miyambo, T.M.; Backer, R.; Engelbrecht, J.; Joubert, F.; van der Merwe, N.A.; van den Berg, N. The Identification and Characterization of Endopolygalacturonases in a South African Isolate of Phytophthora cinnamomi. Microorganisms 2022, 10, 1061. https://DOI.org/10.3390/microorganisms10051061.