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Host range and phylogenetic analysis of Xanthomonas alfalfae causing bacterial leaf spot of alfalfa in Iran

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Abstract

In summer and early autumn 2016, as a part of a country-wide survey, five Gram-negative, yellow-pigmented bacterial isolates were recovered from diseased alfalfa (Medicago sativa cv. Bonab) plants in southern Iran. The isolates were negative for pectolytic activity but showed a weak amylolytic activity, and were sensitive to ≥100 mg/l, ≥0.5 g/l and ≥20 mg/l of copper sulfate, copper oxychloride, and streptomycin, respectively. All isolates were pathogenic on alfalfa as well as on common bean, faba bean, mung bean, soybean and pea. The phenotypic tests and detection with the genus-specific PCR primers established the strains as those belonging to Xanthomonas spp. Phylogenetic analysis, using the hrpB2 gene sequences, showed that all isolates belong to X. alfalfae. Multilocus sequence typing analysis, based on five housekeeping genes (fusA, gapA, gltA, lacF, and lepA) of two representative isolates, identified them as X. alfalfae subsp. alfalfae. The possible impact of this pathogen in the country is discussed, in relation to other recently reported Xanthomonads, including X. euvesicatoria and X. perforans.

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Acknowledgements

This study financially supported by Shiraz University.

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Authors and Affiliations

  1. Department of Plant Protection, College of Agriculture, Shiraz University, Shiraz, 71441-65186, Iran

    Zohreh Yaripour, S. Mohsen Taghavi & Ebrahim Osdaghi

  2. UMR AGIR, INRA, 31326, Castanet-Tolosan, France

    Jay Ram Lamichhane

Authors
  1. Zohreh Yaripour
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  2. S. Mohsen Taghavi
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  3. Ebrahim Osdaghi
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  4. Jay Ram Lamichhane
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Corresponding author

Correspondence to Ebrahim Osdaghi.

Electronic supplementary material

Fig. S1

Phylogeny of the fusA (a), gapA (b), gltA (c), lacF (d), and lepA (e) genes of the representative strains of Xanthomonas alfalfae subsp. alfalfae obtained in this study with other species of Xanthomonas spp. Concatenated phylogenetic tree was constructed based on the sequences of the fusA, gapA, gltA, lacF, and lepA genes in two representative Xanthomonas alfalfae subsp. alfalfae strains, using Neighbor Joining method (f). the percentage of bootstrap values >50% from 2000 samplings was indicated. Phylogeny based on all sequenced genes clustered the strains isolated in this study together with the type strains of X. alfalfae, X. euvesicatoria and X. perforans. None of the individual genes was capable to discriminate Xanthomonas alfalfae, X. euvesicatoria and X. perforans strains. (PDF 25.3 kb)

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Yaripour, Z., Mohsen Taghavi, S., Osdaghi, E. et al. Host range and phylogenetic analysis of Xanthomonas alfalfae causing bacterial leaf spot of alfalfa in Iran. Eur J Plant Pathol 150, 267–274 (2018). https://doi.org/10.1007/s10658-017-1271-0

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