Identification and Characterization of Neodidymelliopsis cynanchi, the Causal Agent of Milkweed Leaf Lesions

Document Type : Original Article

Authors

1 Department of Biodiversity, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran

2 Department of Plant Protection, School of Agriculture, Shiraz University, Shiraz, Iran

Abstract

Calotropis procera is a shrub or small tree growing in tropical regions of the southern provinces in Iran. The economic benefits of the shrub include the use of latex and extract in traditional medicine. In a survey conducted in the southern provinces of Iran, leaf lesion symptoms were observed on C. procera shrubs. The purpose of the current study was to identify and characterize putative pathogens causing these symptoms. Isolations from the symptomatic tissues yielded predominantly a fungus belonging to the Didymellaceae family of Ascomycota. The causal agent was identified as Neodidymelliopsis cynanchi, based on morphological characteristics, and multilocus phylogenetic analysis using ITS, LSU, rpb2, and tub2 sequences. Pathogenicity tests on four-month-old seedlings of C. procera revealed that the isolates caused typical leaf lesions on the host. To the best of our knowledge, this is the first report of N. cynanchi causing disease on C. procera worldwide and the first report of the occurrence of this species in Iran.

Keywords

Main Subjects

References

Ahmad Nejhad, A., Alizadeh Behbahani, B., Hojjati, M., Vasiee, A. and Mehrnia, M. A. 2023. Identification of phytochemical, antioxidant, anticancer and antimicrobial potential of Calotropis procera leaf aqueous extract. Scientific Reports 13(1): 14716.
Ahmadpour, S.A., Mehrabi-Koushki, M. and Farokhinejad, R. 2017. Neodidymelliopsis farokhinejadii, a new fungal species from dead branches of trees in Iran. Sydowia 69: 171–182.
Al-Rowaily, S.L., Abd-ElGawad, A.M., Assaeed, A.M., Elgamal, A.M., Gendy, A.E.N.G.E. et al. 2020. Essential oil of Calotropis procera: Comparative chemical profiles, antimicrobial activity, and allelopathic potential on weeds. Molecules 25(21): 5203.
Aveskamp, M.M., de Gruyter, J. and Crous, P.W. 2008. Biology and recent developments in the systematics of Phoma, a complex genus of major quarantine significance. Fungal Diversity 31: 1–18.
Aveskamp, M.M., de Gruyter, J., Woudenberg, J.H.C., Verkley, G.J.M. and Crous P.W. 2010. Highlights of the Didymellaceae: a polyphasic approach to characterize Phoma and related pleosporalean genera. Studies in Mycology 65: 1–60.
Barreto, R.W., Evans, H.C. and Pomella, A.W. 1999. Fungal pathogens of Calotropis procera (rubber bush), with two new records from Brazil. Australasian Plant Pathology 28: 126–130.
Chen, Q., Hou, L.W., Duan, W.J., Crous, P.W. and Cai, L. 2017. Didymellaceae revisited. Studies in Mycology 87(1): 105–159.
Chen, Q., Jiang, J.R., Zhang, G.Z., Cai, L. and Crous, P.W. 2015. Resolving the Phoma enigma. Studies in Mycology 82: 137–217.
Choi, Y.W., Hyde, K.D. and Ho, W.H. 1999. Single spore isolation of fungi. Fungal Diversity 3: 29–38.
Crous, P.W., Verkley, G.J.M., Groenewald, J.Z. and Samson, R.A. 2009. Fungal biodiversity. CBS Laboratory Manual Series, 1. Westerdijk Fungal Biodiversity Institute, Utrecht, Netherlands.
De Gruyter, J., Aveskamp, M.M., Woudenberg, J.H.C., Verkley, G.J., Groenewald, J.Z. and Crous, P.W. 2009. Molecular phylogeny of Phoma and allied anamorph genera: towards a reclassification of the Phoma complex. Mycological Research 113: 508–519.
Doyle, J.J. and Doyle J.L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19:11-15.
Ershad, D. 2022. Fungi and Fungal Analogues of Iran. Iranian Research Institute of Plant Protection, Tehran, Iran.
Farrar, N., Sadeghi, S.M., Golestaneh, S.R. and Karampour, F. 2020. Values, benefits and use of Calotropis procera in Southern Iran. Iran Nature 5(1): 41–48.
Ghasemi, S., Ghasemi, M., Moradi, N. and Shamili, A. M. 2012. Effect of Calotropis procera leaf extract on seed germination of some plants. Journal of Ornamental and Horticultural Plants 2(1): 27–32
Glass, N.L. and Donaldson, G.C. 1995. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology 61(4): 1323–1330.
Hassan, L.M., Galal, T.M., Farahat, E.A. and El-Midany, M.M. 2015. The biology of Calotropis procera (Aiton) WT. Trees 29: 311–320.
Hou, L.W., Groenewald, J.Z., Pfenning, L.H., Yarden, O., Crous, P.W. and Cai, L. 2020. The phoma-like dilemma. Studies in Mycology 96: 309–396.
Liu, Y.J., Whelen, S. and Hall, B.D. 1999. Phylogenetic relationships among ascomycetes: evidence from an RNA polymerase II subunit. Molecular Biology and Evolution 16: 1799–1808.
Peever, T.L., Canihos, Y., Olsen, L., Ibanez, A., Liu, Y-C. and Timmer, L.W. 1999. Population genetic structure and host specificity of Alternaria spp. causing brown spot of Minneola tangelo and rough lemon in Florida. Phytopathology 89: 851–860.
Posada, D. 2008. jModelTest: phylogenetic model averaging. Molecular biology and evolution 25(7): 1253–1256.
Qian, Z., Zhang, S., Manawasinghe, I.S., Sun, D., Song, J. and Xu, B. 2024. Taxonomic revision of Neodidymelliopsis with N. cynanchi sp. nov., associated with Cynanchum sibiricum leaf spot in Xinjiang, China. New Zealand Journal of Botany 62(2-3): 151–164.
Rahman, M.A. and Wilcock, C.C. 1991. A taxonomic revision of Calotropis (Asclepiadaceae). Nordic Journal of Botany 11: 301–308.
Rambaut, A. 2012. FigTree Version 1.4.0. Available online: http:// tree.bio.ed.ac.uk/software/figtree/ (accessed on 20 February 2024).
Razaghi, P. and Zafari, D. 2018. Characterization of fungi causing lesion blight on Papaver dubium in Iran. Antonie Leeuwenhoek 111: 437–455.
Rehner, S.A. and Samuels, G.J. 1994. Taxonomy and phylogeny of Gliocladium analysed from nuclear large subunit ribosomal DNA sequences. Mycological Research 98: 625–634.
Ronquist, F., Teslenko, M., Van Der Mark, P., Ayres, D.L., Darling, A., et al. 2012. MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology 61: 539–542.
Soleimani, P., Soleimani, M.J. and Hosseini, S. 2018. Phylogenetic relationship and evolution of Neodidymelliopsis isolates collected from Iran. Mycosphere 9(6): 1235–1255.
Sung, G.H., Sung, J.M., Hywel-Jones, N.L. and Spatafora, J. 2007. A multi-gene phylogeny of Clavicipitaceae (Ascomycota, Fungi): identification of localized incongruence using a combinational bootstrap approach. Molecular Phylogenetics and Evolution 44: 1204–1223.
Thompson, J.D., Gibson, T. and Higgins, D.G. 2002. Multiple sequence alignment using ClustalW and ClustalX. Current protocols in bioinformatics 2.3.1–2.3. 22.
Vilgalys, R. and Hester, M. 1990. Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species. Journal of Bacteriology 172: 4238–4246.
White, T.J., Bruns, T., Lee, S., and Taylor, J.W. 1990 Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis PCR Protocols, A Guide to Methods and Applications (M.A. Innis, D.H. Gelfand, J.J. Sninsky & T.J. White eds): 315–322. Academic Press, New York, USA.
Wilkinson, P.M., Thomas-Hall, S., Marney, T.S. and Shivas, R.G. 2005. First record of Passalora calotropidis in Australia and its generic position. Australasian Plant Pathology 34: 95–98.
 
 
Mycologia Iranica
Volume 11, Issue 2
December 2024
Pages 41-49

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History

  • Receive Date: 10 December 2024
  • Revise Date: 17 December 2024
  • Accept Date: 16 December 2024
  • Publish Date: 01 December 2024

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