Molecular Identification of Plenodomus species associated with Brassicaceae plant samples stored in the mycological herbarium LEP

封面

如何引用文章

全文:

开放存取 开放存取
受限制的访问 ##reader.subscriptionAccessGranted##
受限制的访问 订阅存取

详细

Mycological herbaria represent unique and indispensable bioresource collections that play a key role in the study of fungal biodiversity. One of the largest collections of this kind is the herbarium of the All-Russian Institute of Plant Protection (VIZR, LEP), which houses the largest collection of microscopic phytopathogenic fungi in Russia. Over the course of long-term storage, some of the most valuable specimens lose their original properties and become unsuitable for studying by traditional morphological methods. Currently, molecular genetic methods are actively being applied to study herbarium collections, opening new opportunities for the analysis and preservation of biodiversity among phytopathogenic micromycetes. Phoma stem canker and Phoma leaf spot are one of the most widespread and harmfull diseases of cruciferous crops. The causal agents of this disease are closely related fungi, Plenodomus lingam and P. biglobosus. Within these species, two and seven phylogenetic lineages (subclades), respectively, have been identified. Reliable identification of these subclades is only possible using multilocus phylogenetic analysis of nucleotide sequences of the ITS locus and partial regions of actin (act) and β-tubulin (tub2). The aim of this study was to re-identify Plenodomus species in samples of cruciferous plants stored in the Mycological Herbarium LEP for 145–60 years using molecular phylogenetic methods. Nucleotide sequences of the ITS locus were determined in DNA extracted from 16 samples. According to the results of phylogenetic analysis based on ITS sequences, only four samples contained Plenodomus species as the causal agents of Phoma stem canker. In these four samples, in addition to the ITS locus, partial act and tub2 were successfully sequenced. In the phylogenetic tree constructed from nucleotide sequences of all three loci the samples clustered within a clade formed by representative strains of P. lingam subclade ‘brassicae’. Thus, it was reliably confirmed that the causal agent of Phoma stem canker in these four samples was P. lingam ‘brassicae’. Previously in Russia P. lingam ‘brassicae’ has been reliably identified in the Kaliningrad and Leningrad regions, P. biglobosus ‘brassicae’ in the Kaliningrad and Leningrad regions, the Republic of Adygea and Krasnodar Krai. As a result of this study, the presence of the fungus P. lingam ‘brassicae’ was confirmed in herbarium specimens collected in the Leningrad and Rostov regions and in Kazakhstan. To date, the earliest reliable finding of P. lingam in Russia is 114 years old.

全文:

受限制的访问

作者简介

M. Gomzhina

All-Russian Institute of Plant Protection

编辑信件的主要联系方式.
Email: gomzhina91@mail.ru
俄罗斯联邦, St. Petersburg

E. Gasich

All-Russian Institute of Plant Protection

Email: elena_gasich@mail.ru
俄罗斯联邦, St. Petersburg

参考

  1. Dilmaghani A., Balesdent M.H., Didier J. et al. The Leptosphaeria maculans – Leptosphaeria biglobosa species complex in the American continent. Plant Pathol. 2009. V. 58. P. 1044–1058. https://doi.org/10.1111/j.1365-3059.2009.02149.x
  2. Dilmaghani A., Balesdent M.H., Rouxel T. et al. First report of Leptosphaeria biglobosa (blackleg) on Brassica oleracea (cabbage) in Mexico. Plant Dis. 2010. V. 94. P. 791. https://doi.org/10.1094/pdis-94-6-0791c
  3. Doyle J.J., Doyle J.L. Isolation of plant DNA from fresh tissue. Focus. 1990. V. 12. P. 13–15. https://doi.org/10.1007/978-3-642-83962-7_18
  4. Fitt B.D.L., Brun H., Barbetti M.J. et al. World-wide importance of Phoma stem canker (Leptosphaeria maculans and L. biglobosa) on oilseed rape (Brassica napus). Eur. J. Plant Pathol. 2006. V. 114. P. 3–15. https://doi.org/10.1007/s10658-005-2233-5
  5. Fitt B.D.L., Hu B.C., Li Z.Q. et al. Strategies to prevent spread of Leptosphaeria maculans (Phoma stem canker) onto oilseed rape crops in China; costs and benefits. Plant Pathol. 2008. V. 57. P. 652–664. https://doi.org/10.1111/j.1365-3059.2008.01841.x
  6. Gomzhina M.M., Gannibal Ph.B. Phoma-like fungi in the Mycological Herbarium of All-Russian institute of plant protection (LEP). Mikologiya i fitopatologiya. 2020. V. 54 (6). P. 452–459. https://doi.org/10.31857/S0026364820060070
  7. Gomzhina M.M., Gasich E.L. Plenodomus species infecting oilseed rape in Russia. Plant Protection News. 2022. V. 105(3). P. 135–147. https://doi.org/10.31993/2308-6459-2022-105-3-15425
  8. Henderson M.P. The black-leg disease of cabbage caused by Phoma lingam (Tode) Desmaz. Phytopathology. 1918. V. 8. P. 379–431.
  9. King K.M., West J.S. Detection of the Phoma pathogens Plenodomus biglobosus subclades ‘brassicae’ and ‘canadensis’ on wasabi, and ‘canadensis’ in Europe. Eur. J. Plant Pathol. 2022. V. 162. P. 751–756. https://doi.org/10.1007/s10658-021-02428-z
  10. Kumar S., Stecher G., Li M. et al. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. 2018. V. 35. P. 1547–1549. https://doi.org/10.1093/molbev/msy096
  11. Liu Z., Latunde-Dada A.O., Hall A.M. et al. Phoma stem canker disease on oilseed rape (Brassica napus) in China is caused by Leptosphaeria biglobosa ‘brassicae.’ Eur. J. Plant Pathol. 2014. V. 140. P. 841–857. https://doi.org/10.1007/s10658-014-0513-7
  12. Lord E., Leclercq M., Boc A. et al. Armadillo 1.1: An original workflow platform for designing and conducting phylogenetic analysis and simulations. PLOS One. 2012. V. 7 (1). P. e29903. https://doi.org/10.1371/journal.pone.0029903
  13. Luo T., Li G., Yang L. First report of Leptosphaeria biglobosa ‘canadensis’ causing blackleg on oilseed rape (Brassica napus) in China. Plant Dis. 2021. V. 105 (11). P. 3760. https://doi.org/10.1094/PDIS-12-20-2735-PDN
  14. Mendes-Pereira E., Balesdent M.H., Brun H. et al. Molecular phylogeny of the Leptosphaeria maculans – L. biglobosa species complex. Mycol Res. 2003. V. 107. P. 1287–1304. https://doi.org/10.1017/S0953756203008554
  15. Minh B.Q., Schmidt H.A., Chernomor O. et al. IQ-TREE2: New models and efficient methods for phylogenetic inference in the genomic era. Mol. Biol. Evol. 2020. V. 35 (7). P. 1530–1534. https://doi.org/10.1093/molbev/msaa015
  16. Plummer K.M., Dunse K., Howlett B.J. Non-aggressive strains of the blackleg fungus, Leptosphaeria maculans, are present in Australia and can be distinguished from aggressive strains by molecular analysis. Aust J Bot. 1994. V. 42. P. 1–8. https://doi.org/10.1071/BT9940001
  17. Thompson J.D., Gibson T.J., Plewniak F. et al. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997. V. 24. P. 4876–4882. https://doi.org/10.1093/nar/25.24.4876
  18. van de Wouw A.P., Thomas V.L., Cozijnsen A.J. et al. Identification of Leptosphaeria biglobosa ‘canadensis’ on Brassica juncea stubble from northern New South Wales, Australia. Australas. Plant Dis. Notes. 2008. V. 3. P. 124–128. https://doi.org/10.1007/BF03211265
  19. Vincenot L., Balesdent M.H., Li H. et al. Occurrence of a new subclade of Leptosphaeria biglobosa in Western Australia. Phytopathol. 2008. V. 98. P. 321–329. https://doi.org/10.1094/PHYTO-98-3-0321
  20. Voigt K., Cozijnsen A.J., Kroymann J. et al. Phylogenetic relationships between members of the crucifer pathogenic Leptosphaeria maculans species complex as shown by mating type (MAT1–2), actin, and β-tubulin sequences. Mol Phylogenet Evol. 2005. V. 37 (2). P. 541–557. https://doi.org/10.1016/j.ympev.2005.07.006
  21. West J.S., Kharbanda P.D., Barbetti M.J. Epidemiology and management of Leptosphaeria maculans (Phoma stem canker) on oilseed rape in Australia, Canada and Europe. Plant Pathol. 2001. V. 50. P. 10–27. https://doi.org/10.1046/j.1365-3059.2001.00546.x
  22. Zou Z., Zhang X., Parks P. et al. A new subclade of Leptosphaeria biglobosa identified from Brassica rapa. Int. J. Mol. Sci. 2019. V. 20 (7). P. 1668. https://doi.org/10.3390/ijms20071668
  23. Гомжина М.М., Ганнибал Ф.Б. (Gomzhina, Gannibal) Фомоидные грибы на растениях семейства Asteraceae в Микологическом Гербарии ВИЗР LEP // Микология и фитопатология. 2020. Т. 54. № 6. С. 452–459.

补充文件

附件文件
动作
1. JATS XML
下载
2. Fig. 1. Sample LEP 129983.

下载 (193KB)
索引源数据
3. Fig. 2. Sample LEP 129979.

下载 (223KB)
索引源数据
4. Fig. 3. Sample LEP 130090.

下载 (287KB)
索引源数据
5. Fig. 4. Sample LEP 129332.

下载 (295KB)
索引源数据
6. Fig. 5. Combined phylogenetic tree of Plenodomus species and subclades constructed by ML based on ITS, act, and tub2 sequences. Bootstrap support values ​​obtained by ML (≥70), MP (≥70), and Bayesian statistics (≥0.7) are given at the nodes of the phylogram branches, respectively. The sequences of Leptosphaeria doliolum strain CBS130000 are taken as the outgroup. The numbers of the studied samples are highlighted in blue.

下载 (242KB)
索引源数据

版权所有 © Russian Academy of Sciences, 2025