RESEARCH

Exploring the Biological and Molecular Characteristics of Resistance to Fludioxonil in Sclerotinia sclerotiorum From Soybean in China

Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang 453003, China

Henan Engineering Research Center of Biological Pesticide and Fertilizer Development and Synergistic Application, Henan Institute of Science and Technology, Xinxiang 453003, China

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H. Y. Hu

Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang 453003, China

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D. X. Li

Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang 453003, China

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L. G. Tan

Henan Engineering Research Center of Biological Pesticide and Fertilizer Development and Synergistic Application, Henan Institute of Science and Technology, Xinxiang 453003, China

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Q. Zhang

Henan Engineering Research Center of Biological Pesticide and Fertilizer Development and Synergistic Application, Henan Institute of Science and Technology, Xinxiang 453003, China

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H. T. Gao

Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang 453003, China

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H. L. Sun

Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang 453003, China

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X. L. Tian

Henan Engineering Research Center of Biological Pesticide and Fertilizer Development and Synergistic Application, Henan Institute of Science and Technology, Xinxiang 453003, China

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M. W. Shi

Henan Engineering Research Center of Biological Pesticide and Fertilizer Development and Synergistic Application, Henan Institute of Science and Technology, Xinxiang 453003, China

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F. L. Zhang

Henan Engineering Research Center of Biological Pesticide and Fertilizer Development and Synergistic Application, Henan Institute of Science and Technology, Xinxiang 453003, China

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, and

C. W. Li

^†Corresponding author: C. W. Li;

E-mail Address: lichengweiwau@hotmail.com

Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang 453003, China

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Affiliations

Authors and Affiliations

F. Zhou¹²
H. Y. Hu¹
D. X. Li¹
L. G. Tan²
Q. Zhang²
H. T. Gao¹
H. L. Sun¹
X. L. Tian²
M. W. Shi²
F. L. Zhang²
C. W. Li¹^†

¹Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang 453003, China
²Henan Engineering Research Center of Biological Pesticide and Fertilizer Development and Synergistic Application, Henan Institute of Science and Technology, Xinxiang 453003, China

Published Online:12 May 2021https://doi.org/10.1094/PDIS-07-20-1621-RE

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Abstract

Sclerotinia sclerotiorum is one of the most damaging and economically important necrotrophic plant pathogens, infecting more than 400 plant species globally. Although the phenylpyrrole fungicide fludioxonil has high activity against S. sclerotiorum, reports indicate that there is also substantial potential for the development of fungicide resistance. However, the current study investigating five fludioxonil-resistant laboratory mutants found a significant fitness cost associated with fludioxonil resistance resulting in significantly (P < 0.05) reduced mycelial growth and sclerotia formation on potato dextrose agar as well as significantly (P < 0.05) lower pathogenicity on detached tomato leaves, with one mutant, LK-1R, completely losing the capacity to cause infection. In addition, all of the fludioxonil-resistant mutants had significantly (P < 0.05) increased sensitivity to osmotic stress (0.5 M of potassium chloride and 1.0 M of glucose), which is consistent with the proposed fludioxonil target sites within the high osmolarity glycerol stress response mitogen-activated protein kinase (HOG1-MAPK) signaling transduction pathway. Sequence analysis of six genes from this two-component pathway, including SsHk, SsYpd, SsSk1, SsSk2, SsPbs, and SsHog, revealed several mutations that may be associated with fludioxonil resistance. For example, six separate point mutations were found in SsHk that led to changes in the predicted amino acid sequence, including A136G, F249V, G353A, E560K, M610K, and K727R. Similarly, SsPbs had three mutations (D34G, S46L, and L337E), SsSk1 and SsYpd had two (S53G and A795V for SsSk1, and E67G and Y141H for SsYpd), and SsHog and SsSk2 had one each (V220A and S763P, respectively). To our knowledge, these constitute the first reports of amino acid changes in proteins of the HOG1-MAPK pathway being associated with fludioxonil resistance in S. sclerotiorum. This study also showed a positive cross-resistance between fludioxonil and dimethachlone and procymidone, but none with tebuconazole or carbendazim, indicating that the inclusion of tebuconazole within an integrated pest management program could reduce the risk of fludioxonil resistance developing in field populations of S. sclerotiorum and ensure the sustainable production of soybeans in China into the future.

Literature Cited

Amiri, A., Heath, S. M., and Peres, N. A. 2013. Phenotypic characterization of multi-fungicide resistance in Botrytis cinerea isolates from strawberry fields in Florida. Plant Dis. 97:393-401. https://doi.org/10.1094/PDIS-08-12-0748-RE Link, ISI, Google Scholar
Avenot, H. F., and Michailides, T. J. 2015. Detection of isolates of Alternaria alternata with multiple-resistance to fludioxonil, cyprodinil, boscalid and pyraclostrobin in California pistachio orchards. Crop Prot. 78:214-221. https://doi.org/10.1016/j.cropro.2015.09.012 Crossref, ISI, Google Scholar
Bardin, S. D., and Huang, H. C. 2001. Research on biology and control of Sclerotinia diseases in Canada. Can. J. Plant Pathol. 23:88-98. https://doi.org/10.1080/07060660109506914 Crossref, ISI, Google Scholar
Bastien, M., Sonah, H., and Belzile, F. 2014. Genome wide association mapping of Sclerotinia sclerotiorum resistance in soybean with a genotyping-by-sequencing approach. Plant Genome 7:plantgenome2013.10.0030. https://doi.org/10.3835/plantgenome2013.10.0030 Crossref, ISI, Google Scholar
Boland, G. J., and Hall, R. 1994. Index of plant hosts of Sclerotinia sclerotiorum. Can. J. Plant Pathol. 16:93-108. https://doi.org/10.1080/07060669409500766 Crossref, ISI, Google Scholar
Bolton, D. M., Thomma, B. P., and Nelson, B. D. 2006. Sclerotinia sclerotiorum (lib.) de Bray: Biology and molecular traits of a cosmopolitan pathogen. Mol. Plant Pathol. 7:1-16. https://doi.org/10.1111/j.1364-3703.2005.00316.x Crossref, ISI, Google Scholar
Brandhorst, T. T., and Klein, B. S. 2019. Uncertainty surrounding the mechanism and safety of the post-harvest fungicide fludioxonil. Food Chem. Toxicol. 123:561-565. https://doi.org/10.1016/j.fct.2018.11.037 Crossref, ISI, Google Scholar
Cunha, W. G., Tinoco, M. L. P., Pancoti, H. L., Ribeiro, R. E., and Aragão, F. J. L. 2010. High resistance to Sclerotinia sclerotiorum in transgenic soybean plants transformed to express an oxalate decarboxylase gene. Plant Pathol. 59:654-660. https://doi.org/10.1111/j.1365-3059.2010.02279.x Crossref, ISI, Google Scholar
De Miccolis Angelini, R. M., Rotolo, C., and Masiello, M. 2014. Occurrence of fungicide resistance in populations of Botryotinia fuckeliana (Botrytis cinerea) on table grape and strawberry in southern Italy. Pest Manag. Sci. 70:1785-1796. https://doi.org/10.1002/ps.3711 Crossref, ISI, Google Scholar
Dry, I. B., Yuan, K. H., and Hutton, D. G. 2004. Dicarboximide resistance in field isolates of Alternaria alternata is mediated by a mutation in a two-component histidine kinase gene. Fungal Genet. Biol. 41:102-108. https://doi.org/10.1016/j.fgb.2003.09.002 Crossref, ISI, Google Scholar
Duan, Y. B., Ge, C. Y., and Zhou, M. G. 2014. Molecular and biochemical characterization of Sclerotinia sclerotiorum laboratory mutants resistant to dicarboximide and phenylpyrrole fungicides. J. Pest Sci. 87:221-230. https://doi.org/10.1007/s10340-013-0526-6 Crossref, ISI, Google Scholar
Firoz, M. J., Xiao, X., Zhu, F. X., Fu, Y. P., Jiang, D. H., Schnabel, G., and Luo, C. X. 2016. Exploring mechanisms of resistance to dimethachlone in Sclerotinia sclerotiorum. Pest Manag. Sci. 72:770-779. https://doi.org/10.1002/ps.4051 Crossref, ISI, Google Scholar
Gong, C. W., Qin, Y. M., Qu, J. S., and Wang, X. G. 2018. Resistance detection and mechanism of strawberry Botrytis cinerea to fludioxonil in Sichuan Province. Sci. Agric. Sin. 51:4277-4287. Google Scholar
Grabke, A., Fernández-Ortuńo, D., and Schnabel, G. 2013. Fenhexamid resistance in Botrytis cinerea from strawberry fields in the Carolinas is associated with four target gene mutations. Plant Dis. 97:271-276. https://doi.org/10.1094/PDIS-06-12-0587-RE Link, ISI, Google Scholar
Han, X., Zhao, H., Ren, W. C., Lv, C. Y., and Chen, C. J. 2017. Resistance risk assessment for fludioxonil in Bipolaris maydis. Pestic. Biochem. Physiol. 139:32-39. https://doi.org/10.1016/j.pestbp.2017.04.006 Crossref, ISI, Google Scholar
Kuang, J., Wang, J. X., and Zhou, M. G. 2011. Monitoring carbendazim and dimethachlon resistance of Sclerotinia sclerotiorum obtained from the blight stems of rape in Jiangsu Province. Chin. Agric. Sci. Bull. 27:285-291. Google Scholar
Letham, D. B., Huett, D. O., and Trimboli, D. S. 1976. Biology and control of Sclerotinia sclerotiorum in cauliflower and tomato crops in coastal New South Wales. Plant Dis. 60:286-289. ISI, Google Scholar
Lew, R. R. 2010. Turgor and net ion flux responses to activation of the osmotic MAP kinase cascade by fludioxonil in the filamentous fungus Neurospora crassa. Fungal Genet. Biol. 47:721-726. https://doi.org/10.1016/j.fgb.2010.05.007 Crossref, ISI, Google Scholar
Li, H. X., and Xiao, C. L. 2008. Characterization of fludioxonil-resistant and pyrimethanil-resistant phenotypes of Penicillium expansum from apple. Phytopathology 98:427-435. https://doi.org/10.1094/PHYTO-98-4-0427 Link, ISI, Google Scholar
Li, J. L., Kang, T. H., Talab, K., Zhu, F. X., and Li, J. H. 2017. Molecular and biochemical characterization of dimethachlone resistant isolates of Sclerotinia sclerotiorum. Pestic. Biochem. Physiol. 138:15-21. https://doi.org/10.1016/j.pestbp.2017.02.001 Crossref, ISI, Google Scholar
McCaghey, M., Willbur, J., Ranjan, A., Grau, C. R., Chapman, S., Diers, B., Groves, C., Kabbage, M., and Smith, D. L. 2017. Development and evaluation of Glycine max germplasm lines with quantitative resistance to Sclerotinia sclerotiorum. Front. Plant Sci. 8:1495. https://doi.org/10.3389/fpls.2017.01495 Crossref, ISI, Google Scholar
Peters, R. D., Platt, H. W., Drake, K. A., and Coffin, R. H. 2008. First report of fludioxonil-resistant isolates of Fusarium spp. causing potato seed-piece decay. Plant Dis. 92:172. https://doi.org/10.1094/PDIS-92-1-0172A Link, ISI, Google Scholar
Qiu, J. B., Shi, J. R., Xu, J. H., Yu, M. Z., and Qian, Y. 2018. Molecular and characterization, fitness and mycotoxin production of Fusarium asiaticum strains resistant to fludioxonil. Plant Dis. 102:1759-1765. https://doi.org/10.1094/PDIS-11-17-1772-RE Link, ISI, Google Scholar
Ranjan, A., Westrick, N. M., Jain, S., Piotrowski, J. S., Ranjan, M., Kessens, R., Stiegman, L., Grau, C. R., Conley, S. P., Smith, D. L., and Kabbage, M. 2019. Resistance against Sclerotinia sclerotiorum in soybean involves a reprogramming of the phenylpropanoid pathway and up-regulation of antifungal activity targeting ergosterol biosynthesis. Plant Biotechnol. J. 17:1567-1581. https://doi.org/10.1111/pbi.13082 Crossref, ISI, Google Scholar
Ren, W. C., Shao, W. Y., Han, X., Zhou, M., and Chen, C. J. 2016. Molecular and biochemical characterization of laboratory and field mutants of Botrytis cinerea resistant to fludioxonil. Plant Dis. 100:1414-1423. https://doi.org/10.1094/PDIS-11-15-1290-RE Link, ISI, Google Scholar
Ribeiro, M., Chiu, M., Lopes, T., Grimaldi, R., Marsaioli, A., and Goncalves, L. 2017. Synthesis of structured lipids containing behenic acid from fully hydrogenated Crambe abyssinica oil by enzymatic interesterification. J. Food Sci. Technol. 54:1146-1157. https://doi.org/10.1007/s13197-017-2540-9 Crossref, ISI, Google Scholar
Rosslenbroich, H. J., and Stuebler, D. 2000. Botrytis cinerea—History of chemical control and novel fungicides for its management. Crop Prot. 19:557-561. https://doi.org/10.1016/S0261-2194(00)00072-7 Crossref, ISI, Google Scholar
Sang, C. W., Ren, W. C., Wang, J. J., Xu, H., Zhang, Z. H., Zhou, M. G., Chen, C. J., and Wang, K. 2018. Detection and fitness comparison of target-based highly fludioxonil-resistant isolates of Botrytis cinerea from strawberry and cucumber in China. Pestic. Biochem. Physiol. 147:110-118. https://doi.org/10.1016/j.pestbp.2018.01.012 Crossref, ISI, Google Scholar
Tückmantel, S., Greul, J. N., Janning, P., Brockmeyer, A., Grütter, C., Simard, J. R., Gutbrod, O., Beck, M. E., Tietjen, K., Rauh, D., and Schreier, P. H. 2011. Identification of Ustilago maydis Aurora kinase as a novel antifungal target. ACS Chem. Biol. 6:926-933. https://doi.org/10.1021/cb200112y Crossref, ISI, Google Scholar
Wu, D. X., Zhang, R. S., Han, X., Wang, J. X., Zhou, M. G., and Chen, C. J. 2015. Resistance risk assessment for fludioxonil in Stemphylium solani. Ann. Appl. Biol. 167:277-284. https://doi.org/10.1111/aab.12230 Crossref, ISI, Google Scholar
Yoshimi, A., Kojima, K., Takano, Y., and Tanaka, C. 2005. Group III histidine kinase is a positive regulator of Hog1-type mitogen-activated protein kinase in filamentous fungi. Eukaryot. Cell 4:1820-1828. https://doi.org/10.1128/EC.4.11.1820-1828.2005 Crossref, ISI, Google Scholar
Zhang, X., and Ma, X. M. 2014. Advances of mutation sites associated with fungicide resistance in Botrytis cinerea and its molecular detection methods. Curr. Biotechnol. 4:251-257. Google Scholar
Zhao, D., Xu, Y. L., and Li, C. J. 2006. Identification and integrated managements for soybean Sclerotinia sclerotiorum. Chin. J. Soybean Bull. 3:15-16. Google Scholar
Zhao, X., Han, Y. P., Li, Y. H., Liu, D. Y., Sun, M. M., Zhao, Y., Lv, C. M., Li, D. M., Yang, Z. J., Huang, L., Teng, W. L., Qiu, L. J., Zheng, H. K., and Li, W. B. 2015. Loci and candidate gene identification for resistance to Sclerotinia sclerotiorum in soybean (Glycine max L. Merr.) via association and linkage maps. Plant J. 82:245-255. https://doi.org/10.1111/tpj.12810 Crossref, ISI, Google Scholar
Zhou, F., Hu, H. Y., Song, Y. L., Gao, Y. Q., Liu, Q. L., Song, P. W., Chen, E. Y., Yu, Y. A., Li, D. X., and Li, C. W. 2020a. Biological characteristics and molecular mechanism of fludioxonil resistance in Botrytis cinerea from Henan Province of China. Plant Dis. 104:1041-1047. https://doi.org/10.1094/PDIS-08-19-1722-RE Link, ISI, Google Scholar
Zhou, F., Li, D. X., Hu, H. Y., Song, Y. L., Fan, Y. C., Guan, Y. Y., Song, P. W., Wei, Q. C., Yan, H. F., and Li, C. W. 2020b. Biological characteristics and molecular mechanisms of fludioxonil resistance in Fusarium graminearum in China. Plant Dis. 104:2426-2433. https://doi.org/10.1094/PDIS-01-20-0079-RE Link, ISI, Google Scholar
Zhou, F., Lu, F., Zhang, C., Qi, H. X., Wang, X. D., and Zhang, G. Z. 2017. Occurrence of fenhexamid resistance in Botrytis cinerea from greenhouse strawberries in China. J. Phytopathol. 165:455-462. https://doi.org/10.1111/jph.12580 Crossref, ISI, Google Scholar
Zhou, F., Zhang, X. L., Li, J. L., and Zhu, F. X. 2014. Dimethachlon resistance in Sclerotinia sclerotiorum in China. Plant Dis. 98:1221-1226. https://doi.org/10.1094/PDIS-10-13-1072-RE Link, ISI, Google Scholar

Vol. 105, No. 7
July 2021

ISSN:0191-2917
e-ISSN:1943-7692

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Caption

Leaf spot symptoms on Zizyphus mauritiana in the field caused by Colletotrichum spp. (J. Shu et al.). Photo credit: Q. Li. Symptoms of twig dieback of tea plants caused by Fusarium solani (A. K. Pandey et al.). Photo credit: A. K. Pandey.

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Article History

Issue Date: 25 Oct 2021
Published: 12 May 2021
First Look: 12 Oct 2020
Accepted: 8 Oct 2020

Pages: 1936-1941

Information

Funding

Key Scientific and Technological Research Project of Henan Province
Grant/Award Number: 192102110056
Grant/Award Number: 192102110030

First-Level Key Discipline Project of Plant Protection in Henan Province
Grant/Award Number: 107020219001/005

Keywords

The author(s) declare no conflict of interest.