Melanin, DNA replication, and autophagy affect appressorium development in Setosphaeria turcica by regulating glycerol accumulation and metabolism

doi:10.1016/S2095-3119(21)63679-1

Journal of Integrative Agriculture ›› 2022, Vol. 21 ›› Issue (3): 762-773.DOI: 10.1016/S2095-3119(21)63679-1

收稿日期:2021-01-06 接受日期:2021-03-11 出版日期:2022-03-01 发布日期:2021-03-11

Melanin, DNA replication, and autophagy affect appressorium development in Setosphaeria turcica by regulating glycerol accumulation and metabolism

GUO Xiao-yue*, LIU Ning*, LIU Bing-hui, ZHOU Li-hong, CAO Zhi-yan, HAN Jian-min, DONG Jin-gao

State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology/Mycotoxin and Molecular Plant Pathology Laboratory, Hebei Agricultural University, Baoding 071001, P.R.China

Received:2021-01-06 Accepted:2021-03-11 Online:2022-03-01 Published:2021-03-11
About author:GUO Xiao-yue, E-mail: 981320184@qq.com; Correspondence CAO Zhi-yan, Tel: +86-312-7528142, E-mail: caoyan208@126.com; HAN Jian-min, Tel: +86-312-7528876, E-mail: hanjmnd@163.com * These authors contributed equally to this study.
Supported by:
This work was supported by the grants from the National Natural Science Foundation of China (32072370 and 31901827), the China Agriculture Research System of MOF and MARA (CARS-02-25), the Natural Science Foundation of Hebei Province, China (C2020204039 and C2018204059), and the Projects of Overseas Foundation, Hebei Province, China (C20190508).

摘要/Abstract

摘要：

玉米大斑病是世界玉米产区主要的真菌病害之一，引起该病害的真菌为玉米大斑病菌（Setosphaeria turcica），为异宗配合真菌，其无性态为玉米大斑凸脐孺孢（Exserohilum turcicum）。玉米大斑病菌分生孢子萌发后形成高度特化的侵染结构—黑化的附着胞。附着胞通过产生高膨压来穿透植物表皮，其中甘油是产生膨压的主要来源。本研究分析了附着胞侵染玉米叶片的位置，发现大多数萌发的分生孢子通过直接穿透表皮细胞进入叶片内部，无论是通过表皮细胞还是气孔侵染玉米叶片，附着胞都是侵染所必需的。为了进一步确定影响附着胞发育的关键因素，我们分析了黑色素抑制剂（三环唑，TCZ）、DNA复制抑制剂（羟基脲，HU）和自噬抑制剂（3-甲基腺嘌呤，3-MA）等对附着胞膨压和甘油含量的影响。结果表明，在对照和抑制剂处理下，附着胞膨压和甘油浓度在附着胞成熟阶段均达到最高水平，三种抑制剂在附着胞成熟阶段对附着胞膨压的影响最大，糖原和脂质体是产生甘油的主要物质。研究发现，在附着胞的发育过程中，抑制剂会影响糖原和脂质体在分生孢子、芽管和附着胞中的分布。黑色素、DNA复制和自噬通过调节甘油积累和代谢影响玉米大斑病菌附着胞的发育，该研究可为附着胞膨压和甘油含量之间的关系提供新的见解。

Abstract: Setosphaeria turcica (syn. Exserohilum turcicum) is the pathogenic fungus of maize (Zea mays) that causes northern leaf blight, which is a major maize disease worldwide. Melanized appressoria are highly specialized infection structures formed by germinated conidia of S. turcica that infect maize leaves. The appressorium penetrates the plant cuticle by generating turgor, and glycerol is known to be the main source of the turgor. Here, the infection position penetrated by the appressorium on maize leaves was investigated, most of the germinated conidia entered the leaf interior by directly penetrating the epidermal cells, and the appressorium structure was necessary for the infection, whether it occurred through epidermal cells or stomata. Then, to investigate the effects of key factors in the development of the appressorium, we studied the effects of three inhibitors, including a melanin inhibitor (tricyclazole, TCZ), a DNA replication inhibitor (hydroxyurea, HU), and an autophagy inhibitor (3-methyladenine, 3-MA), on appressorium turgor and glycerol content. As results, appressorium turgor pressure and glycerol concentration in the appressorium reached their highest levels at the mature stage of the appressorium under the control and inhibitor treatments. The three inhibitors had the greatest effects on appressorium turgor pressure at this stage. Glycogen and liposomes are the main substances producing glycerol. It was also found inhibitors affected the distribution of glycogen and liposomes, which were detected in the conidia, the germ tube, and the appressorium during appressorium development. This study provides profound insight into the relationship between appressorium turgor pressure and glycerol content, which was affected by the synthesis of melanin, DNA replication, and autophagy in the developing appressorium during a S. turcica infection.

Key words: Melanin , DNA replication , autophagy , appressorium , development , Setosphaeria turcica , glycerol

. [J]. Journal of Integrative Agriculture, 2022, 21(3): 762-773.

GUO Xiao-yue, LIU Ning, LIU Bing-hui, ZHOU Li-hong, CAO Zhi-yan, HAN Jian-min, DONG Jin-gao . Melanin, DNA replication, and autophagy affect appressorium development in Setosphaeria turcica by regulating glycerol accumulation and metabolism[J]. Journal of Integrative Agriculture, 2022, 21(3): 762-773.

参考文献

Bourett T M, Howard R J. 1990. In vitro development of penetration structures in the rice blast fungus M. oryzae. Canadian Journal of Botany, 68, 329–342.
Butler M J, Day A W. 1998. Fungal melanins: A review. Canadian Journal of Microbiology, 44, 1115–1136.
Cao Z Y, Dong J G, Yong S Y, Yao X X. 2007. Characteristics and spectral analysis of melanin in Setosphaeria turcica. Acta Phytopathologica Sinica, 37, 410–417. (in Chinese)
Cao Z Y, Jia H, Zhu X M, Dong J G. 2011. Relationship between DHN melanin and formation of appressorium turgor pressure of Setosphaeria turcica. Scientia Agricultura Sinica, 44, 925–932. (in Chinese)
Dean R A. 1997. Signal pathways and appressorium morphogenesis. Annual Review of Phytopathology, 35, 211–234.
Ebbole D J. 2007. Magnaporthe as a model for understanding host-pathogen interactions. Annual Review of Phytopathology, 45, 437–456.
Eliahu N, Igbaria A, Rose M S, Horwitz B A, Lev S. 2007. Melanin biosynthesis in the maize pathogen Cochliobolus heterostrophus depends on two mitogen-activated protein kinases, Chk1 and Mps1, and the transcription factor Cmr1. Eukaryotic Cell, 6, 421–429.
Foster A J, Ryder L S, Kershaw M J, Talbot N J. 2017. The role of glycerol in the pathogenic lifestyle of the rice blast fungus Magnaporthe oryzae. Environmental Microbiology, 19, 1008–1016.
Gessler N N, Egorova A S, Belozerskaya T A. 2014. Melanin pigments of fungi under extreme environmental conditions (review). Applied Biochemistry and Microbiology, 50, 105–113.
Gu S Q, Li P, Wu M, Hao Z M, Gong X D, Zhang X Y, Tian L, Zhang P, Wang Y, Cao Z Y, Fan Y S, Han J M, Dong J G. 2014. StSTE12 is required for the pathogenicity of Setosphaeria turcica by regulating appressorium development and penetration. Microbiological Research, 169, 817–23.
Henson J M, Butler M J, Day A W. 1999. The dark side of the mycelium: Melanins of phytopathogenic fungi. Annual Review of Phytopathology, 37, 447–471.
Howard R J, Ferrari M A, Roach D H, Money N P. 1991. Penetration of hard substrates by a fungus employing enormous turgors. Proceedings of the National Academy of Sciences of the United States of America, 88, 11281–11284.
Howard R J, Valent B. 1996. Host penetration by the fungal rice blast pathogen M. oryzae. Annual Review of Microbiology, 50, 491–512.
Jong J C D, Mccormack B J, Smirnoff N, Talbot N J. 1997. Glycerol generates turgor in rice blast. Nature, 389, 244–244.
Kimura A. 2001. Peroxisomal metabolic function is required for appressorium-mediated plant infection by Colletotrichum lagenarium. Plant Cell, 13, 1945–1957.
Klionsky D J. 2010. The molecular machinery of autophagy and its role in physiology and disease. Seminars in Cell and Developmental Biology, 21, 663–663.
Koopman R, Schaart G, Hesselink M K. 2001. Optimisation of oil red o staining permits combination with immunofluorescence and automated quantification of lipids. Histochemistry and Cell Biology, 116, 63–68.
Levine B, Klionsky D J. 2004. Development by self-digestion: molecular mechanisms and biological functions of autophagy. Developmental Cell, 6, 463–477.
Liu N, Shen S, Jia H, Yang B B, Guo X Y, Si H L, Cao Z Y, Dong J G. 2019. Heterologous expression of Stlac2, a laccase isozyme of Setosphearia turcica, and the ability of decolorization of malachite green. International Journal of Biological Macromolecules, 138, 21–28.
Liu X H, Lin F C. 2008. Investigation of the biological roles of autophagy in appressorium morphogenesis in Magnaporthe oryzae. Journal of Zhejiang University Science (B), 9, 793–799.
Liu X H, Lu J P, Zhang L, Dong B, Min H, Lin F C. 2007. Involvement of a Magnaporthe oryzae serine/threonine kinase gene, MgATG1, in appressorium turgor and pathogenesis. Eukaryotic Cell, 6, 997–1005.
Lu J P, Liu X H, Feng X X, Min H, Lin F C. 2009. An autophagy gene, MgATG5, is required for cell differentiation and pathogenesis in Magnaporthe oryzae. Current Genetics, 55, 461–473.
Ma S X, Cao K K, Liu N, Meng C, Cao Z Y, Dai D Q, Jia H, Zang J P, Li Z Y, Hao Z M, Gu S Q, Dong J G. 2017. The StLAC2 gene is required for cell wall integrity, DHN-melanin synthesis and the pathogenicity of Setosphaeria turcica. Fungal Biology, 121, 589–601.
Money N P. 1989. Osmotic pressure of aqueous polyethylene glycols: relationship between molecular weight and vapor pressure deficit. Plant Physiology, 91, 766–769.
Nadal M, Gold S E. 2010. The autophagy genes atg8 and atg1 affect morphogenesis and pathogenicity in Ustilago maydis. Molecular Plant Pathology, 11, 463–478.
Nosanchuk J D, Casadevall A. 2006. Impact of melanin on microbial virulence and clinical resistance to antimicrobial compounds. Antimicrobial Agents Chemother, 50, 3519–3528.
Oh Y, Donofrio N, Pan H, Coughlan S, Dean R A. 2008. Transcriptome analysis reveals new insight into appressorium formation and function in the rice blast fungus Magnaporthe oryzae. Genome Biology, 9, 980–986.
Osés-Ruiz M, Sakulkoo W, Littlejohn G R, Martin-Urdiroz M, Talbot N J. 2017. Two independent S-phase checkpoints regulate appressorium-mediated plant infection by the rice blast fungus Magnaporthe oryzae. Proceedings of the National Academy of Sciences of the United States of America, 114, E237–E244.
Peng C, Chen H L, Zhang Y Q, Guo S W. 2011. A review on appressorium initiation and development in Magnaporthe oryzae. Plant Physiology and Biochemistry, 48, 35–44.
Ryder L S, Talbot N J. 2015. Regulation of appressorium development in pathogenic fungi. Current Opinion in Plant Biology, 26, 8–13.
Saunders D G, Aves S J, Talbot N J. 2010. Cell cycle-mediated regulation of plant infection by the rice blast fungus. Plant Cell, 22, 497–507.
Shen S, Hao Z M, Gu S Q, Wang J J, Cao Z Y, Li Z Y, Wang Q, Li P, Hao J, Dong J G. 2013. The catalytic subunit of cAMP-dependent protein kinase a StPKA-c contributes to conidiation and early invasion in the phytopathogenic fungus Setosphaeria turcica. FEMS Microbiology Letters, 343, 135–44.
Talbot N J, Kershaw M J. 2009. The emerging role of autophagy in plant pathogen attack and host defence. Current Opinion in Plant Biology, 12, 444–450.
Thines E, Talbot W N J. 2000. Map kinase and protein kinase A-dependent mobilization of triacylglycerol and glycogen during appressorium turgor generation by Magnaporthe grisea. Plant Cell, 12, 1703–1718.
Tucker S L, Talbot N J. 2001. Surface attachment and pre-penetration stage development by plant pathogenic fungi. Annual Review of Phytopathology, 39, 385–417.
Veneault-Fourrey C, Barooah M, Egan M, Wakley G, Talbot N J. 2006. Autophagic fungal cell death is necessary for infection by the rice blast fungus. Science, 312, 580–583.
Wang M J, Li P, Wu M, Fan Y S, Gu S Q, Dong J G. 2012. Effect of hyperosmotic stress on the growth, development and STK1 expression of Setosphaeria turcica. Scientia Agricultura Sinica, 45, 3965–3970. (in Chinese)
Weber R W S, Pitt D, Webster J. 1998. Teaching techniques for mycology: 3. Amylase secretion by Aspergillus. The Mycologist, 12, 8–9.
Yang J, Kong L, Chen X, Wang D, Qi L, Zhao W, Zhang Y, Liu X, Peng Y L. 2012. A carnitine–acylcarnitine carrier protein, MoCrc1, is essential for pathogenicity in Magnaporthe oryzae. Current Genetics, 58, 139–148.
Zhang C Q, Zhu G N, Ma Z H, Zhou M G. 2006. Isolation, characterization and preliminary genetic analysis of laboratory tricyclazole-resistant mutants of the rice blast fungus, M. oryzae. Journal of Phytopathology, 154, 392–397.
Zhang S R, Hao Z M, Wang L H, Shen S, Cao Z Y, Xin Y Y, Hou M L, Gu S Q, Han J M, Dong J G. 2012. StRas2 regulates morphogenesis, conidiation and appressorium development in Setosphaeria turcica. Microbiological Research, 167, 478–486.
Zeng F L, Meng Y N, Hao Z M, Li P, Zhai W B, Shen S, Cao Z Y, Dong J G. 2020. Setosphaeria turcica ATR turns off appressorium-mediated maize infection and triggers melanin-involved self-protection in response to genotoxic stress. Molecular Plant Pathology, 21, 401–414.

Melanin, DNA replication, and autophagy affect appressorium development in Setosphaeria turcica by regulating glycerol accumulation and metabolism

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