Abstract
Ustiloxin is a kind of 13-membered cyclic peptides found in mature rice false smut generated by Ustilaginoidea virens infecting rice spikelet. So far, six kinds of ustiloxins have been identified from false smut balls (FSBs) in which ustiloxin A is the main component. The toxins can not only inhibit the growth of rice, wheat, and corn, but also poison people and animals. However, so far, there have been few studies of the content of ustiloxin except that in mature FSB. The effect of ustiloxins on the process of infection has not been clarified. In this study, the technique of artificial inoculation coupled with UPLC-ESI–MS was introduced to investigate the content of ustiloxins in the course of infection. The initial formation time of ustiloxin A, B, C, D, F, and G was no later than 5, 5, 9, 7, 7, and 9 days post inoculation (dpi) prior to FSB’s formation, respectively. The content of ustiloxin A per spikelet was increased rapidly from 6.0 ng at 5 dpi to 14,157.1 ng at 25 dpi. Meanwhile, the content of ustiloxin A per dry weight (DW) of the FSBs also peaked at 1321.2 μg/g at 25 dpi. Interestingly, both the contents of ustiloxin A per dry weight and per spikelet were significantly reduced from 25 to 30 dpi. Transcriptome sequencing revealed that a total of 146 transcripts (103 upregulated and 43 downregulated) were significantly changed in rice spikelets after 3-h acute exposure to 100 ng ustiloxin A. In addition, several of the significantly altered genes were validated by RT-qPCR.
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Zhang Y, Zhang K, Fang AF, Han YQ, Yang J, Xue MF, Bao JD, Hu DW, Zhou B, Sun XY, Li SJ, Wen M, Yao N, Ma LJ, Liu YF, Zhang M, Huang F, Luo CX, Zhou LG, Li JQ, Chen ZY, Miao JK, Wang S, Lai JS, Xu JR, Hsiang T, Peng YL, Sun WX (2014) Specific adaptation of Ustilaginoidea virens in occupying host florets revealed by comparative and functional genomics. Nat Commun 5:3849. https://doi.org/10.1038/ncomms4849
Lu DH, Yang XQ, Mao JH, Ye HL, Wang P, Chen YP, He ZQ, Chen F (2009) Characterising the pathogenicity diversity of Ustilaginoidea virens in hybrid rice in China. J Plant Pathol 91:443–451
Tang YX, Jin J, Hu DW, Yong ML, Xu Y, He LP (2013) Elucidation of the infection process of Ustilaginoidea virens (teleomorph: Villosiclava virens) in rice spikelets. Plant Pathol 62:1–8. https://doi.org/10.1111/j.1365-3059.2012.02629.x
Fu XX, Wang XH, Cui YL, Wang AL, Lai DW, Liu Y, Li QX, Wang BM, Zhou LG (2015) A monoclonal antibody-based enzyme-linked immunosorbent assay for detection of ustiloxin A in rice false smut balls and rice samples. Food Chem 181:140–145. https://doi.org/10.1016/j.foodchem.2015.02.068
Koiso Y, Natori M, Iwasaki S, Sato S, Sonoda R, Fujita Y, Yaegashi H, Sato Z (1992) Ustiloxin: a phytotoxin and a mycotoxin from false smuth balls on rice panicles. Tetrahedron Lett 33:4157–4160. https://doi.org/10.1016/S0040-4039(00)74677-6
Shibata S, Ogihara Y, Ohta A (1963) Metabolic products of fungi. XXII. On ustilaginoidins. (2). The structure of ustilaginoidin A. Chem Pharm Bull 11:1179–1182. https://doi.org/10.1248/cpb.11.1179
Lai DW, Meng JJ, Zhang XP, Xu D, Dai JG, Zhou LG (2019) Ustilobisorbicillinol A, a cytotoxic sorbyl-containing aromatic polyketide from Ustilaginoidea virens. Org Lett 21:1311–1314. https://doi.org/10.1021/acs.orglett.8b04101
Wang XH, Wang J, Lai DW, Wang WX, Dai JG, Zhou LG, Liu Y (2017) Ustiloxin G, a new cyclopeptide mycotoxin from rice false smut balls. Toxins 9:54. https://doi.org/10.3390/toxins9020054
Koiso Y, Li Y, Iwasaki S, Hanaka K, Kobayashi T, Sonoda R, Fujita Y, Yaegashi H, Sato Z (1994) Ustiloxins, antimitotic cyclic peptides from false smut balls on rice panicles caused by Ustilaginoidea virens. J Antibiot 47:765–773. https://doi.org/10.7164/antibiotics.47.765
Koiso Y, Morisaki N, Yamashita Y, Mitsui Y, Shirai R, Hashimoto Y, Iwasaki S (1998) Isolation and structure of an antimitotic cyclic peptide, ustiloxin F. J antibiot 51:418–422. https://doi.org/10.7164/antibiotics.51.418
Nakamura K, Izumiyama N, Ohtsubo K, Koiso Y, Iwasaki S (1993) Apoptosis induced in the liver, kidney and urinary bladder of mice by the fungal toxin produced by Ustilaginoidea virens. Mycotoxins 1993:25–30. https://doi.org/10.2520/myco1975.1993.38_25
Hu Z, Dang Y, Liu C, Zhou L, Liu H (2019) Acute exposure to ustiloxin A affects growth and development of early life zebrafish, Danio rerio. Chemosphere 226:851–857. https://doi.org/10.1016/j.chemosphere.2019.04.002
Ludueña RF, Roach MC, Prasad V, Banerjee M, Koiso Y, Li Y, Iwasaki S (1994) Interaction of ustiloxin A with bovine brain tubulin. Biochem Pharmacol 47:1593–1599. https://doi.org/10.1016/0006-2952(94)90537-1
Ranaivoson FM, Gigant B, Berritt S, Joullie M, Knossow M (2012) Structural plasticity of tubulin assembly probed by vinca-domain ligands. Acta Crystallogr D 68:927–934. https://doi.org/10.1107/S0907444912017143
Baidyaroy D, Brosch G, Graessle S, Trojer P, Walton JD (2002) Characterization of inhibitor-resistant histone deacetylase activity in plant-pathogenic fungi. Eukaryot Cell 1:538–547. https://doi.org/10.1128/ec.1.4.538-547.2002
Miller JD, Young JC (1985) Deoxynivalenol in an experimental Fusarium graminearum infection of wheat. Can J Plant Pathol 7:132–134. https://doi.org/10.1080/07060668509501488
Miller JD, Arnison PG (1986) Degradation of deoxynivalenol by suspension cultures of the fusarium head blight resistant wheat cultivar Frontana. Can J Plant Pathol 8:147–150. https://doi.org/10.1080/07060668609501818
Wang XH, Fu XX, Lin FK, Sun WB, Meng JJ, Wang AL, Lai DW, Zhou LG, Liu Y (2016) The contents of ustiloxins A and B along with their distribution in rice false smut balls. Toxins 8:262. https://doi.org/10.3390/toxins8090262
Shan TJ, Sun WB, Wang XH, Fu XX, Sun WX, Zhou LG (2013) Purification of ustiloxins A and B from rice false smut balls by macroporous resins. Molecules 18:8181. https://doi.org/10.3390/molecules18078181
Song JH, Wei W, Lv B, Lin Y, Yin WX, Peng YL, Schnabel G, Huang JB, Jiang DH, Luo CX (2016) Rice false smut fungus hijacks the rice nutrients supply by blocking and mimicking the fertilization of rice ovary. Environ Microbiol 18:3840–3849. https://doi.org/10.1111/1462-2920.13343
Jia Q, Lv B, Guo MY, Luo CX, Zheng L, Hsiang T, Huang JB (2015) Effect of rice growth stage, temperature, relative humidity and wetness duration on infection of rice panicles by Villosiclava virens. Eur J Plant Pathol 141:15–25. https://doi.org/10.1007/s10658-014-0516-4
Fiehn O (2002) Metabolomics-the link between genotypes and phenotypes. Plant Mol Biol 48:155–171. https://doi.org/10.1007/978-94-010-0448-0_11
Langmead B, Salzberg SL (2012) Fast gapped-read alignment with Bowtie 2. Nat Methods 9:357–359. https://doi.org/10.1038/nmeth.1923
Li B, Dewey CN (2011) RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinform 12:323. https://doi.org/10.1186/1471-2105-12-323
Tarazona S, Garcia-Alcalde F, Dopazo J, Ferrer A, Conesa A (2011) Differential expression in RNA-seq: a matter of depth. Genome Res 21:2213–2223. https://doi.org/10.1101/gr.124321.111
Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, Davis AP, Dolinski K, Dwight SS, Eppig JT, Harris MA, Hill DP, Issel-Tarver L, Kasarskis A, Lewis S, Matese JC, Richardson JE, Ringwald M, Rubin GM, Sherlock G (2000) Gene ontology: tool for the unification of biology. Nat Genet 25:25–29. https://doi.org/10.1038/75556
Kanehisa M, Sato Y, Kawashima M, Furumichi M, Tanabe M (2015) KEGG as a reference resource for gene and protein annotation. Nucleic Acids Res 44:D457–D462. https://doi.org/10.1093/nar/gkv1070
Jain M, Nijhawan A, Tyagi AK, Khurana JP (2006) Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. Biochem Bioph Res Co 345:646–651. https://doi.org/10.1016/j.bbrc.2006.04.140
Fan J, Guo XY, Li L, Huang F, Sun WX, Li Y, Huang YY, Xu YJ, Shi J, Lei Y (2015) Infection of Ustilaginoidea virens intercepts rice seed formation but activates grain-filling-related genes. J Integr Plant Biol 57:577–590. https://doi.org/10.1111/jipb.12299
Lanubile A, Logrieco A, Battilani P, Proctor RH, Marocco A (2013) Transcriptional changes in developing maize kernels in response to fumonisin-producing and nonproducing strains of Fusarium verticillioides. Plant Sci 210:183–192. https://doi.org/10.1016/j.plantsci.2013.05.020
Wang WX, Vinocur B, Shoseyov O, Altman A (2004) Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci 9:244–252. https://doi.org/10.1016/j.tplants.2004.03.006
Fang CT, Kuo HH, Hsu SC, Yih LH (2019) HSP70 is required for the proper assembly of pericentriolar material and function of mitotic centrosomes. Cell Div 14:4. https://doi.org/10.1186/s13008-019-0047-7
Suri SS, Dhindsa RS (2008) A heat-activated MAP kinase (HAMK) as a mediator of heat shock response in tobacco cells. Plant Cell Environ 31:218–226. https://doi.org/10.1111/j.1365-3040.2007.01754.x
Li Y, Koiso Y, Kobayashi H, Hashimoto Y, Iwasaki S (1995) Ustiloxins, new antimitotic cyclic peptides: interaction with porcine brain tubulin. Biochem Pharmacol 49:1367–1372. https://doi.org/10.1016/0006-2952(95)00072-8
Chernys JT, Zeevaart JAD (2000) Characterization of the 9-cis-epoxycarotenoid dioxygenase gene family and the regulation of abscisic acid biosynthesis in avocado. Plant Physiol 124:343–354. https://doi.org/10.1104/pp.124.1.343
Sun S, Fan W, Mu Z (2017) The spatio-temporal specificity of PYR1/PYL/RCAR ABA receptors in response to developmental and environmental cues. Plant Signal Behav 12:e1214793. https://doi.org/10.1080/15592324.2016.1214793
Jiang CJ, Nakajima N, Kondo N (1996) Disruption of microtubules by abscisic acid in guard cells of Vicia faba L. Plant Cell Physiol 37:697–701. https://doi.org/10.1093/oxfordjournals.pcp.a029001
Lü B, Gong ZH, Wang J, Zhang JH, Liang JS (2007) Microtubule dynamics in relation to osmotic stress-induced ABA accumulation in Zea mays roots. J Exp Bot 58:2565–2572. https://doi.org/10.1093/jxb/erm107
Yang JC, Zhang JH (2006) Grain filling of cereals under soil drying. New Phytol 169:223–236. https://doi.org/10.1111/j.1469-8137.2005.01597.x
Zhang H, Liu K, Wang ZQ, Liu LJ, Yang JC (2015) Abscisic acid, ethylene and antioxidative systems in rice grains in relation with grain filling subjected to postanthesis soil-drying. Plant Growth Regul 76:135–146. https://doi.org/10.1007/s10725-014-9983-z
Suzuki N, Rivero RM, Shulaev V, Blumwald E, Mittler R (2014) Abiotic and biotic stress combinations. New Phytol 203:32–43. https://doi.org/10.1111/nph.12797
Zhu Y, Qian WQ, Hua J (2010) Temperature modulates plant defense responses through NB-LRR proteins. PLoS Pathog 6:e1000844. https://doi.org/10.1371/journal.ppat.1000844
Liu JZ, Feng LL, Li JM, He ZH (2015) Genetic and epigenetic control of plant heat responses. Front Plant Sci. https://doi.org/10.3389/fpls.2015.00267
Van Loon LC, Rep M, Pieterse CMJ (2006) Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol 44:135–162. https://doi.org/10.1146/annurev.phyto.44.070505.143425
Han YQ, Zhang K, Yang J, Zhang N, Fang AF, Zhang Y, Liu YF, Chen ZY, Hsiang T, Sun WX (2015) Differential expression profiling of the early response to Ustilaginoidea virens between false smut resistant and susceptible rice varieties. BMC Genom 16:955. https://doi.org/10.1186/s12864-015-2193-x
Zhou JM, Trifa Y, Silva H, Pontier D, Lam E, Shah J, Klessig DF (2000) NPR1 differentially interacts with members of the TGA/OBF family of transcription factors that bind an element of the PR-1 gene required for induction by salicylic acid. Mol Plant Microbe In 13:191–202. https://doi.org/10.1094/mpmi.2000.13.2.191
Agrawal GK, Jwa N-S, Rakwal R (2000) A novel rice (Oryza sativa l.) acidic PR1 gene highly responsive to cut, phytohormones, and protein phosphatase inhibitors. Biochem Bioph Res Co 274:157–165. https://doi.org/10.1006/bbrc.2000.3114
Moon SJ, Park HJ, Kim TH, Kang JW, Lee JY, Cho JH, Lee JH, Park DS, Byun MO, Kim BG, Shin DJ (2018) OsTGA2 confers disease resistance to rice against leaf blight by regulating expression levels of disease related genes via interaction with NH1. PLoS ONE 13:e0206910. https://doi.org/10.1371/journal.pone.0206910
Vellosillo T, Martínez M, López MA, Vicente J, Cascón T, Dolan L, Hamberg M, Castresana C (2007) Oxylipins produced by the 9-lipoxygenase pathway in Arabidopsis regulate lateral root development and defense responses through a specific signaling cascade. Plant Cell 19:831–846. https://doi.org/10.1105/tpc.106.046052
Huang JX, Cai MH, Long QZ, Liu LL, Lin QY, Jiang L, Chen SH, Wan JM (2014) OsLOX2, a rice type I lipoxygenase, confers opposite effects on seed germination and longevity. Transgenic Res 23:643–655. https://doi.org/10.1007/s11248-014-9803-2
Iuchi S, Kobayashi M, Taji T, Naramoto M, Seki M, Kato T, Tabata S, Kakubari Y, Yamaguchi-Shinozaki K, Shinozaki K (2001) Regulation of drought tolerance by gene manipulation of 9-cis-epoxycarotenoid dioxygenase, a key enzyme in abscisic acid biosynthesis in Arabidopsis. Plant J 27:325–333. https://doi.org/10.1046/j.1365-313x.2001.01096.x
Nalam VJ, Alam S, Keereetaweep J, Venables B, Burdan D, Lee H, Trick HN, Sarowar S, Makandar R, Shah J (2015) Facilitation of Fusarium graminearum infection by 9-lipoxygenases in Arabidopsis and wheat. Mol Plant Microbe In 28:1142–1152. https://doi.org/10.1094/mpmi-04-15-0096-r
Vicente J, Cascón T, Vicedo B, García-Agustín P, Hamberg M, Castresana C (2012) Role of 9-lipoxygenase and α-dioxygenase oxylipin pathways as modulators of local and systemic defense. Mol Plant 5:914–928. https://doi.org/10.1093/mp/ssr105
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This research was funded by the Special Technical Innovation of Hubei Province (2017ABA146) and the Fundamental Research Funds for the Central Universities (2662018JC048).
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Conceptualization, ZH and LZ; Methodology, LZ and CL; Data Curation, ZH and HL; Validation, JH and HL; Writing-Original Draft Preparation, ZH; Writing-Review & Editing, HL; Supervision, JH and HL; Project Administration, HL; Funding Acquisition, HL.
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Hu, Z., Zheng, L., Huang, J. et al. Ustiloxin A is Produced Early in Experimental Ustilaginoidea virens Infection and Affects Transcription in Rice. Curr Microbiol 77, 2766–2774 (2020). https://doi.org/10.1007/s00284-020-02072-6
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DOI: https://doi.org/10.1007/s00284-020-02072-6