Abstract
Key Message
In woodland strawberry, a brassinosteroid biosynthesis inhibitor propiconazole induced typical brassinosteroid-deficient phenotypes and decreased female fertility due to attenuated female gametophyte development.
Abstract
Brassinosteroids (BRs) play roles in various aspects of plant development. We investigated the physiological roles of BRs in the woodland strawberry, Fragaria vesca. BR-level-dependent phenotypes were observed using a BR biosynthetic inhibitor, propiconazole (PCZ), and the most active natural BR, brassinolide (BL). Endogenous BL and castasterone, the active BRs, were below detectable levels in PCZ-treated woodland strawberry. The plants were typical BR-deficient phenotypes, and all phenotypes were restored by treatment with BL. These observations indicate that PCZ is an effective inhibitor of BR in woodland strawberry. Only one gene for each major step of BR biosynthesis in Arabidopsis is encoded in the woodland strawberry genome. BR biosynthetic genes are highly expressed during the early stage of fruit development. Emasculated flowers treated with BL failed to develop fruit, implying that BR is not involved in parthenocarpic fruit development. Similar to BR-deficient and BR-insensitive Arabidopsis mutants, female fertility was lower in PCZ-treated plants than in mock-treated plants due to failed attraction of the pollen tube to the ovule. In PCZ-treated plants, expression of FveMYB98, the homologous gene for Arabidopsis MYB98 (a marker for synergid cells), was downregulated. Ovules were smaller in PCZ-treated plants than in mock-treated plants, and histological analysis implied that the development of more than half of female gametophytes was arrested at the early stage in PCZ-treated plants. Our findings explain how BRs function during female gametophyte development in woodland strawberry.
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References
Asami T, Min YK, Nagata N et al (2000) Characterization of brassinazol, a triazole-type brassinosteroid biosynthesis inhibitor. Plant Physiol 123:93–100. https://doi.org/10.1104/pp.123.1.93
Asami T, Mizutani M, Fujioka S et al (2001) Selective interaction of triazole derivatives with DWF4, a cytochrome P450 monooxygenase of the brassinosteroid biosynthetic pathway, correlates with brassinosteroid deficiency in planta. J Biol Chem 276:25678–25691. https://doi.org/10.1074/jbc.M103524200
Bajguz A, Chmur M, Gruszka D (2020) Comprehensive overview of the brassinosteroid biosynthesis pathways: substrates, products, inhibitors, and connections. Front Plant Sci 11:1034. https://doi.org/10.3389/fpls.2020.01034
Cai H, Liu L, Huang Y et al (2022) Brassinosteroid signaling regulates female germline specification in Arabidopsis. Curr Biol 32:1102–1114. https://doi.org/10.1016/j.cub.2022.01.022
Chai Y-m, Zhang Q, Tian L et al (2013) Brassinosteroid is involved in strawberry fruit ripening. Plant Growth Regul 69:63–69. https://doi.org/10.1007/s10725-012-9747-6
Choe S, Dikes BP, Fujioka S et al (1998) The DWF4 gene of Arabidopsis encodes a cytochrome P450 that mediates multiple 22α-hydroxylation steps in brassinosteroid biosynthesis. Plant Cell 10:231–243. https://doi.org/10.1105/tpc.10.2.231
Choe S, Fujioka S, Noguchi T et al (2001) Overexpression of DWARF4 in the brassinosteroid biosynthetic pathway results in increased vegetative growth and seed yield in Arabidopsis. Plant J 26:573–582. https://doi.org/10.1046/j.1365-313x.2001.01055.x
Chory J, Nagpal P, Peto CA (1991) Phenotypic and genetic analysis of det2, a new mutant that affects light-regulated seedling development in Arabidopsis. Plant Cell 3:445–459. https://doi.org/10.1105/tpc.3.5.445
Chung Y, Choe S (2013) The regulation of brassinosteroid biosynthesis in Arabidopsis. Crit Rev Plant Sci 32:396–410. https://doi.org/10.1080/07352689.2013.797856
Clouse SD, Langford M, McMorris TCA (1996) A brassinosteroid-insensitive mutant in Arabidopsis thaliana exhibits multiple defects in growth and development. Plant Physiol 111:671–678. https://doi.org/10.1104/pp.111.3.671
Corvalán C, Choe S (2017) Identification of brassinosteroid genes in Brachypodium distachyon. BMC Plant Biol. https://doi.org/10.1186/s12870-016-0965-3
Dziadczyk E, Domaciuk M, Nowak M et al (2011) The development of the female gametophyte in Fragaria × ananassa Duch. cv. Selva. Acta Biol Cracov Series Botanica 53:104–112. https://doi.org/10.2478/v10182-011-0033-0
Fu FQ, Mao WH, Shi K et al (2008) A role of brassinosteroids in early fruit development in cucumber. J Exp Bot 59:2299–2308. https://doi.org/10.1093/jxb/ern093
Fujioka S, Li J, Choi YH et al (1997) The Arabidopsis deetiolated2 mutant is blocked early in brassinosteroid biosynthesis. Plant Cell 9:1951–1962. https://doi.org/10.1105/tpc.9.11.1951
Hartwig T, Corvalan C, Best NB et al (2012) Propiconazole is a specific and accessible brassinosteroid (BR) biosynthesis inhibitor for Arabidopsis and maize. PLOS One 7:e36625. https://doi.org/10.1371/journal.pone.0036625
Hejátko J, Pernisová M, Eneva T et al (2003) The putative sensor histidine kinase CKI1 is involved in female gametophyte development in Arabidopsis. Mol Genet Genom 369:443–453. https://doi.org/10.1007/s00438-003-0858-7
Higashiyama T, Yabe S, Sasaki N et al (2001) Pollen tube attraction by the synergid cell. Science 293:1480–1483. https://doi.org/10.1126/science.1062429
Hollender CA, Geretz AC, Slovin JP et al (2012) Flower and early fruit development in a diploid strawberry, Fragaria vesca. Planta 235:1123–1139. https://doi.org/10.1007/s00425-011-1562-1
Hong Z, Ueguchi-Tanaka M, Umemura K et al (2003) A rice brassinosteroid-deficient mutant, ebisu dwarf (d2), is caused by a loss of function of a new member of cytochrome P450. Plant Cell 15:2900–2910. https://doi.org/10.1105/tpc.014712
Huang H-Y, Jiang W-B, Hu Y-W et al. (2013) BR signal influences Arabidopsis ovule and seed number through regulating related genes expression by BZR1. Mol Plant 6: 456–469. 10/1093/mp/sss070
Jager CE, Symons GM, Nomura T et al (2007) Characterization of two brassinosteroid C-6 oxidase gene in pea. Plant Physiol 143:1894–1904. https://doi.org/10.1104/pp.106.093088
Jia D, Chen L-G, Yin G et al (2020) Brassinosteroids regulate outer ovule integument growth in part via the control of INNER NO OUTER by brassinozole-resistant family transcription factors. J Int Plant Biol 62:1093–1111. https://doi.org/10.1111/jipb.12915
Jiang WB, Huang H-Y, Hu Y-W et al (2013) Brassinosteroid regulates seed size and shape in Arabidopsis. Plant Physiol 162:1965–1977. https://doi.org/10.1104/pp.113.217703
Kang C, Darwish O, Geretz A et al (2013) Genome-scale transcriptomic insights into early-stage fruit development in woodland strawberry Fragaria vesca. Plant Cell 25:1960–1978. https://doi.org/10.1105/tpc.113.111732
Kasahara RD, Portereiko MF, Sandaklie-Nikolava L et al (2005) MYB98 is required for pollen tube guidance and synergid cell differentiation in Arabidopsis. Plant Cell 17:2981–2992. https://doi.org/10.1105/tpc.105.034603
Kim T-W, Hwang J-Y, Kim Y-S et al (2005) Arabidopsis CYP85A2, a cytochrome P450, mediates the Baeyer-Villiger oxidation of castasterone to brassinolide in brassinosteroid biosynthesis. Plant Cell 17:2397–2412. https://doi.org/10.1105/tpc.105.033738
Kurihara D, Mizuta Y, Sato Y et al (2015) ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging. Development 142:4168–4179. https://doi.org/10.1242/dev.127613
Leszczuk A, Domaciuk M, Szczuka E (2018) Unique features of the female gametophyte development of strawberry Fragaria × ananassa Duch. Sci Hort 234:201–209. https://doi.org/10.1016/j.scienta.2018.02.030
Li J, Nam KH, Vafeados D et al (2001) BIN2, a new brassinosteroid-insensitive locus in Arabidopsis. Plant Physiol 127:14–22. https://doi.org/10.1104/pp.127.1.14
Morinaka Y, Sakamoto T, Inukai Y et al (2006) Morphological alteration caused by brassinosteroid insensitivity increases the biomass and grain production of rice. Plant Physiol 141:924–931. https://doi.org/10.1104/pp.106.077081
Nakamura A, Fujioka S, Sunohara H et al (2006) The role of OsBRI1 and its homologous gene, OsBRL1 and OsBRL3, in rice. Plant Physiol 140:580–590. https://doi.org/10.1104/pp.105.072330
Nomura T, Kushiro T, Yokota T et al (2005) The last reaction producing brassinolide is catalyzed by cytochrome P-450s, CYP85A3 in Tomato and CYP85A2 in Arabidopsis. J Biol Chem 280:17873–17879. https://doi.org/10.1074/jbc.M414592200
Oh K, Matsumoto T, Hoshi T et al (2016) In vitro and in vivo evidence for the inhibition brassinosteroid synthesis by propiconazole through interference with side chain hydroxylation. Plant Signal Behav 11:e1158372. https://doi.org/10.1080/15592324.2016.1158372
Okuda S, Tsutsui H, Shiina K et al (2009) Defensin-like polypeptide LUREs are pollen tube attranctants secreted from synergid cells. Nature 458:357–361. https://doi.org/10.1038/nature07882
Pérez-España VH, Sánches-león N, Vielle-Calzada J-P (2011) CYP85A1 is required for the initiation of female gametogenesis in Arabidopsis thaliana. Plant Sign Behav 6:321–326. https://doi.org/10.4161/psb.6.3.13206
Pipattanawong N, Fujishige N, Yamane K et al (1996) Effects of brassinosteroid on vegetative and reproductive growth in two day-neutral strawberries. J Japan Soc Hort Sci 65:651–654. https://doi.org/10.2503/jjshs.65.651
Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochem Biophys Acta 975:384–394. https://doi.org/10.1016/S0005-2728(89)80347-0
Sakurai AF, S, (1993) The current status of physiology and biochemistry of brassinosteroids. Plant Growth Regl 13:147–159. https://doi.org/10.1007/BF00024257
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protocols 3:1101–1108. https://doi.org/10.1038/nprot.2008.73
Sekimata K, Han S-Y, Yoneyama K et al (2002) A specific and potent inhibitor of brassinosteroid biosynthesis possessing a dioxolane ring. J Agric Food Chem 50:3486–3490. https://doi.org/10.1021/jf011716w
Sprunck S, Rademacher S, Vogler F et al (2012) Egg cell-secreted EC1 triggers sperm cell activation during double fertilization. Science 338:1093–1097. https://doi.org/10.1126/science.1223944
Symons GM, Davies C, Shavrukov Y et al (2006) Grapes on steroids. Brassinosteroids are involved in grape berry ripening. Plant Physiol 140:150–158. https://doi.org/10.1104/pp105.070706
Symons GM, Chua Y-J, Ross JJ et al (2012) Hormonal changes during non-climacteric ripening in strawberry. J Exp Bot 63:4741–4750. https://doi.org/10.1093/jxb/ers147
Szekeres M, Németh K, Kocz-Kálmán Z et al (1996) Brassinosteroids rescue the deficiency of CYP90, a cytochrome P450, controlling cell elongation and de-etiolation in Arabidopsis. Cell 85:171–182. https://doi.org/10.1016/S0092-8674(00)81094-6
Wang Z-Y, Nakano T, Chendron J et al (2002) Nuclear-localized BZR1 mediates brassinosteroid-induced growth and feedback suppression of brassinosteroid biosynthesis. Dev Cell 2:505–513. https://doi.org/10.1016/S1534-5807(02)00153-3
Wang L, Liu J, Shen Y et al (2021) Brassinosteroids synthesized by CYP85A/A1 but not CYP85A2 function via a BRI1-like receptor but not via BRI1 in Picea abies. J Exp Bot 72:1748–1763. https://doi.org/10.1093/jxb/eraa557
Yamamuro C, Ihara Y, Wu X et al (2000) Loss of function of a rice brassinosteroid insensitive1 homolog prevents internode elongation and bending of the lamina joint. Plant Cell 12:1591–1605. https://doi.org/10.1105/tpc.12.9.1591
Ye Q, Zhu W, Li L et al (2010) Brassinosteroids control male fertility by regulating the expression of key genes involved in Arabidopsis anther and pollen development. Proc Natl Acad Sci USA 107:6100–6105. https://doi.org/10.1073/pnas.091233107
Zhao B, Li J (2012) Regulation of brassinosteroid biosynthesis and inactivation. J Integr Plant Biol 54:746–759. https://doi.org/10.1111/j.1744-7909.2012.01.168.x
Acknowledgements
We thank Dr. Timo Hytönen, Dr. Takahito Nomura and Dr. Yusuke Jikumaru for their technical advice and Dr. Daisuke Maruyama for his technical advice and critical comments. We thank Ms. Ayaka Saito, Ms. Keiko Nagano, Ms. Keiko Inoue and Ms. Mieko Ito for their technical assistance.
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This study was supported by the Japan Society for The Promotion of Science: Grant-in-Aid for Scientific Research C (A.N., grant No. 15K07296).
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HI and AN conceived and designed research. HI, EY, IA, and AN conducted experiments. EY, TK and MA contributed new analytical tools. HI, IA and AN wrote original draft. HI, TK, MA, YS and AN reviewed and edited manuscript. YS supervised research. All authors read and approved the final manuscript.
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Ishii, H., Ishikawa, A., Yumoto, E. et al. Propiconazole-induced brassinosteroid deficiency reduces female fertility by inhibiting female gametophyte development in woodland strawberry. Plant Cell Rep 42, 587–598 (2023). https://doi.org/10.1007/s00299-023-02981-3
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DOI: https://doi.org/10.1007/s00299-023-02981-3