Abstract
Key message
Hydrogen sulfide positively regulates autophagy and the expression of hypoxia response-related genes under submergence to enhance the submergence tolerance of Arabidopsis.
Abstract
Flooding seriously endangers agricultural production, and it is quite necessary to explore the mechanism of plant response to submergence for improving crop yield. Both hydrogen sulfide (H2S) and autophagy are involved in the plant response to submergence. However, the mechanisms by which H2S and autophagy interact and influence submergence tolerance have not been thoroughly elucidated. Here, we reported that exogenous H2S pretreatment increased the level of endogenous H2S and alleviated plant cell death under submergence. And transgenic lines decreased in the level of endogenous H2S, L-cysteine desulfurase 1 (des1) mutant and 35S::GFP-O-acetyl-L-serine(thiol)lyase A1 (OASA1)/des1-#56/#61, were sensitive to submergence, along with the lower transcript levels of hypoxia response genes, LOB DOMAIN 41 (LBD41) and HYPOXIA RESPONSIVE UNKNOWN PROTEIN 43 (HUP43). Submergence induced the formation of autophagosomes, and the autophagy-related (ATG) mutants (atg4a/4b, atg5, atg7) displayed sensitive phenotypes to submergence. Simultaneously, H2S pretreatment repressed the autophagosome producing under normal conditions, but enhanced this process under submergence by regulating the expression of ATG genes. Moreover, the mutation of DES1 aggravated the sensitivity of des1/atg5 to submergence by reducing the formation of autophagosomes under submergence. Taken together, our results demonstrated that H2S alleviated cell death through regulating autophagy and the expression of hypoxia response genes during submergence in Arabidopsis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Availability of data and materials
Materials to this work will be provided upon request.
References
Alvarez C, Calo L, Romero LC, Garcia I, Gotor C (2010) An O-acetylserine(thiol)lyase homolog with l-cysteine desulfhydrase activity regulates cysteine homeostasis in Arabidopsis. Plant Physiol 152(2):656–669. https://doi.org/10.1104/pp.109.147975
Alvarez C, Garcia I, Moreno I, Perez-Perez ME, Crespo JL, Romero LC, Gotor C (2012) Cysteine-generated sulfide in the cytosol negatively regulates autophagy and modulates the transcriptional profile in Arabidopsis. Plant Cell 24(11):4621–4634. https://doi.org/10.1105/tpc.112.105403
Aroca A, Gotor C (2022) Hydrogen sulfide: a key role in autophagy regulation from plants to mammalians. Antioxidants (basel). https://doi.org/10.3390/antiox11020327
Aroca A, Serna A, Gotor C, Romero LC (2015) S-Sulfhydration: a cysteine posttranslational modification in plant systems. Plant Physiol 168(1):334–342. https://doi.org/10.1104/pp.15.00009
Aroca A, Yruela I, Gotor C, Bassham DC (2021) Persulfidation of ATG18a regulates autophagy under ER stress in Arabidopsis. Proc Natl Acad Sci USA 118(20):e2023604118. https://doi.org/10.1073/pnas.2023604118
Bailey-Serres J, Voesenek LA (2008) Flooding stress: acclimations and genetic diversity. Annu Rev Plant Biol 59:313–339. https://doi.org/10.1146/annurev.arplant.59.032607.092752
Bailey-Serres J, Voesenek LA (2010) Life in the balance: a signaling network controlling survival of flooding. Curr Opin Plant Biol 13(5):489–494. https://doi.org/10.1016/j.pbi.2010.08.002
Bailey-Serres J, Fukao T, Gibbs DJ, Holdsworth MJ, Lee SC, Licausi F, Perata P, Voesenek LA, van Dongen JT (2012) Making sense of low oxygen sensing. Trends Plant Sci 17(3):129–138. https://doi.org/10.1016/j.tplants.2011.12.004
Bassham DC, Laporte M, Marty F, Moriyasu Y, Ohsumi Y, Olsen LJ, Yoshimoto K (2006) Autophagy in development and stress responses of plants. Autophagy 2(1):2–11 https://doi.org/10.4161/auto.2092
Blokhina OB, Chirkova TV, Fagerstedt KV (2001) Anoxic stress leads to hydrogen peroxide formation in plant cells. J Exp Bot 52(359):1179–1190. https://doi.org/10.1093/jexbot/52.359.1179
Calderwood A, Kopriva S (2014) Hydrogen sulfide in plants: From dissipation of excess sulfur to signaling molecule. Nitric Oxide Biol Chem 41:72–78. https://doi.org/10.1016/j.niox.2014.02.005
Chen L, Liao B, Qi H, Xie LJ, Huang L, Tan WJ, Zhai N, Yuan LB, Zhou Y, Yu LJ, Chen QF, Shu WS, Xiao S (2015) Autophagy contributes to regulation of the hypoxia response during submergence in Arabidopsis thaliana. Autophagy 11(12):2233–2246. https://doi.org/10.1080/15548627.2015.1112483
Chen J, Shang YT, Wang WH, Chen XY, He EM, Zheng HL, Shangguan ZP (2016) Hydrogen sulfide-mediated polyamines and sugar changes are involved in hydrogen sulfide-induced drought tolerance in Spinacia oleracea seedlings. Front Plant Sci 7:1173. https://doi.org/10.3389/fpls.2016.01173
Cheng W, Zhang L, Jiao CJ, Su M, Yang T, Zhou L, Peng RY, Wang R, Wang CY (2013) Hydrogen sulfide alleviates hypoxia-induced root tip death in Pisum sativum. Plant Physiol Biochem 70:278–286. https://doi.org/10.1016/j.plaphy.2013.05.042
Contento AL, Xiong Y, Bassham DC (2005) Visualization of autophagy in Arabidopsis using the fluorescent dye monodansylcadaverine and a GFP-AtATG8e fusion protein. Plant J 42(4):598–608. https://doi.org/10.1111/j.1365-313X.2005.02396.x
Doelling JH, Walker JM, Friedman EM, Thompson AR, Vierstra RD (2002) The APG8/12-activating enzyme APG7 is required for proper nutrient recycling and senescence in Arabidopsis thaliana. J Biol Chem 277(36):33105–33114. https://doi.org/10.1074/jbc.M204630200
Fang HH, Liu ZQ, Jin ZP, Zhang LP, Liu DM, Pei YX (2016) An emphasis of hydrogen sulfide-cysteine cycle on enhancing the tolerance to chromium stress in Arabidopsis. Environ Pollut 213:870–877. https://doi.org/10.1016/j.envpol.2016.03.035
Gasch P, Fundinger M, Mullet JT, Lee T, Bailey-Serres J, Mustroph A (2016) Redundant ERF-VII transcription factors bind to an evolutionarily conserved cis-motif to regulate hypoxia-responsive gene expression in Arabidopsis. Plant Cell 28(1):160–180. https://doi.org/10.1105/tpc.15.00866
Guan B, Lin Z, Liu DC, Li CY, Zhou ZQ, Mei FZ, Li JW, Deng XY (2019) Effect of waterlogging-induced autophagy on programmed cell death in Arabidopsis roots. Front Plant Sci 10:468. https://doi.org/10.3389/fpls.2019.00468
Hanada T, Noda NN, Satomi Y, Ichimura Y, Fujioka Y, Takao T, Inagaki F, Ohsumi Y (2007) The Atg12-Atg5 conjugate has a novel E3-like activity for protein lipidation in autophagy. J Biol Chem 282(52):37298–37302. https://doi.org/10.1074/jbc.C700195200
Hanaoka H, Noda T, Shirano Y, Kato T, Hayashi H, Shibata D, Tabata S, Ohsumi Y (2002) Leaf senescence and starvation-induced chlorosis are accelerated by the disruption of an Arabidopsis autophagy gene. Plant Physiol 129(3):1181–1193. https://doi.org/10.1104/pp.011024
Harrison-Lowe NJ, Olsen LJ (2008) Autophagy protein 6 (ATG6) is required for pollen germination in Arabidopsis thaliana. Autophagy 4(3):339–348. https://doi.org/10.4161/auto.5629
Huang L, Yu LJ, Zhang X, Fan B, Wang FZ, Dai YS, Qi H, Zhou Y, Xie LJ, Xiao S (2019) Autophagy regulates glucose-mediated root meristem activity by modulating ROS production in Arabidopsis. Autophagy 15(3):407–422. https://doi.org/10.1080/15548627.2018.1520547
Huang D, Huo J, Liao W (2021) Hydrogen sulfide: Roles in plant abiotic stress response and crosstalk with other signals. Plant Sci 302:110733. https://doi.org/10.1016/j.plantsci.2020.110733
Ichimura Y, Kirisako T, Takao T, Satomi Y, Shimonishi Y, Ishihara N, Mizushima N, Tanida I, Kominami E, Ohsumi M, Noda T, Ohsumi Y (2000) A ubiquitin-like system mediates protein lipidation. Nature 408(6811):488–492. https://doi.org/10.1038/35044114
Inoue Y, Suzuki T, Hattori M, Yoshimoto K, Ohsumi Y, Moriyasu Y (2006) AtATG genes, homologs of yeast autophagy genes, are involved in constitutive autophagy in Arabidopsis root tip cells. Plant Cell Physiol 47(12):1641–1652. https://doi.org/10.1093/pcp/pcl031
Jurado-Flores A, Romero LC, Gotor C (2021) Label-free quantitative proteomic analysis of nitrogen starvation in Arabidopsis root reveals new aspects of H2S signaling by protein persulfidation. Antioxidants-Basel 10(4):508. https://doi.org/10.3390/antiox10040508
Kopriva S (2006) Regulation of sulfate assimilation in Arabidopsis and beyond. Ann Bot Lond 97(4):479–495. https://doi.org/10.1093/aob/mcl006
Kwon SI, Park OK (2008) Autophagy in plants. J Plant Biol 51(5):313–320. https://doi.org/10.1007/BF03036132
Laureano-Marin AM, Moreno I, Romero LC, Gotor C (2016) Negative regulation of autophagy by sulfide is independent of reactive oxygen species. Plant Physiol 171(2):1378–1391. https://doi.org/10.1104/pp.16.00110
Laureano-Marin AM, Aroca A, Perez-Perez ME, Yruela I, Jurado-Flores A, Moreno I, Crespo JL, Romero LC, Gotor C (2020) Abscisic acid-triggered persulfidation of the cys protease ATG4 mediates regulation of autophagy by sulfide. Plant Cell 32(12):3902–3920. https://doi.org/10.1105/tpc.20.00766
Licausi F, van Dongen JT, Giuntoli B, Novi G, Santaniello A, Geigenberger P, Perata P (2010) HRE1 and HRE2, two hypoxia-inducible ethylene response factors, affect anaerobic responses in Arabidopsis thaliana. Plant J 62(2):302–315. https://doi.org/10.1111/j.1365-313X.2010.04149.x
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408. https://doi.org/10.1006/meth.2001.1262
Mousavi SA, Kjeken R, Berg TO, Seglen PO, Berg T, Brech A (2001) Effects of inhibitors of the vacuolar proton pump on hepatic heterophagy and autophagy. Biochim Biophys Acta 1510(1–2):243–257. https://doi.org/10.1016/s0005-2736(00)00354-0
Papenbrock J, Reimenschneider A, Kamp A, Schulz-Vogt HN, Schmidt A (2007) Characterization of cysteine-degrading and H2S-releasing enzymes of higher plants—from the field to the test tube and back. Plant Biol 9(5):582–588. https://doi.org/10.1055/s-2007-965424
Patel S, Dinesh-Kumar SP (2008) Arabidopsis ATG6 is required to limit the pathogen-associated cell death response. Autophagy 4(1):20–27. https://doi.org/10.4161/auto.5056
Paul BD, Snyder SH (2012) H2S signalling through protein sulfhydration and beyond. Nat Rev Mol Cell Biol 13(8):499–507. https://doi.org/10.1038/nrm3391
Peng RY, Bian ZY, Zhou LN, Cheng W, Hai N, Yang CQ, Yang T, Wang XY, Wang CY (2016) Hydrogen sulfide enhances nitric oxide-induced tolerance of hypoxia in maize (Zea mays L.). Plant Cell Rep 35(11):2325–2340. https://doi.org/10.1007/s00299-016-2037-4
Romero LC, Aroca MA, Laureano-Marin AM, Moreno I, Garcia I, Gotor C (2014) Cysteine and cysteine-related signaling pathways in Arabidopsis thaliana. Mol Plant 7(2):264–276. https://doi.org/10.1093/mp/sst168
Sanmartin M, Ordonez A, Sohn EJ, Robert S, Sanchez-Serrano JJ, Surpin MA, Raikhel NV, Rojo E (2007) Divergent functions of VTI12 and VTI11 in trafficking to storage and lytic vacuoles in Arabidopsis. Proc Natl Acad Sci USA 104(9):3645–3650. https://doi.org/10.1073/pnas.0611147104
Schmelzle T, Hall MN (2000) TOR, a central controller of cell growth. Cell 103(2):253–262. https://doi.org/10.1016/s0092-8674(00)00117-3
Slavikova S, Shy G, Yao Y, Glozman R, Levanony H, Pietrokovski S, Elazar Z, Galili G (2005) The autophagy-associated Atg8 gene family operates both under favourable growth conditions and under starvation stresses in Arabidopsis plants. J Exp Bot 56(421):2839–2849. https://doi.org/10.1093/jxb/eri276
Surpin M, Zheng HJ, Morita MT, Saito C, Avila E, Blakeslee JJ, Bandyopadhyay A, Kovaleva V, Carter D, Murphy A, Tasaka M, Raikhel N (2003) The VTI family of SNARE proteins is necessary for plant viability and mediates different protein transport pathways. Plant Cell 15(12):2885–2899. https://doi.org/10.1105/tpc.016121
Suttangkakul A, Li FQ, Chung T, Vierstra RD (2011) The ATG1/ATG13 protein kinase complex is both a regulator and a target of autophagic recycling in Arabidopsis. Plant Cell 23(10):3761–3779. https://doi.org/10.1105/tpc.111.090993
Tai CH, Yoon MY, Kim SK, Rege VD, Nalabolu SR, Kredich NM, Schnackerz KD, Cook PF (1998) Cysteine 42 is important for maintaining an integral active site for O-acetylserine sulfhydrylase resulting in the stabilization of the alpha-aminoacrylate intermediate. Biochemistry 37(30):10597–10604. https://doi.org/10.1021/bi980647k
Thompson AR, Vierstra RD (2005) Autophagic recycling: lessons from yeast help define the process in plants. Curr Opin Plant Biol 8(2):165–173. https://doi.org/10.1016/j.pbi.2005.01.013
Thompson AR, Doelling JH, Suttangkakul A, Vierstra RD (2005) Autophagic nutrient recycling in Arabidopsis directed by the ATG8 and ATG12 conjugation pathways. Plant Physiol 138(4):2097–2110. https://doi.org/10.1104/pp.105.060673
Voesenek LACJ, Sasidharan R (2013) Ethylene- and oxygen signalling-drive plant survival during flooding. Plant Biol 15(3):426–435. https://doi.org/10.1111/plb.12014
Vojtovic D, Luhova L, Petrivalsky M (2021) Something smells bad to plant pathogens: Production of hydrogen sulfide in plants and its role in plant defence responses. J Adv Res 27:199–209. https://doi.org/10.1016/j.jare.2020.09.005
Wang CW, Kim J, Huang WP, Abeliovich H, Stromhaug PE, Dunn WA, Klionsky DJ (2001) Apg2 is a novel protein required for the cytoplasm to vacuole targeting, autophagy, and pexophagy pathways. J Biol Chem 276(32):30442–30451. https://doi.org/10.1074/jbc.M102342200
Wang LK, Feng ZX, Wang X, Wang XW, Zhang XG (2010) DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics 26(1):136–138. https://doi.org/10.1093/bioinformatics/btp612
Xiao YS, Wu XL, Sun MX, Peng FT (2020) Hydrogen sulfide alleviates waterlogging-induced damage in peach seedlings via enhancing antioxidative system and inhibiting ethylene synthesis. Front Plant Sci 11:696. https://doi.org/10.3389/fpls.2020.00696
Xie LJ, Zhou Y, Chen QF, Xiao S (2021) New insights into the role of lipids in plant hypoxia responses. Prog Lipid Res 81:101072. https://doi.org/10.1016/j.plipres.2020.101072 (ARTN101072)
Xie LJ, Chen QF, Chen MX, Yu LJ, Huang L, Chen L, Wang FZ, Xia FN, Zhu TR, Wu JX, Yin J, Liao B, Shi JX, Zhang JH, Aharoni A, Yao N, Shu WS, Xiao S (2015) Unsaturation of very-long-chain ceramides protects plant from hypoxia-induced damages by modulating ethylene signaling in Arabidopsis. Plos Genet 11(3):e1005143. https://doi.org/10.1371/journal.pgen.1005143
Xiong Y, Contento AL, Nguyen PQ, Bassham DC (2007) Degradation of oxidized proteins by autophagy during oxidative stress in Arabidopsis. Plant Physiol 143(1):291–299. https://doi.org/10.1104/pp.106.092106
Yang T, Yuan GQ, Zhang Q, Xuan LJ, Li J, Zhou LN, Shi HH, Wang XY, Wang CY (2021) Transcriptome and metabolome analyses reveal the pivotal role of hydrogen sulfide in promoting submergence tolerance in Arabidopsis. Environ Exp Bot 183:104365. https://doi.org/10.1016/j.envexpbot.2020.104365
Yoshimoto K, Hanaoka H, Sato S, Kato T, Tabata S, Noda T, Ohsumi Y (2004) Processing of ATG8s, ubiquitin-like proteins, and their deconjugation by ATG4s are essential for plant autophagy. Plant Cell 16(11):2967–2983. https://doi.org/10.1105/tpc.104.025395
Yue L, Hu Y, Fu H, Qi L, Sun H (2021) Hydrogen sulfide regulates autophagy in nucleus pulposus cells under hypoxia. JOR Spine 4(4):e1181. https://doi.org/10.1002/jsp2.1181
Zhang B, Shao L, Wang JL, Zhang Y, Guo XS, Peng YJ, Cao YR, Lai ZB (2021) Phosphorylation of ATG18a by BAK1 suppresses autophagy and attenuates plant resistance against necrotrophic pathogens. Autophagy 17(9):2093–2110. https://doi.org/10.1080/15548627.2020.1810426
Acknowledgements
We thank Prof. Yun Xiang (Lanzhou University) for providing the seeds of GFP-ATG8e.
Funding
This work was supported by the National Natural Science Foundation of China (Grant/Award Number: 31670254, 31770199); Fundamental Research Funds for the Central Universities (lzujbky-2021-43); Startup Foundation for Introducing Talent of Lanzhou University (561120206), and Foundation of the Key Laboratory of Cell Activities and Stress Adaptations, Ministry of Education of China (lzujbky-2019-kb05, lzujbky-2021-kb05).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors have no conflict of interests.
Ethical approval
Not applicable.
Consent to participate
Yes.
Consent for publication
All authors read and approved the manuscript.
Additional information
Communicated by Attila Feher.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
299_2022_2872_MOESM1_ESM.docx
Fig. S1 Homozygous plants of atg mutants were identified by PCR. (a-d) Structures of ATG5, ATG7, ATG4a and ATG4b genes were shown with insertion site of T-DNA in mutant plant lines. Exons were indicated by gray arrows, primers for the genetic characterization by small black arrows, and untranslated regions by striped boxes. (e-h) Identification of homozygous atg5, atg7, atg4a and atg4b mutants by PCR. Number 1 of atg5, atg7 and atg4b, and number 3 of atg4a were used for further experiments (DOCX 17 KB)
299_2022_2872_MOESM2_ESM.docx
Fig. S2 Submergence increased the expression level of OASA1 protein. (a, b) Immunoblot analysis showing the expression of GFP-OASA1 in 7-day-old or 26-day-old 35S::GFP-OASA1/des1-#56 under submergence. CBB was used as loading control. GFP-OASA1 normalized to the loading control was shown below to represent the expression level of OASA1 protein (DOCX 16 KB)
299_2022_2872_MOESM3_ESM.xlsx
Fig. S3 Relative expression levels of H2S metabolism-related genes in WT, des1 and 35S::GFP-OASA1/des1-#56 lines. (a) Structure of DES1 gene was shown with insertion site of T-DNA in mutant plant lines. Exons were indicated by gray arrows, primers for the genetic characterization by small black arrows, and untranslated regions by striped boxes. (b, c) qRT-PCR analysis of DES1 or OASA1 expression in WT, des1 or 35S::GFP-OASA1/des1-#56/61 lines. Data represent means ± SE of three independent experiments, and different letters indicate significant differences between the annotated columns (P < 0.05 by Duncan’s test) (XLSX 15 KB)
299_2022_2872_MOESM4_ESM.tif
Fig. S4 Relative expression level of DES1 in WT, des1, des1/atg5 and des1/GFP-ATG8e plants. Data represent means ± SE of three independent experiments, and different letters indicate significant differences between the annotated columns (P < 0.05 by Duncan’s test) (TIF 840 KB)
299_2022_2872_MOESM5_ESM.tif
Fig. S5 Endogenous H2S regulates the expression of ATG8c and ATG18a genes. (a, b) The relative expression of ATG8c and ATG8e genes were analyzed by qRT-PCR in 26-day-old WT and 35S::GFP-OASA1/des1-#56 plants. Data represent means ± SE of three independent experiments, and different letters indicate significant differences between the annotated columns (P < 0.05 by Duncan’s test) (TIF 259 KB)
Rights and permissions
About this article
Cite this article
Xuan, L., Wu, H., Li, J. et al. Hydrogen sulfide reduces cell death through regulating autophagy during submergence in Arabidopsis. Plant Cell Rep 41, 1531–1548 (2022). https://doi.org/10.1007/s00299-022-02872-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-022-02872-z