PpAKR1A, a Novel Aldo-Keto Reductase from Physcomitrella Patens, Plays a Positive Role in Salt Stress
Abstract
:1. Introduction
2. Results
2.1. Expression of PpAKR1A in Response to Salt Stress
2.2. Tissue Specificity and Subcellular Localisation of PpAKR1A
2.3. Response of PpAKR1A Knockout Mutants to Salt Stress
2.4. PpAKR1A Knockout Mutants Exhibit Reduced Tolerance to Methylglyoxal
2.5. Decreased MG Reducing Activity in PpAKR1A Knockout Mutants
2.6. Decreased Activities of ROS-Scavenging Enzymes in PpAKR1A Knockout Mutants
2.7. Bacterially Expressed Recombinant PpAKR1A Protein Effectively Reduced Toxic Aldehydes
3. Discussion
3.1. PpAKR1A Enhances Salt Resistance in P. patens
3.2. The Mechanism of PpAKR1A-Mediated Regulation in Response to Salt Stress
4. Materials and Methods
4.1. Plant Materials and Stress Treatments
4.2. Quantitative Real-Time Reverse Transcription PCR (qRT-PCR) Analysis
4.3. Plasmid Construction
4.4. Transformation of P. patens
4.5. Histochemical Analysis of GUS Activity
4.6. PpAKR1A–GFP Subcellular Localisation Analysis
4.7. Physiological Parameter Measurements
4.8. Oxidative Enzyme Assays
4.9. Cloning and Expression of Recombinant Protein of PpAKR1A
4.10. Purification Protein of the His-Fusion PpAKR1A
4.11. Enzyme Activity and Enzyme Kinetics
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ROS | Reactive oxygen species |
RCS | Reactive carbonyl |
MG | Methylglyoxal |
MDA | Malondialdehyde |
AKR | aldo-keto reductases |
qRT-PCR | real-time polymerase chain reaction |
PCR | polymerase chain reaction |
WT | wild type |
CRISPR/Cas9 | clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 |
CAT | catalase |
SOD | superoxide dismutase |
POD | peroxidase |
IPTG | isopropyl β-D-1-thiogalactopyranoside |
PMSF | phenylmethanesulfonyl fluoride |
SDS-PAGE | sodium dodecylsulphate polyacrylamide gel electrophoresis |
LB | Luria-Bertani |
PpAKR1A | a putative AKR1 from P. patens |
References
- Jez, J.M.; Penning, T.M. The aldo-keto reductase (AKR) superfamily: An update. Chem. Biol. Interact. 2001, 130–132, 499–525. [Google Scholar] [CrossRef]
- Barski, O.A.; Papusha, V.Z.; Kunkel, G.R.; Gabbay, K.H. Regulation of aldehyde reductase expression by STAF and CHOP. Genomics 2004, 83, 119–129. [Google Scholar] [CrossRef]
- Sengupta, D.; Naik, D.; Reddy, A.R. Plant aldo-keto reductases (AKRs) as multi-tasking soldiers involved in diverse plant metabolic processes and stress defense: A structure-function update. J. Plant Physiol. 2015, 179, 40–55. [Google Scholar] [CrossRef] [PubMed]
- Semchyshyn, H.M. Reactive carbonyl species in vivo: generation and dual biological effects. Sci. World J. 2014, 2014, 417842. [Google Scholar] [CrossRef]
- Hoque, T.S.; Hossain, M.A.; Mostofa, M.G.; Burritt, D.J.; Fujita, M.; Tran, L.-S.P. Methylglyoxal: An emerging signaling molecule in plant abiotic stress responses and tolerance. Front Plant Sci. 2016, 7, 1341. [Google Scholar] [CrossRef]
- Li, Z.G. Methylglyoxal and glyoxalase system in plants: old players, new concepts. Bot. Rev. 2016, 82, 183–203. [Google Scholar] [CrossRef]
- Kaur, C.; Sharma, S.; Singla-Pareek, S.L.; Sopory, S.K. Methylglyoxal detoxification in plants: role of glyoxalase pathway. Ind. J. Plant Physiol. 2016, 21, 377–390. [Google Scholar] [CrossRef]
- Sankaranarayanan, S.; Jamshed, M.; Samuel, M.A. Degradation of glyoxalase I in Brassica napus stigma leads to self-incompatibility response. Nat. Plants 2015, 1, 15185. [Google Scholar] [CrossRef]
- Ray, A.; Ray, S.; Mukhopadhyay, S.; Ray, M. Methylglyoxal with glycine or succinate enhances differentiation and shoot morphogenesis in Nicotiana tabacum callus. Biol. Plantarum 2013, 57, 219–223. [Google Scholar] [CrossRef]
- Hoque, T.S.; Uraji, M.; Tuya, A.; Nakamura, Y.; Murata, Y. Methylglyoxal inhibits seed germination and root elongation and up-regulates transcription of stress-responsive genes in ABA-dependent pathway in Arabidopsis. Plant Biol. 2012, 14, 854–858. [Google Scholar] [CrossRef]
- Li, Z.G.; Duan, X.Q.; Min, X.; Zhou, Z.H. Methylglyoxal as a novel signal molecule induces the salt tolerance of wheat by regulating the glyoxalase system, the antioxidant system, and osmolytes. Protoplasma 2017, 254, 1995–2006. [Google Scholar] [CrossRef] [PubMed]
- Mano, J. Reactive carbonyl species: Their production from lipid peroxides, action in environmental stress, and the detoxification mechanism. Plant Physiol. Biochem. 2012, 59, 90–97. [Google Scholar] [CrossRef] [PubMed]
- Mostofa, M.G.; Ghosh, A.; Li, Z.G.; Siddiqui, M.N.; Fujita, M.; Tran, L.P. Methylglyoxal—A signaling molecule in plant abiotic stress responses. Free Radic. Biol. Med. 2018, 122, 96–109. [Google Scholar] [CrossRef] [PubMed]
- Auiyawong, B.; Narawongsanont, R.; Tantitadapitak, C. Characterization of AKR4C15, a novel member of aldo-keto reductase, in comparison with other Rice AKR(s). Protein J. 2017, 36, 257–269. [Google Scholar] [CrossRef] [PubMed]
- Saito, R.; Shimakawa, G.; Nishi, A.; Iwamoto, T.; Sakamoto, K.; Yamamoto, H.; Amako, K.; Makino, A.; Miyake, C. Functional analysis of the AKR4C subfamily of Arabidopsis thaliana: Model structures, substrate specificity, acrolein toxicity, and responses to light and [CO2]. Biosci. Biotechnol. Biochem. 2013, 77, 2038–2045. [Google Scholar] [CrossRef] [PubMed]
- Simpson, P.J.; Tantitadapitak, C.; Reed, A.M.; Mather, O.C.; Bunce, C.M.; White, S.A.; Ride, J.P. Characterization of two novel aldo–Keto reductases from Arabidopsis: Expression patterns, broad substrate specificity, and an open active-site structure suggest a role in toxicant metabolism following stress. J. Mol. Biol. 2009, 392, 465–480. [Google Scholar] [CrossRef] [PubMed]
- Turóczy, Z.; Kis, P.; Török, K.; Cserháti, M.; Lendvai, A.; Dudits, D.; Horváth, G.V. Overproduction of a rice aldo-keto reductase increases oxidative and heat stress tolerance by malondialdehyde and methylglyoxal detoxification. Plant Mol. Biol. 2011, 75, 399–412. [Google Scholar] [CrossRef]
- Jain, D.; Khandal, H.; Khurana, J.P.; Chattopadhyay, D. A pathogenesis related-10 protein CaARP functions as aldo/keto reductase to scavenge cytotoxic aldehydes. Plant Mol. Biol. 2016, 90, 171–187. [Google Scholar] [CrossRef]
- Mudalkar, S.; Sreeharsha, R.V.; Reddy, A.R. A novel aldo-keto reductase from Jatropha curcas L. (JcAKR) plays a crucial role in the detoxification of methylglyoxal, a potent electrophile. J. Plant Physiol. 2016, 195, 39–49. [Google Scholar] [CrossRef]
- Nagy, B.; Majer, P.; Mihály, R.; Pauk, J.; Horváth, G.V. Stress tolerance of transgenic barley accumulating the alfalfa aldose reductase in the cytoplasm and the chloroplast. Phytochemistry 2016, 129, 14–23. [Google Scholar] [CrossRef]
- Singh, P.; Kumar, D.; Sarin, N.B. Multiple abiotic stress tolerance in Vigna mungo is altered by overexpression of ALDRXV4 gene via reactive carbonyl detoxification. Plant Mol. Biol. 2016, 91, 257–273. [Google Scholar] [CrossRef] [PubMed]
- Suekawa, M.; Fujikawa, Y.; Inada, S.; Murano, A.; Esaka, M. Gene expression and promoter analysis of a novel tomato aldo-keto reductase in response to environmental stresses. J. Plant Physiol. 2016, 200, 35–44. [Google Scholar] [CrossRef] [PubMed]
- Frank, W.; Ratnadewi, D.; Reski, R. Physcomitrella patens is highly tolerant against drought, salt and osmotic stress. Planta 2005, 220, 384–394. [Google Scholar] [CrossRef] [PubMed]
- Saavedra, L.; Svensson, J.; Carballo, V.; Izmendi, D.; Welin, B.; Vidal, S. A dehydrin gene in Physcomitrella patens is required for salt and osmotic stress tolerance. Plant J. 2006, 45, 237–249. [Google Scholar] [CrossRef] [PubMed]
- Wang, X.Q.; Yang, P.F.; Liu, Z.; Liu, W.Z.; Hu, Y.; Chen, H.; Kuang, T.Y.; Pei, Z.M.; Shen, S.H.; He, Y.K. Exploring the mechanism of Physcomitrella patens desiccation tolerance through a proteomic strategy. Plant Physiol. 2009, 149, 1739–1750. [Google Scholar] [CrossRef]
- Cove, D.J.; Perroud, P.F.; Charron, A.J.; McDaniel, S.F.; Khandelwal, A.; Quatrano, R.S. The moss Physcomitrella patens: A novel model system for plant development and genomic studies. Cold Spring Harb. Protoc. 2009, 2009, pdb.emo115. [Google Scholar] [CrossRef]
- Rensing, S.A.; Lang, D.; Zimmer, A.D.; Terry, A.; Salamov, A.; Shapiro, H.; Nishiyama, T.; Perroud, P.F.; Lindquist, E.A.; Kamisugi, Y.; et al. The Physcomitrella genome reveals evolutionary insights into the conquest of land by plants. Science 2008, 319, 64–69. [Google Scholar] [CrossRef]
- Zhu, J.K. Abiotic stress signaling and responses in plants. Cell 2016, 167, 313–324. [Google Scholar] [CrossRef]
- Li, B.; Foley, M.E. Cloning and characterization of differentially expressed genes in imbibed dormant and afterripened Arena fatua embryos. Plant Mol. Biol. 1995, 29, 823–831. [Google Scholar] [CrossRef]
- Lee, S.P.; Chen, T.H. Molecular cloning of abscisic acid-responsive mRNAs expressed during the induction of freezing tolerance in bromegrass (Bromus inermis Leyss) suspension culture. Plant Physiol. 1993, 101, 1089–1096. [Google Scholar] [CrossRef]
- Hegedüs, A.; Erdei, S.; Janda, T.; Tóth, E.; Horváth, G.; Dudits, D. Transgenic tobacco plants overproducing alfalfa aldose/aldehyde reductase show higher tolerance to low temperature and cadmium stress. Plant Sci. 2004, 166, 1329–1333. [Google Scholar] [CrossRef]
- Lopez-Obando, M.; Hoffmann, B.; Géry, C.; Guyon-Debast, A.; Téoulé, E.; Rameau, C.; Bonhomme, S.; Nogué, F. Simple and efficient targeting of multiple genes through CRISPR-Cas9 in Physcomitrella patens. G3 Genes Genom. Genet. 2016, 6, 3647–3653. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kanayama, Y.; Mizutani, R.; Yaguchi, S.; Hojo, A.; Ikeda, H.; Nishiyama, M.; Kanahama, K. Characterization of an uncharacterized aldo-keto reductase gene from peach and its role in abiotic stress tolerance. Phytochemistry 2014, 104, 30–36. [Google Scholar] [CrossRef] [PubMed]
- Vemanna, R.S.; Babitha, K.C.; Solanki, J.K.; Amarnatha Reddy, V.; Sarangi, S.K.; Udayakumar, M. Aldo-keto reductase-1 (AKR1) protect cellular enzymes from salt stress by detoxifying reactive cytotoxic compounds. Plant Physiol. Biochem. 2017, 113, 177–186. [Google Scholar] [CrossRef]
- Barclay, K.D.; McKersie, B.D. Peroxidation reactions in plant membranes: Effects of free fatty acids. Lipids 1994, 29, 877–883. [Google Scholar] [CrossRef]
- Mhamdi, A.; Van Breusegem, F. Reactive oxygen species in plant development. Development 2018, 145, dev164376. [Google Scholar] [CrossRef] [Green Version]
- Rai, A.N.; Penna, S. Molecular evolution of plant P5CS gene involved in proline biosynthesis. Mol. Biol. Rep. 2013, 40, 6429–6435. [Google Scholar] [CrossRef]
- Zhu, J.K. Plant salt tolerance. Trends Plant Sci. 2001, 6, 66–71. [Google Scholar] [CrossRef]
- Lade, H.; Paul, D.; Kweon, J.H. Quorum quenching mediated approaches for control of membrane biofouling. Int. J. Biol. Sci. 2014, 10, 550–565. [Google Scholar] [CrossRef]
- Acosta-Motos, J.R.; Hernández, J.A.; Álvarez, S.; Barba-Espín, G.; Sánchez-Blanco, M.J. The long-term resistance mechanisms, critical irrigation threshold and relief capacity shown by Eugenia myrtifolia plants in response to saline reclaimed water. Plant Physiol. Biochem. 2017, 111, 244–256. [Google Scholar] [CrossRef]
- Chang, Q.; Petrash, J.M. Disruption of aldo-keto reductase genes leads to elevated markers of oxidative stress and inositol auxotrophy in Saccharomyces cerevisiae. Biochim. Biophys. Acta 2008, 1783, 237–245. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tan, T.; Sun, Y.; Peng, X.; Wu, G.; Bao, F.; He, Y.; Zhou, H.; Lin, H. ABSCISIC ACID INSENSITIVE3 is involved in cold response and freezing tolerance regulation in Physcomitrella patens. Front. Plant Sci. 2017, 8, 1599. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sun, X.; Yuan, S.; Lin, H.H. Salicylic acid decreases the levels of dehydrin-like proteins in Tibetan hulless barley leaves under water stress. Z. Naturforsch. C J. Biosci. 2006, 61, 245–250. [Google Scholar] [CrossRef] [PubMed]
- Miller, G.; Suzuki, N.; Ciftci-Yilmaz, S.; Mittler, R. Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell Environ. 2010, 33, 453–467. [Google Scholar] [CrossRef] [PubMed]
- Mittler, R.; Vanderauwera, S.; Gollery, M.; Van Breusegem, F. Reactive oxygen gene network of plants. Trends Plant Sci. 2004, 9, 490–498. [Google Scholar] [CrossRef] [PubMed]
- Mhamdi, A.; Queval, G.; Chaouch, S.; Vanderauwera, S.; Van Breusegem, F.; Noctor, G. Catalase function in plants: A focus on Arabidopsis mutants as stress-mimic models. J. Exp. Bot. 2010, 61, 4197–4220. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Apel, K.; Hirt, H. Reactive oxygen species: Metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant Biol. 2004, 55, 373–399. [Google Scholar] [CrossRef] [Green Version]
- Yadav, S.K.; Singla-Pareek, S.L.; Ray, M.; Reddy, M.K.; Sopory, S.K. Methylglyoxal levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem. Biophys. Res. Commun. 2005, 337, 61–67. [Google Scholar] [CrossRef]
- Singla-Pareek, S.L.; Yadav, S.K.; Pareek, A.; Reddy, M.K.; Sopory, S.K. Transgenic tobacco overexpressing glyoxalase pathway enzymes grow and set viable seeds in zinc-spiked soils. Plant Physiol. 2006, 140, 613–623. [Google Scholar] [CrossRef] [Green Version]
- Kaur, C.; Ghosh, A.; Pareek, A.; Sopory, S.K.; Singla-Pareek, S.L. Glyoxalases and stress tolerance in plants. Biochem. Soc. Trans. 2014, 42, 485–490. [Google Scholar] [CrossRef]
- Paulus, C.; Köllner, B.; Jacobsen, H.J. Physiological and biochemical characterization of glyoxalase I, a general marker for cell proliferation, from a soybean cell suspension. Planta 1993, 189, 561–566. [Google Scholar] [CrossRef] [PubMed]
- Wang, X.; Zhou, S.; Chen, L.; Quatrano, R.S.; He, Y. Phospho-proteomic analysis of developmental reprogramming in the moss Physcomitrella patens. J. Proteomics 2014, 108, 284–294. [Google Scholar] [CrossRef] [PubMed]
- Khandelwal, A.; Cho, S.H.; Marella, H.; Sakata, Y.; Perroud, P.F.; Pan, A.; Quatrano, R.S. Role of ABA and ABI3 in desiccation tolerance. Science 2010, 327, 546. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Landberg, K.; Pederson, E.R.; Viaene, T.; Bozorg, B.; Friml, J.; Jönsson, H.; Thelander, M.; Sundberg, E. The MOSS Physcomitrella patens reproductive organ development is highly organized, affected by the two SHI/STY genes and by the level of active auxin in the SHI/STY expression domain. Plant Physiol. 2013, 162, 1406–1419. [Google Scholar] [CrossRef] [Green Version]
- Haro, R.; Fraile-Escanciano, A.; González-Melendi, P.; Rodríguez-Navarro, A. The potassium transporters HAK2 and HAK3 localize to endomembranes in Physcomitrella patens. HAK2 is required in some stress conditions. Plant Cell Physiol. 2013, 54, 1441–1454. [Google Scholar] [CrossRef] [Green Version]
- Xu, F.; Zhang, D.W.; Zhu, F.; Tang, H.; Lv, X.; Cheng, J.; Xie, H.F.; Lin, H.H. A novel role for cyanide in the control of cucumber (Cucumis sativus L.) seedlings response to environmental stress. Plant Cell Environ. 2012, 35, 1983–1997. [Google Scholar] [CrossRef]
- Li, Q.; Zhang, X.; Lv, Q.; Zhu, D.; Qiu, T.; Xu, Y.; Bao, F.; He, Y.; Hu, Y. Physcomitrella Patens dehydrins (PpDHNA and PpDHNC) confer salinity and drought tolerance to transgenic Arabidopsis plants. Front. Plant Sci. 2017, 8, 1316. [Google Scholar] [CrossRef] [Green Version]
- Bates, L.S.; Waldren, R.P.; Teare, I.D. Rapid determination of free proline for water-stress studies. Plant Soil 1973, 39, 205–207. [Google Scholar] [CrossRef]
- Narawongsanont, R.; Kabinpong, S.; Auiyawong, B.; Tantitadapitak, C. Cloning and characterization of AKR4C14, a rice aldo-keto reductase, from Thai Jasmine rice. Protein J. 2012, 31, 35–42. [Google Scholar] [CrossRef]
Substrate | Km (mM) | Kcat (min−1) | kcat/Km (min−1 mM−1) |
---|---|---|---|
Aldehydes | |||
Methylglyoxal | 0.2262 ± 0.063 | 51.6 ± 3.3 | 228.6 |
Glyoxal | 1.526 ± 0.237 | 3.78 ± 2.46 | 3.78 |
Acrolein | 6.2 ± 1.114 | 33.42 ± 1.86 | 5.34 |
Sugars | |||
Glucose | nd | nd | nd |
Xylose | nd | nd | nd |
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Chen, L.; Bao, F.; Tang, S.; Zuo, E.; Lv, Q.; Zhang, D.; Hu, Y.; Wang, X.; He, Y. PpAKR1A, a Novel Aldo-Keto Reductase from Physcomitrella Patens, Plays a Positive Role in Salt Stress. Int. J. Mol. Sci. 2019, 20, 5723. https://doi.org/10.3390/ijms20225723
Chen L, Bao F, Tang S, Zuo E, Lv Q, Zhang D, Hu Y, Wang X, He Y. PpAKR1A, a Novel Aldo-Keto Reductase from Physcomitrella Patens, Plays a Positive Role in Salt Stress. International Journal of Molecular Sciences. 2019; 20(22):5723. https://doi.org/10.3390/ijms20225723
Chicago/Turabian StyleChen, Lu, Fang Bao, Shuxuan Tang, Enhui Zuo, Qiang Lv, Dongyang Zhang, Yong Hu, Xiaoqin Wang, and Yikun He. 2019. "PpAKR1A, a Novel Aldo-Keto Reductase from Physcomitrella Patens, Plays a Positive Role in Salt Stress" International Journal of Molecular Sciences 20, no. 22: 5723. https://doi.org/10.3390/ijms20225723