Abstract
Cold shock domain proteins (CSPs), initially identified in Escherichia coli, have been demonstrated to play a positive role in cold resistance. Previous studies in wheat, rice, and Arabidopsis have indicated the functional conservation of CSPs in cold resistance between bacteria and higher plants. However, the biological functions of PbrCSPs in pear pollen tubes, which represent the fragile reproductive organs highly sensitive to low temperature, remain largely unknown. In this study, a total of 22 CSPs were identified in the seven Rosaceae species, with a focus on characterizing four PbrCSPs in pear (Pyrus bretschneideri Rehder). All four PbrCSPs were structurally conserved and responsive to the abiotic stresses, such as cold, high osmotic, and abscisic acid (ABA) treatments. PbrCSP1, which is specifically expressed in pear pollen tubes, was selected for further research. PbrCSP1 was localized in both the cytoplasm and nucleus. Suppressing the expression of PbrCSP1 significantly inhibited the pollen tube growth in vitro. Conversely, overexpression of PbrCSP1 promoted the growth of pear pollen tubes under the normal condition and, notably, under the cold environment at 4 °C. These findings highlight an essential role of PbrCSP1 in facilitating the normal growth and enhancing cold resistance in pear pollen tubes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data and materials used in this study are publicly available.
References
Bae WH, Xia B, Inouye M, Severinov K (2000) Escherichia coli CspA-family RNA chaperones are transcription antiterminators. Proc Natl Acad Sci U S A 97(14):7784–7789. https://doi.org/10.1073/pnas.97.14.7784
Chaikam V, Karlson D (2008) Functional characterization of two cold shock domain proteins from Oryza sativa. Plant Cell Environ 31(7):995–1006. https://doi.org/10.1111/j.1365-3040.2008.01811.x
Chaikam V, Karlson DT (2010) Comparison of structure, function and regulation of plant cold shock domain proteins to bacterial and animal cold shock domain proteins. BMB Rep 43(1):1–8. https://doi.org/10.5483/bmbrep.2010.43.1.001
Chen J, Li XY, Wang DQ, Li LT, Zhou HS, Liu Z, Wu J, Wang P, Jiang XT, Fabrice MR et al (2015) Identification and testing of reference genes for gene expression analysis in pollen of Pyrus bretschneideri. Sci Hort 190:43–56. https://doi.org/10.1016/j.scienta.2015.04.010
Chen J, Wang P, de Graaf BHJ, Zhang H, Jiao H, Tang C, Zhang S, Wu J (2018) Phosphatidic acid counteracts S-RNase signaling in pollen by stabilizing the actin cytoskeleton. Plant Cell 30(5):1023–1039. https://doi.org/10.1105/tpc.18.00021
Chen J, Jing Y, Zhang X, Li L, Wang P, Zhang S, Zhou H, Wu J (2016) Evolutionary and expression analysis provides evidence for the plant glutamate-like receptors family is involved in woody growth-related function. Sci Rep, 6. https://doi.org/10.1038/srep32013
Choi MJ, Park YR, Park SJ, Kang H (2015) Stress-responsive expression patterns and functional characterization of cold shock domain proteins in cabbage (Brassica rapa) under abiotic stress conditions. Plant Physiol Biochem 96:132–140. https://doi.org/10.1016/j.plaphy.2015.07.027
Fu Y, Wu G, Yang Z (2001) Rop GTPase-dependent dynamics of tip-localized F-actin controls tip growth in pollen tubes. J Cell Biol 152:1019–1032. https://doi.org/10.1083/jcb.152.5.1019
Fusaro AF, Bocca SN, Ramos RLB, Barroco RM, Magioli C, Jorge VC, Coutinho TC, Rangel-Lima CM, De Rycke R, Inze D et al (2007) AtGRP2, a cold-induced nucleo-cytoplasmic RNA-binding protein, has a role in flower and seed development. Planta 225(6):1339–1351. https://doi.org/10.1007/s00425-006-0444-4
Gao YB, Wang CL, Wu JY, Zhou HS, Jiang XT, Wu J, Zhang SL (2014) Low temperature inhibits pollen tube growth by disruption of both tip-localized reactive oxygen species and endocytosis in Pyrus bretschneideri Rehd. Plant Physiol Biochem 74:255–262. https://doi.org/10.1016/j.plaphy.2013.11.018
Graumann PL, Marahiel MA (1998) A superfamily of proteins that contain the cold-shock domain. Trends Biochem Sci 23(8):286–290. https://doi.org/10.1016/s0968-0004(98)01255-9
Jiang WN, Hou Y, Inouye M (1997) CspA, the major cold-shock protein of Escherichia coli, is an RNA chaperone. J Biol Chem 272(1):196–202. https://doi.org/10.1074/jbc.272.1.196
Karlson D, Nakaminami K, Toyomasu T, Imai R (2002) A cold-regulated nucleic acid-binding protein of winter wheat shares a domain with bacterial cold shock proteins. J Biol Chem 277(38):35248–35256. https://doi.org/10.1074/jbc.M205774200
Kim JS, Park SJ, Kwak KJ, Kim YO, Kim JY, Song J, Jang B, Jung CH, Kang H (2007) Cold shock domain proteins and glycine-rich RNA-binding proteins from Arabidopsis thaliana can promote the cold adaptation process in Escherichia coli. Nucleic Acids Res 35(2):506–516. https://doi.org/10.1093/nar/gkl1076
Kim MH, Sasaki K, Imai R (2009) Cold shock domain protein 3 regulates freezing tolerance in Arabidopsis thaliana. J Biol Chem 284(35):23454–23460. https://doi.org/10.1074/jbc.M109.025791
Kim MH, Sato S, Sasaki K, Saburi W, Matsui H, Imai R (2013) COLD SHOCK DOMAIN PROTEIN 3 is involved in salt and drought stress tolerance in Arabidopsis. FEBS Open Bio 3:438–442. https://doi.org/10.1016/j.fob.2013.10.003
Li C, Hou N, Fang N, He J, Ma Z, Ma F, Guan Q, Li X (2021) Cold shock protein 3 plays a negative role in apple drought tolerance by regulating oxidative stress response. Plant Physiol Biochem 168:83–92. https://doi.org/10.1016/j.plaphy.2021.10.003
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
Mordovkina D, Lyabin DN, Smolin EA, Sogorina EM, Ovchinnikov LP, Eliseeva I (2020) Y-box binding proteins in mRNP assembly, translation, and stability control. Biomolecules, 10(4). https://doi.org/10.3390/biom10040591
Moutinho A, Camacho L, Haley A, Pais MS, Trewavas A, Malho R (2001) Antisense perturbation of protein function in living pollen tubes. Sex Plant Reprod 14(1–2):101–104. https://doi.org/10.1007/s004970100086
Nakaminami K, Sasaki K, Kajita S, Takeda H, Karlson D, Ohgi K, Imai R (2005) Heat stable ssDNA/RNA-binding activity of a wheat cold shock domain protein. FEBS Lett 579(21):4887–4891. https://doi.org/10.1016/j.febslet.2005.07.074
Nakaminami K, Karlson DT, Imai R (2006) Functional conservation of cold shock domains in bacteria and higher plants. Proc Natl Acad Sci U S A 103(26):10122–10127. https://doi.org/10.1073/pnas.0603168103
Park SJ, Kwak KJ, Oh TR, Kim YO, Kang H (2009) Cold shock domain proteins affect seed germination and growth of Arabidopsis thaliana under abiotic stress conditions. Plant Cell Physiol 50(4):869–878. https://doi.org/10.1093/pcp/pcp037
Phadtare S, Inouye M, Severinov K (2002) The nucleic acid melting activity of Escherichia coli CspE is critical for transcription antitermination and cold acclimation of cells. J Biol Chem 277(9):7239–7245. https://doi.org/10.1074/jbc.M111496200
Radkova M, Vitamvas P, Sasaki K, Imai R (2014) Development- and cold-regulated accumulation of cold shock domain proteins in wheat. Plant Physiol Biochem 77:44–48. https://doi.org/10.1016/j.plaphy.2014.01.004
Sasaki K, Imai R (2011) Pleiotropic roles of cold shock domain proteins in plants. Front Plant Sci 2:116. https://doi.org/10.3389/fpls.2011.00116
Sasaki K, Kim MH, Imai R (2007) Arabidopsis COLD SHOCK DOMAIN PROTEIN2 is a RNA chaperone that is regulated by cold and developmental signals. Biochem Biophys Res Commun 364(3):633–638. https://doi.org/10.1016/j.bbrc.2007.10.059
Sasaki K, Kim MH, Imai R (2013) Arabidopsis cold shock domain protein 2 is a negative regulator of cold acclimation. New Phytol 198(1):95–102. https://doi.org/10.1111/nph.12118
Sparkes IA, Runions J, Kearns A, Hawes C (2006) Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nat Protoc 1(4):2019–2025. https://doi.org/10.1038/nprot.2006.286
Tang C, Wang P, Zhu X et al (2023) Acetylation of inorganic pyrophosphatase by S-RNase signaling induces pollen tube tip swelling by repressing pectin methylesterase. Plant Cell 35(9):3544–3565. https://doi.org/10.1093/plcell/koad162
Wang N, Yamanaka K, Inouye M (1999) CspI, the ninth member of the CspA family of Escherichia coli, is induced upon cold shock. J Bacteriol 181(5):1603–1609. https://doi.org/10.1128/JB.181.5.1603-1609.1999
Wu J, Wang Z, Shi Z et al (2013) The genome of the pear (Pyrus bretschneideri Rehd.). Genome Res 23(2):396–408. https://doi.org/10.1101/gr.144311.112
Xia B, Ke HP, Inouye M (2001) Acquirement of cold sensitivity by quadruple deletion of the cspA family and its suppression by PNPase S1 domain in Escherichia coli. Mol Microbiol 40(1):179–188. https://doi.org/10.1046/j.1365-2958.2001.02372.x
Xie Q, Wang P, Liu X, Yuan L, Wang L, Zhang C, Li Y, Xing H, Zhi L, Yue Z et al (2014) LNK1 and LNK2 are transcriptional coactivators in the Arabidopsis circadian oscillator. Plant Cell 26(7):2843–2857. https://doi.org/10.1105/tpc.114.126573
Xie Y, Chen P, Yan Y, Bao C, Li X, Wang L, Shen X, Li H, Liu X, Niu C et al (2018) An atypical R2R3 MYB transcription factor increases cold hardiness by CBF-dependent and CBF-independent pathways in apple. New Phytol 218(1):201–218. https://doi.org/10.1111/nph.14952
Acknowledgements
We thank the Bioinformatics Center of Nanjing Agricultural University for supporting bioinformatic analysis. We thank Dr. Yuehua Ma (Central laboratory of College of Horticulture, Nanjing Agricultural University) for assistance in using laser scanning confocal microscope LSM800. We thank Dr. Barend H.J. de Graaf (Cardiff University) for offering us the vector NTP303:GFP.
Funding
This work was financially supported through grants from the open funds of the Jiangsu Agricultural Science and Technology Innovation Fund (CX(22)3161), National Natural Science Foundation of China (32172543, 32102358), the guidance foundation of Hainan Institute of Nanjing Agricultural University (NAUSY-MS08), Fundamental Research Fund for the Central Universities (YDZX2023019), National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops (Horti-KF-2023–05), Xinjiang Tianshan Innovation Team Foundation (2023D14015), Ningbo Key Laboratory of Characteristic Horticultural Crops in Quality Adjustment and Resistance Breeding (NBYYL2023001), Priority Academic Program Development of Jiangsu Higher Education Institutions, and Earmarked Fund for China Agriculture Research System (CARS-28), Natural Science Foundation of Jiangsu Province (BK20210394). This study was supported by the High-performance Computing Platform of the Bioinformatics Center, Nanjing Agricultural University.
Author information
Authors and Affiliations
Contributions
All authors contributed to the research planning and experimental designs. Juyou Wu, Shaoling Zhang, and Peng Wang initiated and supervised the project. Xiaoxuan Zhu, Chao Tang, and Ting Zhang performed the experiments and analyzed the data. Xiaoxuan Zhu and Chao Tang wrote the manuscript. All authors read and revised the manuscript.
Corresponding authors
Ethics declarations
Ethics approval
All methods in this research were carried out in accordance with relevant guidelines. No specific permits are required for sample collection in this study.
Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Additional information
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.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhu, X., Tang, C., Zhang, T. et al. PbrCSP1, a pollen tube–specific cold shock domain protein, is essential for the growth and cold resistance of pear pollen tubes. Mol Breeding 44, 18 (2024). https://doi.org/10.1007/s11032-024-01457-w
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11032-024-01457-w