木薯MeRS40蛋白参与调控植物盐胁迫应答机制研究

doi:10.1016/j.jia.2023.04.003

Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (5): 1396-1411.DOI: 10.1016/j.jia.2023.04.003

木薯MeRS40蛋白参与调控植物盐胁迫应答机制研究

收稿日期:2022-08-31 接受日期:2023-02-23 出版日期:2023-05-20 发布日期:2023-02-23

Cassava MeRS40 is required for the regulation of plant salt tolerance

MA Xiao-wen¹, MA Qiu-xiang², MA Mu-qing¹, CHEN Yan-hang^{1, 3}, GU Jin-bao^{1, 3}, LI Yang^{1, 3}, HU Qing¹, LUO Qing-wen³, WEN Ming-fu³, ZHANG Peng², LI Cong^{1, 3#}, WANG Zhen-yu^{1, 3#}

¹Institute of Nanfan & Seed Industry, Guangdong Academy of Sciences, Guangzhou 510316, P.R.China
²National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 200032, P.R.China
³Zhanjiang Research Center, Institute of Nanfan & Seed Industry, Guangdong Academy of Sciences, Zhanjiang 524300, P.R.China

Received:2022-08-31 Accepted:2023-02-23 Online:2023-05-20 Published:2023-02-23
About author:MA Xiao-wen, E-mail: ma-xiaowen@outlook.com; #Correspondence WANG Zhen-yu, E-mail: wangzy80@126.com; LI Cong, E-mail: licong202103@163.com
Supported by:
This work was supported by grants from the Talent Program of Guangdong Academy of Sciences, China (2021GDASYL-20210103038, 2020GDASYL-2020102011, and 2021GDASYL-20210103036), the National Natural Science Foundation of China (32171292 and 32100294), the Guangdong Pearl River Talents Program, China (2021CX02N173), the China Postdoctoral Science Foundation (2020M682629), and the Zhanjiang Plan for Navigation, China (211207157080997).

摘要/Abstract

摘要：

盐胁迫下选择性剪接可调控丝氨酸/精氨酸丰富(SR)蛋白的表达和异构体的形成。前期研究鉴定了木薯SR蛋白家族中的两个亚家族SCL和 SR，这两个亚家族参与调控植物非生物胁迫的响应，然而SR蛋白家族中的其他亚家族是否也在转录后水平上调控植物盐胁迫应答有待探究。本研究通过11个物种RS亚家族的同源性比对找到37个基因，并系统性的分析了RS40 和 RS31基因在非生物胁迫条件下的表达情况。进一步蛋白结构域分析表明植物RS亚家族在非生物胁迫响应中其作用可能是保守的。在拟南芥中过表达MeRS40基因可通过维持活性氧的动态平衡和调控盐胁迫响应基因的表达进而提高植物的耐盐性。然而，在木薯中过表达MeRS40基因则通过负调节自身的pre-mRNA来抑制其内源性基因表达，从而降低转基因木薯的耐盐性。此外，MeRS40蛋白与木薯MeU1-70Ks（MeU1-70Ka 和 MeU1-70Kb）蛋白在体内和体外互作。因此，我们的研究为木薯SR蛋白参与调控盐胁迫应答提供了新的理论基础和探索方向。

Abstract:

Soil salinity affects the expression of serine/arginine-rich (SR) genes and isoforms by alternative splicing, which in turn regulates the adaptation of plants to stress. We previously identified the cassava spliceosomal component 35 like (SCL) and SR subfamilies, belonging to the SR protein family, which are extensively involved in responses to abiotic stresses. However, the post-transcriptional regulatory mechanism of cassava arginine/serine-rich (RS) subfamily in response to salt stress remains to be explored. In the current study, we identified 37 genes of the RS subfamily from 11 plant species and systematically investigated the transcript levels of the RS40 and RS31 genes under diverse abiotic stress conditions. Subsequently, an analysis of the conserved protein domains revealed that plant RS subfamily genes were likely to preserve their conserved molecular functions and played critical functional roles in responses to abiotic stresses. Importantly, we found that overexpression of MeRS40 in Arabidopsis enhanced salt tolerance by maintaining reactive oxygen species homeostasis and up-regulating the salt-responsive genes. However, overexpression of MeRS40 gene in cassava reduced salt tolerance due to the depression of its endogenous gene expression by negative autoregulation of its own pre-mRNA. Moreover, the MeRS40 protein interacted with MeU1-70Ks (MeU1-70Ka and MeU1-70Kb) in vivo and in vitro, respectively. Therefore, our ﬁndings highlight the critical role of cassava SR proteins in responses to salt stress in plants.

Key words: cassava , alternative splicing , serine/arginine-rich proteins , salt stress

MA Xiao-wen, MA Qiu-xiang, MA Mu-qing, CHEN Yan-hang, GU Jin-bao, LI Yang, HU Qing, LUO Qing-wen, WEN Ming-fu, ZHANG Peng, LI Cong, WANG Zhen-yu. 木薯MeRS40蛋白参与调控植物盐胁迫应答机制研究[J]. Journal of Integrative Agriculture, 2023, 22(5): 1396-1411.

MA Xiao-wen, MA Qiu-xiang, MA Mu-qing, CHEN Yan-hang, GU Jin-bao, LI Yang, HU Qing, LUO Qing-wen, WEN Ming-fu, ZHANG Peng, LI Cong, WANG Zhen-yu.

Cassava MeRS40 is required for the regulation of plant salt tolerance [J]. Journal of Integrative Agriculture, 2023, 22(5): 1396-1411.

参考文献

Albaqami M, Laluk K, Reddy A S N. 2019. The Arabidopsis splicing regulator SR45 confers salt tolerance in a splice isoform-dependent manner. Plant Molecular Biology, 100, 379–390.

Ali G S, Prasad K V, Hanumappa M, Reddy A S. 2008. Analyses of in vivo interaction and mobility of two spliceosomal proteins using FRAP and BiFC. PLoS ONE, 3, e1953.

An D, Yang J, Zhang P. 2012. Transcriptome profiling of low temperature-treated cassava apical shoots showed dynamic responses of tropical plant to cold stress. BMC Genomics, 13, 64.

Barta A, Kalyna M, Reddy A S. 2010. Implementing a rational and consistent nomenclature for serine/arginine-rich protein splicing factors (SR proteins) in plants. The Plant Cell, 22, 2926–2929.

Berg M G, Singh L N, Younis I, Liu Q, Pinto A M, Kaida D, Zhang Z, Cho S, Sherrill-Mix S, Wan L, Dreyfuss G. 2012. U1 snRNP determines mRNA length and regulates isoform expression. Cell, 150, 53–64.

Bourgeois C F, Lejeune F, Stevenin J. 2004. Broad specificity of SR (serine/arginine) proteins in the regulation of alternative splicing of pre-messenger RNA. Progress in Nucleic Acid Research and Molecular Biology, 78, 37–88.

Bredeson J V, Lyons J B, Prochnik S E, Wu G A, Ha C M, Edsinger-Gonzales E, Grimwood J, Schmutz J, Rabbi I Y, Egesi C, Nauluvula P, Lebot V, Ndunguru J, Mkamilo G, Bart R S, Setter T L, Gleadow R M, Kulakow P, Ferguson M E, Rounsley S, et al. 2016. Sequencing wild and cultivated cassava and related species reveals extensive interspecific hybridization and genetic diversity. Nature Biotechnology, 34, 562–570.

Bull S E, Owiti J A, Niklaus M, Beeching J R, Gruissem W, Vanderschuren H. 2009. Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava. Nature Protocols, 4, 1845–1854.

Carvalho R F, Carvalho S D, Duque P. 2010. The plant-specific SR45 protein negatively regulates glucose and ABA signaling during early seedling development in Arabidopsis. Plant Physiology, 154, 772–783.

Carvalho R F, Szakonyi D, Simpson C G, Barbosa I C, Brown J W, Baena-Gonzalez E, Duque P. 2016. The Arabidopsis SR45 splicing factor, a negative regulator of sugar signaling, modulates SNF1-related protein kinase 1 stability. The Plant Cell, 28, 1910–1925.

Chen L, Yang D, Zhang Y, Wu L, Zhang Y, Ye L, Pan C, He Y, Huang L, Ruan Y L, Lu G. 2018. Evidence for a specific and critical role of mitogen-activated protein kinase 20 in uni-to-binucleate transition of microgametogenesis in tomato. The New Phytologist, 219, 176–194.

Chen T, Cui P, Chen H, Ali S, Zhang S, Xiong L. 2013. A KH-domain RNA-binding protein interacts with FIERY2/CTD phosphatase-like 1 and splicing factors and is important for pre-mRNA splicing in Arabidopsis. PLoS Genetics, 9, e1003875.

Chen Y, Weng X, Zhou X, Gu J, Hu Q, Luo Q, Wen M, Li C, Wang Z Y. 2022. Overexpression of cassava RSZ21b enhances drought tolerance in Arabidopsis. Journal of Plant Physiology, 268, 153574.

Clough S J, Bent A F. 1998. Floral dip: A simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. The Plant Journal, 16, 735–743.

Duque P. 2011. A role for SR proteins in plant stress responses. Plant Signaling & Behavior, 6, 49–54.

Feng J, Li J, Gao Z, Lu Y, Yu J, Zheng Q, Yan S, Zhang W, He H, Ma L, Zhu Z. 2015. SKIP confers osmotic tolerance during salt stress by controlling alternative gene splicing in Arabidopsis. Molecular Plant, 8, 1038–1052.

Fica S M, Nagai K. 2017. Cryo-electron microscopy snapshots of the spliceosome: Structural insights into a dynamic ribonucleoprotein machine. Nature Structural & Molecular Biology, 24, 791–799.

Golovkin M, Reddy A S. 1999. An SC35-like protein and a novel serine/arginine-rich protein interact with Arabidopsis U1-70K protein. The Journal of Biological Chemistry, 274, 36428–36438.

Graveley B R. 2000. Sorting out the complexity of SR protein functions. RNA, 6, 1197–1211.

Graveley B R, Hertel K J, Maniatis T. 1999. SR proteins are ‘locators’ of the RNA splicing machinery. Current Biology, 9, R6–R13.

Gu J, Ma S, Zhang Y, Wang D, Cao S, Wang Z Y. 2020. Genome-wide identification of cassava serine/arginine-rich proteins: Insights into alternative splicing of pre-mRNAs and response to abiotic stress. Plant & Cell Physiology, 61, 178–191.

Gu J, Xia Z, Luo Y, Jiang X, Qian B, Xie H, Zhu J K, Xiong L, Zhu J, Wang Z Y. 2018. Spliceosomal protein U1A is involved in alternative splicing and salt stress tolerance in Arabidopsis thaliana. Nucleic Acids Research, 46, 1777–1792.

Hu Q, Chen Y, Zhao Y, Gu J, Ma M, Li H, Li C, Wang Z Y. 2021. Overexpression of SCL30A from cassava (Manihot esculenta) negatively regulates salt tolerance in Arabidopsis. Functional Plant Biology, 48, 1213–1224.

Isshiki M, Tsumoto A, Shimamoto K. 2006. The serine/arginine-rich protein family in rice plays important roles in constitutive and alternative splicing of pre-mRNA. Plant Cell, 18, 146–158.

Jansson C, Westerbergh A, Zhang J, Hu X, Sun C. 2009. Cassava, a potential biofuel crop in (the) People’s Republic of China. Applied Energy, 86, S95–S99.

Johnson T L, Vilardell J. 2012. Regulated pre-mRNA splicing: The ghostwriter of the eukaryotic genome. Biochimica et Biophysica Acta, 1819, 538–545.

Kaida D, Berg M G, Younis I, Kasim M, Singh L N, Wan L, Dreyfuss G. 2010. U1 snRNP protects pre-mRNAs from premature cleavage and polyadenylation. Nature, 468, 664–668.

Kalyna M, Lopato S, Voronin V, Barta A. 2006. Evolutionary conservation and regulation of particular alternative splicing events in plant SR proteins. Nucleic Acids Research, 34, 4395–4405.

Kastner B, Will C L, Stark H, Luhrmann R. 2019. Structural insights into nuclear pre-mRNA splicing in higher eukaryotes. Cold Spring Harbor Perspectives in Biology, 11, a032417.

Kumar K, Sinha S K, Maity U, Kirti P B, Kumar K R R. 2022. Insights into established and emerging roles of SR protein family in plants and animals. Wiley Interdiscip Reviews: RNA, e1763.

Kumar S, Stecher G, Tamura K. 2016. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33, 1870–1874.

Letunic I, Khedkar S, Bork P. 2021. SMART: Recent updates, new developments and status in 2020. Nucleic Acids Research, 49, D458–D460.

Li Y, Guo Q, Liu P, Huang J, Zhang S, Yang G, Wu C, Zheng C, Yan K. 2021. Dual roles of the serine/arginine-rich splicing factor SR45a in promoting and interacting with nuclear cap-binding complex to modulate the salt stress response in Arabidopsis. The New Phytologist, 230, 641–655.

Lopato S, Forstner C, Kalyna M, Hilscher J, Langhammer U, Indrapichate K, Lorkovic Z J, Barta A. 2002. Network of interactions of a novel plant-specific Arg/Ser-rich protein, atRSZ33, with atSC35-like splicing factors. The Journal of Biological Chemistry, 277, 39989–39998.

Lopato S, Kalyna M, Dorner S, Kobayashi R, Krainer A R, Barta A. 1999. AtSRp30, one of two SF2/ASF-like proteins from Arabidopsis thaliana, regulates splicing of specific plant genes. Genes & Development, 13, 987–1001.

Lorković Z J, Barta A. 2002. Genome analysis: RNA recognition motif (RRM) and K homology (KH) domain RNA-binding proteins from the flowering plant Arabidopsis thaliana. Nucleic Acids Research, 30, 623–635.

Magnani E, Bartling L, Hake S. 2006. From gateway to multisite gateway in one recombination event. BMC Molecular Biology, 7, 46.

Matera A G,Wang Z. 2014. A day in the life of the spliceosome. Molecular Cell Biology, 15, 108–121.

McCallum E J, Anjanappa R B, Gruissem W. 2017. Tackling agriculturally relevant diseases in the staple crop cassava (Manihot esculenta). Current Opinion in Plant Biology, 38, 50–58.

Morante N, Sánchez T, Ceballos H, Calle F, Pérez J C, Egesi C, Cuambe C E, Escobar A F, Ortiz D, Chávez A L, Fregene M. 2010. Tolerance to postharvest physiological deterioration in cassava roots. Crop Science, 50, 1333–1338.

Morton M, AlTamimi N, Butt H, Reddy A S N, Mahfouz M. 2019. Serine/Arginine-rich protein family of splicing regulators: New approaches to study splice isoform functions. Plant Science, 283, 127–134.

Muthusamy M, Yoon E K, Kim J A, Jeong M J, Lee S I. 2020. Brassica rapa SR45a regulates drought tolerance via the alternative splicing of target genes. Genes, 11, 2073–4425.

Palusa S G, Ali G S, Reddy A S. 2007. Alternative splicing of pre-mRNAs of Arabidopsis serine/arginine-rich proteins: regulation by hormones and stresses. The Plant Journal, 49, 1091–1107.

Plaschka C, Newman A J, Nagai K. 2019. Structural basis of nuclear pre-mRNA splicing: lessons from yeast. Cold Spring Harbor Perspectives in Biology, 11, a032391.

Quesada V, Macknight R, Dean C, Simpson G G. 2003. Autoregulation of FCA pre-mRNA processing controls Arabidopsis flowering time. The EMBO Journal, 22, 3142–3152.

Rauch H B, Patrick T L, Klusman K M, Battistuzzi F U, Mei W, Brendel V P, Lal S K. 2013. Discovery and expression analysis of alternative splicing events conserved among plant SR proteins. Molecular Biology and Evolution, 31, 605–613.

Reddy A S, Shad Ali G. 2011. Plant serine/arginine-rich proteins: Roles in precursor messenger RNA splicing, plant development, and stress responses. Wiley Interdisciplinary Reviews: RNA, 2, 875–889.

Schmal C, Reimann P, Staiger D. 2013. A circadian clock-regulated toggle switch explains AtGRP7 and AtGRP8 oscillations in Arabidopsis thaliana. PLoS Computational Biology, 9, e1002986.

Schoning J C, Streitner C, Page D R, Hennig S, Uchida K, Wolf E, Furuya M, Staiger D. 2007. Auto-regulation of the circadian slave oscillator component AtGRP7 and regulation of its targets is impaired by a single RNA recognition motif point mutation. The Plant Journal, 52, 1119–1130.

So B R, Di C, Cai Z, Venters C C, Guo J, Oh J M, Arai C, Dreyfuss G. 2019. A complex of U1 snRNP with cleavage and polyadenylation factors controls telescripting, regulating mRNA transcription in human cells. Molecular Cell, 76, 590–599.

Sperschneider J, Catanzariti A M, DeBoer K, Petre B, Gardiner D M, Singh K B, Dodds P N, Taylor J M. 2017. LOCALIZER: Subcellular localization prediction of both plant and effector proteins in the plant cell. Scientific Reports, 7, 44598.

Tanabe N, Kimura A, Yoshimura K, Shigeoka S. 2009. Plant-specific SR-related protein atSR45a interacts with spliceosomal proteins in plant nucleus. Plant Molecular Biology, 70, 241–252.

Tanabe N, Yoshimura K, Kimura A, Yabuta Y, Shigeoka S. 2007. Differential expression of alternatively spliced mRNAs of Arabidopsis SR protein homologs, atSR30 and atSR45a, in response to environmental stress. Plant & Cell Physiology, 48, 1036–1049.

Thomas J, Palusa S G, Prasad K V, Ali G S, Surabhi G K, Ben-Hur A, Abdel-Ghany S E, Reddy A S. 2012. Identification of an intronic splicing regulatory element involved in auto-regulation of alternative splicing of SCL33 pre-mRNA. The Plant Journal, 72, 935–946.

Thompson J D, Gibson T J, Plewniak F, Jeanmougin F, Higgins D G. 1997. The CLUSTAL_X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25, 4876–4882.

Thordal-Christensen H, Zhang Z, Wei Y, Collinge D B. 1997. Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley–powdery mildew interaction. The Plant Journal, 11, 1187–1194.

Weng X, Zhou X, Xie S, Gu J, Wang Z Y. 2021. Identification of cassava alternative splicing-related genes and functional characterization of MeSCL30 involvement in drought stress. Plant Physiology and Biochemistry, 160, 130–142.

Wu Z, Zhu D, Lin X, Miao J, Gu L, Deng X, Yang Q, Sun K, Zhu D, Cao X, Tsuge T, Dean C, Aoyama T, Gu H, Qu L J. 2016. RNA binding proteins RZ-1B and RZ-1C play critical roles in regulating pre-mRNA splicing and gene expression during development in Arabidopsis. The Plant Cell, 28, 55–73.

Yan Q, Xia X, Sun Z, Fang Y. 2017. Depletion of Arabidopsis SC35 and SC35-like serine/arginine-rich proteins affects the transcription and splicing of a subset of genes. PLoS Genetics, 13, e1006663.

Yoshimura K, Mori T, Yokoyama K, Koike Y, Tanabe N, Sato N, Takahashi H, Maruta T, Shigeoka S. 2011. Identification of alternative splicing events regulated by an Arabidopsis serine/arginine-like protein, atSR45a, in response to high-light stress using a tiling array. Plant & Cell Physiology, 52, 1786–1805.

Zhang J, Sun Y, Zhou Z, Zhang Y, Yang Y, Zan X, Li X, Wan J, Gao X, Chen R, Huang Z, Li L, Xu Z. 2022. OsSCL30 overexpression reduces the tolerance of rice seedlings to low temperature, drought and salt. Scientific Reports, 12, 8385.

Zhang W, Du B, Liu D, Qi X. 2014. Splicing factor SR34b mutation reduces cadmium tolerance in Arabidopsis by regulating iron-regulated transporter 1 gene. Biochemical and Biophysical Research Communications, 455, 312–317.

Zhu J K. 2003. Regulation of ion homeostasis under salt stress. Current Opinion in Plant Biology, 6, 441–445.

Zhu J K. 2016. Abiotic stress signaling and responses in plants. Cell, 167, 313–324.

木薯MeRS40蛋白参与调控植物盐胁迫应答机制研究

Cassava MeRS40 is required for the regulation of plant salt tolerance

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