Snakin-2 interacts with cytosolic glyceraldehyde-3-phosphate dehydrogenase 1 to inhibit sprout growth in potato tubers

Abstract

The potato tuber is the main nutrient supply and reproductive organ; however, tuber sprouting can reduce its commercial value. Snakin-2 (StSN2) was first reported as an antimicrobial peptide that positively regulates potato disease resistance. Our recent study suggested StSN2 overexpression inhibited sprout growth, while the sprouting process was accelerated in StSN2 RNAi lines. Cytoplasmic glyceraldehyde-3-phosphate dehydrogenase 1 (StGAPC1) was identified as a candidate protein that interacts with StSN2 in co-immunoprecipitation/mass spectrometry experiments. Here, we report that the expression levels of StSN2 and StGAPC1 decreased during sprouting compared with dormancy. Coexpression of StSN2 and StGAPC1 in bud eyes and apical buds was verified by immunofluorescence analysis of paraffin sections. In addition, interaction of StSN2 and StGAPC1 was confirmed by yeast two- hybrid, co-immunoprecipitation, and split luciferase complementation assays. Overexpression of StGAPC1 depressed sprout growth, which is similar to the function of StSN2 , and StSN2 - and StGAPC1 -overexpressing lines showed decreased glucose, fructose, and galactose contents. The interaction of StSN2 and StGAPC1 enhanced StGAPC1 activity and decreased its oxidative modification to inhibit sprout growth. Our results suggest that StSN2 plays a regulatory role in tuber sprout growth through interaction with StGAPC1.

References

[1] Sonnewald S, Sonnewald U . Regulation of potato tuber sprouting. Planta. 2014; 239: 27-38.

[2] Debast S, Nunes-Nesi A, Hajirezaei MR et al. Altering trehalose-6-phosphate content in transgenic potato tubers affects tuber growth and alters responsiveness to hormones during sprouting. Plant Physiol. 2011; 156: 1754-71.

[3] Pasare SA, Ducreux LJM, Morris WL et al. The role of the potato (Solanum tuberosum) CCD8 gene in stolon and tuber development. New Phytol. 2013; 198: 1108-20.

[4] Morris WL, Alamar MC, Lopez-Cobollo RM et al. A member of the TERMINAL FLOWER 1/CENTRORADIALIS gene family controls sprout growth in potato tubers. J Exp Bot. 2018; 70: 835-43.

[5] Hartmann A, Senning M, Hedden P et al. Reactivation of meristem activity and sprout growth in potato tubers require both cytokinin and gibberellin. Plant Physiol. 2011; 155: 776-96.

[6] Nahirñak V, Almasia NI, Hopp HE et al. Snakin/GASA proteins: involvement in hormone crosstalk and redox homeostasis. Plant Signal Behavior. 2012; 7: 1004-8.

[7] Zhong CM, Xu H, Ye S et al. Gibberellic Acid-Stimulated Arabidopsis6 serves as an integrator of gibberellin, abscisic acid, and glucose signaling during seed germination in Arabidopsis. Plant Physiol. 2015; 169: 2288-303.

[8] Berrocal-Lobo M, Segura A, Moreno M et al. Snakin-2, an antimicrobial peptide from potato whose gene is locally induced by wounding and responds to pathogen infection. Plant Physiol. 2002; 128: 951-61.

[9] Mohan S, Meiyalaghan S, Latimer JM et al. GSL2 overexpression confers resistance to Pectobacterium atrosepticum in potato. Theor Appl Genet. 2014; 127: 677-89.

[10] Li LQ, Zou X, Deng MS et al. Comparative morphology, transcription, and proteomics study revealing the key molecular mechanism of camphor on the potato tuber sprouting effect. Int J Mol Sci. 2017; 18: e2280.

[11] Liu B, Zhang N, Zhao S et al. Proteomic changes during tuber dormancy release process revealed by iTRAQ quantitative proteomics in potato. Plant Physiol Biochem. 2015; 86: 181-90.

[12] Deng MS, Peng J, Zhang J et al. The cysteine-rich peptide snakin-2 negatively regulates tubers sprouting through modulating lignin biosynthesis and H₂O₂ accumulation in potato . Int J Mol Sci. 2021; 22: 2287.

[13] Yang Y, Kwon HB, Peng HP et al. Stress responses and metabolic regulation of glyceraldehyde-3-phosphate dehydrogenase genes in Arabidopsis. Plant Physiol. 1993; 101: 209-16.

[14] Sirover MA . Subcellular dynamics of multifunctional protein regulation: mechanisms of GAPDH intracellular translocation. J Cell Biochemistry. 2012; 113: 2193-200.

[15] Bertomeu JM, Cascales-Miñana B, Mulet JM et al. Plastidial glyceraldehyde-3-phosphate dehydrogenase deficiency leads to altered root development and affects the sugar and amino acid balance in Arabidopsis. Plant Physiol. 2009; 151: 541-58.

[16] Liu TF, Fang H, Liu J et al. Cytosolic glyceraldehyde-3-phosphate dehydrogenases play crucial roles in controlling cold-induced sweetening and apical dominance of potato (Solanum tuberosum L.) tubers. Plant Cell Environ. 2017; 40: 3043-54.

[17] Wang XY, Li J, Zou X et al. Ectopic expression of AtCIPK23 enhances tolerance against low-K⁺ stress in transgenic potato . Am J Potato Res. 2010; 88: 153-9.

[18] Hajirezaei MR, Takahata Y, Trethewey RN et al. Impact of elevated cytosolic and apoplastic invertase activity on carbon metabolism during potato tuber development. J Exp Bot. 2000; 51: 439-45.

[19] Bethke PC, Sabba R, Bussan AJ et al. Tuber water and pressure potentials decrease and sucrose contents increase in response to moderate drought and heat stress. Am J Potato Res. 2009; 86: 519-32.

[20] Rius SP, Casati P, Iglesias AA et al. Characterization of Arabidopsis lines deficient in GAPC-1, a cytosolic NAD-dependent glyceraldehyde-3-phosphate dehydrogenase. Plant Physiol. 2008; 148: 1655-67.

[21] Xu J, Li HD, Chen LQ et al. A protein kinase, interacting with two calcineurin b-like proteins, regulates K+ transporter AKT1 in Arabidopsis. Cell. 2006; 125: 1347-60.

[22] Sonnewald U. Control of potato tuber sprouting. Trends Plant Sci. 2001; 6: 333-5.

[23] Rubinovich L, Ruthstein S, Weiss D et al. The Arabidopsis cysteine-rich GASA5 is a redox-active metalloprotein that suppresses gibberellin responses. Mol Plant. 2014; 7: 244-7.

[24] Roxrud I, Lid SE, Fletcher JC et al. GASA4, one of the 14-member Arabidopsis GASA family of small polypeptides, regulates flowering and seed development. Plant Cell Physiol. 2007; 48: 471-83.

[25] Nahirñak V, Almasia NI, Fernandez PV et al. Potato Snakin-1 gene silencing affects cell division, primary metabolism, and cell wall composition. Plant Physiol. 2012; 158: 252-63.

[26] Annadana S, Mlynárová L, Udayakumar M et al. The potato Lhca3.St.1 promoter confers high and stable transgene expression in chrysanthemum, in contrast to CaMV-based promoters. Mol Breeding. 2001; 8: 335-44.

[27] Nahirñak V, Rivarola M, Almasia NI et al. Snakin-1 affects reactive oxygen species and ascorbic acid levels and hormone balance in potato. PLoS One. 2019; 14: 1-18.

[28] Hajirezaei MR, Biemelt S, Peisker M et al. The influence of cytosolic phosphorylating glyceraldehyde 3-phosphate dehydrogenase (GAPC) on potato tuber metabolism. J Exp Bot. 2006; 57: 2363-77.

[29] Wiberley-Bradford AE, Bethke PC . Suppression of the vacuolar invertase gene delays senescent sweetening in chipping potatoes. J Sci Food Agric. 2018; 98: 354-60.

[30] Deryabin AN, Trunova TI . The physiological and biochemical mechanisms providing the increased constitutive cold resistance in the potato plants, expressing the yeast SUC2 gene encoding apoplastic invertase. J Stress Physiol Biochem. 2016; 12: 39-52.

[31] Ceserani T, Trofka A, Gandotra N et al. VH1/BRL2 receptor-like kinase interacts with vascular-specific adaptor proteins VIT and VIK to influence leaf venation. Plant J. 2009; 57: 1000-14.

[32] Wang L, Wang Z, Xu Y et al. OsGSR1 is involved in crosstalk between gibberellins and brassinosteroids in rice. Plant J. 2009; 57: 498-510.

[33] Backhausen JE, Vetter S, Baalmann E et al. NAD-dependent malate dehydrogenase and glyceraldehyde 3-phosphate dehydrogenase isoenzymes play an important role in dark metabolism of various plastid types. Planta. 1998; 205: 359-66.

[34] Piattoni C, Ferrero DML, Dellaferrera I et al. Cytosolic glyceraldehyde-3-phosphate dehydrogenase is phosphorylated during seed development. Front Plant Sci. 2017; 8: 522.

[35] Schneider M, Knuesting J, Birkholz O et al. Cytosolic GAPDH as a redox-dependent regulator of energy metabolism. BMC Plant Biol. 2018; 18: 184.

[36] Harris PWR, Yang SH, Molina A et al. Plant antimicrobial peptides Snakin-1 and Snakin-2: chemical synthesis and insights into the disulfide connectivity. Chemistry European Journal. 2014; 20: 5102-10.

[37] Rubinovich L, Weiss D . The Arabidopsis cysteine-rich protein GASA4 promotes GA responses and exhibits redox activity in bacteria and in planta. Plant J. 2010; 64: 1018-27.

[38] Sun S, Wang H, Yu H et al. GASA14 regulates leaf expansion and abiotic-stress resistance by modulating reactive oxygen species accumulation. J Exp Bot. 2013; 64: 1637-47.

[39] Baek D, Jin Y, Jeong JC et al. Suppression of reactive oxygen species by glyceraldehyde-3-phosphate dehydrogenase. Phytochemistry. 2008; 69: 333-8.

[40] Liu BL, Zhao S, Tan F et al. Changes in ROS production and antioxidant capacity during tuber sprouting in potato. Food Chem. 2017; 237: 205-13.

[41] Wawer I, Bucholc M, Astier J et al. Regulation of Nicotiana tabacum osmotic stress-activated protein kinase and its cellular partner GAPDH by nitric oxide in response to salinity. Biochem J. 2010; 429: 73-83.

[42] Kim SC, Guo L, Wang X et al. Phosphatidic acid binds to cytosolic glyceraldehyde-3-phosphate dehydrogenase and promotes its cleavage in Arabidopsis. J Biol Chem. 2013; 288: 11834-44.

[43] Aroca A, Schneider M, Scheibe R et al. Hydrogen sulfide regulates the cytosolic/nuclear partitioning of glyceraldehyde-3-phosphate dehydrogenase by enhancing its nuclear localization. Plant Cell Physiol. 2017; 58: 983-92.

[44] Vescovi M, Zaffagnini M, Festa M et al. Nuclear accumulation of cytosolic glyceraldehyde-3-phosphate dehydrogenase in cadmium stressed Arabidopsis roots. Plant Physiol. 2013; 162: 333-246.

[45] Zhang H, Zhao Y, Zhou DX et al. Rice NAD⁺-dependent histone deacetylase OsSRT1 represses glycolysis and regulates the moonlighting function of GAPDH as a transcriptional activator of glycolytic genes . Nucleic Acids Res. 2017; 45: 12241-55.

[46] Kim SC, Guo L, Wang X et al. Nuclear moonlighting of cytosolic glyceraldehyde-3-phosphate dehydrogenase regulates Arabidopsis response to heat stress. Nat Commun. 2020; 11: 3439.

[47] Zaffagnini M, Fermani S, Costa A et al. Plant cytoplasmic GAPDH: redox post-translational modifications and moonlighting properties. Front Plant Sci. 2013; 4: 450.

[48] Peralta DA, Araya A, Busi MV et al. The E3 ubiquitin-ligase SEVEN IN ABSENTIA like 7 mono-ubiquitinates glyceraldehyde-3-phosphate dehydrogenase 1 isoform in vitro and is required for its nuclear localization in Arabidopsis thaliana. Int J Biochem Cell Biol. 2016; 70: 48-56.