, Volume 248, Issue 3, pp 675–690 | Cite as

Roles of γ-aminobutyric acid on salinity-responsive genes at transcriptomic level in poplar: involving in abscisic acid and ethylene-signalling pathways

  • Jing Ji
  • Jianyun Yue
  • Tiantian Xie
  • Wei Chen
  • Changjian Du
  • Ermei Chang
  • Lanzhen Chen
  • Zeping JiangEmail author
  • Shengqing ShiEmail author
Original Article


Main conclusion

γ-Aminobutyric acid (GABA) affected ABA and ethylene metabolic genes and signal components in salt-treated poplar, indicating its potential role in signal pathways of ABA and ethylene during salt stress.

GABA is a small signalling molecule that accumulates rapidly in plants exposed to various stresses. However, the relationship between GABA and other signalling molecules, such as hormones, remains unclear. Here, in the poplar woody plant under 200-mM NaCl conditions, the application of low (0.25 mM) and high (10 mM) exogenous GABA, compared to 0 mM, affected the accumulation of hydrogen peroxide and hormones, including ABA and ethylene, in different manners. Transcriptomic analysis demonstrated that 1025 differentially expressed genes (DEGs; |log2Ratio| ≥ 1.5) were widely affected by exogenous GABA under salt stress. A clustering analysis revealed that GABA could rescue or promote the effects of salt stress on gene expression. Among them, 146 genes involved in six hormone-signalling pathways were enriched, including 22 ABA- and 50 ethylene-related genes. Quantitative expression of selected genes involved in hormone-related pathways showed that ABA metabolic genes (ABAG, ABAH2, and ABAH4), ethylene biosynthetic genes (ACO1, ACO2, ACO5, ACOH1, ACS1, and ACS7) and receptor genes (PYL1, PYL2, PYL4, and PYL6) were regulated by exogenous GABA, even at a 0.1 mM level. The production of ABA was negatively correlated with ABAH expression levels at different GABA concentrations. The increase of endogenous GABA, resulting from inhibitor (succinyl phosphonate) of α-ketoglutarate dehydrogenase, affected the PYLs levels. Thus, GABA may be involved in ABA- and ethylene-signalling pathways. Our data provide a better understanding of GABA’s roles in the plant responses to environmental stresses.


GABA Hormone Populus Salt stress Signal transduction 



Abscisate beta-glucosyltransferase


Abscisic acid 8-hydroxylase


1-Aminocyclopropane-1-carboxylate oxidase


1-Aminocyclopropene-1-carboxylate synthase


γ-Aminobutyric acid





We acknowledge the support from the Fundamental Research Funds for the Central Non-profit Research Institution of CAF (CAFYBB2014ZX001-3), and the National Natural Science Foundation of China (31100490). We thank Prof. Dr. Mengzhu Lu for the good suggestion during analyzing data, and Mrs. Lingyu Zheng for the help in parts of physiological measurements.

Supplementary material

425_2018_2915_MOESM1_ESM.tif (33.7 mb)
Fig. S1 Effects of exogenous GABA on superoxide dismutase (SOD) and peroxidase (POD) activities in poplar under 200-mM NaCl for 0, 6, and 24 h. SOD activities in leaves (a) and roots (b). POD activities in leaves (c) and roots (d). Vertical bars represent the mean ± SD of at least three replicated experiments. Data were analyzed using t test in the SPASS software 19 version. * indicates statistically significant differences between samples with and without GABA (P < 0.05) (TIFF 34510 kb)
425_2018_2915_MOESM2_ESM.tif (32.2 mb)
Fig. S2 Verification of six selected differentially expressed genes by qRT-PCR. Comparison of RNA-seq data with qRT-PCR data (red line). The normalized expression level (reads per kilobase per million reads) of the relative qRT-PCR expression level is indicated on the y-axis to the left. RNA-seq is shown on the y-axis to the right. Internal reference gene: Actin (POPTR_0019s02630g). Three biological replicates were performed (TIFF 32964 kb)
425_2018_2915_MOESM3_ESM.tif (12.5 mb)
Fig. S3 Endogenous GABA levels in poplar roots under the treatments of succinyl phosphonate (0, 50, 100, and 150 µM). Data were analyzed using t test in the SPASS software 19 version. * indicates statistically significant differences between samples with and without treatment (P < 0.05) (TIFF 12827 kb)
425_2018_2915_MOESM4_ESM.docx (20 kb)
Online Resource S1 Detailed experimental methods for RNA-seq and qRT-PCR (DOCX 19 kb)
425_2018_2915_MOESM5_ESM.docx (18 kb)
Table S1 Primers for qRT-PCR (DOCX 17 kb)
425_2018_2915_MOESM6_ESM.xlsx (14 kb)
Table S2 Summary of sequencing data (XLSX 13 kb)
425_2018_2915_MOESM7_ESM.xlsx (12 kb)
Table S3 Pearson correlation analysis of FPKM values between replicates for the treatments of poplar under 200-mM NaCl with the application of GABA (XLSX 11 kb)
425_2018_2915_MOESM8_ESM.xlsx (565 kb)
Table S4 Read counts for differentially expressed genes regulated by GABA in the roots of poplar under 200-mM NaCl (XLSX 564 kb)
425_2018_2915_MOESM9_ESM.xlsx (604 kb)
Table S5 Differentially expressed genes in poplar roots under 200-mM NaCl with the application of GABA (XLSX 604 kb)
425_2018_2915_MOESM10_ESM.xlsx (133 kb)
Table S6 Differentially expressed genes for clustering (XLSX 132 kb)
425_2018_2915_MOESM11_ESM.xlsx (108 kb)
Table S7 GO enrichments for differentially expressed genes regulated by GABA in poplar roots under 200-mM NaCl (XLSX 107 kb)
425_2018_2915_MOESM12_ESM.xlsx (68 kb)
Table S8 KEGG pathways for differentially expressed genes regulated by GABA in poplar roots under 200-mM NaCl (XLSX 67 kb)
425_2018_2915_MOESM13_ESM.xlsx (82 kb)
Table S9 Differentially expressed genes involved in hormone synthesis and signal transduction (XLSX 82 kb)
425_2018_2915_MOESM14_ESM.docx (22 kb)
Table S10 qRT-PCR values of selected ABA- and ethylene-related genes regulated by 0-, 0.25-, and 10-mM GABA in poplar roots under 0- and 200-mM NaCl for 6 h. Fold changes were calculated by comparing with 0-mM GABA without salinity. Internal reference gene: UBQL (POPTR_0005s22060g) (DOCX 21 kb)


  1. Achard P, Cheng H, De Grauwe L, Decat J, Schoutteten H, Moritz T, Van Der Straeten D, Peng J, Harberd NP (2006) Integration of plant responses to environmentally activated phytohormonal signals. Science 311:91–94CrossRefGoogle Scholar
  2. Aghdam MS, Naderi R, Jannatizadeh A, Babalar M, Sarcheshmeh MA, Faradonbe MZ (2016) Impact of exogenous GABA treatments on endogenous GABA metabolism in anthurium cut flowers in response to postharvest chilling temperature. Plant Physiol Biochem 106:11–15CrossRefGoogle Scholar
  3. Araújo WL, Nunes-Nesi A, Trenkamp S, Bunik VI, Fernie AR (2008) Inhibition of 2-oxoglutarate dehydrogenase in potato tuber suggests the enzyme is limiting for respiration and confirms its importance in nitrogen assimilation. Plant Physiol 148:1782–1796CrossRefGoogle Scholar
  4. Araújo WL, Tohge T, Nunes-Nesi A, Daloso DM, Nimick M, Krahnert I, Bunik VI, Moorhead GB, Fernie AR (2012) Phosphonate analogs of 2-oxoglutarate perturb metabolism and gene expression in illuminated Arabidopsis leaves. Front Plant Sci 3:114PubMedPubMedCentralGoogle Scholar
  5. Bao H, Chen X, Lv S, Jiang P, Feng J, Fan P, Nie L, Li Y (2015) Virus-induced gene silencing reveals control of reactive oxygen species accumulation and salt tolerance in tomato by γ-aminobutyric acid metabolic pathway. Plant Cell Environ 38:600–613CrossRefGoogle Scholar
  6. Batushansky A, Kirma M, Grillich N, Toubiana D, Pham PA, Balbo I, Fromm H, Galili G, Fernie AR, Fait A (2014) Combined transcriptomics and metabolomics of Arabidopsis thaliana seedlings exposed to exogenous GABA suggest its role in plants is predominantly metabolic. Mol Plant 7:1065–1068CrossRefGoogle Scholar
  7. Batushansky A, Kirma M, Grillich N, Pham PA, Rentsch D, Galili G, Fernie AR, Fait A (2015) The transporter GAT1 plays an important role in GABA-mediated carbon-nitrogen interactions in Arabidopsis. Front Plant Sci 6:785CrossRefGoogle Scholar
  8. Beuve N, Rispail N, Laine P, Cliquet J-B, Ourry A, Le Deunef E (2004) Putative role of γ-aminobutyric acid (GABA) as a long-distance signal in up-regulation of nitrate uptake in Brassica napus L. Plant Cell Environ 27:1035–1046CrossRefGoogle Scholar
  9. Bouché N, Fromm H (2004) GABA in plants: just a metabolite? Trends Plant Sci 9:110–115CrossRefGoogle Scholar
  10. Bouché N, Fait A, Bouchez D, Møller SG, Fromm H (2003) Mitochondrial succinic-semialdehyde dehydrogenase of the gamma-aminobutyrate shunt is required to restrict levels of reactive oxygen intermediates in plants. Proc Natl Acad Sci USA 100:6843–6848CrossRefGoogle Scholar
  11. Bown AW, Shelp BJ (2016) Plant GABA: not just a metabolite. Trends Plant Sci 21:811–813CrossRefGoogle Scholar
  12. Bown AW, Macgregor KB, Shelp BJ (2006) Gamma-aminobutyrate: defense against invertebrate pests. Trends Plant Sci 11:424–427CrossRefGoogle Scholar
  13. Chaiwanon J, Wang W, Zhu JY, Oh E, Wang ZY (2016) Information integration and communication in plant growth regulation. Cell 164:1257–1268CrossRefGoogle Scholar
  14. Cheng T, Chen J, Ef AA, Wang P, Wang G, Hu X, Shi J (2015) Quantitative proteomics analysis reveals that S-nitrosoglutathione reductase (GSNOR) and nitric oxide signaling enhance poplar defense against chilling stress. Planta 242:1361–1390CrossRefGoogle Scholar
  15. Cheng T, Shi J, Dong Y, Ma Y, Peng Y, Hu X, Chen J (2017) Hydrogen sulfide enhances poplar tolerance to high-temperature stress by increasing S-nitrosoglutathione reductase (GSNOR) activity and reducing reactive oxygen/nitrogen damage. Plant Growth Regul 9:1–13Google Scholar
  16. Cutler SR, Rodriguez PL, Finkelstein RR, Abrams SR (2010) Abscisic acid: emergence of a core signalling network. Annu Rev Plant Biol 61:651–679CrossRefGoogle Scholar
  17. Datta R, Kumar D, Sultana A, Hazra S, Bhattacharyya D, Chattopadhyay S (2015) Glutathione regulates 1-aminocyclopropane-1-carboxylate synthase transcription via WRKY33 and 1-aminocyclopropane-1-carboxylate oxidase by modulating messenger RNA stability to induce ethylene synthesis during stress. Plant Physiol 169:2963–2981PubMedPubMedCentralGoogle Scholar
  18. De Angeli A, Zhang J, Meyer S, Martinoia E (2013) AtALMT9 is a malate-activated vacuolar chloride channel required for stomatal opening in Arabidopsis. Nat Commun 4:1804CrossRefGoogle Scholar
  19. De Diego N, Sampedro MC, Barrio RJ, Saiz-Fernández I, Moncaleán P, Lacuesta M (2013) Solute accumulation and elastic modulus changes in six radiata pine breeds exposed to drought. Tree Physiol 33:69–80CrossRefGoogle Scholar
  20. Durley RC, Simpson GM (1982) Leaf analysis for abscisic, phaseic and 3-indolylacetic acids by high-performance liquid chromatography. J Chromatogr A 236:181–188CrossRefGoogle Scholar
  21. Fait A, Fromm H, Walter D, Galili G, Fernie AR (2008) Highway or byway: the metabolic role of the GABA shunt in plants. Trends Plant Sci 13:14–19CrossRefGoogle Scholar
  22. Fuchs S, Tischer SV, Wunschel C, Christmann A, Grill E (2014) Abscisic acid sensor RCAR7/PYL13, specific regulator of protein phosphatase coreceptors. Proc Natl Acad Sci USA 111:5741–5746CrossRefGoogle Scholar
  23. Gilliham M, Tyerman SD (2016) Linking metabolism to membrane signalling: the GABA–malate connection. Trends Plant Sci 21:295–301CrossRefGoogle Scholar
  24. Granger AJ, Mulder N, Saunders A, Sabatini BL (2016) Cotransmission of acetylcholine and GABA. Neuropharmacology 100:40–46CrossRefGoogle Scholar
  25. Kathiresan A, Tung P, Chinnappa CC, Reid DM (1997) γ-Aminobutyric acid stimulates ethylene biosynthesis in sunflower. Plant Physiol 11:129–135CrossRefGoogle Scholar
  26. Kinnersley AM, Lin F (2000) Receptor modifiers indicate that 4-aminobutyric acid (GABA) is a potential modulator of ion transport in plants. Plant Growth Regul 32:65–76CrossRefGoogle Scholar
  27. Kinnersley AM, Turano FJ (2000) Gamma-aminobutyric acid (GABA) and plant responses to stress. Crit Rev Plant Sci 19:479–509CrossRefGoogle Scholar
  28. Kobayashi M, Ohura I, Kawakita K, Yokota N, Fujiwara M, Shimamoto K, Doke N, Yoshioka H (2007) Calcium dependent protein kinases regulate the production of reactive oxygen species by potato NADPH oxidase. Plant Cell 19:1065–1080CrossRefGoogle Scholar
  29. Kumar D, Hazra S, Datta R, Chattopadhyay S (2016) Transcriptome analysis of Arabidopsis mutants suggests a crosstalk between ABA, ethylene and GSH against combined cold and osmotic stress. Sci Rep 6:36867CrossRefGoogle Scholar
  30. Li Z, Yu J, Peng Y, Huang B (2017) Metabolic pathways regulated by abscisic acid, salicylic acid and γ-aminobutyric acid in association with improved drought tolerance in creeping bentgrass (Agrostis stolonifera). Physiol Plant 159:42–58CrossRefGoogle Scholar
  31. Liu Z, Yan JP, Li DK, Luo Q, Yan Q, Liu ZB, Ye LM, Wang JM, Li XF, Yang Y (2015) UDP-glucosyltransferase 71c5, a major glucosyltransferase, mediates abscisic acid homeostasis in Arabidopsis. Plant Physiol 167:1659–1670CrossRefGoogle Scholar
  32. Ma Y, Szostkiewicz I, Korte A, Moes D, Yang Y, Christmann A, Grill E (2009) Regulators of PP2C phosphatase activity function as abscisic acid sensors. Science 324:1064–1068PubMedGoogle Scholar
  33. Mekonnen DW, Flügge UI, Ludewig F (2016) Gamma-aminobutyric acid depletion affects stomata closure and drought tolerance of Arabidopsis thaliana. Plant Sci 245:25–34CrossRefGoogle Scholar
  34. Meyer S, Mumm P, Imes D, Endler A, Weder B, Al-Rasheid KA, Geiger D, Marten I, Martinoia E, Hedrich R (2010) AtALMT12 represents an R-type anion channel required for stomatal movement in Arabidopsis guard cells. Plant J 63:1054–1062CrossRefGoogle Scholar
  35. Michaeli S, Fromm H (2015) Closing the loop on the GABA shunt in plants: are GABA metabolism and signaling entwined? Front Plant Sci 6:419CrossRefGoogle Scholar
  36. Miller G, Suzuki N, Ciftci-Yilmaz S, Mittler R (2010) Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell Environ 33:453–467CrossRefGoogle Scholar
  37. Molina-Rueda JJ, Pascual MB, Pissarra J, Gallardo F (2015) A putative role for γ-aminobutyric acid (GABA) in vascular development in pine seedlings. Planta 241:257–267CrossRefGoogle Scholar
  38. Palanivelu R, Brass L, Edlund AF, Preuss D (2003) Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABA levels. Cell 114:47–59CrossRefGoogle Scholar
  39. Papacek M, Christmann A, Grill E (2017) Interaction network of ABA receptors in grey poplar. Plant J 92:199–210CrossRefGoogle Scholar
  40. Ramesh SA, Tyerman SD, Xu B, Bose J, Kaur S, Conn V, Domingos P, Ullah S, Wege S, Shabala S, Feijó JA, Ryan PR, Gilliham M (2015) GABA signalling modulates plant growth by directly regulating the activity of plant-specific anion transporters. Nat Commun 6:7879CrossRefGoogle Scholar
  41. Ramesh SA, Tyerman SD, Gilliham M, Xu B (2017) γ-Aminobutyric acid (GABA) signalling in plants. Cell Mol Life Sci 74:1577–1603CrossRefGoogle Scholar
  42. Renault H, Roussel V, El Amrani A, Arzel M, Renault D, Bouchereau A, Deleu C (2010) The Arabidopsis pop2-1 mutant reveals the involvement of GABA transaminase in salt stress tolerance. BMC Plant Biol 10:20CrossRefGoogle Scholar
  43. Renault H, El Amrani A, Berger A, Mouille G, Soubigou-Taconnat L, Bouchereau A, Deleu C (2013) γ-Aminobutyric acid transaminase deficiency impairs central carbon metabolism and leads to cell wall defects during salt stress in Arabidopsis roots. Plant Cell Environ 36:1009–1018CrossRefGoogle Scholar
  44. Roberts MR (2007) Does GABA act as a signal in plants? Hints from molecular studies. Plant Signal Behav 2:408–409CrossRefGoogle Scholar
  45. Roelfsema M, Hedrich R, Geiger D (2012) Anion channels: master switches of stress responses. Trends Plant Sci 17:221–229CrossRefGoogle Scholar
  46. Santiago J, Dupeux F, Round A, Antoni R, Park SY, Jamin M, Cutler SR, Rodriguez PL, Márquez JA (2009) The abscisic acid receptor PYR1 in complex with abscisic acid. Nature 462:665–668CrossRefGoogle Scholar
  47. Shelp BJ, Bown AW, Mclean M (1999) Metabolism and functions of γ-aminobutyric acid. Trends Plant Sci 4:446–452CrossRefGoogle Scholar
  48. Shi S-Q, Shi Z, Jiang Z-P, Qi L-W, Sun X-M, Li C-X, Liu J-F, Xiao W-F, Zhang S-G (2010) Effects of exogenous GABA on gene expression of Caragana intermedia roots under NaCl stress: regulatory roles for H2O2 and ethylene production. Plant Cell Environ 33:149–162CrossRefGoogle Scholar
  49. Shulga A, Rivera C (2013) Interplay between thyroxin, BDNF and GABA in injured neurons. Neuroscience 239:241–252CrossRefGoogle Scholar
  50. Steward FC, Thompson JF, Dent CE (1949) γ-Aminobutyric acid: a constituent of the potato tuber? Science 110:439–440Google Scholar
  51. Umezawa T, Okamoto M, Kushiro T, Nambara E, Oono Y, Seki M, Kobayashi M, Koshiba T, Kamiya Y, Shinozaki K (2006) CYP707A3, a major ABA 8′-hydroxylase involved in dehydration and rehydration response in Arabidopsis thaliana. Plant J 46:171–182CrossRefGoogle Scholar
  52. Wang KL, Li H, Ecker JR (2002) Ethylene biosynthesis and signalling networks. Plant Cell 14(Suppl):S131–S151CrossRefGoogle Scholar
  53. Yang R, Hui Q, Gu Z (2016) Effects of ABA and CaCl2 on GABA accumulation in fava bean germinating under hypoxia-NaCl stress. Biosci Biotechnol Biochem 80:540–546CrossRefGoogle Scholar
  54. Zarei A, Trobacher CP, Shelp BJ (2016) Arabidopsis aldehyde dehydrogenase 10 family members confer salt tolerance through putrescine-derived 4-aminobutyrate (GABA) production. Sci Rep 6:35115CrossRefGoogle Scholar
  55. Žárský V (2015) Signal transduction: GABA receptor found in plants. Nat Plants 1:15115CrossRefGoogle Scholar
  56. Zhang G, Bown AW (1997) The rapid determination of γ-aminobutyric acid. Phytochemistry 44:1007–1009CrossRefGoogle Scholar
  57. Zhou J, Wang J, Li X, Xia XJ, Zhou YH, Shi K, Chen Z, Yu JQ (2014) H2O2 mediates the crosstalk of brassinosteroid and abscisic acid in tomato responses to heat and oxidative stresses. J Exp Bot 65:4371–4383CrossRefGoogle Scholar
  58. Zhu T, Deng X, Zhou X, Zhu L, Zou L, Li P, Zhang D, Lin H (2016) Ethylene and hydrogen peroxide are involved in brassinosteroid-induced salt tolerance in tomato. Sci Rep 6:35392CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of State Forestry AdministrationResearch Institute of Forestry, Chinese Academy of ForestryBeijingChina
  2. 2.Institute of Forest Ecology, Environment and ProtectionChinese Academy of ForestryBeijingChina
  3. 3.Institute of Apicultural ResearchChinese Academy of Agricultural SciencesBeijingChina
  4. 4.Risk Assessment Laboratory for Bee ProductsQuality and Safety of Ministry of AgricultureBeijingChina

Personalised recommendations