Expression of AtGA2ox1 enhances drought tolerance in maize

  • Ziqi Chen
  • Yang Liu
  • Yuejia Yin
  • Qing Liu
  • Nan Li
  • Xia Li
  • Wenzhu He
  • Dongyun Hao
  • Xiangguo LiuEmail author
  • Changhong GuoEmail author
Original Paper


Drought is a major limiting factor to maize (Zea mays L.) yield. Plant hormones, including gibberellins (GAs), play important roles in plant response to drought stress. In previous studies, significant reductions in GAs levels have been reported under drought stress. In maize, GA content is correlated to drought tolerance, but the molecular mechanism remains unclear. In the present study, AtG2ox1, a member of the GA2ox family with a clear function, was used to create GA deficiency maize. The transgenic maize had a higher chlorophyll content and faster growth rate, when compared to the wild type (WT) plants, under drought stress in a greenhouse. The physiological and biochemical test results revealed that transgenic maize had decreased levels of GA1 and malondialdehyde (MDA), and increased content of proline and soluble sugars, and antioxidant enzyme activities, when compared to the WT. Furthermore, the transcriptomic analysis revealed that some differentially expressed genes involved in transcription factors correlated to drought stress and abiotic stress responses, and that signaling was enriched. All these results reveal the possible molecular mechanism of GA regulation in drought tolerance, in which the overexpression of AtGA2ox1 altered the expression of multiple genes correlated to the internal antioxidant system and maintenance of cell osmotic potential. The present study demonstrates that the overexpression of AtGA2ox1 could control GA content and improve drought tolerance in transgenic maize. Furthermore, this strategy represents a novel approach to address drought tolerance in maize breeding.


AtGA2ox1 Gibberellin Drought tolerance Maize 



This work was supported by the Agricultural Science and Technology Innovation Program of Jilin Province (CXGC2017ZY026) and the National Natural Science Foundation of China (No. 31771879).

Author contribution

ZC and YL contributed equally to the study. CG and XL designed the experiments. ZC, YY, YL, QL, NL, WH and DH performed the experiments. ZC and XL analyzed the data. ZC and XL wrote the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest in this work.

Supplementary material

10725_2019_526_MOESM1_ESM.docx (50 kb)
Supplementary material 1 (DOCX 49 kb)
10725_2019_526_MOESM2_ESM.docx (16 kb)
Supplementary material 2 (DOCX 15 kb)
10725_2019_526_MOESM3_ESM.docx (16 kb)
Supplementary material 3 (DOCX 16 kb)
10725_2019_526_MOESM4_ESM.docx (34 kb)
Supplementary material 4 (DOCX 34 kb)
10725_2019_526_MOESM5_ESM.docx (16 kb)
Supplementary material 5 (DOCX 16 kb)


  1. Achard P, Renou JP, Berthomé R, Harberd NP, Genschik P (2008a) Plant DELLAs restrain growth and promote survival of adversity by reducing the levels of reactive oxygen species. Curr Biol 18(9):656–660Google Scholar
  2. Achard P, Gong F, Cheminant S, Alioua M, Hedden P, Genschik P (2008b) The cold-inducible CBF1 factor-dependent signaling pathway modulates the accumulation of the growth-repressing DELLA proteins via its effect on gibberellin metabolism. Plant Cell 20(8):2117–2129Google Scholar
  3. Agharkar M, Lomba P, Altpeter F, Zhang H, Lange T (2007) Stable expression of AtGA2ox1 in a low-input turfgrass (Paspalum notatum Flugge) reduces bioactive gibberellin levels and improves turf quality under field conditions. Plant Biotechnol J 5(6):791–801Google Scholar
  4. Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39(1):205–207Google Scholar
  5. Beers RF, Sizer IW (1952) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195(1):133–140Google Scholar
  6. Biemelt S, Tschiersch H, Sonnewald U (2004) Impact of altered gibberellin metabolism on biomass accumulation, lignin biosynthesis, and photosynthesis in transgenic tobacco plants. Plant Physiol 135(1):254–265Google Scholar
  7. Chen J, Nolan TM, Ye H, Zhang M, Tong H, Xin P et al (2017) Arabidopsis WRKY46, WRKY54, and WRKY70 transcription factors are involved in brassinosteroid-regulated plant growth and drought responses. Plant Cell 29(6):1425–1439Google Scholar
  8. Colebrook EH, Thomas SG, Phillips AL, Hedden P (2014) The role of gibberellin signalling in plant responses to abiotic stress. J Exp Biol 217(1):67–75Google Scholar
  9. Dijkstra C, Adams E, Bhattacharya A, Page AF, Anthony P, Kourmpetli S et al (2008) Over-expression of a gibberellin 2-oxidase gene from Phaseolus coccineus L. enhances gibberellin inactivation and induces dwarfism in Solanum species. Plant Cell Rep 27(3):463–470Google Scholar
  10. Edmeades GO, Cooper M, Lafitte R, Zinselmeier C, Ribaut JM, Habben JE et al (2000) Abiotic stresses and staple crops. In: Crop science: progress and prospects (no. CIS-4429). CIMMYTGoogle Scholar
  11. FAO (2010) FAOSTAT statistical database of the Food and Agriculture Organization of the United Nations. FAO, Rome. Accessed 14 July 2011
  12. Gallego-Bartolome J, Alabadi D, Blazquez MA (2011) DELLA-induced early transcriptional changes during etiolated development in Arabidopsis thaliana. PLoS ONE 6:e23918Google Scholar
  13. Gilbert HS, Stump DD, Roth EF Jr (1984) A method to correct for errors caused by generation of interfering compounds during erythrocyte lipid peroxidation. Anal Biochem 137(2):282–286Google Scholar
  14. Guo M, Liu JH, Ma X, Luo DX, Gong ZH, Lu MH (2016) The plant heat stress transcription factors (HSFs): structure, regulation, and function in response to abiotic stresses. Front Plant Sci 7:114Google Scholar
  15. Han F, Zhu B (2011) Evolutionary analysis of three gibberellin oxidase genes in rice, Arabidopsis, and soybean. Gene 473(1):23–35Google Scholar
  16. Hedden P, Phillips AL (2000) Gibberellin metabolism: new insights revealed by the genes. Trends Plant Sci 5(12):523–530Google Scholar
  17. Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207(4):604–611Google Scholar
  18. Huang J, Tang D, Shen Y, Qin B, Hong L, You A, Cheng Z (2010a) Activation of gibberellin 2-oxidase 6 decreases active gibberellin levels and creates a dominant semi-dwarf phenotype in rice (Oryza sativa L.). J Genet Genomics 37(1):23–36Google Scholar
  19. Huang XS, Liu JH, Chen XJ (2010b) Overexpression of PtrABF gene, a bZIP transcription factor isolated from Poncirus trifoliata, enhances dehydration and drought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes. BMC Plant Biol 10(1):230Google Scholar
  20. Ishida Y, Hiei Y, Komari T (2007) Agrobacterium-mediated transformation of maize. Nat Protoc 2(7):1614Google Scholar
  21. Lee DJ, Zeevaart JA (2005) Molecular cloning of GA 2-oxidase3 from spinach and its ectopic expression in Nicotiana sylvestris. Plant Physiol 138(1):243–254Google Scholar
  22. Lee J, He K, Stolc V, Lee H, Figueroa P, Gao Y, Tongprasit W, Zhao H, Lee I, Deng XW (2007) Analysis of transcription factor HY5 genomic binding sites revealed its hierarchical role in light regulation of development. Plant Cell 19:731–749Google Scholar
  23. Liu HC, Liao HT, Charng YY (2011) The role of class A1 heat shock factors (HSFA1s) in response to heat and other stresses in Arabidopsis. Plant Cell Environ 34(5):738–751Google Scholar
  24. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408Google Scholar
  25. Lo SF, Yang SY, Chen KT, Hsing YI, Zeevaart JA, Chen LJ, Yu SM (2008) A novel class of gibberellin 2-oxidases control semidwarfism, tillering, and root development in rice. Plant Cell 20(10):2603–2618Google Scholar
  26. Lo SF, Ho THD, Liu YL, Jiang MJ, Hsieh KT, Chen KT, Yu LC et al (2017) Ectopic expression of specific GA2 oxidase mutants promotes yield and stress tolerance in rice. Plant Biotechnol J 15(7):850–864Google Scholar
  27. Ma H, Wang C, Yang B, Cheng H, Wang Z, Mijiti A et al (2016) CarHSFB2, a class B heat shock transcription factor, is involved in different developmental processes and various stress responses in chickpea (Cicer arietinum L.). Plant Mol Biol Report 34(1):1–14Google Scholar
  28. Magome H, Yamaguchi S, Hanada A, Kamiya Y, Oda K (2004) Dwarf and delayed-flowering 1, a novel Arabidopsis mutant deficient in gibberellin biosynthesis because of overexpression of a putative AP2 transcription factor. Plant J 37(5):720–729Google Scholar
  29. Magome H, Yamaguchi S, Hanada A, Kamiya Y, Oda K (2008) The DDF1 transcriptional activator upregulates expression of a gibberellin-deactivating gene, GA2ox7, under high-salinity stress in Arabidopsis. Plant J 56(4):613–626Google Scholar
  30. Moore K, Roberts LJ (1998) Measurement of lipid peroxidation. Free Radic Res 28(6):659–671Google Scholar
  31. Nelissen H, Sun XH, Rymen B, Jikumaru Y, Kojima M, Takebayashi Y et al (2018) The reduction in maize leaf growth under mild drought affects the transition between cell division and cell expansion and cannot be restored by elevated gibberellic acid levels. Plant Biotechnol J 16(2):615–627Google Scholar
  32. Ogawa D, Yamaguchi K, Nishiuchi T (2007) High-level overexpression of the Arabidopsis HsfA2 gene confers not only increased thermotolerance but also salt/osmotic stress tolerance and enhanced callus growth. J Exp Bot 58(12):3373–3383Google Scholar
  33. Oh E, Yamaguchi S, Hu J, Yusuke J, Jung B, Paik I et al (2007) PIL5, a phytochrome-interacting bHLH protein, regulates gibberellin responsiveness by binding directly to the GAI and RGA promoters in Arabidopsis seeds. Plant Cell 19(4):1192–1208Google Scholar
  34. Park HY, Seok HY, Woo DH, Lee SY, Tarte VN, Lee EH et al (2011) AtERF71/HRE2 transcription factor mediates osmotic stress response as well as hypoxia response in Arabidopsis. Biochem Biophys Res Commun 414(1):135–141Google Scholar
  35. Qi T, Huang H, Wu D, Yan J, Qi Y, Song S, Xie D (2014) Arabidopsis DELLA and JAZ proteins bind the WD-Repeat/bHLH/MYB complex to modulate gibberellin and jasmonate signaling synergy. Plant Cell 26:1118–1133Google Scholar
  36. Sakamoto T, Miura K, Itoh H, Tatsumi T, Ueguchi-Tanaka M, Ishiyama K et al (2004) An overview of gibberellin metabolism enzyme genes and their related mutants in rice. Plant Physiol 134(4):1642–1653Google Scholar
  37. Schomburg FM, Bizzell CM, Lee DJ, Zeevaart JA, Amasino RM (2003) Overexpression of a novel class of gibberellin 2-oxidases decreases gibberellin levels and creates dwarf plants. Plant Cell 15(1):151–163Google Scholar
  38. Shan X, Li Y, Jiang Y, Jiang Z, Hao W, Yuan Y (2013) Transcriptome profile analysis of maize seedlings in response to high-salinity, drought and cold stresses by deep sequencing. Plant Mol Biol Report 31(6):1485–1491Google Scholar
  39. Shan C, Mei Z, Duan J, Chen H, Feng H, Cai W (2014) OsGA2ox5, a gibberellin metabolism enzyme, is involved in plant growth, the root gravity response and salt stress. PLoS ONE 9(1):e87110Google Scholar
  40. Sharma P, Jha AB, Dubey RS, Pessarakli M (2012) Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. J Bot. Google Scholar
  41. Shim JS, Jung C, Lee S, Min K, Lee YW, Choi Y et al (2013) AtMYB 44 regulates WRKY70 expression and modulates antagonistic interaction between salicylic acid and jasmonic acid signaling. Plant J 73(3):483–495Google Scholar
  42. Somasundaram R, Jaleel CA, Abraham SS, Azooz MM, Panneerselvam R (2009) Role of paclobutrazol and ABA in drought stress amelioration in Sesamum indicum L. Glob J Mol Sci 4(2):56–62Google Scholar
  43. Song J, Guo B, Song F, Peng H, Yao Y, Zhang Y, Sun Q, Ni Z (2011) Genome-wide identification of gibberellins metabolic enzyme genes and expression profiling analysis during seed germination in maize. Gene 482(1–2):34–42Google Scholar
  44. Sperdouli I, Moustakas M (2012) Interaction of proline, sugars, and anthocyanins during photosynthetic acclimation of Arabidopsis thaliana to drought stress. Plant Physiol 169:577–585Google Scholar
  45. Spitzer T, Míša P, Bílovský J, Kazda J (2015) Management of maize stand height using growth regulators. Plant Prot Sci 51(4):223–230Google Scholar
  46. Storey JD, Tibshirani R (2003) Statistical significance for genomewide studies. Proc Natl Acad Sci USA 100(16):9440–9445Google Scholar
  47. Thirumalaikumar VP, Devkar V, Mehterov N, Ali S, Ozgur R, Turkan I et al (2018) NAC transcription factor JUNGBRUNNEN 1 enhances drought tolerance in tomato. Plant Biotechnol J 16(2):354–366Google Scholar
  48. Thomas SG, Phillips AL, Hedden P (1999) Molecular cloning and functional expression of gibberellin 2-oxidases, multifunctional enzymes involved in gibberellin deactivation. Proc Natl Acad Sci USA 96(8):4698–4703Google Scholar
  49. Upadhyaya A, Sankhla D, Davis TD, Sankhla N, Smith BN (1985) Effect of paclobutrazol on the activities of some enzymes of activated oxygen metabolism and lipid peroxidation in senescing soybean leaves. J Plant Physiol 121(5):453–461Google Scholar
  50. Urbanová T, Tarkowská D, Novák O, Hedden P, Strnad M (2013) Analysis of gibberellins as free acids by ultra performance liquid chromatography–tandem mass spectrometry. Talanta 112:85–94Google Scholar
  51. Vettakkorumakankav NN, Falk D, Saxena P, Fletcher RA (1999) A crucial role for gibberellins in stress protection of plants. Plant Cell Physiol 40(5):542–548Google Scholar
  52. Wang C, Yang A, Yin H, Zhang J (2008) Influence of water stress on endogenous hormone contents and cell damage of maize seedlings. J Integr Plant Biol 50(4):427–434Google Scholar
  53. Wu A, Allu AD, Garapati P, Siddiqui H, Dortay H, Zanor MI et al (2012) JUNGBRUNNEN1, a reactive oxygen species-responsive NAC transcription factor, regulates longevity in Arabidopsis. Plant Cell 24(2):482–506Google Scholar
  54. Yamaguchi S (2008) Gibberellin metabolism and its regulation. Annu Rev Plant Biol 59:225–251Google Scholar
  55. Yang D, Wang N, Yan X, Shi J, Zhang M, Wang Z, Yuan H (2014) Microencapsulation of seed-coating tebuconazole and its effects on physiology and biochemistry of maize seedlings. Colloids Surf B 114:241–246Google Scholar
  56. Yemm EW, Willis AJ (1954) The estimation of carbohydrates in plant extracts by anthrone. Biochem J 57(3):508Google Scholar
  57. Yokotani N, Ichikawa T, Kondou Y, Matsui M, Hirochika H, Iwabuchi M, Oda K (2008) Expression of rice heat stress transcription factor OsHsfA2e enhances tolerance to environmental stresses in transgenic Arabidopsis. Planta 227(5):957–967Google Scholar
  58. Zhong T, Zhang L, Sun S, Zeng H, Han L (2014) Effect of localized reduction of gibberellins in different tobacco organs on drought stress tolerance and recovery. Plant Biotechnol Rep 8(5):399–408Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Ziqi Chen
    • 1
  • Yang Liu
    • 2
  • Yuejia Yin
    • 2
  • Qing Liu
    • 2
  • Nan Li
    • 2
  • Xia Li
    • 3
  • Wenzhu He
    • 4
  • Dongyun Hao
    • 1
    • 2
  • Xiangguo Liu
    • 2
    Email author
  • Changhong Guo
    • 1
    Email author
  1. 1.Key Laboratory of Molecular Cytogenetics and Genetic Breeding of Heilongjiang Province, College of Life Science and TechnologyHarbin Normal UniversityHarbinChina
  2. 2.Jilin Provincial Key Laboratory of Agricultural Biotechnology, Agro-Biotechnology InstituteJilin Academy of Agricultural SciencesChangchunChina
  3. 3.Changchun Academy of Agricultural ScienceChangchunChina
  4. 4.Crop Research Institute of Sichuan Academy of Agricultural SciencesChengduChina

Personalised recommendations