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GA-DELLA pathway is involved in regulation of nitrogen deficiency-induced anthocyanin accumulation

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Abstract

Key message

DELLA proteins positively regulate nitrogen deficiency-induced anthocyanin accumulation through directly interaction with PAP1 to enhance its transcriptional activity on anthocyanin biosynthetic gene expressions.

Abstract

Plants can survive a limiting nitrogen supply by undergoing adaptive responses, including induction of anthocyanin production. However, the detailed mechanism is still unclear. In this study, we found that this process was impaired and enhanced, respectively, by exogenous GA3 (an active form of GAs) and paclobutrazol (PAC, a specific GA biosynthesis inhibitor) in Arabidopsis seedlings. Consistently, the nitrogen deficiency-induced transcript levels of several key genes involved in anthocyanin biosynthesis, including F3′H, DFR, LDOX, and UF3GT, were decreased and enhanced by exogenous GA3 and PAC, respectively. Moreover, the nitrogen deficiency-induced anthocyanin accumulation and biosynthesis gene expressions were impaired in the loss-of-function mutant gai-t6/rga-t2/rgl1-1/rgl2-1/rgl3-1 (della) but enhanced in the GA-insensitive mutant gai, suggesting that DELLA proteins, known as repressors of GA signaling, are necessary for fully induction of nitrogen deficiency-driven anthocyanin biosynthesis. Using yeast two-hybrid (Y2H) assay, pull-down assay, and luciferase complementation assay, it was found that RGA, a DELLA of Arabidopsis, could strongly interact with PAP1, a known regulatory transcription factor positively involved in anthocyanin biosynthesis. Furthermore, transient expression assays indicated that RGA and GAI could enhance the transcriptional activities of PAP1 on its downstream genes, including F3′H and DFR. Taken together, this study suggests that DELLAs are necessary regulators for nitrogen deficiency-induced anthocyanin accumulation through interaction with PAP1 and enhancement of PAP1’s transcriptional activity on its target genes. GA-DELLA-involved anthocyanin accumulation is important for plant adaptation to nitrogen deficiency.

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Abbreviations

CHI:

Chalcone isomerase

CHS:

Chalcone synthase

della :

Gai-t6/rga-t2/rgl1-1/rgl2-1/rgl3-1

DFR:

Dihydroflavanol reductase

EGL3:

Enhancer of glabra 3

F3′H:

Flavonoid 3′-hydroxylase

GA:

Gibberellin

GA2ox:

Gibberellin 2-oxidase

GA3ox:

Gibberellin 3-oxidase

GA20ox:

Gibberellin 20-oxidase

GAI:

GA-insensitive

GL3:

Glabra 3

HN:

High nitrate

LBG:

Late biosynthetic gene

LDOX:

Leucoanthocyanidin dioxygenase

LN:

Low nitrate

MBW:

MYB-bHLH-WD40

PAP1:

Production of anthocyanin pigment 1

PAP2:

Production of anthocyanin pigment 2

qRT-PCR:

Quantitative reverse transcription PCR

RGA:

Repressor of ga1-3

TT8:

Transparent testa 8

TTG1:

Transparent testa glabra 1

UF3GT:

UDP-flavonoid 3-glucosyl transferase

Y2H:

Yeast two-hybrid

References

  • Achard P, Liao LL, Jiang CF, Desnos T, Bartlett J, Fu XD, Harberd NP (2007) DELLAs contribute to plant photomorphogenesis. Plant Physiol 143:1163–1172

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Baudry A, Caboche M, Lepiniec L (2006) TT8 controls its own expression in a feedback regulation involving TTG1 and homologous MYB and bHLH factors, allowing a strong and cell-specific accumulation of flavonoids in Arabidopsis thaliana. Plant J 46:768–779

    Article  CAS  PubMed  Google Scholar 

  • Chen HM, Zou Y, Shang YL, Lin HQ, Wang YJ, Cai R, Tang XY, Zhou JM (2008) Firefly luciferase complementation imaging assay for protein-protein interactions in plants. Plant Physiol 146:368–376

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Das PK, Shin DH, Choi SB, Park YI (2012a) Sugar-hormone cross-talk in anthocyanin biosynthesis. Mol Cells 34:501–507

  • Das PK, Shin DH, Choi SB, Yoo SD, Choi G, Park YI (2012b) Cytokinins enhance sugar-induced anthocyanin biosynthesis in Arabidopsis. Mol Cells 34:93–101

  • Davière JM, Achard P (2013) Gibberellin signaling in plants. Development 140:1147–1151

    Article  PubMed  Google Scholar 

  • Davière JM, de Lucas M, Prat S (2008) Transcriptional factor interaction: a central step in DELLA function. Curr Opin Genet Dev 18:295–303

    Article  PubMed  Google Scholar 

  • Diaz C, Saliba-Colombani V, Loudet O, Belluomo P, Moreau L, Daniel-Vedele F, Morot-Gaudry JF, Masclaux-Daubresse C (2006) Leaf yellowing and anthocyanin accumulation are two genetically independent strategies in response to nitrogen limitation in Arabidopsis thaliana. Plant Cell Physiol 47:74–83

    Article  CAS  PubMed  Google Scholar 

  • Dubos C, Le Gourrierec J, Baudry A, Huep G, Lanet E, Debeaujon I, Routaboul JM, Alboresi A, Weisshaar B, Lepiniec L (2008) MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana. Plant J 55:940–953

    Article  CAS  PubMed  Google Scholar 

  • Feng SH, Martinez C, Gusmaroli G, Wang Y, Zhou JL, Wang F, Chen LY, Yu L, Iglesias-Pedraz JM, Kircher S, Schäfer E, Fu XD, Fan LM, Deng XW (2008) Coordinated regulation of Arabidopsis thaliana development by light and gibberellins. Nature 451:475–479

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Feyissa DN, Løvdal T, Olsen KM, Slimestad R, Lillo C (2009) The endogenous GL3, but not EGL3, gene is necessary for anthocyanin accumulation as induced by nitrogen depletion in Arabidopsis rosette stage leaves. Planta 230:747–754

    Article  CAS  PubMed  Google Scholar 

  • Frink CR, Waggoner PE, Ausubel JH (1999) Nitrogen fertilizer: retrospect and prospect. Proc Natl Acad Sci USA 96:1175–1180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gonzalez A, Zhao MZ, Leavitt JM, Lloyd AM (2008) Regulation of the anthocyanin biosynthetic pathway by the TTG1/bHLH/Myb transcriptional complex in Arabidopsis seedlings. Plant J 53:814–827

    Article  CAS  PubMed  Google Scholar 

  • Gould KS (2004) Nature’s Swiss army knife: the diverse protective roles of anthocyanins in leaves. J Biomed Biotechnol 2004:314–320

    Article  PubMed  PubMed Central  Google Scholar 

  • Hauvermale AL, Ariizumi T, Steber CM (2012) Gibberellin signaling: a theme and variations on DELLA repression. Plant Physiol 160:83–92

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hellens RP, Allan AC, Friel EN, Bolitho K, Grafton K, Templeton MD, Karunairetnam S, Gleave AP, Laing WA (2005) Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants. Plant Methods 1:13

    Article  PubMed  PubMed Central  Google Scholar 

  • Jeong SW, Das PK, Jeoung SC, Song JY, Lee HK, Kim YK, Kim WJ, Park YI, Yoo SD, Choi SB, Choi G (2010) Ethylene suppression of sugar-induced anthocyanin pigmentation in Arabidopsis. Plant Physiol 154:1514–1531

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jiang CF, Gao XH, Liao LL, Harberd NP, Fu XD (2007) Phosphate starvation root architecture and anthocyanin accumulation responses are modulated by the gibberellin-DELLA signaling pathway in Arabidopsis. Plant Physiol 145:1460–1470

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lea US, Slimestad R, Smedvig P, Lillo C (2007) Nitrogen deficiency enhances expression of specific MYB and bHLH transcription factors and accumulation of end products in the flavonoid pathway. Planta 225:1245–1253

    Article  CAS  PubMed  Google Scholar 

  • Lee J, He K, Stolc V, Lee H, Figueroa P, Gao Y, Tongprasit W, Zhao HY, Lee I, Deng XW (2007) Analysis of transcription factor HY5 genomic binding sites revealed its hierarchical role in light regulation of development. Plant Cell 19:731–749

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li QF, Wang CM, Jiang L, Li S, Sun SS, He JX (2012) An interaction between BZR1 and DELLAs mediates direct signaling crosstalk between brassinosteroids and gibberellins in Arabidopsis. Sci Signal 5:ra72

    PubMed  Google Scholar 

  • Li Y, den Ende WV, Rolland F (2014) Sucrose induction of anthocyanin biosynthesis is mediated by DELLA. Mol Plant 7:570–572

    Article  CAS  PubMed  Google Scholar 

  • Liu ZJ, Zhang YQ, Wang JF, Li P, Zhao CZ, Chen YD, Bi YR (2015) Phytochrome-interacting factors PIF4 and PIF5 negatively regulate anthocyanin biosynthesis under red light in Arabidopsis seedlings. Plant Sci 238:64–72

    Article  CAS  PubMed  Google Scholar 

  • Loreti E, Povero G, Novi G, Solfanelli C, Alpi A, Perata P (2008) Gibberellins, jasmonate and abscisic acid modulate the sucrose-induced expression of anthocyanin biosynthetic genes in Arabidopsis. New Phytol 179:1004–1016

    Article  CAS  PubMed  Google Scholar 

  • Maier A, Schrader A, Kokkelink L, Falke C, Welter B, Iniesto E, Rubio V, Uhrig JF, Hulskamp M, Hoecker U (2013) Light and the E3 ubiquitin ligase COP1/SPA control the protein stability of the MYB transcription factors PAP1 and PAP2 involved in anthocyanin accumulation in Arabidopsis. Plant J 74:638–651

    Article  CAS  PubMed  Google Scholar 

  • Martin T, Oswald O, Graham IA (2002) Arabidopsis seedling growth, storage lipid mobilization, and photosynthetic gene expression are regulated by carbon: nitrogen availability. Plant Physiol 128:472–481

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Peng MS, Hudson D, Schofield A, Tsao R, Yang R, Gu HL, Bi YM, Rothstein SJ (2008) Adaptation of Arabidopsis to nitrogen limitation involves induction of anthocyanin synthesis which is controlled by the NLA gene. J Exp Bot 59:2933–2944

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Petroni K, Tonelli C (2011) Recent advances on the regulation of anthocyanin synthesis in reproductive organs. Plant Sci 181:219–229

    Article  CAS  PubMed  Google Scholar 

  • Plackett AR, Powers SJ, Fernandez-Garcia N, Urbanova T, Takebayashi Y, Seo M, Jikumaru Y, Benlloch R, Nilsson O, Ruiz-Rivero O, Phillips AL, Wilson ZA, Thomas SG, Hedden P (2012) Analysis of the developmental roles of the Arabidopsis gibberellin 20-oxidases demonstrates that GA20ox1, -2, and -3 are the dominant paralogs. Plant Cell 24:941–960

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Qi TC, Song SS, Ren QC, Wu DW, Huang H, Chen Y, Fan M, Peng W, Ren CM, Xie DX (2011) The jasmonate-ZIM-domain proteins interact with the WD-repeat/bHLH/MYB complexes to regulate jasmonate-mediated anthocyanin accumulation and trichome initiation in Arabidopsis thaliana. Plant Cell 23:1795–1814

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Qi TC, Huang H, Wu DW, Yan JB, Qi YJ, Song SS, Xie DX (2014) Arabidopsis DELLA and JAZ proteins bind the WD-repeat/bHLH/MYB complex to modulate gibberellin and jasmonate signaling synergy. Plant Cell 26:1118–1133

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rieu I, Eriksson S, Powers SJ, Gong F, Griffiths J, Woolley L, Benlloch R, Nilsson O, Thomas SG, Hedden P, Phillips AL (2008a) Genetic analysis reveals that C19-GA 2-oxidation is a major gibberellin inactivation pathway in Arabidopsis. Plant Cell 20:2420–2436

  • Rieu I, Ruiz-Rivero O, Fernandez-Garcia N, Griffiths J, Powers SJ, Gong F, Linhartova T, Eriksson S, Nilsson O, Thomas SG, Phillips AL, Hedden P (2008b) The gibberellin biosynthetic genes AtGA20ox1 and AtGA20ox2 act, partially redundantly, to promote growth and development throughout the Arabidopsis life cycle. Plant J 53:488–504

  • Shin DH, Choi M, Kim K, Bang G, Cho M, Choi SB, Choi G, Park YI (2013) HY5 regulates anthocyanin biosynthesis by inducing the transcriptional activation of the MYB75/PAP1 transcription factor in Arabidopsis. FEBS Lett 587:1543–1547

    Article  CAS  PubMed  Google Scholar 

  • Sun TP (2011) The molecular mechanism and evolution of the GA-GID1-DELLA signaling module in plants. Curr Biol 21:R338–R345

    Article  CAS  PubMed  Google Scholar 

  • Teng S, Keurentjes J, Bentsink L, Koornneef M, Smeekens S (2005) Sucrose-specific induction of anthocyanin biosynthesis in Arabidopsis requires the MYB75/PAP1 gene. Plant Physiol 139:1840–1852

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vandenbussche F, Habricot Y, Condiff AS, Maldiney R, Van der Straeten D, Ahmad M (2007) HY5 is a point of convergence between cryptochrome and cytokinin signalling pathways in Arabidopsis thaliana. Plant J 49:428–441

    Article  CAS  PubMed  Google Scholar 

  • Xie Y, Tan HJ, Ma ZX, Huang JR (2016) DELLA proteins promote anthocyanin biosynthesis via sequestering MYBL2 and JAZ suppressors of the MYB/bHLH/WD40 complex in Arabidopsis thaliana. Mol Plant 9:711–721

    Article  CAS  PubMed  Google Scholar 

  • Yamaguchi S (2008) Gibberellin metabolism and its regulation. Annu Rev Plant Biol 59:225–251

    Article  CAS  PubMed  Google Scholar 

  • Yoo SD, Cho YH, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2:1565–1572

    Article  CAS  PubMed  Google Scholar 

  • Zhang YQ, Liu ZJ, Liu RZ, Hao HL, Bi YR (2011a) Gibberellins negatively regulate low temperature-induced anthocyanin accumulation in a HY5/HYH-dependent manner. Plant Signal Behav 6:632–634

  • Zhang ZL, Ogawa M, Fleet CM, Zentella R, Hu J, Heo JO, Lim J, Kamiya Y, Yamaguchi S, Sun TP (2011b) Scarecrow-like 3 promotes gibberellin signaling by antagonizing master growth repressor DELLA in Arabidopsis. Proc Natl Acad Sci USA 108:2160–2165

  • Zhang YQ, Liu ZJ, Chen YD, He JX, Bi YR (2015) PHYTOCHROME-INTERACTING FACTOR 5 (PIF5) positively regulates dark-induced senescence and chlorophyll degradation in Arabidopsis. Plant Sci 237:57–68

    Article  CAS  PubMed  Google Scholar 

  • Zhou LL, Shi MZ, Xie DY (2012) Regulation of anthocyanin biosynthesis by nitrogen in TTG1-GL3/TT8-PAP1-programmed red cells of Arabidopsis thaliana. Planta 236:825–837

    Article  CAS  PubMed  Google Scholar 

  • Zimmermann IM, Heim MA, Weisshaar B, Uhrig JF (2004) Comprehensive identification of Arabidopsis thaliana MYB transcription factors interacting with R/B-like BHLH proteins. Plant J 40:22–34

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Dr. Nicholas Harberd for providing gai and gai-t6/rga-24 mutant seeds, Dr. Xing-Wang Deng for providing 35S::TAP-RGAd17 seeds. The vectors pSAT6-EYFP-C1 (CD3-1103), pUC19-cLUC (CD3-1702), and pUC19-nLUC (CD3-1701) are order from ABRC. This work was supported by the Fundamental Research Funds for the Central Universities (lzujbky-2015-91) and National Science Foundation of China (31422047).

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Authors and Affiliations

  1. Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, People’s Republic of China

    Yongqiang Zhang, Zhongjuan Liu, Jianping Liu, Sheng Lin, Wenxiong Lin & Weifeng Xu

  2. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University, Fuzhou, 350002, People’s Republic of China

    Yongqiang Zhang, Zhongjuan Liu, Jianping Liu, Sheng Lin, Wenxiong Lin & Weifeng Xu

  3. Key Laboratory of Cell Activities and Stress Adaptations, Ministry of Education, School of Life Sciences, Lanzhou University, Lanzhou, 730000, People’s Republic of China

    Yongqiang Zhang

  4. State Key Laboratory of Grassland Agro-Ecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, 730000, People’s Republic of China

    Jianfeng Wang

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  1. Yongqiang Zhang
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  2. Zhongjuan Liu
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Communicated by Baochun Li.

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Zhang, Y., Liu, Z., Liu, J. et al. GA-DELLA pathway is involved in regulation of nitrogen deficiency-induced anthocyanin accumulation. Plant Cell Rep 36, 557–569 (2017). https://doi.org/10.1007/s00299-017-2102-7

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