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EGY1 plays a role in regulation of endodermal plastid size and number that are involved in ethylene-dependent gravitropism of light-grown Arabidopsis hypocotyls

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Abstract

Egy1 was isolated as an ethylene-dependent gravitropism-deficient Arabidopsis mutant. Molecular studies reveal that EGY1 gene encodes a 59-kDa plastid-targeted metalloprotease. It is actively expressed in hypocotyl tissue and targets to endodermal and cortex plastid. Its protein level is up-regulated by both ethylene and light. CAB protein accumulation and chlorophyll level is severely reduced in hypocotyls and endodermal cells, respectively. Sucrose is able to restore the severely reduced starch and lipid contents as well as the deficient endodermal plastid size found in light-grown egy1 hypocotyls yet it fails to rescue the reduced plastid number and chlorophyll level in egy1 endodermal cells. The loss-of-function egy1 mutation results in a smaller size (1.9 ± 0.3 μm in diameter) and less number (5 ± 1) of plastids in endodermal cells, which are nearly 50% of the wild-type. EGY1 is specially required for the development of full-size endodermal plastid in seedlings that are grown on sucrose-free media under light. It plays a direct role in controlling the light-induced chlorophyll production, grana formation and plastid replication in endodermal cell. However, it plays an indirect role in regulation of endodermal plastid size. It is likely that the ethylene-dependent gravitropism-deficient phenotype of egy1 hypocotyls may result from the smaller size and less number of endodermal plastids. Gravicurvature assays performed on ethylene-insensitive mutants, etr1-1, etr2-1, ers2-1, ein4-1 and ein2-5, have clearly demonstrated the necessary role for ethylene in vigorous gravitropism of light-grown hypocotyls. The degree of ethylene-dependent gravicurvature is positively correlated with the combined state of endodermal plastid mass and number. Neither ethylene nor EGY1-regulated full-size endodermal plastid is sufficient for promotion of vigorous hypocotyl gravitropism. Presence of 4 full-size plastids per endodermal cell together with ethylene pretreatment of hypocotyls becomes sufficient to trigger vigorous gravicurvature in light-grown seedlings. A model is therefore proposed to address the role of EGY1 in regulation of endodermal plastid size and number as well as the stimulatory effect of ethylene on hypocotyl gravitropism.

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References

  • Baluska F, Hasenstein KH (1997) Root cytoskeleton: its role in perception of and response to gravity. Planta 203(suppl):S69–S78

    Article  PubMed  CAS  Google Scholar 

  • Behringer FJ, Lomax TL (1999) Genetic analysis of the roles of phytochromes A and B1 in the reversed gravitropic response of the lz-2 tomato mutant. Plant Cell Environ 22:551–558

    Article  PubMed  CAS  Google Scholar 

  • Brown MS, Goldstein JL (1997) The SREBP pathway: regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor. Cell 89:331–340

    Article  PubMed  CAS  Google Scholar 

  • Canizares-Maclas P, Hernandez-Garciadiego L, Gomez-Ruiz H (2001) An automated flow injection analysis procedure for the determination of reducing sugars by DNSA method. J Food Sci 66:407–411

    Article  Google Scholar 

  • Caspar T, Pickard BG (1989) Gravitropism in a starchless mutant of Arabidopsis: implications for the starch-statolith theory of gravity sensing. Planta 177:185–197

    Article  PubMed  CAS  Google Scholar 

  • Chen R, Guan C, Boonsirichai K, Masson PH (2002) Complex physiological and molecular processes underlying root gravitropism. Plant Mol Biol 49:305–317

    Article  PubMed  CAS  Google Scholar 

  • Chen G, Bi YR, Li N (2005) EGY1 encodes a membrane-associated and ATP-independent metalloprotease that is required for chloroplast development. Plant J 41:364–375

    Article  PubMed  CAS  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  PubMed  CAS  Google Scholar 

  • DiLaurenzio L, WysockaDiller J, Malamy JE, Pysh L, Helariutta Y, Freshour G, Hahn MG, Feldmann KA, Benfey PN (1996) The SCARECROW gene regulates an asymmetric cell division that is essential for generating the radial organization of the Arabidopsis root. Cell 86(3):423–433

    Article  CAS  Google Scholar 

  • Edelmann HG (2002) Ethylene perception generates gravicompetence in gravi-incompetent leaves of rye seedlings. J Exp Bot 53:1825–1828

    Article  PubMed  CAS  Google Scholar 

  • Estelle M (1996) Plant tropisms: the ins and outs of auxin. Curr Biol 6:1589–1591

    Article  PubMed  CAS  Google Scholar 

  • Fasano JM, Swanson SJ, Blancaflor EB, Dowd PE, Kao TH, Gilroy S (2001) Changes in root cap pH are required for the gravity response of the Arabidopsis root. Plant Cell 13:907–921

    Article  PubMed  CAS  Google Scholar 

  • Fitzelle KJ, Kiss JZ (2001) Restoration of gravitropic sensitivity in starch-deficient mutants of Arabidopsis by hypergravity. J Exp Bot 52:265–275

    Article  PubMed  CAS  Google Scholar 

  • Fluhr R, Mattoo AK (1996) Ethylene: Biosynthesis and perception. Crit Rev Plant Sci 12:479–523

    Article  Google Scholar 

  • Friedman H, Meir S, Rosenberger I, Halevy AH, Kaufman PB, Philosoph-Hadas S (1998) Inhibition of the gravitropic response of snapdragon spikes by the calcium-channel blocker lanthanum chloride. Plant Physiol 118:483–492

    Article  PubMed  CAS  Google Scholar 

  • Friml J, Wisniewska J, Benkova E, Mendgen K, Palme K (2002) Lateral relocation of auxin efflux regulator PIN3 mediates tropism in Arabidopsis. Nature 415:806–809

    PubMed  Google Scholar 

  • Fukaki H, Wysocka-Diller J, Kato T, Fujisawa H, Benfey PN, Tasaka M (1998) Genetic evidence that the endodermis is essential for shoot gravitropism in Arabidopsis thaliana. Plant J 14:425–430

    Article  PubMed  CAS  Google Scholar 

  • Ge L, Liu JZ, Wong WS, Hsiao WLW, Chong K, Xu ZK, Yang SF, Kung SD, Li N (2000) Identification of a novel multiple environmental factor-responsive 1-aminocyclopropane-1-carboxylate synthase gene, NT-ACS2, from tobacco. Plant Cell Environ 23:1169–1182

    Article  CAS  Google Scholar 

  • Golan A, Tepper M, Soudry E, Horwitz BA, Gepstein S (1996) Cytokinin, acting through ethylene, restores gravitropism to Arabidopsis seedlings grown under red light. Plant Physiol 112:901–904

    Article  PubMed  CAS  Google Scholar 

  • Guisinger MM, Kiss JZ (1999) The influence of microgravity and spaceflight on columella cell ultrastructure in starch-deficient mutants of Arabidopsis. Am J Bot 86(10):1357–1366

    Article  PubMed  CAS  Google Scholar 

  • Guzman P, Ecker JR (1990) Exploiting the triple response of Arabidopsis to identify ethylene-related mutants. Plant Cell 2:513–523

    Article  PubMed  CAS  Google Scholar 

  • Hangarter RP (1997) Gravity, light and plant form. Plant Cell Environ 20:796–800

    Article  PubMed  CAS  Google Scholar 

  • Hua J, Meyerowitz EM (1998) Ethylene responses are negatively regulated by a receptor gene family in Arabidopsis thaliana. Cell 94:261–271

    Article  PubMed  CAS  Google Scholar 

  • Kato T, Morita MT, Fukaki H, Yamauchi Y, Uehara M, Niihama M, Tasaka M (2002) SGR2, a phospholipase-like protein, and ZIG/SGR4, a SNARE, are involved in the shoot gravitropism of Arabidopsis. Plant Cell 14:33–46

    Article  PubMed  CAS  Google Scholar 

  • Kiss JZ, Edelmann RE (1999) Spaceflight experiments with Arabidopsis starch-deficient mutants support a statolith-based model for graviperception. Adv Space Res 24:755–762

    Article  PubMed  CAS  Google Scholar 

  • Kiss JZ, Hertel R, Sack FD (1989) Amyloplasts are necessary for full gravitropic sensitivity in roots of Arabidopsis thaliana. Planta 177:198–206

    Article  PubMed  CAS  Google Scholar 

  • Kiss JZ, Guisinger MM, Miller AJ, Stackhouse KS (1997) Reduced gravitropism in hypocotyls of starch-deficient mutants of Arabidopsis. Plant Cell Physiol 38:518–525

    PubMed  CAS  Google Scholar 

  • Kiss JZ, Edelmann RE, Wood PC (1999) Gravitropism of hypocotyls of wild-type and starch-deficient Arabidopsis seedlings in spaceflight studies. Planta 209:96–103

    Article  PubMed  CAS  Google Scholar 

  • Liscum E, Hangarter RP (1993) Genetic evidence that the red-absorbing form of phytochrome b modulates gravitropism in Arabidopsis thaliana. Plant Physiol 103:15–19

    PubMed  CAS  Google Scholar 

  • Lu BW, Chan WK, Zhang H, Li N (1999) Ethylene as a dual function modulator of the negative gravitropism of arabidopsis inflorescence stem. Plant Biol, July 24–28, Baltimore, MD, USA

  • Lu BW, Pei LK, Chan WK, Zhang H, Zhu G, Li JY, Li N (2001) The dual effects of ethylene on the negative gravicurvature of arabidopsis inflorescence, an intriguing action model for the plant hormone ethylene. Chin Sci Bull 46:279–283

    CAS  Google Scholar 

  • Lu BW, Yu HY, Pei LK, Wong MY, Li N (2002) Prolonged exposure to ethylene stimulates the negative gravitropic responses of Arabidopsis inflorescence stems and hypocotyls. Funct Plant Biol 29:987–997

    Article  CAS  Google Scholar 

  • MacCleery SA, Kiss JZ (1999) Plastid sedimentation kinetics in roots of wild-type and starch-deficient mutants of Arabidopsis. Plant Physiol 120(1):183–192

    Article  PubMed  CAS  Google Scholar 

  • Madlung A, Behringer FJ, Lomax TL (1999) Ethylene plays multiple nonprimary roles in modulating the gravitropic response in tomato. Plant Physiol 120(3):897–906

    Article  PubMed  CAS  Google Scholar 

  • Masuda CA, Xavier MA, Mattos KA, Galina A, Montero-Lomeli M (2001) Phosphoglucomutase is an in vivo lithium target in yeast. J Biol Chem 276:37794–37801

    PubMed  CAS  Google Scholar 

  • Millenaar FF, Cox MC, van Berkel YE, Welschen RA, Pierik R, Voesenek LA, Peeters AJ (2005) Ethylene-induced differential growth of petioles in Arabidopsis. Analyzing natural variation, response kinetics, and regulation. Plant Physiol 137:998–1008

    Article  PubMed  CAS  Google Scholar 

  • Morita MT, Tasaka M (2004) Gravity sensing and signaling. Curr Opin Plant Biol 7:712–718

    Article  PubMed  CAS  Google Scholar 

  • Morita MT, Sakaguchi K, Kiyose S, Taira K, Kato T, Nakamura M, Tasaka M (2006) A C2H2-type zinc finger protein, SGR5, is involved in early events of gravitropism in Arabidopsis inflorescence stems. Plant J 47:619–628

    Article  PubMed  CAS  Google Scholar 

  • Paciorek T, Zazimalova E, Ruthardt N, Petrasek J, Stierhof YD, Kleine-Vehn J, Morris DA, Emans N, Jurgens G, Geldner N, Friml J (2005) Auxin inhibits endocytosis and promotes its own efflux from cells. Nature 435:1251–1256

    Article  PubMed  CAS  Google Scholar 

  • Perera IY, Hung CY, Brady S, Muday GK, Boss WF (2006) A universal role for inositol 1,4,5-trisphosphate-mediated signaling in plant gravitropism. Plant Physiol 140:746–760

    Article  PubMed  CAS  Google Scholar 

  • Pucher GW, Leavenworth CS, Vickery HB (1948) Determination of starch in plant tissues. Anal Chem 20:850–853

    Article  CAS  Google Scholar 

  • Robertson EJ, Pyke KA, Leech RM (1995) Arc6, an extreme chloroplast division mutant of Arabidopsis also alters proplastid proliferation and morphology in shoot and root apices. J Cell Sci 108:2937–2944

    PubMed  CAS  Google Scholar 

  • Sack FD (1991) Plant gravity sensing. Int Rev Cytol 127:193–252

    PubMed  CAS  Google Scholar 

  • Saito C, Morita MT, Kato T, Tasaka M (2005) Amyloplasts and vacuolar membrane dynamics in the living graviperceptive cell of the Arabidopsis inflorescence stem. Plant Cell 17:548–558

    Article  PubMed  CAS  Google Scholar 

  • Sakamoto W, Tamura T, Hanba-Tomita Y, Murata M, Sodmergen (2002) The VAR1 locus of Arabidopsis encodes a chloroplastic FtsH and is responsible for leaf variegation in the mutant alleles. Genes Cells 7:769–780

    Article  PubMed  CAS  Google Scholar 

  • Schepens I, Duek P, Fankhauser C (2004) Phytochrome-mediated light signalling in Arabidopsis. Curr Opin Plant Biol 7:564–569

    Article  PubMed  CAS  Google Scholar 

  • Sinclair W, Trewavas AJ (1997) Calcium in gravitropism. A re-examination. Planta 203(suppl 1):S85–S90

    Article  PubMed  CAS  Google Scholar 

  • Swarup R, Parry G, Graham N, Allen T, Bennett M (2002) Auxin cross-talk: integration of signalling pathways to control plant development. Plant Mol Biol 49:411–426

    Article  PubMed  CAS  Google Scholar 

  • Tasaka M, Kato T, Fukaki H (2001) Genetic regulation of gravitropism in higher plants. Int Rev Cytol 206:135–154

    PubMed  CAS  Google Scholar 

  • Telewski FW (2006) A unified hypothesis of mechanoperception in plants. Am J Bot 93:1466–1476

    Google Scholar 

  • Thomson WW, Whatley JM (1980). Development of non-green plastids. Annu Rev Plant Physiol 31:375–394

    Article  Google Scholar 

  • Thum KE, Kim M, Christopher DA, Mullet JE (2001) Cryptochrome 1, cryptochrome 2, and phytochrome a co-activate the chloroplast psbD blue light-responsive promoter. Plant Cell 13:2747–2460

    Article  PubMed  CAS  Google Scholar 

  • Turgeon R (2006) Phloem loading: how leaves gain their independence. Bioscience 56:15–24

    Article  Google Scholar 

  • Vitha S, Froehlich JE, Koksharova O, Pyke KA, van Erp H, Osteryoung KW (2003) ARC6 is a J-domain plastid division protein and an evolutionary descendant of the cyanobacterial cell division protein Ftn2. Plant Cell 15:1918–1933

    Article  PubMed  CAS  Google Scholar 

  • Wang NN, Shih MC, Li N (2005) The GUS reporter-aided analysis of the promoter activities of Arabidopsis ACC synthase genes AtACS4, AtACS5, and AtACS7 induced by hormones and stresses. J Exp Bot 56:909–920

    Article  PubMed  CAS  Google Scholar 

  • Wayne R, Staves MP (1997) A down-to-earth model of gravisensing. Gravit Space Biol Bull 10(2):57–64

    PubMed  CAS  Google Scholar 

  • Wheeler RM, Salisbury FB (1980) Gravitropism in plant stems may require ethylene. Science 209:1126–1127

    Article  PubMed  Google Scholar 

  • Wysocka-Diller JW, Helariutta Y, Fukaki H, Malamy JE, Benfey PN (2000) Molecular analysis of SCARECROW function reveals a radial patterning mechanism common to root and shoot. Development 127:595–603

    PubMed  CAS  Google Scholar 

  • Xiao Y, Huang Y, Chen ZY (2005) Distribution, depletion and recovery of docosahexaenoic acid are region-specific in rat brain. Br J Nutr 94:544–550

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto K, Pyke KA, Kiss JZ (2002) Reduced gravitropism in inflorescence stems and hypocotyls, but not roots, of Arabidopsis mutants with large plastids. Plant Physiol 114:627–636

    Article  CAS  Google Scholar 

  • Yoder TL, Zheng HQ, Todd P, Staehelin LA (2001) Amyloplast sedimentation dynamics in maize columella cells support a new model for the gravity-sensing apparatus of roots. Plant Physiol 125:1045–1060

    Article  PubMed  CAS  Google Scholar 

  • Zobel RW (1973) Some physiological characteristics of the ethylene-requiring tomato mutant diageotropica. Plant Physiol 52:385–389

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

Authors like to express sincere thanks to Drs. Chentao LIN, Joseph Ecker and Elliot Meyerowitz for providing Arabidopsis mutants. The authors also like to extend their gratitude to Drs. Zhenyu CHEN and Ying XIAO for offering the GC columns and the useful instructions in analysis of lipid and many thanks to Ms. Kaying MA for her help in management of GC machine. This work was supported by grants, DAG04/05.SC08, DAG05/06.SC09, HKUST6105/01M, HKUST6276/03M and HKUST6413/06M awarded to Ning LI from the Research Grant Council of Hong Kong.

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Guo, D., Gao, X., Li, H. et al. EGY1 plays a role in regulation of endodermal plastid size and number that are involved in ethylene-dependent gravitropism of light-grown Arabidopsis hypocotyls. Plant Mol Biol 66, 345–360 (2008). https://doi.org/10.1007/s11103-007-9273-5

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  • DOI: https://doi.org/10.1007/s11103-007-9273-5

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