Skip to main content
SpringerLink
Log in

Silencing of a BYPASS1 homolog results in root-independent pleiotrophic developmental defects in Nicotiana benthamiana

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The Arabidopsis bypass1 mutant (bps1) exhibits defective shoot and root growth that is associated with constitutive production of a root-derived carotenoid-related signal (Van Norman et al., Curr Biol 14:1739–1746, 2004). Since the identity of the signal and the function of BPS1 are still unknown, we investigated effects of BPS1 depletion in Nicotiana benthamiana to elucidate BPS1 function in plant growth and development. The predicted protein of NbBPS1, a BPS1 homolog of N. benthamiana, contains a central transmembrane domain, and a NbBPS1:GFP fusion protein was mainly associated with the endoplasmic reticulum. Virus-induced gene silencing (VIGS) of NbBPS1 resulted in pleiotrophic phenotypes, including growth retardation and abnormal leaf development. At the cellular level, the plants exhibited hyperproliferation of the cambial cells and defective xylem differentiation during stem vascular development. Hyperactivity of the cambium was associated with an elevated auxin and cytokinin response. In contrast, the leaves had reduced numbers of cells with increased cell size and elevated endoreduplication. Cell death in NbBPS1 VIGS leaves started with vacuole collapse, followed by degeneration of the organelles. Interestingly, these phenotypes were mainly caused by silencing of NbBPS1 in the aerial parts of the plants, different from the case of the Arabidopsis bps1 mutant. These results suggest that NbBPS1 plays a role in the control of cell division and differentiation in the cambium of N. benthamiana, and BPS homologs may have a diverse function in different tissues and in different species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Ahn J-W, Kim M, Lim JH, Kim G-T, Pai H-S (2004) Phytocalpain controls the proliferation and differentiation fates of cells in plant organ development. Plant J 38:969–981

    Article  PubMed  CAS  Google Scholar 

  • Aloni R, Schwalm K, Langhans M, Ullrich CI (2003) Gradual shifts in sites of free-auxin production during leaf-primordium development and their role in vascular differentiation and leaf morphogenesis in Arabidopsis. Planta 216:841–853

    PubMed  CAS  Google Scholar 

  • Baima S, Possenti M, Matteucci A, Wisman E, Altamura MM, Ruberti I, Morelli G (2001) The Arabidopsis ATHB-8 HD-zip protein acts as a differentiation-promoting transcription factor of the vascular meristems. Plant Physiol 126:643–655

    Article  PubMed  CAS  Google Scholar 

  • Bennett T, Sieberer T, Willett B, Booker J, Luschnig C, Leyser O (2006) The Arabidopsis MAX pathway controls shoot branching by regulating auxin transport. Curr Biol 16:553–563

    Article  PubMed  CAS  Google Scholar 

  • Booker J, Auldridge M, Wills S, McCarty D, Klee H, Leyser O (2004) MAX3/CCD7 is a carotenoid cleavage dioxygenase required for the synthesis of a novel plant signaling molecule. Curr Biol 14:1232–1238

    Article  PubMed  CAS  Google Scholar 

  • Bouvier F, Suire C, Mutterer J, Camara B (2003) Oxidative remodeling of chromoplast carotenoids: identification of the carotenoid dioxygenase CsCCD and CsZCD genes involved in Crocus secondary metabolite biogenesis. Plant Cell 15:47–62

    Article  PubMed  CAS  Google Scholar 

  • Cho HS, Lee SS, Kim KD, Kim SJ, Hwang I, Lim JS, Park YI, Pai H-S (2004) DNA gyrase is involved in chloroplast nucleoid partitioning. Plant Cell 16:2665–2682

    Article  PubMed  CAS  Google Scholar 

  • Cho M, Lee OR, Ganguly A, Cho HT (2007) Auxin-signaling: short and long. J Plant Biol 50:79–89

    Article  CAS  Google Scholar 

  • Christensen SK, Dagenais N, Chory J, Weigel D (2000) Regulation of auxin response by the protein kinase PINOID. Cell 100:469–478

    Article  PubMed  CAS  Google Scholar 

  • D’Agostino IB, Deruere J, Kieber JJ (2000) Characterization of the response of the Arabidopsis response regulator gene family to cytokinin. Plant Physiol 124:1706–1717

    Article  PubMed  CAS  Google Scholar 

  • De Veylder L, Beeckman T, Beemster GT, Krols L, Terras F, Landrieu I, van der Schueren E, Maes S, Naudts M, Inzé D (2001) Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Plant Cell 13:1653–1668

    Article  PubMed  Google Scholar 

  • den Boer BG, Murray JA (2000) Triggering the cell cycle in plants. Trends Cell Biol 10:245–250

    Article  Google Scholar 

  • Dewitte W, Murray JA (2003) The plant cell cycle. Annu Rev Plant Biol 54:235–264

    Article  PubMed  CAS  Google Scholar 

  • Donnelly PM, Bonetta D, Tsukaya H, Dengler RE, Dengler NG (1999) Cell cycling and cell enlargement in developing leaves of Arabidopsis. Dev Biol 215:407–419

    Article  PubMed  CAS  Google Scholar 

  • Foo E, Turnbull CG, Beveridge CA (2001) Long-distance signaling and the control of branching in the rms1 mutant of pea. Plant Physiol 126:203–209

    Article  PubMed  CAS  Google Scholar 

  • Francis D (2007) The plant cell cycle – 15 years on. New Phytol 174:261–278

    Article  PubMed  CAS  Google Scholar 

  • Fukuda H (2004) Signals that control plant vascular cell differentiation. Nat Rev Mol Cell Biol 5:379–391

    Article  PubMed  CAS  Google Scholar 

  • Fukuda H, Komamine A (1980) Direct evidence for cytodifferentiation to tracheary elements without intervening mitosis in a culture of single cells isolated from the mesophyll of Zinnia elegans. Plant Physiol 65:61–64

    Article  PubMed  CAS  Google Scholar 

  • Gälweiler L, Guan C, Müller A, Wisman E, Mendgen K, Yephremov A, Palme K (1998) Regulation of polar auxin transport by AtPIN1 in Arabidopsis vascular tissue. Science 282:2226–2230

    Article  PubMed  Google Scholar 

  • Gegas VC, Doonan JH (2006) Expression of cell cycle genes in shoot apical meristems. Plant Mol Biol 60:947–961

    Article  PubMed  CAS  Google Scholar 

  • Groover A, Jones AM (1999) Tracheary element differentiation uses a novel mechanism coordinating programmed cell death and secondary cell wall synthesis. Plant Physiol 119:375–384

    Article  PubMed  CAS  Google Scholar 

  • Gutierrez C (2005) Coupling cell proliferation and development in plants. Nat Cell Biol 7:535–541

    Article  PubMed  CAS  Google Scholar 

  • Hardtke CS, Berleth T (1998) The Arabidopsis gene MONOPTEROS encodes a transcription factor mediating embryo axis formation and vascular development. EMBO J 17:1405–1411

    Article  PubMed  CAS  Google Scholar 

  • Joubès J, Chevalier C (2000) Endoreduplication in higher plants. Plant Mol Biol 43:735–745

    Article  PubMed  Google Scholar 

  • Jung JH, Park CM (2007) Vascular development in plants: specification of xylem and phloem tissues. J Plant Biol 50:301–305

    CAS  Google Scholar 

  • Kessler S, Sinha N (2004) Shaping up: the genetic control of leaf shape. Curr Opin Plant Biol 7:65–72

    Article  PubMed  CAS  Google Scholar 

  • Kim M, Lim J-H, Ahn CS, Park K, Kim GT, Kim WT, Pai H-S (2006) Mitochondria-associated hexokinases play a role in the control of programmed cell death in Nicotiana benthamiana. Plant Cell 18:2341–2355

    Article  PubMed  CAS  Google Scholar 

  • Kosugi S, Ohashi Y (2003) Constitutive E2F expression in tobacco plants exhibits altered cell cycle control and morphological change in a cell type-specific manner. Plant Physiol 132:2012–2022

    Article  PubMed  CAS  Google Scholar 

  • Lee YJ, Kim DH, Kim Y-W, Hwang I (2001) Identification of a signal that distinguishes between the chloroplast outer envelope membrane and the endomembrane system in vivo. Plant Cell 13:2175–2190

    Article  PubMed  CAS  Google Scholar 

  • Liu JX, Srivastava R, Che P, Howell SH (2007) An endoplasmic reticulum stress response in Arabidopsis is mediated by proteolytic processing and nuclear relocation of a membrane-associated transcription factor, bZIP28. Plant Cell 19:4111–4119

    Article  PubMed  CAS  Google Scholar 

  • Mähönen AP, Bonke M, Kauppinen L, Riikonen M, Benfey PN, Helariutta Y (2000) A novel two-component hybrid molecule regulates vascular morphogenesis of the Arabidopsis root. Genes Dev 14:2938–2943

    Article  PubMed  Google Scholar 

  • Mähönen AP, Bishopp A, Higuchi M, Nieminen KM, Kinoshita K, Tormakangas K, Ikeda Y, Oka A, Kakimoto T, Helariutta Y (2006) Cytokinin signaling and its inhibitor AHP6 regulate cell fate during vascular development. Science 311:94–98

    Article  PubMed  CAS  Google Scholar 

  • Nieminen KM, Kauppinen L, Helariutta Y (2004) A weed for wood? Arabidopsis as a genetic model for xylem development. Plant Physiol 135:653–659

    Article  PubMed  CAS  Google Scholar 

  • Obara K, Kuriyama H, Fukuda H (2001) Direct evidence of active and rapid nuclear degradation triggered by vacuole rupture during programmed cell death in Zinnia. Plant Physiology 125:615–626

    Article  PubMed  CAS  Google Scholar 

  • Park J-A, Ahn J-W, Kim Y-K, Kim SJ, Kim J-K, Kim WT, Pai H-S (2005) Retinoblastoma protein regulates cell proliferation, differentiation, and endoreduplication in plants. Plant J 42:153–163

    Article  PubMed  CAS  Google Scholar 

  • Parker G, Schofield R, Sundberg B, Turner S (2003) Isolation of COV1, a gene involved in the regulation of vascular patterning in the stem of Arabidopsis. Development 130:2139–2148

    Article  PubMed  CAS  Google Scholar 

  • Pineau C, Freydier A, Ranocha P, Jauneau A, Turner S, Lemonnier G, Renou JP, Tarkowski P, Sandberg G, Jouanin L, Sundberg B, Boudet AM, Goffner D, Pichon M (2005) hca: An Arabidopsis mutant exhibiting unusual cambial activity and altered vascular patterning. Plant J 44:271–289

    Article  PubMed  CAS  Google Scholar 

  • Ramirez-Parra E, Desvoyes B, Gutierrez C (2005) Balance between cell division and differentiation during plant development. Int J Dev Biol 49:467–477

    Article  PubMed  Google Scholar 

  • Ratcliff F, Martin-Hernansez AM, Baulcombe DC (2001) Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J 25:237–245

    Article  PubMed  CAS  Google Scholar 

  • Scarpella E, Meijer AH (2004) Pattern formation in the vascular system of monocot and dicot plant species. New Phytol 164:209–242

    Article  CAS  Google Scholar 

  • Simkin AJ, Schwartz SH, Auldridge M, Taylor MG, Klee HJ (2004) The tomato carotenoid cleavage dioxygenase 1 genes contribute to the formation of the flavor volatiles beta-ionone, pseudoionone, and geranylacetone. Plant J 40:882–892

    Article  PubMed  CAS  Google Scholar 

  • Sorefan K, Booker J, Haurogné K, Goussot M, Bainbridge K, Foo E, Chatfield S, Ward S, Beveridge C, Rameau C, Leyser O (2003) MAX4 and RMS1 are orthologous dioxygenase-like genes that regulate shoot branching in Arabidopsis and pea. Genes Dev 17:1469–1474

    Article  PubMed  CAS  Google Scholar 

  • Stirnberg P, van de Sande K, Leyser HMO (2002) MAX1 and MAX2 control shoot lateral branching in Arabidopsis. Development 129:1131–1141

    PubMed  CAS  Google Scholar 

  • Sugimoto-Shirasu K, Roberts K (2003) “Big it up”: endoreduplication and cell-size control in plants. Curr Opin Plant Biol 6:544–553

    Article  PubMed  CAS  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  PubMed  CAS  Google Scholar 

  • Ulmasov T, Murfett J, Hagen G, Guilfoyle TJ (1997) Aux/IAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements. Plant Cell 9:1963–1971

    Article  PubMed  CAS  Google Scholar 

  • Valentine T, Shaw J, Blok VC, Phillips MS, Oparka KJ, Lacomme C (2004) Efficient virus-induced gene silencing in roots using a modified tobacco rattle virus vector. Plant Physiol 136:3999–4009

    Article  PubMed  CAS  Google Scholar 

  • Van Norman JM, Sieburth LE (2007) Dissecting the biosynthetic pathway for the bypass1 root-derived signal. Plant J 49:619–628

    Article  PubMed  CAS  Google Scholar 

  • Van Norman JM, Frederick RL, Sieburth LE (2004) BYPASS1 negatively regulates a root-derived signal that controls plant architecture. Curr Biol 14:1739–1746

    Article  PubMed  CAS  Google Scholar 

  • Ye ZH (2002) Vascular tissue differentiation and pattern formation in plants. Annu Rev Plant Biol 53:183–202

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The authors wish to thank Dr. Tom J. Guilfoyle (University of Missouri, USA) for providing the DR::GUS construct, and Drs. Jeong Ho Roh and Hye Kyung Rhee (National Horticultural Research Institute, Korea) for providing a technical help for flow cytometry. This research was supported by grants from the Plant Diversity Research Center of the 21st Century Frontier Research Program, a KOSEF grant (M10749000002-07N4900-00210), and the Plant Signaling Network Research Center (at Korea University) of the Science Research Center Program, all of which are funded by the Ministry of Science and Technology of Korea.

Author information

Authors and Affiliations

  1. Department of Biology, Yonsei University, Seoul, 120-749, Korea

    Yong Won Kang, Woo Taek Kim & Hyun-Sook Pai

  2. Plant Genome Research Center, Korea Research Institute of Bioscience and Biotechnology, Taejon, 305-333, Korea

    Ryong Nam Kim & Hye Sun Cho

  3. Department of Plant Sciences, Seoul National University, Seoul, 151-742, Korea

    Doil Choi

Authors
  1. Yong Won Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ryong Nam Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hye Sun Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Woo Taek Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Doil Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hyun-Sook Pai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Doil Choi or Hyun-Sook Pai.

Additional information

Y. W. Kang and R. N. Kim contributed equally to this work.

GenBank accession numbers: EU243336 (NbBPS1) and EU243335 (NbBPS2).

Electronic supplementary material

11103_2008_9384_MOESM1_ESM.ppt

Supplementary Fig. 1 Phenotype variability in the NbBPS1 VIGS lines and silencing of NbBPS1 in the stem. (a) Relative phenotype severity. The average number of abnormal leaves of TRV:BPS1(U) lines was approximately 57% of that of TRV:BPS1(C) lines at 25 DAI (7.3 leaves; n = 50). (b) Semiquantitative RT-PCR analysis of the NbBPS1 mRNA levels in the stem using BPS1-C2 primers. RT-PCR showed significantly reduced PCR products in both lines relative to the TRV:GFP lines, indicating that the endogenous level of NbBPS1 transcripts is reduced in the stem tissue

Supplementary Fig. 2 Effects of fluridone treatment on the NbBPS1 VIGS phenotype. (a) Fluridone (FL) treatment fully rescued the growth arrest and partially rescued the abnormal leaf morphology of the NbBPS1 VIGS plants at 22 DAI. VIGS plants were irrigated with 0.1% ethanol with or without fluoridone (0, 0.1 and 10 μM) from 2 DAI to 22 DAI. (b) Effects of fluridone on plant height of the NbBPS1 VIGS plants at 22 DAI. Relative height (%) was calculated comparing to the height of TRV control treated with 0.1% ethanol (PPT 1735 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kang, Y.W., Kim, R.N., Cho, H.S. et al. Silencing of a BYPASS1 homolog results in root-independent pleiotrophic developmental defects in Nicotiana benthamiana . Plant Mol Biol 68, 423–437 (2008). https://doi.org/10.1007/s11103-008-9384-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11103-008-9384-7

Keywords

Search

Navigation