Over-expression of OsPIN2 leads to increased tiller numbers, angle and shorter plant height through suppression of OsLAZY1.

Crop architecture parameters such as tiller number, angle and plant height are important agronomic traits that have been considered for breeding programmes. Auxin distribution within the plant has long been recognized to alter architecture. The rice (Oryza sativa L.) genome contains 12 putative PIN genes encoding auxin efflux transporters, including four PIN1 and one PIN2 genes. Here, we report that over-expression of OsPIN2 through a transgenic approach in rice (Japonica cv. Nipponbare) led to a shorter plant height, more tillers and a larger tiller angle when compared with wild type (WT). The expression patterns of the auxin reporter DR5::GUS and quantification of auxin distribution showed that OsPIN2 over-expression increased auxin transport from the shoot to the root-shoot junction, resulting in a non-tissue-specific accumulation of more free auxin at the root-shoot junction relative to WT. Over-expression of OsPIN2 enhanced auxin transport from shoots to roots, but did not alter the polar auxin pattern in the roots. Transgenic plants were less sensitive to N-1-naphthylphthalamic acid, an auxin transport inhibitor, than WT in their root growth. OsPIN2-over-expressing plants had suppressed the expression of a gravitropism-related gene OsLazy1 in the shoots, but unaltered expression of OsPIN1b and OsTAC1, which were reported as tiller angle controllers in rice. The data suggest that OsPIN2 has a distinct auxin-dependent regulation pathway together with OsPIN1b and OsTAC1 controlling rice shoot architecture. Altering OsPIN2 expression by genetic transformation can be directly used for modifying rice architecture.

[1]  Zerihun Tadele,et al.  PIN Proteins Perform a Rate-Limiting Function in Cellular Auxin Efflux , 2006, Science.

[2]  B. Bartel,et al.  Auxin: regulation, action, and interaction. , 2005, Annals of botany.

[3]  Ian T. Baldwin,et al.  Use of real-time PCR for determining copy number and zygosity in transgenic plants , 2004, Plant Cell Reports.

[4]  G. Muday,et al.  Auxin Transport and the Integration of Gravitropic Growth , 2008 .

[5]  T. Kagawa,et al.  Phototropin and light-signaling in phototropism. , 2006, Current opinion in plant biology.

[6]  P. Masson,et al.  The arabidopsis thaliana AGRAVITROPIC 1 gene encodes a component of the polar-auxin-transport efflux carrier. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R Chen,et al.  Arabidopsis thalianaのAGRAVITROPIC 1遺伝子は極性オーキシン輸送の流出キャリアの構成員をコード化する , 1998 .

[8]  J. Friml,et al.  PIN Polar Targeting1 , 2008, Plant Physiology.

[9]  R. Firn,et al.  Gravitropic sign reversal—a fundamental feature of the gravitropic perception or response mechanisms in some plant organs , 1994 .

[10]  Litao Yang,et al.  Estimating the copy number of transgenes in transformed rice by real-time quantitative PCR , 2005, Plant Cell Reports.

[11]  Yonghong Wang,et al.  The Plant Architecture of Rice (Oryza sativa) , 2005, Plant Molecular Biology.

[12]  Qian Qian,et al.  Regulation of OsSPL14 by OsmiR156 defines ideal plant architecture in rice , 2010, Nature Genetics.

[13]  G. Muday,et al.  Inhibition of auxin movement from the shoot into the root inhibits lateral root development in Arabidopsis. , 1998, Plant physiology.

[14]  D. Luo,et al.  Genetic control of rice plant architecture under domestication , 2008, Nature Genetics.

[15]  S. Mccouch,et al.  Transgressive Segregation of Tiller Angle in Rice Caused by Complementary Gene Action , 1998 .

[16]  Ben Scheres,et al.  Polar PIN Localization Directs Auxin Flow in Plants , 2006, Science.

[17]  W. Zhai,et al.  TAC1, a major quantitative trait locus controlling tiller angle in rice. , 2007, The Plant journal : for cell and molecular biology.

[18]  P. Waterhouse,et al.  Agrobacterium-mediated transformation of Australian rice cultivars Jarrah and Amaroo using modified promoters and selectable markers , 2000 .

[19]  P. McSteen Auxin and monocot development. , 2010, Cold Spring Harbor perspectives in biology.

[20]  A. Murphy,et al.  Multidrug Resistance–like Genes of Arabidopsis Required for Auxin Transport and Auxin-Mediated Development Article, publication date, and citation information can be found at www.aspb.org/cgi/doi/10.1105/tpc.010350. , 2001, The Plant Cell Online.

[21]  Q. Shen,et al.  Effects of different nitrogen forms on the growth and cytokinin content in xylem sap of tomato (Lycopersicon esculentum Mill.) seedlings , 2009, Plant and Soil.

[22]  Q. Qian,et al.  LAZY1 controls rice shoot gravitropism through regulating polar auxin transport , 2007, Cell Research.

[23]  Klaus Palme,et al.  AtPIN2 defines a locus of Arabidopsis for root gravitropism control , 1998, The EMBO journal.

[24]  C. Bell,et al.  Requirement of the Auxin Polar Transport System in Early Stages of Arabidopsis Floral Bud Formation. , 1991, The Plant cell.

[25]  Yi-Ling Lin,et al.  Expression of bioactive human interferon-gamma in transgenic rice cell suspension cultures , 2004, Transgenic Research.

[26]  Przemyslaw Prusinkiewicz,et al.  Control of bud activation by an auxin transport switch , 2009, Proceedings of the National Academy of Sciences.

[27]  A. Müller,et al.  Regulation of polar auxin transport by AtPIN1 in Arabidopsis vascular tissue. , 1998, Science.

[28]  C. Luschnig Auxin transport: Why plants like to think BIG , 2001, Current Biology.

[29]  Yinggen Ke,et al.  Effect of Polar Auxin Transport on Rice Root Development , 2003 .

[30]  T. Baskin,et al.  Gravitropism of Arabidopsis thaliana Roots Requires the Polarization of PIN2 toward the Root Tip in Meristematic Cortical Cells[C][W] , 2010, Plant Cell.

[31]  Q. Shen,et al.  Two rice phosphate transporters, OsPht1;2 and OsPht1;6, have different functions and kinetic properties in uptake and translocation. , 2009, The Plant journal : for cell and molecular biology.

[32]  Klaus Palme,et al.  The PIN auxin efflux facilitator network controls growth and patterning in Arabidopsis roots , 2005, Nature.

[33]  Wenzislava Ckurshumova,et al.  Auxin Signaling in Arabidopsis Leaf Vascular Development1 , 2003, Plant Physiology.

[34]  K. Palme,et al.  Auxin transport and gravitational research: perspectives , 2006, Protoplasma.

[35]  Klaus Palme,et al.  Lateral relocation of auxin efflux regulator PIN3 mediates tropism in Arabidopsis , 2002, Nature.

[36]  S. Shimizu-Sato,et al.  Auxin–cytokinin interactions in the control of shoot branching , 2009, Plant Molecular Biology.

[37]  Ping Wu,et al.  Expression of PIN genes in rice (Oryza sativa L.): tissue specificity and regulation by hormones. , 2009, Molecular plant.

[38]  H. Miyagawa,et al.  Metabolism of indole-3-acetic acid in rice: identification and characterization of N-beta-D-glucopyranosyl indole-3-acetic acid and its conjugates. , 2007, Phytochemistry.

[39]  Klaus Palme,et al.  AtPIN4 Mediates Sink-Driven Auxin Gradients and Root Patterning in Arabidopsis , 2002, Cell.

[40]  Shubin Sun,et al.  Conservation and divergence of both phosphate- and mycorrhiza-regulated physiological responses and expression patterns of phosphate transporters in solanaceous species. , 2007, The New phytologist.

[41]  T. Yoshihara,et al.  Identification of the gravitropism-related rice gene LAZY1 and elucidation of LAZY1-dependent and -independent gravity signaling pathways. , 2007, Plant & cell physiology.

[42]  R. Reski,et al.  Overexpression of the Arabidopsis Gene UPRIGHT ROSETTE Reveals a Homeostatic Control for Indole-3-Acetic Acid1[C][W] , 2010, Plant Physiology.

[43]  A. Bajguz,et al.  Conjugates of auxin and cytokinin. , 2009, Phytochemistry.

[44]  Shubin Sun,et al.  Proton pump OsA8 is linked to phosphorus uptake and translocation in rice. , 2009, Journal of experimental botany.

[45]  Mariusz Kowalczyk,et al.  Biosynthesis, conjugation, catabolism and homeostasis of indole-3-acetic acid in Arabidopsis thaliana , 2002, Plant Molecular Biology.

[46]  T. Sakai,et al.  The ABC subfamily B auxin transporter AtABCB19 is involved in the inhibitory effects of N-1-naphthyphthalamic acid on the phototropic and gravitropic responses of Arabidopsis hypocotyls. , 2008, Plant & cell physiology.

[47]  J. Friml,et al.  PIN phosphorylation is sufficient to mediate PIN polarity and direct auxin transport , 2009, Proceedings of the National Academy of Sciences.

[48]  Ping Wu,et al.  A PIN1 family gene, OsPIN1, involved in auxin-dependent adventitious root emergence and tillering in rice. , 2005, Plant & cell physiology.

[49]  I. Blilou,et al.  The PIN auxin efflux facilitators: evolutionary and functional perspectives. , 2005, Trends in plant science.

[50]  Michael Sauer,et al.  Efflux-dependent auxin gradients establish the apical–basal axis of Arabidopsis , 2003, Nature.

[51]  E. Blancaflor,et al.  Complex regulation of Arabidopsis AGR1/PIN2-mediated root gravitropic response and basipetal auxin transport by cantharidin-sensitive protein phosphatases. , 2005, The Plant journal : for cell and molecular biology.

[52]  D. Crosby,et al.  Indole‐3‐acetic Acid , 2003 .

[53]  D. Luth,et al.  Activity of a maize ubiquitin promoter in transgenic rice , 1993, Plant Molecular Biology.

[54]  Jiayang Li,et al.  Increased Expression of MAP KINASE KINASE7 Causes Deficiency in Polar Auxin Transport and Leads to Plant Architectural Abnormality in Arabidopsis[W] , 2005, The Plant Cell Online.

[55]  K. Feldmann,et al.  Arabidopsis AUX1 Gene: A Permease-Like Regulator of Root Gravitropism , 1996, Science.

[56]  Hideyuki Takahashi,et al.  Lazy gene (la) responsible for both an agravitropism of seedlings and lazy habit of tiller growth in rice (Oryza sativa L.) , 1996, Journal of Plant Research.

[57]  S. Neill,et al.  OsEXPA4 and OsRWC3 are involved in asymmetric growth during gravitropic bending of rice leaf sheath bases , 2007 .

[58]  A. Murphy,et al.  The ABC of auxin transport: The role of p‐glycoproteins in plant development , 2006, FEBS letters.

[59]  Célia Baroux,et al.  Cellular efflux of auxin catalyzed by the Arabidopsis MDR/PGP transporter AtPGP1. , 2005, The Plant journal : for cell and molecular biology.

[60]  G. Hagen,et al.  Aux/IAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements. , 1997, The Plant cell.

[61]  PAT: waking up a lazy sleeping beauty , 2007, Cell Research.