Identification and overexpression of gibberellin 2‐oxidase ( GA 2ox ) in switchgrass ( P anicum virgatum L.) for improved plant architecture and reduced biomass recalcitrance

Wuddineh, Wegi A.; Mazarei, Mitra; Zhang, Jiyi; Poovaiah, Charleson R.; Mann, David G. J.; Ziebell, Angela; Sykes, Robert W.; Davis, Mark F.; Udvardi, Michael K.; Stewart, Jr, Charles Neal

doi:10.1111/pbi.12287

Identification and overexpression of gibberellin 2‐oxidase ( GA 2ox ) in switchgrass ( P anicum virgatum L.) for improved plant architecture and reduced biomass recalcitrance

Journal Article · Fri Nov 14 00:00:00 EST 2014 · Plant Biotechnology Journal

DOI:https://doi.org/10.1111/pbi.12287· OSTI ID:1401831

Wuddineh, Wegi A. ^[1]; Mazarei, Mitra ^[1]; Zhang, Jiyi ^[2]; Poovaiah, Charleson R. ^[1]; Mann, David G. J. ^[1]; Ziebell, Angela ^[3]; Sykes, Robert W. ^[3]; Davis, Mark F. ^[3]; Udvardi, Michael K. ^[2]; Stewart, Jr, Charles Neal ^[1]

Department of Plant Sciences University of Tennessee Knoxville TN 37996 USA, Bioenergy Science Center Oak Ridge National Laboratory Oak Ridge TN 37831 USA
Bioenergy Science Center Oak Ridge National Laboratory Oak Ridge TN 37831 USA, Plant Biology Division Samuel Roberts Noble Foundation Ardmore OK 73401 USA
Bioenergy Science Center Oak Ridge National Laboratory Oak Ridge TN 37831 USA, National Renewable Energy Laboratory Golden CO 80401 USA

Summary

Gibberellin 2‐oxidases ( GA 2oxs) are a group of 2‐oxoglutarate‐dependent dioxygenases that catalyse the deactivation of bioactive GA or its precursors through 2β‐hydroxylation reaction. In this study, putatively novel switchgrass C ₂₀ GA 2ox genes were identified with the aim of genetically engineering switchgrass for improved architecture and reduced biomass recalcitrance for biofuel. Three C ₂₀ GA 2ox genes showed differential regulation patterns among tissues including roots, seedlings and reproductive parts. Using a transgenic approach, we showed that overexpression of two C20 GA2ox genes, that is PvGA2ox5 and PvGA2ox9 , resulted in characteristic GA‐deficient phenotypes with dark‐green leaves and modified plant architecture. The changes in plant morphology appeared to be associated with GA2ox transcript abundance. Exogenous application of GA rescued the GA ‐deficient phenotypes in transgenic lines. Transgenic semi‐dwarf lines displayed increased tillering and reduced lignin content, and the syringyl/guaiacyl lignin monomer ratio accompanied by the reduced expression of lignin biosynthetic genes compared to nontransgenic plants. A moderate increase in the level of glucose release in these transgenic lines might be attributed to reduced biomass recalcitrance as a result of reduced lignin content and lignin composition. Our results suggest that overexpression of GA 2ox genes in switchgrass is a feasible strategy to improve plant architecture and reduce biomass recalcitrance for biofuel.

View Accepted Manuscript (Publisher)

Sponsoring Organization:: USDOE

OSTI ID:: 1401831

Alternate ID(s):: OSTI ID: 1220794

Journal Information:: Plant Biotechnology Journal, Journal Name: Plant Biotechnology Journal Journal Issue: 5 Vol. 13; ISSN 1467-7644

Publisher:: Wiley-BlackwellCopyright Statement

Country of Publication:: United Kingdom

Language:: English

References (61)

Gibberellin: inhibitor of an inhibitor of...? Harberd, Nicholas P.; King, Kathryn E.; Carol, Pierre BioEssays, Vol. 20, Issue 12 https://doi.org/10.1002/(SICI)1521-1878(199812)20:12<1001::AID-BIES6>3.0.CO;2-O	journal	December 1998
Influence of antibiotic cefotaxime on somatic embryogenesis and plant regeneration in indica rice Grewal, Deepinder; Gill, Raman; Gosal, Satbir Singh Biotechnology Journal, Vol. 1, Issue 10 https://doi.org/10.1002/biot.200600139	journal	October 2006
Engineering of a high-throughput screening system to identify cellulosic biomass, pretreatments, and enzyme formulations that enhance sugar release Studer, Michael H.; DeMartini, Jaclyn D.; Brethauer, Simone Biotechnology and Bioengineering, Vol. 105, Issue 2, p. 231-238 https://doi.org/10.1002/bit.22527	journal	February 2010
Gibberellin Phytohormone Metabolism Peters, Reuben J. Isoprenoid Synthesis in Plants and Microorganisms https://doi.org/10.1007/978-1-4614-4063-5_16	book	January 2012
High-Throughput Screening of Plant Cell-Wall Composition Using Pyrolysis Molecular Beam Mass Spectroscopy Sykes, Robert; Yung, Matthew; Novaes, Evandro Biofuels https://doi.org/10.1007/978-1-60761-214-8_12	book	January 2009
Over-expression of a gibberellin 2-oxidase gene from Phaseolus coccineus L. enhances gibberellin inactivation and induces dwarfism in Solanum species Dijkstra, C.; Adams, E.; Bhattacharya, A. Plant Cell Reports, Vol. 27, Issue 3 https://doi.org/10.1007/s00299-007-0471-z	journal	November 2007
Engineering gibberellin metabolism in Solanum nigrum L. by ectopic expression of gibberellin oxidase genes Bhattacharya, A.; Ward, D. A.; Hedden, P. Plant Cell Reports, Vol. 31, Issue 5 https://doi.org/10.1007/s00299-011-1214-8	journal	January 2012
Genetic improvement of C4 grasses as cellulosic biofuel feedstocks Jakob, Katrin; Zhou, Fasong; Paterson, Andrew H. In Vitro Cellular & Developmental Biology - Plant, Vol. 45, Issue 3 https://doi.org/10.1007/s11627-009-9214-x	journal	May 2009
An Improved Tissue Culture System for Embryogenic Callus Production and Plant Regeneration in Switchgrass (Panicum virgatum L.) Burris, Jason N.; Mann, David G. J.; Joyce, Blake L. BioEnergy Research, Vol. 2, Issue 4 https://doi.org/10.1007/s12155-009-9048-8	journal	October 2009
Developmental Control of Lignification in Stems of Lowland Switchgrass Variety Alamo and the Effects on Saccharification Efficiency Shen, Hui; Fu, Chunxiang; Xiao, Xirong BioEnergy Research, Vol. 2, Issue 4 https://doi.org/10.1007/s12155-009-9058-6	journal	October 2009
Expression of gibberellin 2-oxidase 4 from Arabidopsis under the control of a senescence-associated promoter results in a dominant semi-dwarf plant with normal flowering Lee, Dong Hee; Lee, In Chul; Kim, Kook Jin Journal of Plant Biology, Vol. 57, Issue 2 https://doi.org/10.1007/s12374-013-0528-1	journal	March 2014
Gibberellin metabolism: new insights revealed by the genes Hedden, Peter; Phillips, Andrew L. Trends in Plant Science, Vol. 5, Issue 12 https://doi.org/10.1016/S1360-1385(00)01790-8	journal	December 2000
Activation of gibberellin 2-oxidase 6 decreases active gibberellin levels and creates a dominant semi-dwarf phenotype in rice (Oryza sativa L.) Huang, Jian; Tang, Ding; Shen, Yi Journal of Genetics and Genomics, Vol. 37, Issue 1 https://doi.org/10.1016/S1673-8527(09)60022-9	journal	January 2010
High throughput Agrobacterium-mediated switchgrass transformation Li, Ruyu; Qu, Rongda Biomass and Bioenergy, Vol. 35, Issue 3 https://doi.org/10.1016/j.biombioe.2010.11.025	journal	March 2011
Shedding light on gibberellic acid signalling Schwechheimer, Claus; Willige, Björn Christopher Current Opinion in Plant Biology, Vol. 12, Issue 1 https://doi.org/10.1016/j.pbi.2008.09.004	journal	February 2009
Plants to power: bioenergy to fuel the future Yuan, Joshua S.; Tiller, Kelly H.; Al-Ahmad, Hani Trends in Plant Science, Vol. 13, Issue 8 https://doi.org/10.1016/j.tplants.2008.06.001	journal	August 2008
Gibberellin control of stamen development: a fertile field Plackett, Andrew R. G.; Thomas, Stephen G.; Wilson, Zoe A. Trends in Plant Science, Vol. 16, Issue 10 https://doi.org/10.1016/j.tplants.2011.06.007	journal	October 2011
Promotion of Cell Division in Tissue Cultures by Gibberellic Acid Digby, J.; Thomas, T. H.; Wareing, P. F. Nature, Vol. 203, Issue 4944 https://doi.org/10.1038/203547b0	journal	August 1964
‘Green revolution’ genes encode mutant gibberellin response modulators Peng, Jinrong; Richards, Donald E.; Hartley, Nigel M. Nature, Vol. 400, Issue 6741 https://doi.org/10.1038/22307	journal	July 1999
A mutant gibberellin-synthesis gene in rice Sasaki, A.; Ashikari, M.; Ueguchi-Tanaka, M. Nature, Vol. 416, Issue 6882 https://doi.org/10.1038/416701a	journal	April 2002
Genetic manipulation of gibberellin metabolism in transgenic rice Sakamoto, Tomoaki; Morinaka, Yoichi; Ishiyama, Kanako Nature Biotechnology, Vol. 21, Issue 8 https://doi.org/10.1038/nbt847	journal	July 2003
A protocol for Agrobacterium-mediated transformation in rice Nishimura, Asuka; Aichi, Ikuko; Matsuoka, Makoto Nature Protocols, Vol. 1, Issue 6 https://doi.org/10.1038/nprot.2006.469	journal	December 2006
Gibberellin biosynthesis and its regulation Hedden, Peter; Thomas, Stephen G. Biochemical Journal, Vol. 444, Issue 1 https://doi.org/10.1042/BJ20120245	journal	April 2012
Gibberellin 2-oxidation and the SLN gene of Pisum sativum Lester, Diane R.; Ross, John J.; Smith, Jennifer J. The Plant Journal, Vol. 19, Issue 1 https://doi.org/10.1046/j.1365-313X.1999.00501.x	journal	July 1999
Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass Fu, Chunxiang; Mielenz, Jonathan R.; Xiao, Xirong Proceedings of the National Academy of Sciences, Vol. 108, Issue 9, p. 3803-3808 https://doi.org/10.1073/pnas.1100310108	journal	February 2011
Molecular cloning and functional expression of gibberellin 2- oxidases, multifunctional enzymes involved in gibberellin deactivation Thomas, S. G.; Phillips, A. L.; Hedden, P. Proceedings of the National Academy of Sciences, Vol. 96, Issue 8 https://doi.org/10.1073/pnas.96.8.4698	journal	April 1999
The CYP88A cytochrome P450, ent-kaurenoic acid oxidase, catalyzes three steps of the gibberellin biosynthesis pathway Helliwell, C. A.; Chandler, P. M.; Poole, A. Proceedings of the National Academy of Sciences, Vol. 98, Issue 4 https://doi.org/10.1073/pnas.98.4.2065	journal	February 2001
Manipulation of plant architecture to enhance lignocellulosic biomass Stamm, P.; Verma, V.; Ramamoorthy, R. AoB Plants, Vol. 2012, Issue 0 https://doi.org/10.1093/aobpla/pls026	journal	January 2012
Gibberellins control Arabidopsis hypocotyl growth via regulation of cellular elongation Cowling, R. J.; Harberd, N. P. Journal of Experimental Botany, Vol. 50, Issue 337 https://doi.org/10.1093/jxb/50.337.1351	journal	August 1999
Decreased shoot stature and grain -amylase activity following ectopic expression of a gibberellin 2-oxidase gene in transgenic wheat Appleford, N. E. J.; Wilkinson, M. D.; Ma, Q. Journal of Experimental Botany, Vol. 58, Issue 12 https://doi.org/10.1093/jxb/erm166	journal	July 2007
Identification and genetic characterization of a gibberellin 2-oxidase gene that controls tree stature and reproductive growth in plum El-Sharkawy, I.; El Kayal, W.; Prasath, D. Journal of Experimental Botany, Vol. 63, Issue 3 https://doi.org/10.1093/jxb/err345	journal	November 2011
MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0 Tamura, Koichiro; Stecher, Glen; Peterson, Daniel Molecular Biology and Evolution, Vol. 30, Issue 12 https://doi.org/10.1093/molbev/mst197	journal	October 2013
Expression of Two Genes Encoding Gibberellin 2- and 3-oxidases in Developing Seeds of Phaseoluscoccineus Solfanelli, Cinzia; Ceron, Francesca; Paolicchi, Fabio Plant and Cell Physiology, Vol. 46, Issue 7 https://doi.org/10.1093/pcp/pci124	journal	July 2005
Gibberellin 20-Oxidase Gene OsGA20ox3 Regulates Plant Stature and Disease Development in Rice Qin, Xue; Liu, Jun Hua; Zhao, Wen Sheng Molecular Plant-Microbe Interactions, Vol. 26, Issue 2 https://doi.org/10.1094/MPMI-05-12-0138-R	journal	February 2013
Gibberellin Produced in the Cotyledon Is Required for Cell Division during Tissue Reunion in the Cortex of Cut Cucumber and Tomato Hypocotyls Asahina, Masashi; Iwai, Hiroaki; Kikuchi, Akira Plant Physiology, Vol. 129, Issue 1 https://doi.org/10.1104/pp.010886	journal	March 2002
Activation Tagging of a Dominant Gibberellin Catabolism Gene ( GA 2-oxidase ) from Poplar That Regulates Tree Stature Busov, Victor B.; Meilan, Richard; Pearce, David W. Plant Physiology, Vol. 132, Issue 3 https://doi.org/10.1104/pp.103.020354	journal	June 2003
Impact of Altered Gibberellin Metabolism on Biomass Accumulation, Lignin Biosynthesis, and Photosynthesis in Transgenic Tobacco Plants Biemelt, Sophia; Tschiersch, Henning; Sonnewald, Uwe Plant Physiology, Vol. 135, Issue 1 https://doi.org/10.1104/pp.103.036988	journal	April 2004
Molecular Cloning of GA 2-Oxidase3 from Spinach and Its Ectopic Expression in Nicotiana sylvestris Lee, Dong Ju; Zeevaart, Jan A. D. Plant Physiology, Vol. 138, Issue 1 https://doi.org/10.1104/pp.104.056499	journal	April 2005
Ectopic Expression of Pumpkin Gibberellin Oxidases Alters Gibberellin Biosynthesis and Development of Transgenic Arabidopsis Plants Radi, Abeer; Lange, Theo; Niki, Tomoya Plant Physiology, Vol. 140, Issue 2 https://doi.org/10.1104/pp.105.073668	journal	December 2005
The SLENDER Gene of Pea Encodes a Gibberellin 2-Oxidase Martin, David N.; Proebsting, William M.; Hedden, Peter Plant Physiology, Vol. 121, Issue 3 https://doi.org/10.1104/pp.121.3.775	journal	November 1999
Expression of a Gibberellin 2-Oxidase Gene around the Shoot Apex Is Related to Phase Transition in Rice Sakamoto, Tomoaki; Kobayashi, Masatomo; Itoh, Hironori Plant Physiology, Vol. 125, Issue 3 https://doi.org/10.1104/pp.125.3.1508	journal	March 2001
Overexpression of a Novel Class of Gibberellin 2-Oxidases Decreases Gibberellin Levels and Creates Dwarf Plants Schomburg, Fritz M.; Bizzell, Colleen M.; Lee, Dong Ju The Plant Cell, Vol. 15, Issue 1 https://doi.org/10.1105/tpc.005975	journal	December 2002
Gibberellin Signaling: Biosynthesis, Catabolism, and Response Pathways Olszewski, Neil; Sun, Tai-ping; Gubler, Frank The Plant Cell, Vol. 14, Issue suppl 1 https://doi.org/10.1105/tpc.010476	journal	May 2002
Genetic Analysis Reveals That C ₁₉ -GA 2-Oxidation Is a Major Gibberellin Inactivation Pathway in Arabidopsis Rieu, Ivo; Eriksson, Sven; Powers, Stephen J. The Plant Cell, Vol. 20, Issue 9 https://doi.org/10.1105/tpc.108.058818	journal	September 2008
A Novel Class of Gibberellin 2-Oxidases Control Semidwarfism, Tillering, and Root Development in Rice Lo, Shuen-Fang; Yang, Show-Ya; Chen, Ku-Ting The Plant Cell, Vol. 20, Issue 10 https://doi.org/10.1105/tpc.108.060913	journal	October 2008
The gibberellin biosynthetic genes AtGA20ox1 and AtGA20ox2 act, partially redundantly, to promote growth and development throughout the Arabidopsis life cycle: GA20ox function in Arabidopsis Rieu, Ivo; Ruiz-Rivero, Omar; Fernandez-Garcia, Nieves The Plant Journal, Vol. 53, Issue 3 https://doi.org/10.1111/j.1365-313X.2007.03356.x	journal	October 2007
A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures Murashige, Toshio; Skoog, Folke Physiologia Plantarum, Vol. 15, Issue 3, p. 473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x	journal	July 1962
Stable expression of AtGA2ox1 in a low-input turfgrass (Paspalum notatum Flugge) reduces bioactive gibberellin levels and improves turf quality under field conditions Agharkar, Mrinalini; Lomba, Paula; Altpeter, Fredy Plant Biotechnology Journal, Vol. 5, Issue 6 https://doi.org/10.1111/j.1467-7652.2007.00284.x	journal	November 2007
Gateway-compatible vectors for high-throughput gene functional analysis in switchgrass (Panicum virgatum L.) and other monocot species: Gateway-compatible vectors for monocot transformation Mann, David G. J.; LaFayette, Peter R.; Abercrombie, Laura L. Plant Biotechnology Journal, Vol. 10, Issue 2 https://doi.org/10.1111/j.1467-7652.2011.00658.x	journal	September 2011
Silencing of 4-coumarate:coenzyme A ligase in switchgrass leads to reduced lignin content and improved fermentable sugar yields for biofuel production Xu, Bin; Escamilla-Treviño, Luis L.; Sathitsuksanoh, Noppadon New Phytologist, Vol. 192, Issue 3 https://doi.org/10.1111/j.1469-8137.2011.03830.x	journal	July 2011
Tissue-specific expression of Populus C19 GA 2-oxidases differentially regulate above- and below-ground biomass growth through control of bioactive GA concentrations Gou, Jiqing; Ma, Cathleen; Kadmiel, Mahita New Phytologist, Vol. 192, Issue 3 https://doi.org/10.1111/j.1469-8137.2011.03837.x	journal	August 2011
Functional characterization of the switchgrass (Panicum virgatum) R2R3-MYB transcription factor PvMYB4 for improvement of lignocellulosic feedstocks Shen, Hui; He, Xianzhi; Poovaiah, Charleson R. New Phytologist, Vol. 193, Issue 1, p. 121-136 https://doi.org/10.1111/j.1469-8137.2011.03922.x	journal	October 2011
Two-year field analysis of reduced recalcitrance transgenic switchgrass Baxter, Holly L.; Mazarei, Mitra; Labbe, Nicole Plant Biotechnology Journal, Vol. 12, Issue 7 https://doi.org/10.1111/pbi.12195	journal	April 2014
Molecular Mechanism of Gibberellin Signaling in Plants Sun, Tai-ping; Gubler, Frank Annual Review of Plant Biology, Vol. 55, Issue 1 https://doi.org/10.1146/annurev.arplant.55.031903.141753	journal	June 2004
Gibberellin Metabolism and its Regulation Yamaguchi, Shinjiro Annual Review of Plant Biology, Vol. 59, Issue 1 https://doi.org/10.1146/annurev.arplant.59.032607.092804	journal	June 2008
Enhanced characteristics of genetically modified switchgrass (Panicum virgatum L.) for high biofuel production Shen, Hui; Poovaiah, Charleson R.; Ziebell, Angela Biotechnology for Biofuels, Vol. 6, Issue 1 https://doi.org/10.1186/1754-6834-6-71	journal	January 2013
Gibberellin signaling in plants Daviere, J. -M.; Achard, P. Development, Vol. 140, Issue 6 https://doi.org/10.1242/dev.087650	journal	February 2013
Rice Snl6, a Cinnamoyl-CoA Reductase-Like Gene Family Member, Is Required for NH1-Mediated Immunity to Xanthomonas oryzae pv. oryzae Bart, Rebecca S.; Chern, Mawsheng; Vega-Sánchez, Miguel E. PLoS Genetics, Vol. 6, Issue 9, Article No. e1001123 https://doi.org/10.1371/journal.pgen.1001123	journal	September 2010
OsGA2ox5, a Gibberellin Metabolism Enzyme, Is Involved in Plant Growth, the Root Gravity Response and Salt Stress Shan, Chi; Mei, Zhiling; Duan, Jianli PLoS ONE, Vol. 9, Issue 1 https://doi.org/10.1371/journal.pone.0087110	journal	January 2014
Over-expression of the AtGA2ox8 gene decreases the biomass accumulation and lignification in rapeseed (Brassica napus L.) Zhao, Xiao-ying; Zhu, Deng-feng; Zhou, Bo Journal of Zhejiang University SCIENCE B, Vol. 11, Issue 7 https://doi.org/10.1631/jzus.B1000161	journal	July 2010
Describing and Quantifying Growth Stages of Perennial Forage Grasses Moore, K. J.; Moser, L. E.; Vogel, K. P. Agronomy Journal, Vol. 83, Issue 6 https://doi.org/10.2134/agronj1991.00021962008300060027x	journal	January 1991

Similar Records

Identification and overexpression of a knotted1-like transcription factor in switchgrass (Panicum virgatum L.) for lignocellulosic feedstock improvement

Journal Article · Wed Apr 27 20:00:00 EDT 2016 · Frontiers in Plant Science · OSTI ID:1254112

Gene regulatory networks for lignin biosynthesis in switchgrass (Panicum virgatum )

Journal Article · Sun Sep 16 20:00:00 EDT 2018 · Plant Biotechnology Journal · OSTI ID:1471144

Identification and overexpression of gibberellin 2‐oxidase ( GA 2ox ) in switchgrass ( P anicum virgatum L.) for improved plant architecture and reduced biomass recalcitrance

Citation Formats

References (61)

Similar Records

Related Subjects