Doctor Robert Sharwood

Robert Sharwood Dr Sharwood’s research focuses on the molecular engineering of higher plant chloroplasts to improve many facets of plant productivity. Chloroplasts harbour the key biochemical reactions of photosynthesis, a process that underpins all life on earth. With climate change and population growth posing significant threats to national and global food security, it is becoming increasingly urgent to bioengineer the world’s critical crop plants to enhance growth under the current and future environments. This significant problem has augmented concerted efforts to engineer chloroplasts in higher plants to improve photosynthetic efficiency.

Dr Sharwood completed his PhD in Plant Science at the Australian National University. His key contributions to the Rubisco engineering field include:

  • Demonstrating that a hybrid Rubisco comprising of sunflower large and tobacco small subunits is not kinetically impaired and can be adequately regulated by tobacco Rubisco activase;
  • Developing a novel fusion strategy for engineering the Rubisco large and small subunits simultaneously without perturbing their kinetics.

Dr Sharwood’s initial postdoctoral research resulted in development of tobacco master line that now paves the way for innovative research into Rubisco biochemistry within higher plants.

During his recent position at the Boyce Thompson Institute, Cornell University, he focused on understanding the mechanism(s) of post-transcriptional regulation of chloroplast gene expression. This has required the use of cutting edge genetic technology such as Virus Induced Gene Silencing (VIGS) to study the functions of nucleus-encoded ribonucleases that are imported to the chloroplast.

Dr Sharwood’s research achievements have impacted this field by advancing the understanding of chloroplast RNA processing and turnover events. This has provided critical information for plant engineers to successfully transfer and express genes of interest into the chloroplast either to enhance plant productivity or for important biotechnological applications such as biofuels and biopharming.

Key contributions that have made this research internationally recognised include:

  • Defining a new paradigm for chloroplast gene regulation, RNA surveillance, which is mediated by the bi-functional ribonuclease RNase J (submitted to RNA; IF 5.2, ERA A);
  • Demonstrating a role for a chloroplast non-coding RNA in regulating chloroplast gene expression; and
  • Characterised the role of the non-specific endoribonculease (CSP41) in chloroplast RNA metabolism.

Areas of Research/Teaching Expertise

Molecular Biology, Plant Biochemistry, Plant Bioengineering, and Plant Physiology.

Grants / Current Projects

  • Dissecting the roles of pyrophosphate hydrolase (PPH) and poly(A) polymerase (PAP) in chloroplast RNA metabolism
    Partner/Funding Body: UWS Seed Grant
    Period: 2011
  • Elucidating the role of the unique ribonuclease, RNase J, in chloroplast RNA processing
    Co-Researchers: David Stern (Boyce Thompson Institute - USA) and Gadi Schuster (Technion University, Israel)
  • Interrogating the diversity of C4 photosynthesis among Australian C4 grasses
    Co-Researchers: Dr Oula Ghannoum
  • Genetic Engineering of the a new C4 model plant – Setaria veridis
    Co-Researchers: Dr Tom Brutnell and Dr Joyce van Eck at the Boyce Thompson Institute -USA
  • Development of a second-generation tobacco masterline for Rubisco engineering
    Co-Researchers: Associate Professor Spencer Whitney at the Australian National University

Award

  • 2013 Science and Innovation Award for Young People in Agriculture, Fisheries and Forestry – Recipient of the Cotton Research and Development Corporation Award

Selected Publications

Book Chapters

Whitney SM, Sharwood RE, (2011) ‘Plastid transformation for Rubisco engineering and protocols for assessing expression’. In: Methods in Molecular Biology: Chloroplast Biotechnology: Methods and Protocols, Edited by: P Maliga, Humana Press Inc., Totowa, NY. In Press (Accepted 25.02.11)

Sharwood RE, Whitney SM, (2010) ‘Engineering the sunflower Rubisco subunits into tobacco chloroplasts – New considerations’. Chapter 19 In: The Chloroplast: Biochemistry, Molecular Biology and Bioengineering: Advances in Photoysnthesis  (http://www.vlpbp.org/Chloroplast%20Book1.html). Edited by C.A. Rebeiz: Springer, Netherlands. Volume 31, 285-306.

Journal Articles

Sharwood RE, Halpert M, Luro S, Schuster G, Stern DB, (2011) 'Chloroplast RNase J compensates for inefficient transcription termination by removal of antisense RNA', RNA, vol.17, no.12, pp 2165-2176

Sharwood RE, Hotto AM, Bollenbach TB, Stern DB (2011) ‘Overaccumulation of the chloroplast antisense RNA AS5 is correlated with decreased abundance of 5S rRNA in vivo and inefficient maturation in vitro’, RNA, vol.17, no.2, pp 230-243. [Ranked in the upper quartile Q1 for the Biochemistry and Molecular Biology Category]

Whitney SM, Sharwood RE, Orr D, White SJ, Alonso H, Galmés J, (2011) 'Isoleucine 309 acts as a C4 catalytic switch that increases ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) carboxylation rate in flaveria', Proceedings of the National Academy of Sciences of the United States of America, Vol.108, no.35, pp 14688-14693

Whitney SM, Kane HJ, Houtz RL, Sharwood RE, (2009) ‘Rubisco oligomers composed of small and large subunits assemble in tobacco plastids and have higher affinities for CO2 and O2’, Plant Physiology, vol.149, no.4, pp 1887-1895 [Ranked 7 /173 in the Plant Sciences Category; 6 Citations]

Bollenbach TJ, Sharwood RE, Gutierrez R, Lerbs-Mache S, Stern DB, (2009) ‘The RNA-binding proteins CSP41a and CSP41b may regulate transcription and translation of chloroplast-encoded RNAs in Arabidopsis’, Plant Molecular Biology, vol.69, no.5, pp 541-52 [Ranked 15/173 in the Plant Sciences Category; 8 Citations]

Sharwood RE, von Caemmerer S, Maliga P, Whitney SM, (2008) ‘The catalytic properties of hybrid Rubisco comprising tobacco small and sunflower large subunits mirror the kinetically equivalent source Rubiscos and can support tobacco growth’, Plant Physiology, vol.146, no.1, pp 83-96 [Ranked 7 /173 in the Plant Sciences Category; 17 Citations]

Whitney SM, Sharwood RE, (2008) ‘Construction of a tobacco master line to improve Rubisco engineering in chloroplasts’,  Journal of Experimental Botany, vol.59, no.7, pp 1909-1921 [Ranked 14/173 in the Plant Sciences Category; 9 Citations]

Hendrickson L, Sharwood RE, Ludwig M, Whitney SM, Badger MR, von Caemmerer S, (2008) ‘The effects of Rubisco activase on C4 photosynthesis and metabolism at high temperature’, Journal of Experimental Botany, vol.59, no.7, pp 1789-1798 [Ranked 14/173 in the Plant Sciences Category; 5 Citations]

Whitney SM, Sharwood RE, (2007) ‘Linked Rubisco subunits can assemble into functional oligomers without impeding catalytic performance’, Journal of Biological Chemistry , vol.282, no.6, pp 3809-3818 [Ranked in the upper quartile Q1 in the Biochemistry and Molecular Biology category; 12 Citations]

Baker RT, Catanzariti AM, Karunasekara Y, Soboleva TA, Sharwood R, Whitney S, Board, PG, (2005) ‘Using deubiquitylating enzymes as research tools’, Methods in Enzymology, vol.398, pp 540-554. [23 Citations]

Conference Proceedings

Sharwood RE, Whitney SM, Andrews TJ, (2001) ‘A comparison of the effectiveness of the promoters and 5’ regions of two plastid genes in directing the synthesis of Rubisco small subunits in tobacco plastids’, 12th International Congress for Photosynthesis; CSIRO Publishing, www.publish.csiro.au/issue/1342.htm

International Conference Presentations

‘Dissecting the role of a novel bi-functional ribonuclease in chloroplast RNA metabolism’, selected speaker and poster presentation at the FASEB Summer Conferences – Post-transcriptional Control of Gene Expression: Mechanisms of mRNA Decay – Carefree Arizona USA (2010)

‘Synthesis and assembly of foreign Rubiscos transplanted into the chloroplasts of higher plants’, poster presentation at the Gordon Research Conference – CO2 assimilation in Plants: Genome to Biome – Maine USA (2008)

‘Improving the throughput for generating and kinetically screening mutated tobacco Rubiscos’, selected speaker and poster presentation at the 14th International Congress of Photosynthesis – Glasgow United Kingdom (2007)

‘A transplastomic method for screening mutated ‘Form I’ Rubisco’s in heteroplasmic tobacco transformants’, selected speaker and poster presentation at the Satellite Meeting – Research Frontiers with Rubisco, the “Elixir of Life” in the Biosphere – Harpenden United Kingdom (2007)

‘The effects of Rubisco activase on C4 photosynthesis and metabolism at high temperature’, Poster presentation at the Satellite Meeting – Research Frontiers with Rubisco, the “Elixir of Life” in the Biosphere – Harpenden United Kingdom (2007)

‘Transplantation of the Rubisco large subunit from the C4 plant Amaranthus edulis into the chloroplasts of tobacco’, poster presentation at Plant Biology 2007 – Chicago USA (2007)

‘Engineering the Sunflower Rubisco subunits into tobacco chloroplasts: New Considerations’ selected speaker presentation at the International Symposium on Chloroplast Bioengineering – University of Illinois (Urbana-Champaign) USA (2005)

‘Transplanting Sunflower Rubisco Subunits into Tobacco Chloroplasts – Strategies and Consequences’, selected speaker presentation at the 13th International Congress for Photosynthesis – Montreal Canada (2004)

‘A comparison of the effectiveness of the promoters and 5’ regions of two plastid genes in directing the synthesis of Rubisco small subunits in tobacco plastids’, poster presentation at the 12th International Congress for Photosynthesis – Brisbane Australia (2001)

Australian Conference Presentations

‘The assembly of foreign Rubisco hexdecamers within chloroplasts of higher plants by linking the large and small subunits’, selected speaker presentation at COMBIO (Combined Biochemistry and Molecular Biology Conference) Brisbane (2006)

‘Analysis of Photosynthesis and Growth by Tobacco Plants harbouring a hybrid Rubisco enzyme composed of Sunflower Large Subunits and Tobacco Small Subunits’, poster presentation at COMBIO (Combined Biochemistry and Molecular Biology Conference) Adelaide (2005)

‘Transplantation of the Sunflower Rubisco Subunits into Tobacco Chloroplasts’ selected speaker presentation at the National Institute of Bioscience- Bringing Bioscience Together*, Canberra (2004) *Participated in Conference Organisation – Preparation of scientific program, abstract booklets and general conference management

‘Expression and Assembly of a ‘Green’ Form I Rubisco from Synechococcus PCC6301 within tobacco plastids’, poster presentation at COMBIO (Combined Biochemistry and Molecular Biology Conference) Melbourne (2003) Awarded student poster prize

‘Evaluation of the effectiveness of the rbcL and psbA promoter and 5’regions in directing the synthesis of Rubisco small subunits in tobacco plastids’, poster presentation at COMBIO (Combined Biochemistry and Molecular Biology Conference) Canberra (2001)

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