The Australian Plant Nutriomics Network

The Groups > Professor Barry Rolfe

Professor Barry G. Rolfe
Professor and Head of Genomic Interactions Group
Research School of Biological Sciences,
The Australian National University
GPO Box 475 , Canberra ACT 2601

Personal Website address:
Email: rolfe@rsbs.anu.edu.au

Research Aims

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The Big Picture

To understand how to control meristem differentiation and hence the general architecture of the plant and root organogenesis

The Short-Term Goals

Chemical signal exchanges between the Rhizobium and the legume
Rhizobium symbiosis and the influence of quorum sensing signals
Proteomic analysis of differentiation signals of somatic embryogenesis of Medicago truncatula
Plant growth promotion by rhizosphere bacteria
Metabolomic analysis of Rhizobium inoculated Medicago truncatula

Group (lab) Members and areas of research

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Dr Michael Djordjevic proteomic research, plant peptides and flavonoid compounds
Dr Jeremy Weinman proteomics, bioinformatics and plant development
Dr Hancai Chen quorum sensing signals and their effects on bacterial eukaryotes
Dr Nijat Imin proteomics and plant development
Dr Tursun Kerim proteomics and plant development and protein bioinformatics
Dr Charles Hocart chemical and mass spectroscopy analysis of small molecules
Dr Angel Morris cell biology analysis of nodulation
Dr Georg Weiller bioinformatics and proteomics
Dr Kerong Zhang isolation of peptides and small proteins
Dr Ulrike Mathesius symbiosis and proteomics
Dr Puddy Hall proteomic analysis of root tip, lateral root and nodule meristems
Mr Giel van Noorden proteomic and physiological analysis of the sunn mutant
Mr Greg Bodulovic legal, political and sociological analysis of the GMO debate
Mr Joko Prayitno proteomic and physiological analysis of the skl mutant
Ms Femke de Jong Proteomic analysis of somatic embryogenesis
Ms Jiau (Jean) Wen bioinformatic analysis of legume systems
Ms Peta Holmes proteomic analysis of root tip, lateral root and nodule meristems
Ms Marie Oakes chemical and mass spectroscopy analysis of legume sap
Ms Elena Gartner legume growth and microscopy analysis

Funding

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Current

ARC Centre of Excellence 2003-2007 $10 million
ANU block grant and special support for the ARC Centre
USDA shared grant Prof WD Bauer 2001-2005
ARC LIEF grants for Mass spectroscopy equipment

Techniques in the Laboratory

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Currently active

proteomic analysis, 2-DE, mini-gels and LC/MS
gene cloning and analysis
quantitative real time PCR
protein isolation and characterisation
bioinformatics analysis of proteins and genomes
legume protoplast culture and analysis
HPLC and GC/MS, LC/MS analysis

Main areas of expertise

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Top three areas of knowledge

Molecular genetic analysis
Proteomic analysis of bacterial and plant systems
Legume developmental biology and nodule formation

Top three technical skills

Microbial genetics and physiology
Induction and regulation of nodulation of legumes
Research and Development of biotechnology

Statement of Most Significant Contributions To This Research Field

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I have assembled and maintained a focused group undertaking long-term basic research using molecular biology techniques to analyse Plant Microbe interactions. Crucial to my success in this competitive research field has been my capacity to harness newly emerging technologies.

In the 1970's I began a transposon mutagenesis program of Rhizobium and my group developed the first systematic nomenclature describing symbiotic mutant phenotypes, contributing to the more rapid genetic analysis of the Rhizobium -legume symbiosis. John Shine and I then developed the "circuit experiment", using transposon mutations to tag, clone, and verify by complementation individual sym -genes. This led to sequencing the first nodulation and nitrogenase genes of several chosen rhizobia. My group pioneered isolation and characterisation of legume molecules which control Rhizobium nodulation gene expression.

During this period, I examined the basis of Rhizobium host-specificity and its possible extension to the non-leguminous plants rice and wheat. I was first to prove Rhizobium colonisation within the roots of rice.

In the late 1980s my group expanded into plant molecular biology, developing a series of specific transgenic clovers to analyse signal transduction pathways, phytohormone interactions, root cortical cell division and nodule primordia formation in legumes.

In 1994 I initiated my group's proteomic analysis of Plant Microbe interactions, developing proteome reference maps for both Rhizobium and the legume Medicago truncatula. I obtained a DISR grant to set up a collaboration with the Australian Proteome Analysis Facility in Sydney that joined the international consortia undertaking genomic and post-genomic analyses of the model symbiotic partners, S. meliloti and M. truncatula. From 1996, with support first from the RIRDC and later from the CRC for Sustainable Rice Production, my group established an efficient method for the isolation of rice anthers at specific developmental stages. Our analyses produced the first proteome reference maps of rice anthers of the commercial Australian rice cultivar Doongara. These proteome reference maps contain over 4,000 rice anther proteins expressed during the early stages of microspore development. Although plant proteomics is in its early days my group has played a major role in developing a proteomic capability for plant biology in Australia .

Other Evidence of Impact and Contributions to the Field

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Awarded the W.B. Clarke Medal (in Botany) (1989) by the Royal Society of New South Wales for contributions to biological sciences.
Attracted over 6.54 million dollars in refereed external funding for research since 1979.
Published 210 refereed papers, 18 invited book chapters, 7 invited review articles and 153 published conference papers.
Co-inventor: 6 patents in plant-microbe interactions and chemical compounds produced.
Appointed to the second Australian group of scientists to assess China 's efforts on Biological Nitrogen Fixation, sponsored by the Australian Academy of Science and the Academy Sinica (1978 and 1983).
Visiting Professor, UCLA , USA (1975); Visiting Professor, Department of Botany, University of Texas , Austin , USA (1983).
French Embassy award of Visiting Scientist to CNRS, Gif , France (1998) and (1999).
Invited Grant Reviewer for the US National Science Foundation (since 1977), US Department of Agriculture (since 1982); Invited Reviewer, Australian Research Grants Committee.
Presented invited lectures on his BNF research in universities and research institutes in France, Netherlands, Philippines, USA, Australia and the various IS-MPMI conferences (since 1982).
Elected Secretary of the Australian Society of Nitrogen Fixation studies (1984-89).
Elected Secretary of the International Society for Molecular Plant-Microbe Interactions (IS-MPMI) (1992-94); Vice President (1994-96); President of the IS-MPMI (1996-1999).
President of the Australian Electrophoresis Society (2000-2001); Vice-President (1999-2000).
Currently serves on the International Boards and review panels of:

(a) the International Congress on Nitrogen Fixation;

(b) the International Symposium on the Molecular Plant-Microbe Interactions;

(c) European Nitrogen Fixation Biennial Conference;

(d) the Editorial board of the Journal Symbiosis; and

(e) is Associate Editor-in-Chief of the Plant Physiology and Biochemistry Journal.
Over thirty years, has supervised over forty successful PhD and MSc graduate students in Canada , USA and Australia .

Publications - Last 5 Years

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Internationally Refereed Journals

Imin, N., de Jong F., Mathesius U., van Noorden G., Saeed N A., Nolan K E., Ding X., Rose R J., Rolfe, B G . Proteome reference maps of Medicago truncatula Embryogenic cell cultures generated from single protoplasts. Proteomics (In Press).
Imin, N., Kerim, T, Rolfe, B G , Weinman, J J. Effect of early stress on the maturation of rice anthers. Proteomics (In Press).
M. A. Djordjevic, S.H. Natera, H-C Chen, G. Weiller, C. Menzel, G. Van Noorden, J. Weinman, S. Taylor, K. Guo and B. G. Rolfe . "Proteome analysis of Sinorhizobium meliloti". (in press)
Teplitski M., Chen H., Rajamani S., Gao M., Merighi M., Sayre R., Robinson J B., Rolfe B G ., Bauer W D. Chlamydomonas secretes compounds that mimic bacterial signals and interfere with quorum sensing regulation in bacteria. Plant Physiology (In Press).
Weiller, G F, Caraux, G, and Sylvester, N. The modal distribution of protein isoelectric points reflects amino acid properties rather than sequence evolution. Proteomics ( in Press).
Rolfe, B G , Mathesius U. , Djordjevic M A., Gresshoff P M. (2003) 11th International Congress on Molecular Plant-Microbe Interactions held at St. Petersburg - A report. Indian Journal of Experimental Biology 41: 1205-1208.
Mathesius U., Mulders S, Gao M, Teplitski M, Caetano-Anolles G, Rolfe B G , Bauer WD. (2003) "Extensive and specific responses of a eukaryote to bacterial quorum-sensing signals" Procedings National Academy Sciences 100: 1444-1449.
Chen H., Teplitski M., Robinson J B., Rolfe B G ., Bauer W D. (2003) Proteomic analysis of wild-type Sinorhizobium meliloti responses to N -Acyl homoserine Lactone quorum-sensing signals and the transition to stationary phase. J Bacteriology 185: 5029-5036.
Rolfe B G ., Mathesius U., Djordjevic M., Weinman J., Hocart C., Weiller G., Bauer W D. (2003) Proteomic analysis of legume-microbe interactions. Comparative and Functional Genomics 4: 225-228.
Djordjevic M A., Chen H C., Natera S., van Noorden G., Menzel C., Taylor S., Renard C., Geiger O., the Sinorhizobium DNA sequencing Consortium and Weiller G F. (2003) A global analysis of protein expression profiles in Sinorhizobium meliloti : Discovery of new genes for nodule occupancy and stress adaptation. Molecular Plant-Microbe Interactions 16: 508-524.
Kerim, T, Imin, N, Weinman, J J, Rolfe, B G . (2003) Proteome analysis of male gametophyte development in rice anthers. Proteomics 3: 738-751.
Kerim, T, Imin, N, Weinman, J J, Rolfe, B G. (2003) Proteomic analysis reveals developmentally expressed rice homologues of grass group II pollen allergens. Functional Plant Biology 30: 843-852.
Djordjevic MA, Mathesius U, Weinman J.J., Rolfe B.G. (2002) "Two overlapping pathways for triggering nodule development in white clover". pp53-58 In Biology of plant-microbe interactions. Vol 2. (de Wit PJGM, Bisseling T Stiekemea WJ eds) International society for plant-microbe interactions, St Paul Minnesota .
Mathesius U., N. Imin, H.C. Chen, M.A. Djordjevic, J.J. Weinman, S.H.A. Natera, A.C. Morris, T. Kerim, S. Paul, C. Menzel, G. F.Weiller, B. G.Rolfe. Evaluation of proteome reference maps for cross-species identification of proteins by peptide mass fingerprinting. Proteomics (2002) 2: 1288-1303.
Rolfe, B.G ., Weinman J.J. (2001) Rice cultivars and endophytic bacteria Rural Industries Research and Development Corporation Publication, ISBN 0 642 58394 3 20 page booklet.
Mathesius U., Keijzer, G., Natera, S. H. A., Weinman J.J., Djordjevic M.A., Rolfe, B.G. (2001) "Establishment of a root proteome reference map for the model legume Medicago truncatula using the expressed sequence tag database for peptide mass fingerprinting". Proteomics 1: 1424-1440.
Imin, N., Kerim, T., Weinman J.J., Rolfe, B.G . (2001) "Characterisation of rice anther proteins expressed at the young microspore stage". Proteomics 1: 1149-1161.
Perrine , F. M., Prayitno, J., Weinman J.J., Dazzo, F. B., Rolfe, B.G . (2001) " Rhizobium plasmids are involved in the inhibition or stimulation of rice growth and development". Aust. J. Plant Physiology 28: 923-937.
Mathesius U, Keijzers G, Natera S, Weinman J, Djordjevic M and Rolfe B . (2001) Establishment of a root proteome reference map for the model legume Medicago truncatula using the expressed sequence tage database for peptide mass fingerprinting. Proteomics 1 : 1424-1440.
Perrine F, Prayitno J, Weinman J, Dazzo F and Rolfe B. Rhizobium plasmids are involved in the inhibition or stimulation of rice growth and development. (2001) Australian Journal of Plant Physiology 28 : 923-937.
Yanni Y, Rizk R, Abd El Fattah F, Squartini A, Corich V, Giacomini A, de Bruijn F, Rademaker J, Maya-Flores J, Ostrom P, Vega-Hernandez M, Hollingsworth R, Martinez-Molina E, Mateos P, Velazquez E, Wopereis J, Triplett E, Umali-Garcia M, Anarna J, Rolfe B , Ladha J, Hill J, Mujoo R, Ng P and Dazzo F. (2001) The beneficial plant growth-promoting association of Rhizobium leguminosarum bv. trifolii with rice roots. Australian Journal of Plant Physiology 28(9) : 845-870.
Biswas, J. C., Ladha, J. K., Dazzo, F. B., Yanni, Y. G., Rolfe, B. G. (2000) Rhizobial inoculation influences seedling vigor and yield of rice. Agron. J. 92: 880-886.
Mathesius U, Weinman J. J., Rolfe B.G., Djordjevic M. A. (2000) "Rhizobia can induce nodules in white clover by "hijacking" mature cortical cells activated during lateral root development". Molecular Plant-Microbe Interactions, 13: 170-182.
Mathesius U. , Charon C., Rolfe, B.G., Kondorosi A., Crespi M. (2000) "Temporal and spatial order of events during the induction of cortical cell divisions in white clover by Rhizobium leguminosarum bv. trifolii inoculation or localized cytokinin addition". Molecular Plant-Microbe Interactions, 13: 617-628.
Guerreiro N., Ksenzenko V. N., Djordjevic M.A., Ivashina T. V., Rolfe, B.G. (2000) "Elevated levels of synthesis of over 20 proteins results after mutation of the Rhizobium leguminosarum Exopolysaccharide synthesis gene pssA" . Journal of Bacteriology 182: 4521-4532.
de Majnik, J., Weinman, J. J., Djordjevic, M. A., Rolfe, B.G., Tanner, G. J., Joseph, R. G., Larkin, P. J. (2000) "Anthocyanin regulatory gene expression in transgenic white clover can result in an altered pattern of pigmentation". Australian Journal of Plant Physiology 27: 659-667.
Chen H., Higgins J., Oresnik I. J., Hynes M. F., Natera S., Djordjevic M. A., Weinman J. J., Rolfe, B.G. (2000a) "Proteome analysis demonstrates complex replicon and luteolin interactions in pSyma-cured derivatives of Sinorhizobium meliloti strain 2011". Electrophoresis 21: 3833-3842.
Chen, H., Higgins, J., Kondorosi, E., Kondorosi, A., Djordjevic, M. A., Weinman, J. J., Rolfe, B.G. (2000b) "Identification of nolR- regulated proteins in Sinorhizobium meliloti using proteome analysis". Electrophoresis 21: 3823-3832.
Guerreiro N., Djordjevic M.A., Rolfe B.G . (1999) "Proteome analysis of the model microsymbiont Sinorhizobium meliloti: Isolation and characterisation of novel proteins". Electrophoresis 20: 818-825.
Ovtsyna A.O., Rademaker G.-J., Esser E., Weinman J.J., Rolfe B.G., Tikhonovich I.A., Lugtenberg B.J.J., Thomas-Oates J.E., Spaink H.P. (1999) "Comparison of characteristics of the nodX genes from various Rhizobium leguminosarum strains". Molecular Plant-Microbe Interactions, 12: 252-258.
Hutangura P., Mathesius U., Jones M.G.K., Rolfe B.G. (1999) "Auxin induction is a trigger for root gall formation caused by root-knot nematodes in white clover and is associated with the activation of the flavonoid pathway". Australian Journal of Plant Physiology 26: 221-231.
Prayitno J., Stefaniak J., McIver J., Weinman J.J., Dazzo F.B., Ladha,J.K., Barraquio W., Yanni Y.G., Rolfe B.G . (1999) "Interactions of rice seedlings with bacteria isolated from rice roots". Australian Journal of Plant Physiology 26: 521-535.
Worland S., Guerreiro N., Yip L., Djordjevic M.A., Djordjevic S.P., Weinman, J.J., Rolfe, B.G. (1999) " Rhizobium purine auxotrophs, perturbed in nodulation, have multiple changes in protein synthesis". Australian Journal of Plant Physiology 26: 511-519.

Book Chapters

Rolfe, B G , Djordjevic M A., Chen H-C, Hocart C, Weiller G, Mathesius U. Proteomic Analysis of Bacterial Systems. Methods In Micrbial Ecology ( in Press).
Mathesius U., Imin N., Natera S H A., Rolfe B G . (2003) Proteomics as a Functional Genomics Tool. Methods in Molecular Biology 236: Plant Functional Genomics: Methods and Protocols, Edit Grotewold E. pp 395-413, Humana Press, Inc., Totowa , NJ , USA .
Rolfe B.G., Mathesius U., Djordjevic M.A., Weinman J.J., Guerreiro N., Natera S., Morris A.C. (2000) "The use of the Genus Trifolium for the study of plant-microbe interactions, root development and plant defence responses" pp 643-656 In: Prokaryotic Nitrogen Fixation: A model system for analysis of a biological process, E. W. Triplett (Ed), Horizon Scientific Press, Wymondham, UK.
Rolfe B.G., Mathesius U., Prayitno J., Perrine F., Weinman J.J., Stefaniak J., Djordjevic M.A., Guerreiro N., Dazzo F. B. (2000) " Rhizobium nodulation and interaction with legumes and nonlegumes", pp 291-309 In: The quest for nitrogen fixation in rice, Proceedings of the Third Working Group Meeting on assessing opportunities for nitrogen fixation in rice, Ladha,J.K., Reddy P.M. (Eds.) Los Banos, Laguna, Philippines, Makati City, International Rice Research Institute, Philippines .
Dazzo F.B., Yanni Y.G., Rizk R., de Bruijn F.J., Rademaker J., Squartini A., Rolfe B.G., et al., (2000) "Progress in multinational collaborative studies on the beneficial association between Rhizobium leguminosarum bv. trifolii and rice", pp 167-190 In: The quest for nitrogen fixation in rice, Proceedings of the Third Working Group Meeting on assessing opportunities for nitrogen fixation in rice, Ladha,J.K., Reddy P.M. (Eds.) Los Banos, Laguna, Philippines, Makati City, International Rice Research Institute, Philippines.

Ten Career-Best Publications

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Redmond J.W., Batley M., Djordjevic M.A., Innes R.W., Kuempel P.L., Rolfe B.G. (1986) "Flavones induce expression of nodulation genes in Rhizobium" . Nature 323: 632-636.
Djordjevic M.A., Redmond J.W., Batley M., Rolfe B.G . (1987) "Clovers secrete specific phenolic compounds which either stimulate or repress nod gene expression in Rhizobium trifolii ". The EMBO Journal 6: 1173-1179.
McIver J., Djordjevic M.A., Weinman J.J., Bender GL, Rolfe B.G. (1989) "Extension of host range of Rhizobium leguminosarum bv. trifolii caused by point mutations in nodD that result in alterations in regulatory function and recognition of inducer molecules". Molecular Plant-Microbe Interactions 2: 97-106.
Lawson C.G.R., Djordjevic M.A., Weinman J.J., Rolfe B.G. (1994) " Rhizobium inoculation and physical wounding results in the rapid induction of the same chalcone synthase copy in Trifolium subterraneum". Molecular Plant-Microbe Interactions 7: 498-507.
Guerreiro N., Redmond J.W., Rolfe B.G., Djordjevic M.A. (1997) "New Rhizobium leguminosarum flavonoid-induced proteins revealed by proteome analysis of differentially displayed proteins". Molecular Plant-Microbe Interactions 10: 506-516.
Mathesius U., Schlaman H.R.M., Spaink H.P., Sautter C., Rolfe B.G., Djordjevic M.A. (1998) "Auxin transport inhibition precedes root nodule formation in white clover roots and is regulated by flavonoids and derivatives of chitin oligosaccharides". The Plant Journal 14: 23-43.
Mathesius U., Charon C., Rolfe B.G., Kondorosi A., Crespi M. (2000) "Temporal and spatial order of events during the induction of cortical cell divisions in white clover by Rhizobium leguminosarum bv. trifolii inoculation or localized cytokinin addition". Molecular Plant-Microbe Interactions 13: 617-628.
Guerreiro N., Ksenzenko V. N., Djordjevic M.A., Ivashina T. V., Rolfe, B.G. (2000) "Elevated levels of synthesis of over 20 proteins results after mutation of the Rhizobium leguminosarum Exopolysaccharide synthesis gene pssA" . Journal of Bacteriology 182: 4521-4532.
Imin, N., Kerim, T., Weinman J.J., Rolfe, B.G. (2001) "Characterisation of rice anther proteins expressed at the young microspore stage". Proteomics 1: 1149-1161.
Mathesius U., Mulders S, Gao M, Teplitski M, Caetano-Anolles G, Rolfe B G, Bauer WD. (2003) "Extensive and specific responses of a eukaryote to bacterial quorum-sensing signals" Procedings National Academy Sciences 100: 1444-1449.

Additional career-best publications:

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Rolfe B. G., Gresshoff P.M., Shine J. (1980) "Rapid screening for symbiotic mutants of Rhizobium and white clover" Plant Science Letters 19: 277-284.
Djordjevic S.P., Chen, H., Batley M., Redmond J.W., Rolfe B.G. (1987) "Restoration of nitrogen-fixation ability of exopolysaccharide synthesis mutants of Rhizobium strains of NGR234 and R. trifolii by the addition of purified exopolysaccharide" Journal of Bacteriology 169: 53-60.
Rolfe B.G. and Gresshoff P. M. (1988) "Genetic analysis of legume nodule initiation". Annual Review of Plant Physiology and Plant Molecular Biology 39: 297-319.
Guerreiro N., Djordjevic M.A., Rolfe B.G . (1999) "Proteome analysis of the model microsymbiont Sinorhizobium meliloti: Isolation and characterisation of novel proteins". Electrophoresis 20: 818-825..
Mathesius U., Keijzer, G., Natera, S. H. A., Weinman J.J., Djordjevic M.A., Rolfe, B.G. (2001) "Establishment of a root proteome reference map for the model legume Medicago truncatula using the expressed sequence tag database for peptide mass fingerprinting". Proteomics 1: 1424-1440.
Mathesius U, Weinman J. J., Rolfe B.G., Djordjevic M. A. (2000) "Rhizobia can induce nodules in white clover by "hijacking" mature cortical cells activated during lateral root development". Molecular Plant-Microbe Interactions , 13: 170-182.