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  Effects of natural organic substances on sulphur nutrition (absorption, transport, metabolisation)

The aim of this project is to examine sulphate transporter gene expression in wheat in relation to nutrient availability. Expression of members of the sulphate transporter gene family will analysed in component plant parts in response to different nutrient regimes (with and without defined organic additives) and related to plant tissue nutrient status as assessed by chemical analysis.

  Optimising nutrient use in cereals

Nitrogen (N) is a major input determining productivity, with considerable financial and environmental costs. N is required for canopy formation for efficient carbon capture, thus determining yield. N use efficiency (NUE) (yield/available N) is the product of N uptake efficiency (N taken up/N available) and N utilization efficiency (yield/N taken up). Component traits will be de-convoluted and component genes identified, exploiting variation in alleles and expression profiles in modern wheat cultivars. Roles of nitrate transporters in the roots in controlling uptake processes will be determined: cellular nitrate pools will be measured and compared in soil grown roots of selected cultivars; nitrate influx will be measured on roots growing in soil; the wasteful loss of nitrate from root cells through the efflux pathways will be assessed; key nitrate transporters involved in sensing N availability and uptake will be characterised. Traits associated with NUtE (N and carbon assimilation and partitioning) will be de-convoluted, mapped and data made available for modelling studies. N is utilised for canopy production/photosynthesis which subsequently determines yield and this same N is utilised for grain formation, obviating the need for further uptake. Timing of senescence is critical to extend the photosynthetically active period and to maximise N transfer to the grain. We will investigate variation in associated parameters (canopy architecture, photosynthetic capacity, N storage and remobilisation) screening germplasm, functional staygreen mutants and transgenic lines. Key genes, both those involved in the pathways themselves or in the control of the pathways (transcription factors, signalling pathways) which underpin NUE traits will be identified using mapping approaches and transcriptomics. Cell-specific gene expression of glutamine synthetase and other candidate genes will be examined.

  Optimising nutritional quality of crops

1. Production of transgenic plants with enhanced amino acid synthesis
The primary mechanism for increased nutritional quality of crops will be through the application of gene technology and the production of transgenic plant materials. In order to develop crops with increased amino acid content, unique combinations of optimised inputs, enhanced biosynthesis and increased sink strength will be used. A large number of transgenic lines will be generated and analysed. Expected achievements are:
- Production of plants with enhanced lysine production and decreased catabolism
- Plants with constitutively expressed sulfate uptake and/or over-expressed delivery to sink tissue (RES)
- Production of plants with enhanced glutathione synthesis
- Production of plants with enhanced flux to methionine and SMM, and hence delivery of reduced-sulfur to sink tissues
- Production of plants with synergistically combined manipulations, such as:
+ Increased sulfate uptake and sulfur-assimilatory pathway (RES)
+ Increased amino acid biosynthesis and sink protein production (lysine- and sulfur-amino acids)
+ Increased sulfur delivery to sink organs and sink proteins (RES)
2. Transfer of technology to produce crop plants with increased nutritional value
3. Increased knowledge on pathways controlling plant amino acid biosynthesis
4. Knowledge on consequences of transgenic manipulation of targeted aspects of metabolism.

  Regulation of sulphate transporter gene expression and sulphur metabolism in cereals, source-sink interactions and sulphur supply to grain tissues

S-transport has potential application, both in maximising uptake from the soil and for manipulating allocation within the plant, for example in optimising allocation to the grain. Additionally S-transport is a useful model system to evaluate the effectiveness of genetic manipulation of nutrient uptake systems.

Previous work has resulted in the cloning of sulphate transporter genes in a tropical legume and in barley. These were the first plant sulphate transporters (STs) to be isolated. Two functionally distinct types were found in Stylosanthes, but only one, a high affinity root uptake system was isolated from barley. A major goal remains in the isolation of the lower affinity types in cereals (an example of which was successfully cloned in the Stylosanthes work). These are hypothesised to be responsible for internal cycling of sulphate, and particularly for S-resource allocation to the grain. Emphasis in the project is on cereals, and on wheat in particular, however some underpinning work is performed with Arabidopsis. Our specific goal is to clone the transporters responsible for delivery of S to the grain, a target that is particularly appropriate for transgenic manipulation.

Our long term aims are:
1. Cloning and analysis of components of S-uptake/assimilatory pathway including the transporters and the regulatory elements
2. Manipulation of S-metabolism to favour S-allocation to harvested parts of the plant for crop improvement

The immediate goal for this project is:
The isolation of the full complement of sulphate transporters in wheat and determination of patterns of expression of uptake and partitioning in relation to nutrition and S-assimilate partitioning in plants.

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• Malcolm J. Hawkesford, Saroj Parmar and Peter Buchner (2012). Mineral composition analysis: measuring anions in plant tissues and their uptake.  Ed.  Frans Maathuis. "Methods in Molecular Biology, Humana Press, USA'


• Emmanuelle Cabannes, Peter Buchner, and Malcolm J. Hawkesford (2012). Identification and sequence analysis of sulfate/selenate transporters in selenium hyper- and non-accumulating astragalus plant species.  “Proceedings of the 8th Sulfur Workshop” held in Creswick, Australia 2010.


• Hideki Takahashi, Peter Buchner, Naoko Yoshimoto, Malcolm J. Hawkesford and Shin-Han Shiu. (2012). Evolutionary Relationships and Functional Diversity of Plant Sulfate Transporters. Front Plant Sci. Vol 2, Article 119


•  Emmanuelle Cabannes,  Peter Buchner, Martin R. Broadley and Malcolm J. Hawkesford (2011) A comparison of sulfate and selenium accumulation in relation to expression of sulfate transporter genes in Astragalus species. Plant Physiololgy 157, 2227-22239


• Peter R Shewry, Jacqueline L Stroud, Zhao Fang-Jie, Peter Buchner, Fumie Shinmachi, Stephen P McGrath, Joel Abecassis, Malcolm J Hawkesford (2010). Impacts of sulphur nutrition on selenium and molybdenum concentrations in wheat grain. Journal of Cereal Science 52, 111-113 


• Fumie Shinmachi , Peter Buchner , Jacqueline L. Stroud , Saroj Parmar , Fang-Jie Zhao , Steve P. McGrath , and Malcolm J. Hawkesford (2010). Influence of sulfur deficiency on the expression of specific sulfate transporters and the distribution of sulfur, selenium and molybdenum in wheat. Plant Physiology 153, 327-336. 


• Peter Buchner, Saroj Parmar, Anne Kriegel, Magali Carpentier and Malcolm J. Hawkesford (2010). The sulfate transporter family in wheat: tissue-specific gene expression in relation to nutrition. Molecular Plant, 3, 374-389.


• Peter Buchner and Malcolm Hawkesford (2009). The wheat sulphate transporter family in relation to other cereals. Sirko et al. (eds), Sulfur Metabolism in higher plants, pp. 163-167. Markgraf Publisher, Germany.


• Emmanuelle Cabannes, Peter Buchner, Martin R. Broadley, Philip J. White and Malcolm Hawkesford (2009). Sulfate/Selenate transporters in selenium accumulating plants. Sirko et al. (eds), Sulfur Metabolism in higher plants, pp. 175-178. Markgraf Publisher, Germany.


• A. Koralewska, C.E.E. Stuiver, F.S. Posthumus, M. Shahbaz, P. Buchner, M.J. Hawkesford, L.J. De Kok  (2009). Whole plant regulation of sulfate uptake and distribution in brassica species. 18th Symposium of the International Scientific Centre of Fertilizers, Italy 


• Aleksandra Koralewska, Peter Buchner, C. Elisabeth E. Stuiver, Freek S. Posthumus, Stanislav Kopriva, Malcolm J. Hawkesford and Luit J. De Kok (2009). Expression and activity of sulfate transporters and APS reductase in curly kale in response to sulphate deprivation and re-supply. Journal of Plant Physiology, 166(2),168-79 


• Peter Buchner Plant Sulfate Transporter - in Plant membrane and vacuolar transporters. Editors: P K Jaiwal, R P Singh & O P Dhankher,  Publisher: CAB International, UK, June 2008. 


• Baxter I, Muthukumar B, Park HC, Buchner P, Lahner B, Danku J, Zhao K, Lee J, Hawkesford MJ, Guerinot ML, Salt DE (2008). Variation in Molybdenum Content Across Broadly Distributed Populations of Arabidopsis thaliana is Controlled By a Mitochondrial Molybdenum Transporter (MOT1), PLOS Genetics 4, 1-13


• Dimitris L. Bouranis, Peter Buchner, Styliani N. Chorianopoulou,Laura Hopkins, Vassilis E. Protonotarios,, Vassilis F. Siyiannis,and Malcolm J. Hawkesford (2008). Responses to Sulfur Limitation in Maize. N.A. Khan, S.Singh, S.Umar (eds.), Sulfur Assimilation and Abiotic Stress in Plants. Springer-Verlag Berlin Heidelberg.


• Koralewska, F. S. Posthumus, C. E. E. Stuiver, P. Buchner, M. J. Hawkesford, and L. J. De Kok (2007). The Characteristic High Sulfate Content in Brassica oleracea is controlled by the Expression and Activity of Sulfate Transporters. Plant Biology 9, 654-661.


• S. Parmar, P. Buchner and M. J. Hawkesford (2007). Leaf developmental stage affects sulfate depletion and specific sulfate transporter expression during sulfur deprivation in Brassica napus L. Plant Biology 9, 647-653. (Impact factor 2.059)


• Sonia Plaza; Kathryn L. Tearall, Fang-Jie Zhao, Peter Buchner, Steve P. McGrath, Malcolm J. Hawkesford (2007) Expression and functional analysis of metal transporter genes in two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens. Journal of Experimental Botany 58, 1717-1728.


• Malcolm J Hawkesford, Jonathan R Howarth and Peter Buchner (2006). Control of sulfur uptake, assimilation and metabolism. In: Control of Primary Metabolism in Plants, Ed. Plaxton & McManu. TechBook, New Dehli.


•  MJ Hawkesford, P Buchner, J R Howarth and C Lu (2005). Understanding the Regulation of sulphur nutrition: from sulphate transporter genes to the field. Phyton 45, 57-67.


• P Buchner and M J Hawkesford (2005). Root sulphate uptake and distribution is mediated by multiple sulphate transporter isoforms. Aspects of Applied Biology 73, 119-131.


• Kataoka T, Watanabe-Takahashi A, Hayashi N, Ohnishi M, Mimura T, Buchner P, Hawkesford MJ, Yamaya T, Takahashi H (2004). Vacuolar sulfate transporters play critical roles in root-to-shoot transport of sulfate in Arabidopsis. The Plant Cell 16, 2693-2704.


• Buchner P, Stuiver CEE, Westermann S, Wirtz M, Hell R, Hawkesford MJ, deKok LJ (2004). Regulation of sulfate uptake and expression of sulfate transporter genes in Brassica oleracea l. as affected by atmospheric h2s and pedospheric sulfate nutrition. Plant Physiology 136, 3396-3408.


• Buchner P, Takahashi H, Hawkesford MJ (2004). Plant sulphate transport � a highly regulated cascade of uptake, intracellular and long distance transport. Journal of Experimental Botany 55, 1785-1798.


• Buchner P, Prosser I, Hawkesford MJ (2004). Phylogeny and expression of paralogous and orthologous sulphate transporter genes in diploid and hexaploid wheats. Genome 47 (3), 526-534.


• Casteleden CK, Aoki N, Gillespie VJ, MacRae EA, Quich WP, Buchner P, Foyer CH, Furbank RT, Lunn JE (2004). Evolution and function of sucrose-phosphate synthase gene families in wheat (Triticum aestivum L.) and other grasses. Plant Physiology 135, 1753-1764.


• Gomez, LD, Vanacker H, Buchner P, Noctor G, and Foyer CH (2004). Intercellular distribution of glutathione synthesis in maize leaves and its response to short-term chilling. Plant Physiology� 134, 1662-1672.


• MJ Hawkesford, P Buchner, L Hopkins and J Howarth (2003). Sulphate uptake and transport. In: Sulfur in Plants, eds YP Abrol and A Ahmad. Kluwer Academic Press, Dordrecht, The Netherland. pp 71-86.


• Hawkesford MJ, Buchner P, Hopkins L, Howarth JR. (2003). The plant sulphate transporter family: specialized function and integration with whole plant nutrition. In: Davidian J-C, Grill D, de Kok LJ, Stulen I, Hawkesford MJ, Schnug E, Rennenberg H. Eds. Sulfur transport and assimilation in plants. Leiden, The Netherlands: Backhuys Publishers, 1-10.


• Buchner P, Rochat C, Wuilleme S, and Boutin J-P. 2002. Characterization of a tissue-specific and developmantally regulated β-1.3-glucanase gene in pea (Pisum sativum). Plant Molecular Biology 49, 171-186.


• Buchner P. (2001). Differential cloning. In: Hawkesford MJ, Buchner P. Eds. Molecular analysis of plant adaptation to environment. Plant Ecophysiology Series - Dordrecht, The Netherland, Kluwer Academic pubishers, 17-42.


• Czihal A, Conrad B, Buchner P, Brevis R, Farouk AF, Manteuffel R, Adler K, Wobus U, Hofemeister J, B�umlein H. (1999). Gene Farming in Plant: Expression of a heatstable Bacillus Amylase in transgenic legume seeds. Journal Plant Physiology 155, 183-189.


• Buchner, P. and Boutin (1998). A MADS box transcription factor of the AP1/AGL9 subfamily is also expressed in the seed coat of pea (Pisum saivum) during development. Plant Molecular Biology 38, 1253-1255.


• Buchner, P., Poret, M., Rochat, C. (1998). Cloning and characterization of a cDNA (Accession No. AJ001071) encoding a second sucrose synthase sene in Pea (Pisum sativum L.). (PGR98-105) Plant Physiology 117, 719.


• Albrecht, T., Greve, B., Pusch, K., Kossmann, J., Buchner, P., Wobus, U., Steup, M. (1998). Homodimers and heterodimers of Pho1-type phosphorylase isoforms in Solanum tuberosum L. as revealed by sequence specific antibodies. European. Journal of Biochemistry 251, 343-252. 


• Buchner, P., Borisjuk, L., Wobus, U. (1996). Glucan Phosphorylases in Vicia faba L.: cloning, structural analysis and expression patterns of cytosolic and plastidic forms in relation to starch. Planta 199, 64-73.


• Buchner, Peter (1996). Molecular studies of a-1,4 glucan phosphorylases in Vicia faba L. and transgenic Vicia narbonensis L. PhD-Thesis, University of Halle-Wittenberg, Germany.


• Weber, H., Heim, U., Borisjuk, L., Buchner, P., Wobus, U. (1995). Seed coat-associated invertases of Fava bean control both unloading and storage functions: Cloning of cDNAs and cell type-specific expression. The Plant Cell, Vol. 7, 1835-1846.


• Weber H., Buchner, P., Borisjuk, L., Wobus, U. (1995). Sucrose phosphate synthase and sucrose synthase are both involved in sucrose metabolism in developing Vicia faba L.: Expression pattern, metabolic regumation and implications for seed development. Plant Journal 195, 352-361.


• Wobus, U., Weber, H., Borisjuk, L., Panitz, R., Heim, U., Buchner, P., Baumlein, H., and Weschke, W. (1994). Seed development: Developmental and metabolic regulation of storage compound synthesis. 2th Korea-Germany Joint Symposium in Plant Biotechnology, pp. 67-77.


• Buchner, P., Weber, H., Heim, U., Borisjuk, LM., Wobus, U. (1994). alpha-ADP-Glucose Pyrophosphorylase and a-Glucan Phosphorylases in Vicia faba: tissue specific expression in relation to starch synthesis during seed development. Botanikertagung 94, p. 276.