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Drought_PhD Booklet 2012 v2

Drought_PhD Booklet 2012 v2

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Drought Research Group

PhD research areas 2012

The drought issue
South Australia is the driest state in the driest continent on the planet. Despite the adverse environmental conditions, there is still an opportunity for significant genetic gain in essential crops such as wheat and barley. Plant tolerance to drought is a complex trait. Breeding is further complicated since several types of abiotic stress, such as high temperatures, high irradiance and nutrient toxicities or deficiencies can challenge crop plants simultaneously. The task is particularly challenging in wheat and barley because of the size and complexity of their genomes. However, recent technological advances and the imperative to ensure sustainable food production is driving research programs to genetically improve these crops.

Our aim at ACPFG
Improve the drought tolerance of wheat and barley varieties by discovering and using new genes and alleles. The aim is not to ‘convert wheat to a cactus’ but to allow wheat and barley to continue to grow and yield grain under water limited conditions.

Our strategy
Our research programs tackle this problem in a multi-disciplinary approach, considering interaction between multiple stresses and integrating the physiological dissection of drought tolerance traits by using genetic and genomics tools, such as quantitative trait loci (QTL), microarrays and transgenics.

Cloned gene sequences are introduced into our target crop species by either biolistics (wheat) or Agrobacterium-mediated transformation (barley). The following projects have been initiated: – Detailed physiological analysis of drought tolerant and intolerant wheat lines under controlled environment and field conditions Genetic analysis of drought tolerance under field and growth room conditions in three large populations of wheat Genetic dissection of root development and architecture under normal and drought conditions Development of metabolite and transcript profiles of parental lines under water limited conditions – – – • Reverse genetics: from gene to phenotype We seek to isolate gene sequences important for conferring drought tolerance from both model and crop species.The teams • Forward genetics: from phenotype to gene We observe and analyse the characteristics of drought tolerance in wheat and barley and seek to identify the genes responsible of the tolerance. Project areas include: – Bioinformatics and the identification of drought related gene sequences Transcription factors and the regulation of drought-stress responses Development of commercially viable drought tolerant GM wheat and barley – – . Transgenic plants are then assayed under controlled and field conditions for drought tolerance.

mCherry) Bioinformatics Plant Physiology High throughput phenotyping by imaging Chlorophyll content Stomatal conductance Photosynthesis Plant and soil water potentials Canopy temperature Root and shoot anatomy Root morphology Genetics QTL mapping Molecular markers Positional cloning Marker assisted selection .Our collaborators BBG University of Adelaide (South Australia) DPI La Trobe University (Victoria) INRA Clermont-Ferrand (France) SCRI Dundee (Scotland) IPK Gatersleben (Germany) CIMMYT (Mexico) Pioneer Hi-Bred International/DuPont (USA) INRA Montpellier (France) University of Bologna (Italy) The techniques Molecular biology DNA and RNA extractions PCRs and restriction digests Transcript profiling by microarrays and sequencing Metabolite and protein profiling Confocal microscopy BAC library screening Biotechnology Cloning Biolistics and Agrobacterium-mediated plant transformation Fluorescent reporters (GFP.

University of Adelaide) . Peter Langridge Dr Sergiy Lopato Prof Diane Mather Dr Boris Parent Dr Bujun Shi Dr Ryan Whitford Adelaide node of the ACPFG at the Plant Genomics Centre (Waite Campus.Potential PhD supervisors: Dr Ute Baumann Dr Omid Eini Dr Delphine Fleury Dr Chunyuan Huang Dr Nataliya Kovalchuk Prof.

Initiate positional cloning of heat tolerance genes via candidate genes. Comparative mapping and sequence analysis for targeted marker generation and candidate gene identification. there is an urgent need for reproducible growth chamber assays for heat tolerance that are relevant to the field. Use new or established populations to map chromosome regions (QTLs) controlling variation in tolerance. and this situation will worsen with climate change. there is virtually nothing known about how much wheat genotypes naturally vary for tolerance to heat-induced sterility effects. DArT and SNP markers. In vitro pollen viability/tube-growth assays. SSR. AIMS  Use a growth chamber to characterize the precise developmental stages where durum and bread wheat are most sensitive to heat-induced sterility. Therefore.com.PhD projects: Project title: Genetic and physiological characterization of tolerance to heatinduced floret sterility in wheat SUPERVISOR: Dr Nick Collins CONTACT DETAILS: nick. Chlorophyll fluorescence measurements (Fv/Fm). Characterize the biological basis for the tolerance in various sources.au BACKGROUND Brief periods of heat stress severely impact on wheat production. However. Screen local and exotic durum and bread wheat varieties for variation in tolerance to heatinduced sterility. Gene transcript quantification and tissue-localization by qRT-PCR. and because natural heat events are unpredictable in severity and timing.collins@acpfg. Besides this.     TECHNIQUES TO BE USED        Growth chamber assays for heat tolerance. molecular markers linked to genes controlling heat tolerance. Anecdotal reports indicate that Australian durum wheat varieties are more prone to this form of heat damage than bread wheats. it is difficult for breeders to select for heat tolerance in the field. One type of heat damage is the floret sterility (decrease grain set) following heat events at around or just before pollination. because heat affects wheat in different ways depending on the growth stage. or cloned heat tolerance genes for transformation breeding. . however this difference needs to be formally tested. Use this information to design tolerance assays targeting sterility effects of heat. Characterize tiller stages by examining developing spikes by microscopy (stage of meiosis or development of female reproductive structures).

Saini HS.) cultivars by pollen-based parameters. Assessment of cold and heat tolerance of winter-grown canola (Brassica napus L. Journal of Agronomy & Crop Science 194:225-236. . Australian Journal of Plant Physiology 11:243-253. Cell and Environment 31:11-38. Feher A (2008). Heat treatment of wheat plants using a growth chamber. Reddy KR (2008). Sedgley M. Aspinall D (1984). Baldwin B.REFERENCES Barnabas B. Jager K. Developmental anatomy in wheat of male sterility induced by heat stress. Brand D. Kakani VG. Singh SK. water deficit or abscisic acid. The effect of drought and heat stress on reproductive processes in cereals. Plant. Plants are grown in the greenhouse before and after a brief heat treatment applied at a specific developmental stage.

fleury@acpfg. for which we already have large populations of recombinant inbred lines.langridge@acpfg. SNP). We have identified several QTLs of wheat controlling yield in dry environment. Prof. high throughput phenotyping in the Plant Accelerator facility. Different mechanisms of adaptation to cyclic water stress in two South Australian bread wheat cultivars.com. Peter Langridge CONTACT DETAILS: delphine. AIMS AND SIGNIFICANCE : Identify and validate candidate gene underlying yield QTL TECHNIQUES TO BE USED : Molecular markers (SSR. genome sequence analysis.au BACKGROUND The key objective of the drought program of ACPFG is to generate detailed knowledge of the mechanisms of drought adaptation under the Mediterranean type growing conditions. Journal of Experimental Botany 59. Jefferies S. with a view to developing plants tolerant to multiple components: osmotic & oxidative stress. This type of drought is characterised by water deficit at the late stages of crop development. Journal of Experimental Botany 61: 3211-3222 . peter.au. 2010.com. usually during flowering and grain filling. 3327-3346. Condon AG. statistical analysis. Schnurbusch T. Fleury D. measurement of physiological traits related to yield and plant water status. We are now increasing the number of markers for fine-mapping of each region to the resolution of each local map to <1 cM using sequence-based markers. 2008. dehydration. Tester M. heat. Kuchel H.Project title: Positional cloning of QTL for drought tolerance in wheat SUPERVISORS: Dr Delphine Fleury. Genetic and genomic tools to improve drought tolerance in wheat. The availability of new genomics resources. Langridge P. Langridge P. enables now to make tremendous progress in gene cloning in wheat. Four QTL are targeted for map-based cloning. particularly the next-generation sequencing data. functional analysis of genes REFERENCES Izanloo A. We can now envisage identifying and cloning genes that control yield under drought conditions in wheat.

pp 66-75. These traits are usually controlled by few major genes.au. 1996.whitford@acpfg. and Langridge P. male and female are enclosed in spikelet. Dr Ryan Whitford CONTACT DETAILS: delphine. Hybrid plants obtained by intercrossing inbred lines show an increase in biomass and production.au BACKGROUND One of the Green revolution technologies was the hybrid seeds.com. TECHNIQUES TO BE USED : Molecular markers. . phenotyping of floral development. flowering time isn’t synchronised between male and female parental lines. statistical analysis. Science 327: 818-822. anthers and styles are short. inter-crossing plants is difficult. However due to its selfpollinated nature. Hybrid wheat: advances and challenges. Chasmogamic species are characterized by open flowers and exposed stamens and styles that facilitate inter-pollination. Past studies showed that yield increase is possible to achieve in hybrid wheat compared to the conventional inbred lines.Project title: Genetic study of floral architecture for hybrid wheat system SUPERVISORS: Dr Delphine Fleury. genetic mapping and QTL software REFERENCES Tester M. This heterotic effect is particularly strong in out-breeding species such as maize. Breeding technologies to increase crop production in a changing world. AIMS AND SIGNIFICANCE : Identify QTL and genes controlling flower architecture of wheat for increasing hybridization rate in hybrid seed production. genome sequence analysis. Jordaan JP. Heritability of flower architecture is medium to high suggesting that progress could be made in improving the cross-pollinating ability of parental lines. In: MP Reynolds. One of the factors that impair out-crossing in wheat is its flower architecture and biology: the spike is compact. 2010. The aim of this project is to identify wheat loci and genes that will increase chasmogamy and facilitate inter-crossing using hybridization systems.fleury@acpfg. CIMMYT. Mexico. S Rajaram and A McNab (eds) Increasing yield potential in wheat: breaking the barriers.com. ryan.

The new root technique has been trialled for high-throughput root phenotyping of field-grown cultivars. Langridge P (2007) A study of the role of root morphological traits in growth of barley in zinc-deficient soil. Hall S. Wiebkin S. 2775-2784 McKay A. The current techniques for root phenotyping are labour-intensive and error-prone. We have developed a novel root phenoptyping technique in collaboration with researchers in SARDI. Prof. it improves efficiency and accuracy of root phenotyping. J.au BACKGROUND Root systems determine the ability of a plant to capture available water and nutrients. Huang CY. peter.com. This new technique provides accurate measurements of living root cells. Most of past research had concentrated on above-ground characteristics of plants. Bot.com. 58. DNA sequencing and gene analysis REFERENCES Genc Y. while below-ground characteristics were mostly neglected.Project title: QTL mapping of root traits associated with efficient wheat root systems in drought environments SUPERVISORS: Dr Chunyuan Huang.au. Peter Langridge CONTACT DETAILS: chunyuan.htm#TopOfPage) Root analysis software (WinRhizo) . AIMS AND SIGNIFICANCE : Identifying QTLs associated with efficient wheat root systems in drought environments. Li G. Exp. and improving wheat tolerance to drought TECHNIQUES TO BE USED : Measurement of root traits using a range of techniques(WinRHIZO). using quantitative real time PCR.huang@acpfg. genetic mapping and QTL software. (http://www. and therefore.au/au/asa/2008/plenary/biotechnology/5945_mckay. Coventry S. Root proliferation is sensitive to environmental conditions. Huang CY (2008) Studying root development in soil using DNA technology: idea to impact. Our results show that restrained root growth is important for high grain yield in Mediterranean drought environments.org. Q-PCR. especially for field-grown plants.langridge@acpfg. Riley IT.regional. The QTL mapping of restrained root systems will open opportunities for genetic improvement of crop tolerance to drought. Hartley D. Herdina.

Field trials for root phenotyping using DNA technique .Transgenic barley roots express GFP. indicating new living roots in green (red arrow) and the dead roots (white arrow).

2010). Shinozaki et al. 4. Participation in field trials. TECHNIQUES TO BE USED 1. Kovalchuk et al. Plant Biotechnol J.com. . 2006 25(12):1263-1274.au BACKGROUND: Drought is among the major environmental factors limiting crop productivity worldwide. AIM OF THE PROJECT : To characterise and test different types of drought inducible promoters from wheat using transient and stable expression of promoter-reporter gene constructs in wheat and barley.kovalchuk@acpfg. REFERENCES: Agarwal PK et al. omid.au.. Defensin promoters as potential tools for engineering disease resistance in cereal grains.com. 2008. We have successfully used wheat and barley cDNA libraries for a yeast one-hybrid (Y1H) screen for cDNAs encoding transcription factors (TFs) upregulated by drought (Lopato et al..au. Isolation of plant transcription factors using a modified yeast one-hybrid system..com. Gene networks involved in drought stress response and tolerance J. 2006. in press).. 3. Careful analysis of transgenic wheat and barley development.. Lopato S et al. Bot. 2006). Some TFs have been over-expressed in transgenic barley and wheat under constitutive promoters and generated transgenic plants demonstrate increased drought tolerance. Mapping of stress specific cis-elements in some of the promoters and isolation of up-stream transcription factors using Y1H system will be inclusive to this project. Rai et al. 2. identification of stress responsive cis-elements. Shinozaki K and Yamaguchi-Shinozaki K. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Biotechnol J. 2007). Analysis of promoter structure. (2008) 6(5):465-76. 2010 8(1):47-64 Li M et al.. expression of TFs under constitutive promoters often leads to undesirable developmental phenotypes (Morran et al. Dr Omid Eini. 2009. Plant Methods 2006. Transgenic Res. Plant Cell Rep. Exp. An important response to this stress is the temporal and spatial modulation of transcription of specific sets of genes. 2007 58:221-7. Several wheat promoters will be analysed for spatial and temporal expression in transgenic wheat and/or barley in the absence of stress and then under drought and several other abiotic stresses (Li et al. These transgenics express drought tolerance TF under several different drought inducible and spike specific promoters. Spatial and temporal expression of endosperm transfer cell-specific promoters in transgenic rice and barley. Compare transgenic wheat and barley exhibiting differing expression patterns of the same drought tolerance TF for survival and yield under drought stress. The transcriptional response of plants to drought stress is controlled by numerous transcription factors (Agarwal et al. 2:3-17 Rai M et al. To overcome this problem we prepared a collection of drought inducible and tissue-specific promoters. and isolation of upstream transcription regulators using the Y1H screen.lopato@acpfg. sergiy. Comparative functional analysis of three abiotic stress-inducible promoters in transgenic rice.Project title: Analysis of drought inducible promoters from wheat SUPERVISORS: Dr Nataliya Kovalchuk. Unfortunately..eini@acpfg. which enables the plant to rapidly adapt to altered environmental conditions.. Dr Sergiy Lopato CONTACT DETAILS: nataliya. 2009 18(5):787-99. Kovalchuk N et al. which we are currently testing in transgenic wheat and barley..

control 14 days no water Line BW8-2 Line BW8-6 Line BW8-10 7 days after rewatering 14 days after rewatering Fig. 1. Trangenic wheat lines with drought inducible over-expression of DREB/CBF demonstrate improved survival under stringent drought conditions .

2006 25(12):1263-1274 . Li et al. We have used wheat and maize spike and developing grain subjected to drought stress to make cDNA libraries for Y2H screens (Lopato et al. and to characterise transgenic wheat and barley plants with constitutive and drought inducible over-expression of two NF-Y subunits. – Participation in field trials.lopato@acpfg..com. Prof. 1). Dr Omid Eini. Nuclear Factor Y (NF-Y) is a trimeric complex that binds to the CCAAT box of plant promoters.Project title: Isolation and characterization of genes involved in the formation and regulation of drought responsive NF-Y transcription complex(es) SUPERVISORS: Dr Sergiy Lopato. Shinozaki et al. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants.. characterisation of these genes. At least 37 genes for NF-Y subunits have been identified in wheat.eini@acpfg. and preparation of constructs for wheat and barley transformation.. over-expression of NF-YB and NF-YA genes in transgenic Arabidopsis and maize plants can confer drought tolerance without negatively affecting plant development (Nelson et al. developing grain and root subjected to drought stress – Analysis of expression of isolated genes using quantitative PCR (Q-PCR). 2006). 2007).. Plant Cell Rep. Furthermore. and at least some of these are up-regulated by drought (Stephenson et al. Plants respond to drought by the temporal and spatial transcriptional modulation genes . which may include NF-Y subunits or other types of factors that cooperate with NF-Y trimeric complexes during drought stress (unpublished data). it is unclear which of these subunits participate in particular complexes. TECHNIQUES TO BE USED The project will involve further Y2H screens for NF-Y interacting factors.. However. Specific techniques will include: – Isolation of genes relevant to NF-Y complexes using PCR-based cloning and the Y2H screen of cDNA libraries prepared from spike. 2008). with each subunit being required for DNA binding (Fig.au. and whether subunits in a complex can be substituted without affecting functionality. Peter Langridge CONTACT DETAILS: sergiy.a response orchestrated by various transcription factors (Agarwal et al. Dr Maria Hrmova. 2007. 2007). REFERENCES: Agarwal PK et al. Some of the TFs identified have already been over-expressed in transgenic barley and wheat under constitutive and inducible promoters.au BACKGROUND: Drought stress is a major constraint to the production and yield stability of wheat in Australia. Two transgenics overexpressing NF-Y genes are already available for drought tolerance evaluation. omid. We are now screening these libraries to identify interacting partners of NF-Y subunits. – Modelling of NF-Y protein-protein and protein-DNA interactions (optional) – Analysis of transgenic wheat and barley plants with constitutive and inducible overexpression of two NF-Y factors for survival and performance under drought stress.com. The project may also involve structural modelling and experimental verification of protein-protein and protein-DNA interactions involving NF-Y trimeric complexes. AIM OF THE PROJECT : To isolate wheat genes encoding drought related members of the NF-Y complex using PCR-based cloning and a yeast two-hybrid (Y2H) screen. 2006.

HAP (Heme Activator Protein) Complex or CBF (CCAAT Binding Factor) or NF-Y (Nuclear Factor Y) Complex . 2007 58:221-7 Stephenson TJ. 1. Heard JE. Zhu JK.Li WX. Bot. Donnarummo MG. Ratcliffe OJ. Wu J. Zhu J. Langridge P. Shinozaki K and Yamaguchi-Shinozaki K. Shirley N. Plant Cell. Collet C. Repetti PP. Borisjuk L. Castiglioni PP. He XJ. Genome-wide identification and expression analysis of the NF-Y family of transcription factors in Triticum aestivum. Parsley K. Bazanova N. Gene networks involved in drought stress response and tolerance J. Lopato S.20(8):2238-51.The Arabidopsis NFYA5 transcription factor is regulated transcriptionally and posttranscriptionally to promote drought resistance. Lu XY. Krolikowski KA. McIntyre CL. Systematic identification of factors involved in post-transcriptional processes in wheat grain. Canales RD. Kumimoto RW. Gutterson N. Exp. Plant Mol Biol. Iida K. Bensen RJ. Hinchey BS. Cui X. Xue GP. Warner DC. Dotson SB. 2006 62(4-5):637-53 Nelson DE. Plant nuclear factor Y (NF-Y) B subunits confer drought tolerance and lead to improved corn yields on water-limited acres. Jin H. Proc Natl Acad Sci U S A. Maszle DR. 2007 65(1-2):77-92. Wu JM. Creelman RA. Milligan AS. 2007 104(42):16450-5. Oono Y. HAP2/ CBFB/ NF-YA HAP3/ CBFA/ NF-YB HAP5/ CBFC/ NF-YC HAP3/ HAP2/ HAP5/ CBFA/ CBFB/ CBFC/ NF-YB NF-YA NF-YC C C A A T C C A A T Fig. Plant Mol Biol. Anstrom DC. 2008 Aug. Adams TR.

au BACKGROUND: Drought is a major constraint to the production and yield stability of wheat in Australia. temporal and stress inducible patterns of expression. AIM OF THE PROJECT : To isolate members of several families of root specific and drought inducible transcription factors using yeast one-hybrid (Y1H) or two-hybrid (Y2H) screens. – Isolation of full length genes and promoters for root specific TFs.Project title: Isolation and characterization of root specific and drought inducible transcription factors from wheat roots subjected to drought and salt stress SUPERVISORS: Dr Sergiy Lopato. Gantet P. and to characterise these genes for their spatial. root length density. 2006. in press). We have made cDNA libraries from wheat and barley spike and developing grain subjected to drought stress and successfully used these in Y1H (Fig. 2007).. 1) and Y2H screens for transcription factors (TFs) up-regulated by drought (Lopato et al. which enables the plant roots to adapt to altered environmental conditions. Over-expression of some of these TFs in transgenic barley and wheat under constitutive and inducible promoters was found to provide improved survival under severe drought conditions (Morran et al. particularly in a drought specific manner (Hirsch and Oldroyd. the model cereal. Root characteristics.lopato@acpfg. Genetic control of root development in rice. very little is known about the transcription factors that control root growth. Coudert et al. Shinozaki et al. The transcriptional response of plants to drought stress is controlled by various transcription factors (Agarwal et al. and assessment of TF expression under well watered or drought conditions. chunyuan.huang@acpfg. – Preparation of constructs for transformation of wheat and barley REFERENCES: Agarwal PK et al. are critical for the exploitation of available soil water and for healthy above-ground growth. Trends Plant Sci... together with PCR-based cloning. Specific activities include: – Isolation of several transcription factors using Y1H and Y2H screens of root cDNA libraries and PCR-based cloning – Analysis of expression of isolated TFs using quantitative PCR (Q-PCR) on RNA prepared from stressed plants. 2009. Plant Cell Rep. 2006a.com. 2006b). Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Courtois B. However. as well as PCRbased cloning. 2010 15(4):219-26 .com.. Prof. and the number of thick roots. Périn C. Khong NG.. We have also made several cDNA libraries from wheat and maize roots under drought and salt stress. An important response to drought is the temporal and spatial transcriptional modulation of specific genes. TECHNIQUES TO BE USED The project will include isolation of TFs from root libraries using Y1H and Y2H screens. Peter Langridge CONTACT DETAILS: sergiy. 2010).au. and initial tests suggest that these libraries provide a good source of root specific and stress inducible TF genes. 2006 25(12):1263-1274 Coudert Y. Dr Chunyuan Huang. Analysis of promoter sequences. especially root length. Cloning of root specific genes and promoters and mapping of stress specific and root specific cis-elements in selected promoters will be part of this project.

1. Parsley K. 2006 62(4-5):637-53 Shinozaki K and Yamaguchi-Shinozaki K. Schematic representation of the selection of positive clones during Y1H screen. Plant Signal Behav. 2009 4(8):698-700 Lopato S et al.Hirsch S. Plant Methods 2006. Bazanova N. Gene networks involved in drought stress response and tolerance J. Langridge P. Oldroyd GE.. Bot. 2007 58:221-7 Isolation of TFs using Y1H screen Library protein AD Transcription if hybrid protein interacts with bait sequence TATA HIS3 selection gene Transcription if non-specific protein interacts with bait sequence Bait sequence GAL4 TATA MEL1 selection gene Non-specific protein AD Selection for positive growth on CM –His –Leu 3-AT and X-GAL plates Fig. Shirley N. 2:3-17 Lopato S. Exp. Milligan AS. Plant Mol Biol. . GRAS-domain transcription factors that regulate plant development. Systematic identification of factors involved in post-transcriptional processes in wheat grain. Isolation of plant transcription factors using a modified yeast one-hybrid system. Borisjuk L.

Science 301: 336–8. some of which are highly expressed. TECHNIQUES TO BE USED : Most of molecular biology techniques such as expression cloning. Cell 140:111-22. while others are lowly expressed under drought. REFERENCES Baulcombe D (2007). Carrington J. molecular genetic techniques and a range of bioinformatics approaches. The expected results would represent a significant advance in cereal miRNA research and the effort to enhance crop drought tolerance. is severely limited by drought stress. Science 315: 199–200. miRNAs function very much like animal miRNAs. Role of microRNAs in plant and animal development. northern blot hybridization and western blotting. but play additional roles in development and adaptive responses to stresses. Weigel D. To investigate the role of miRNAs to drought stress in barley and wheat.com. Their productivity. Southern blot hybridization. To develop barley and wheat tolerant to drought stress.au. In plants. Prof Peter Langridge CONTACT DETAILS: bujun. Although 32 miRNAs from wheat have been reported. Seumel GI.com. and express miRNA in transgenic barley or wheat to improve drought tolerance. MicroRNAs (miRNAs) are one of the two key players in this process and have become a hot topic in molecular biology.au BACKGROUND The molecular biology of gene silencing represents one of the most important scientific advance in recent years. Transcriptional control of gene expression by microRNAs. Reski R. cell biology techniques. AIMS AND SIGNIFICANCE : Determine miRNA function in drought stress responses. High throughput sequencing of these libraries reveals a number of candidate miRNAs. Ambros V (2003). miRNAs are a distinct class of small non-coding RNAs of 18-25 nucleotides in length and conserved in eukaryotic organisms. polymerase chain reaction (PCR). Ossowski S. peter. Amplified silencing. let alone their role in drought stress.shi@acpfg. In animals. identifying key regulators of stress tolerance is crucial. however.Project title: Determination of microRNAs involved in drought tolerance in barley or wheat SUPERVISORS: Dr Bu-Jun Shi. we have constructed 2 small RNA libraries from barley and 6 small RNA libraries from wheat under normal and drought conditions. Frank W (2010). Arif MA. . Wheat and barley are two most important and widely consumed cereals in the world. miRNAs regulate 68% of total genes and participate in the regulation of almost every cellular process. Other techniques include biochemical techniques.langridge@acpfg. Khraiwesh B. Expression of some miRNAs is induced by drought. To date. suggesting a role for miRNAs in drought stress. their function has not yet been experimentally confirmed. no barley miRNA has been identified. The next step is to design experiments to confirm which miRNAs are truly associated with drought stress and investigate options for enhancing stress tolerance.

The mechanism of Micro RNA generation and function Genomic DNA Transcription Pri-miRNA Enzymatic process Pre-miRNA Process further miRNA-miRNA* duplex ( ~21 nt) Unwind Single-stranded RISC assembly Bind to target mRNA Cleave target mRNA Less miRNA(X) More miRNA(X) Barley growth under drought .

International students can apply for these ACPFG scholarships only if they have received a university tuition fee waiver (or if they receive a stipend which covers the tuition fee).html Contact Education manager: Dr Monica Ogierman Email: monica.edu.au/scholarship/default. Further scholarships are also available through the: University of Adelaide http://www. Food and Wine University of Adelaide Waite Campus Plant Genomics Centre Hartley Grove.asp) University of Melbourne http://www. Students applying for ACPFG PhD scholarships must be Australian citizens.ogierman@acpfg.edu.000 tax free per year.au/research/scholarships.adelaide.unimelb.com. plus support to attend an international conference and for professional development.com.edu. Urrbrae Postal: PMB1 Glen Osmond SA 5064 http://www.futurestudents.Scholarships ACPFG PhD scholarships consist of $25. or New Zealand Citizens.unisa. permanent residents.au/graduatecentre/scholarships/postgrad/ University of South Australia http://www.acpfg.au .au Phone: (08) 8303 6725 Fax: (08) 8303 7102 Australian Centre for Plant Functional Genomics (ACPFG) School of Agriculture.

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