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Transgenic strategies for improvement of drought tolerance of cereals to reduce the consequences of water limitations caused by climate change János Györgyey,

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Presentation on theme: "Transgenic strategies for improvement of drought tolerance of cereals to reduce the consequences of water limitations caused by climate change János Györgyey,"— Presentation transcript:

1 Transgenic strategies for improvement of drought tolerance of cereals to reduce the consequences of water limitations caused by climate change János Györgyey, Gábor V. Horváth, Dénes Dudits Institute of Plant Biology, Biological Research Centre, Hungarian Academy of Sciences, Buchanan, Gruissem, Jones: Biochemistry & Molecular Biology of Plants; chapter 22, and results of the Cell cycle and Stress Adaptation Group

2 N. Sreenivasulu et al. / Gene 388 (2007) 1–13

3 Umezawa et al 2006, 17: Strategies for the genetic engineering of drought tolerance.

4 Buchanan, Gruissem, Jones: Biochemistry & Molecular Biology of Plants; Fig 22.3



7 Series of responses to drought stress ABA peak Stoma closure Reduced photosynthetic activity Block of cell division, elongation Activation of protective (stress) genes (DRE, ABRE elements) e.g. ALR Accumulation of osmolytes Long term adaptation

8 Fig.1 A schematic representation of cellular signal transduction Pathways between stress signal perception and gene expression and the cis- and trans-elements involved in stress responsive gene expression. DREB1/CBF and DREB2 distinguish two different signal transduction pathways in response to cold and drought stresses, respectively. DRE: drought responsive element, ABRE: abscisic acid responsive binding element, MYBRS: MYB recognition site, MYCRS: MYC recognition site, bZIP: basic-domain leucine-zipper Agarwal et al. Plant Cell Rep (2006)25:1263–1274







15 Osmotic stress of wheat plantlets in hydroponics UntreatedPEG-treated: 0 day(10days old plantlets) 100mOsm 200mOsm 400mOsm 2.days 4.days 7.days 9.days 11.days 14.days Sampling

16 Rice chip – app unigene - hybridised with PEG-treated / untreated Kobomugi root samples (day 9) - color flip repeat - app spots gave measurable data in both case >2x induction: more than 1100 spots >5x induction: 345 spots >2x repression: more than 400 spots >5x repression: 77 spots


18 Relative transcript level of four selected genes exhibiting induction during osmotic stress Q-PCR approved




22 EXPERIMENTAL SYSTEM FOR EXPOSURE OF WHEAT PLANTLETS TO LONG TERM DROUGHT STRESS IN EXPANDED PERLITE 0 day (16 day old plantlets) 1. week Sampling Normal irrigation 2. week 3. week 4. week Reduced irrigation (30%)

23 68.97 grain yield (% of control) stomata conductance (mmol H 2 O m -2 s -1 ) CO 2 fixation (cpmx10 -5 ) fructose accumulation (nmol g -1 ) lipid peroxidation (MDA pmol cm CuZn SOD (% of control) Increase in root weight (g/plant) 13 É. Sárvári et al. Kobomugi Plainsmann

24 Growth rate of two genotypes under water limitation (30% water supply) Kobomugi Plainsman V

25 P5CS mRNA in shoots

26 Transcript level changes in wheat roots during drought adaptation measured on barley macroarray Plainsmann (adapting, having good yield) Kobomugi (fast responding survivor) During 4 weeks not changed 7378 Change in 1-2 weeks up down Change in 3-4 weeks up down (percentage of clones)

27 9% 11% 12% 34% 19% 6% 29% Functional classification of genes upregulated in one genotype only Gene expression Signal transd. Transport Cytosceleton and cell wall Cell growth and division Metabolism Kobomugi Plainsman Stress and defense Protein synthesis Protein degradation

28 putative cyclin-dependent kinase B1-1 expansin EXPB2 calmodulin-binding heat-shock protein xyloglucan endotransglycosylase Xet3 protein caffeic acid O-methyltransferase putative cellulose synthase catalytic subunit betaine aldehyde dehydrogenase 16 Cluster analysis Kobomugi Plainsman

29 Kobomugi Plainsman 18

30 Divergent drought adaptation strategies of the two genotypes are refelected in their transcript profiles. Long term adaptation is dependent on moderate changes in the expression of large set of genes in a coordinated manner. Transient gene activation is characteristic to Kobomugi, while genes of the more adaptive Plainsmann genoptype exhibit prolonged upregulation. Based on the yield performance and photosynthetic activity, Kobomugi represents escaper strategy while Plainsmann cultivar is capable to maintain physiological functions in harmony with gene expression reprogramming. Conclusions

31 Promoter elements in rice orthologue of ODA1 gene No. of motif Name of motif Function 13ABREABA-responsive binding site 4C-repeat DREDrought response element 1CE1ABRE motif linked cis-element 3CGTCAJasmonate response cis element 3GCN4Endosperm specific expression 14HSEHeat shock response, HSF binding 2LTRLow temperature response 2TCA elemSalicilic acid response 3TGACGJasmonate response cis element 6WUNWound response 1As1Root specific expression Osmotic and Drought Adaptation induced clone Protein function is not known, similar to LEA family and WSI18

32 Relative transcript levels in roots of Kobomugi during acute drought stress (desiccation)

33 Standard system for water limitation Soil sand:perlite=2:1 Control plants 70-80% soil water saturation Moderately stressed plants 30-40% soil water saturation Watering daily, weight measurment

34 Relative transcript levels in shoots of Kobomugi after two weeks of moderate drought stress (limited water supply)

35 Azsuka Bioryza H Sandora Marilla - cv. Sandora - control (100%), stressed (20%) water capacity samples (at 8, 14, 18) - 22 k rice oligo-chip Daily change in transcript profile during water limitation in roots of rice

36 Info 1Info 2 F-box domain, putative11682.m04419 oxidoreductase, short chain dehydrogenase/reductase family11670.m04026 AT5g64600/MUB3_ m00842 jmjC domain, putative11668.m04545 putative chelatase subunit11669.m03601 Protein kinase domain, putative11686.m00607 Similar to C-x8-C-x5-C-x3-H type Zinc finger protein, putative11669.m01922 And 7 genes with unknown function Genes induced during the day in rice roots under water limitation

37 Info 1Info 2 Hpt domain, putative11674.m04480 Af10-protein11667.m05271 putative reductase11669.m05442 O-methyltransferase, putative11670.m00047 UDP-glucoronosyl and UDP-glucosyl transferase11673.m03060 Similar to GMFP m02628 profilin a11676.m01466 hypothetical protein, (thylakoid membrane phosphoprotein 14 kda, chloroplast precursor, putative, expressed )11667.m05484 expressed protein, (putrescine-binding periplasmic protein, putative, expressed )11670.m05010 putative transport protein particle component11669.m02901 pectate lyase precursor (ec )11668.m01124 putative oxidase11669.m03600 receptor-like protein kinase, putative11674.m01711 Genes repressed during the day in rice roots under water limitation




41 Genetic engineering of the glyoxalase pathway in tobacco leads to enhanced salinity tolerance Singla-Pareek et al. PNAS 100, (2003)

42 Wide range of substrate specificity Highly conserved structure (NADH or NADPH binding region, catalytic tetrad) Occurrence: from bacteria to Homo sapiens polyol pathway detoxification of reactive aldehydes About the aldose reductase superfamily in general:



45 Effects of MsALR overproduction on tobacco plants: protection against lipid peroxidation under chemical and drought stresses (Oberschall et al. 2000) protection during drought and UV-B stresses (Hideg et al. 2003) transgenic plants showed higher tolerance to low temperature and cadmium stress (Hegedűs et al. 2004) increased tolerance to the effects of high temperature and high light intensity (Horváth and Hideg, unpublished)

46 Development of first shoots on AAR medium Transgenic plantlets in soil Regeneration of the ALR transformants and growth of mature plants Fertile ALR spikes

47 PAR ( mol m -2 s -1 ) control 4310-b control 4310-b PAR ( mol m -2 s -1 ) Electrontransport (a. u.) E. Hideg et al.

48 J. Pauk et al.


50 Thank you for your attention

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