1. Drought tolerance Shaping the future for African cereal crops MM O’Kennedy, J. Becker, R. Chikwamba ACGT forum, 12 October 2012

2. Slide 2 CSIR Research Impact Areas Materials Photonics Robotics Nanotechnology NATURAL ENVIRONMENT INDUSTRY BUILT ENVIRONMENT DEFENCE & SECURITY HEALTH ENERGY Enabling technologies

3. Slide 3 CSIR Campus CSIR campus where Biosciences is located Emerging Health Technologies Protein Technologies & Bioprocessing Natural Products & Agroprocessing

4. Slide 4 Innovation “Chasm” Products & Services Knowledge Basic Research Applied Research Bioprocess Development Piloting Commercial Manufacture Commercial Manufacture Commercial Implementation Regional Manufacturing Hubs e.g. Biovac, OBP Our Vision is to be the leading organization in South Africa that translates Biosciences research outputs into market ready products and processes HEIs Small Companies Imported Technologies CSIR Bioprocess Development and Piloting Centre at CSIR

5. Slide 5 © CSIR 2012 www.csir.co.za Strategy  Drought threatens the food security of millions  Genomics approaches to produce cereals more resilient to drought  Unravel the regulatory mechanisms in drought tolerant cereals  Drought tolerance is the ability to loose 30-50% water for short periods; Drought resistance is the ability to loose water for prolonged periods of time Approach: Sorghum is a wealth of genetic make-up to understand pathways involved in adaptation identify transcription factors (TFs) and microRNAs (miRNAs) that govern a subset of drought stress related genes Identify ortologues in maize and pearl millet Cloning genes and regulatory elements Validation in maize

6. Slide 6 © CSIR 2012 www.csir.co.za Absolute water content (Abs WC) fw-dw (water lost)/fw X 100 = % absolute water content on a fw basis Developing a growthroom seedling screen

7. Slide 7 © CSIR 2012 www.csir.co.za Temperature and humidity in growthroom Humidity during drought simulations?

8. Slide 8 © CSIR 2012 www.csir.co.za Establishing a seedling drought screen Controls: 58 ICSV112 susceptible 59 - P898012 tolerant 58 and 8 clearly drought susceptible! Accessions 6, 35 and 36 performs better than 59 (drought tolerant control) Random design, but no border pots during simulation of drought Stress 5, 6, 35 and 36, plus controls for greenhouse screens Critical threshold – below 30% Abs WC 1/3 of original % AWC P = 0.023 P = 0.097 P = 0.022P = 0.006P = 0.72 P = 0.14

9. Slide 9 © CSIR 2012 www.csir.co.za Establishing a seedling drought screen (continue) P-value a = < 0.05 b = > 0.05

10. Slide 10 © CSIR 2012 www.csir.co.za Re-watering time points (photographic record) 10 days of simulated drought stress (4 pots of each accession) – 11 days after re-watering commenced Accession 35 & 36 thrives even in comparison to the drought tolerant breeding line 59 (P898012), whereas 58 is clearly the drought susceptible 5 6 8 35 36 58 59 Critical threshold – below 30% Abs WC; 1/3 of original % Abs WC

11. Slide 11 © CSIR 2012 www.csir.co.za Rooting of selected sorghum accessions (photographic record) S T

12. Slide 12 © CSIR 2012 www.csir.co.za Sorghum for food, feed and fuel 5 5 CSIR identified 4 distinct African sorghum landraces (Sorghum bicolor (L.) Moench), with drought tolerance superior to a drought tolerant breeding (accessions 5, 6, 35 & 36) 6 35 36 Capacity  Growthroom screening methodology  Custom designed Agilent sorghum microarray

13. Slide 13 © CSIR 2012 www.csir.co.za Drought initiative – transcriptome profiling Custom designed Agilent sorghum microarray (shoots & roots) Spectral reflectance as stress indicator (Dr Moses Cho, NRE) 9 plants per time point

14. Slide 14 © CSIR 2012 www.csir.co.za Drought initiative – transcriptome profiling Custom designed Agilent sorghum microarray Direct comparitive profiling  Venn diagram  Heat maps of candidate genes and TFs  Shoot  Roots  Functional classification of differentially expressed transcripts  Several TFs families: DREB1&2, bZIP, KNOX & WUSCHEL  Unique: candidate genes related to sustaining yield during water deficit  qRT-PCR and protein profiling

15. Slide 15 © CSIR 2012 www.csir.co.za Gene Discovery Confirmed Transgenics Selfing or hybridization Lines selfed to homozygosity Field testing Vector Construction Discovery (CSIR) Development (ARC; Seed company) Trait efficacy demonstrated Complementary space Competitors Water efficient maize for Africa (WEMA) (Monsanto, ARC) Bt cspB gene (help bacteria deal with cold stress by disentangling RNA; RNA folds abnormally when the plant is water stressed) FP7 DROPS (Drought tolerant plants 2010-2015; Syngenta&Pioneer) Maize for African Soils (IMAS) (Pioneer, CIMMYT, KARI & ARC) Drought initiative value chain Way forward:NRF PDP programme (Natrisha Devnarain) GDARD funding

16. CSIR Biosciences Rachel Chikwamba John Becker (ACGT) Maretha O’Kennedy Natrisha Devnarain (NRF PDP) Technical assistance: Mulalo Nemutanzhela, Stella Manganye & Taolo Shai Thank you!