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

Drought tolerance

Shaping the future for African cereal crops

MM O’Kennedy, J. Becker, R. Chikwamba

ACGT forum, 12 October 2012

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© 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


Absolute water content (Abs WC)fw-dw (water lost)/fw X 100 = % absolute water content on a fw basis

Developing a growthroom seedling screen


Temperature and humidity in growthroom

Humidity during drought simulations?


Establishing a seedling drought screen

Controls: 58 ICSV112 susceptible59 - P898012 tolerant

58 and 8 clearly drought susceptible!Accessions 6, 35 and 36 performs better than 59 (drought tolerant control)

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5 6 8 10 12 19 23 24 29 31 32 33 35 36 41 44 50 58 59

% R

WC

Sorghum accessions

Day 1

Day 6

Day 7

Day 8

Day 9

Day 10

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 = 0.023P = 0.097 P = 0.022 P = 0.006P = 0.72 P = 0.14


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5 6 8 10 12 19 23 24 29 31 32 33 35 36 41 44 50 58 59

Sorghum accessions

% R

WC

Time point day 10

0

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1 2 3 4 7 9 11 13 14 15 17 18 20 27 28 30 37 38 39 40 42 43 45 46 47 48 49 58 59 5 6 35 36

Sorghum accessions

% R

WC

Time point day 10

b a a a

b b b b b a a

B

A

Establishing a seedling drought screen (continue)

P-valuea = < 0.05b = > 0.05


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


Rooting of selected sorghum accessions (photographic record)

ST


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)

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35 36Capacity Growthroom screening methodology Custom designed Agilent sorghum microarray


Drought initiative – transcriptome profiling

% Soil moisture content

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Day 1 Day 6 Day 9 RI

Timepoints

% s

oil

mo

istu

re c

on

ten

t

35C

36C

59C

35S

36S

59S

Chlorophyll content

2527293133353739414345

Day 1 Day 6 Day 9 RI

Timepoints

Ch

loro

ph

yll

con

ten

t

35C

36C

59C

35S

36S

59S

Landrace 36

7072747678808284868890

1 6 9 RI

Days (timepoints)

% A

bs

FW

36C

36S

Drought tolerant P898012

7072747678808284868890

1 6 9 RI

Days (timepoints)

% A

bs

FW

59C

59S

Custom designed Agilent sorghum microarray (shoots & roots)

Spectral reflectance as stress indicator (Dr Moses Cho, NRE)

9 plants per time point


Drought initiative – transcriptome profilingCustom designed Agilent sorghum microarray

Direct comparitive profiling

6-36 9-36

RI-36

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22 356

278

6-36 9-36

RI-36

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22 356

278

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


Gene Discovery

Confirmed Transgenics

Selfing or hybridization

Lines selfed to homozygosity

Field testingVector 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

CSIR Biosciences

Rachel Chikwamba

John Becker (ACGT)

Maretha O’Kennedy

Natrisha Devnarain (NRF PDP)

Technical assistance: Mulalo Nemutanzhela, Stella Manganye & Taolo Shai

Thank you!

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

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