Phenotyping for tolerance to abiotic stresses

A member of CGIAR consortium www.iita.org

PHENOTYPING FOR TOLERANCE TO ABIOTIC STRESSES IN YAM, MAIZE,

BANANA AND COWPEA

Workshop on Implementation of IITA’s Genetic Improvement

Strategy

IITA-HQ, Ibadan Nouhoun Belko 09 September 2015 Postdoc Cowpea Agro-Physio


Climate change will increase intensify & frequency of drought


Global low soil P availability is a primary constraint to life on earth

Dominance of red and light-gray colors, indicating soil P deficiency for the growth of many cultivated species Importance of P availability as a primary limitation to agricultural productivity in terrestrial environments (from Jaramillo-Velastagui, J. Lynch 2011).


N balance deficiency – Issue of accessibility & affordability in SSA


Soil Fertility

Yiel

d

Traditional genotypes lodge at high fertility

But no yield gain at low fertility (*)

Can we develop genotypes with superior yield at all fertility levels?

Benefit from 20th century green revolution

Potential Benefit from 21st century green revolution

Dwarf genotypes respond to high fertility


Objectives: Know environment, Understand mechanisms, high genetic gainHypothesis: Mine available nutrients, Tap water, Save water, Secure reproductionApproaches: Direct (yield) and Indirect (root - shoot phenes important for specific stress)Water Phosphorus

5%

18%

11%

23%

10cm

20cm

30cm

4ppm

2ppm

0.5ppm

0.25ppm40cm

Stomata response differences to VPD relate to plant hydraulics


Setting up Screening Protocol for N and K Use Efficiency in Yam

SELECT PARENTS- NUE Trials, Uromi- Performance in Low Fertility Environment, Mokwa

Genotype parents Using SSR/SNP Markers

Generate Populations

Phenotype two Populations In Low N and K Plotsand NUE Trials (field and SH)

Genotype Populations Using polymorphic SSR/SNP Markers

Find Marker - TraitAssociation for NUE of N and K or other agronomic traits

Validate Favourable Markers/QTL (s) using other populations generated with the selected parents

A. Lopez-Montes and R. Bhattacharjee


Screening Maize for Tolerance to Drought

S. K. Meseka and A. MenkirDrought tolerance: Ikenne, Forestry Zone Field experiments: irrigated & non-irrigated Blocks Planting during the second week of November Sprinkler irrigation system supply 20mm/week Drought stress imposed from 35 DAP until harvest Agronomic traits measured

Results from 2000 to 2015 Genetic bases of tolerance to drought understood 2000+ improved lines developed & shared in WCA NARS & seed companies released varieties/hybrids Farmers adopted improved drought tolerant materials


Screening Maize for Tolerance Drought & Heat

S. K. Meseka and A. MenkirDrought & heat tolerance: Kadawa, Kano State Field experiments: only one block used Planting during second week of

February Gravity irrigation (furrow) every four

days Irrigation stop in April for 21d then ok

once a week Agronomic traits measured

Results from 2013 to 2015 Genetic bases of tolerance not well

understood 400+ varieties/ hybrids & 1500 inbred

lines screened New inbred lines tolerant to drought and

heat stress with high grain yield identified

Month

Temperature Co

RHMax Min

February 34 21 16

March 39 26 13

April 42 28 15.5

May 40 27 17

June 35 24 21.8


High N Low N

Low N site developmentSelection & depletion of nitrogen Planting high population density Removal of all crop residuesLow N block (20 kg N/ha) High N (90 kg N/ha)

Screening Maize for Tolerance to low N

S. K. Meseka and A. MenkirLow nitrogen tolerance: Mokwa, Niger State Inbred lines combined tolerance to Low N/Drought Non-additive genetic effects for grain yield provided basis for the exploitation of heterosis Hybrids combining high grain yield with tolerance to drought/low N developed and disseminated to NARS

Results from 1995 to 2015 Genetic bases of inheritance

understood Several low N efficient maize lines

developed and disseminated to NARS NARS in WCA released several

improved low N efficient varieties/ hybrids


Drought stress research conducted in forestry zone with incidence of rainfall at flowering/grain filling stages

Lack of modern high throughput phenotyping tools & plant growth facilities for screening large populations

Limited knowledge of genetic bases of combined drought and heat stress tolerance in maize plants

Absence of fully irrigated block for comparison in screening for tolerance to combined drought & heat

Traits measured did not include the Plant Root System – relevant indirect trait in water & nutrient uptake

Measurement of most Physiological Traits are time consuming and not practical with large breeding populations

CHALLENGES AND RESOURCE NEEDS


High throughputgenotyping

Environmental data

Field drought evaluation site + Root phenotyping in Mokwa

Greenhouse drought evaluation, modern tools, collaboration with CERAAS

Marker trait associationMarker assisted

selection

OPPORTUNITIES AND WAY FORWARDInitiation breeding for tolerance to flood and soil acidity/salinity with climate change

http://www.google.com.ng/url?url=http://www.elegran.com/blog/2013/07/the-greenhouse-effect-bright-farms&rct=j&frm=1&q=&esrc=s&sa=U&ved=0CBcQwW4wAWoVChMIk5mU-s7YxwIVbgfbCh1xQAJm&usg=AFQjCNEaLET21p1is3oFIWlWLucT2Mu2qg

http://www.google.com.ng/url?url=http://passel.unl.edu/pages/informationmodule.php?idinformationmodule=1075412493&topicorder=9&maxto=12&rct=j&frm=1&q=&esrc=s&sa=U&ved=0CB0QwW4wBGoVChMI-sL21s_YxwIVYmvbCh3D5w2u&usg=AFQjCNFwijX86GaoaS8KIbexSWL6S9_Txg


Musa Phenotypic Response to Drought in 4 Env.

Prof. Swennen Rony and collaborators


Williams

Cachaco

Lep Chang Kut

Osmotic stress during in vitro growth


Green-house screening for leaf area and transpiration efficiency response to drought

Leaf

are

a af

ter

18

wee

ksTr

ansp

irat

ion

effic

ienc

y


4 varieties Pahang (AA) Guyod (AA) Cachaco (ABB) Nakitengwa (AAAh)2 water treatments Irrigation No irrigation10 replicates

Field screening for pseudostem height and leaf area response to drought between 10 - 40 DAIS


Field high throughput phenotyping for Gs and ΔT response to drought

Evolution of ΔT across daytime

Transp ~ evapo cooling Detected by IR imaging

Stress

Optimal

CONCLUSION AND WAY FORWARD - Screening tools use in breeding

program - The B & A genomes sources of

resistance - AA genome screening with LIPI

partners- Segregating populations: QTLs

800 seeds - Sequencing all hybrids


Screening for tolerance to low soil P and rock P use efficiency in Cowpea

K. Suzuki, C. Fatokun, O. Boukar

Pot trials for comparison of shoot growth response to different P applications Genotype with 28 entries P application with 3 levels (0 and 30 mg P/kg of KH2PO4 and 90 mg P/kg of Togolese Rock P) In a split plot design with 2 replications

N uptake(g/pot)

Shoot dryweight at 8WAS (g/pot)

Chlorophyllcontent at 5 WAP

Chlorophyllcontent at 7 WAP

Plant height at5 WAP (cm)

Plant height at7 WAP (cm)

Nodulenumber

Nodule dryweight (g/pot)

Root dryweight (g/pot)

P uptake (g/pot) 0.888** 0.817** 0.342** 0.232ns 0.308** 0.219ns 0.548** 0.098ns -0.056ns

N uptake (g/pot) 0.845** 0.299** 0.221ns 0.388** 0.319** 0.541** 0.137ns 0.045ns

Shoot dry weight at8 WAS (g/pot) 0.283ns 0.348** 0.490** 0.327** 0.575** 0.021ns 0.011ns

Shoot dry weight at 8 WAP has significant correlation with P and N uptake.


Variations in shoot growth response to low soil P and rock P in Cowpea

Shoot DW under 90 mg P/kg Rock P

Shoot DW under 0 mg P/kg KH2PO4

Decrease in shoot biomass in 90 mg P/kg Rock P relative to 30 mg P/kg KH2PO4

Decrease in shoot biomass in 0 mg P/kg relative to 30 mg P/kg KH2PO4Based on their shoot

biomass production under both 0 mg P/kg KH2PO4 and 90 mg P/kg Rock P: Iron Bean, IT87D-941-1,

IT90K-284-2, IT95K-1543 and IT97K-499-38 were consistently low P tolerant and rock P efficient lines

Tvu-7778, Sanzi and IT97K-499-35 were the most low P sensitive and rock P un-efficient lines


Pot trials for evaluation of optimum dose of rock P for cowpea production Selected lines: Iron bean, IT97K-499-38, IT87D-941-1, Dan Ila

and IT97K-499-35 Rock P application with 5 levels: 0, 30, 60, 90 and 120 mg P/kg In a split plot design with 10 replications

Genotypic difference in shoot growth response to different doses of rock P application

0.01.02.03.04.05.06.07.08.09.0

10.0

0 30 60 90 120

aa a

aIron bean at 4 WAP

0.01.02.03.04.05.06.07.08.09.0

10.0

0 30 60 90 120

abaaa

bb

IT97K-499-35 at 4 WAP

Shoo

t D

W

(g/p

lant

)


Genotypic difference in grain yield and nodule number under different doses of rock P

Rock P application (mg P/kg)


0.002.004.006.008.00

10.0012.0014.0016.00

0 30 60 90 120

Gra

in y

ield

(g

/pla

nt)


aab ab

aIron Bean

0.002.004.006.008.00

10.0012.0014.0016.00

0 30 60 90 120

ababaa

bb

IT97K-499-35

0

10

20

30

40

50

60

70

0 30 60 90 120

Nod

ule

num

ber

(-)

Iron Bean

0

10

20

30

40

50

60

70

0 30 60 90 120



Gra

in y

ield

(g

/pla

nt)

Nod

ule

num

ber

(-)

IT97K-499-35

60+ mg/kg of rock P appear to be optimum for cowpea shoot production

30+ mg/kg of rock P allow significant increase in grain yield in rock P efficient lines

No clear pattern of rock P applications effects on nodules number across genotypes

Iron Bean and IT97K-499-35 are potential parents contrasting for tolerance to low P and rock P use efficiency and could therefore be used in cowpea breeding program.


Effects of plant genotype roots and rock P applications on rhizosphere soil pH

4.604.704.804.905.005.105.205.305.405.505.60

IT97

K-4

99-3

8

Iron

bea

n

IT97

K-4

99-3

5

Dan

Ila

IT87

D-9

41-1

no-p

lant

Rhi

zosp

here

soil

pH

Cowpea lines

a a

b

c

aa

4.84.94.95.05.05.15.15.25.25.35.3

0 30 60 90 120

Rhi

zosp

here

soil

pH

Rock P application amounts (mg P /kg)

abab

ab

b

a

P uptake by cowpea roots decreases pH in rhizosphere soil

Rhizosphere soil pH increases with increase in rock P applications WAY FORWARD

Elucidate P uptake mechanisms in cowpea Elucidate mechanisms of improved rock P

solubility & uptake


Dry Seed Weight (g/plant) Stressed plants: 0 to 15.4 g/plant Non-stressed plants: 1.5 to 58.4 g/plant

Number of days to flowering Drought escape strategy: Flowering time reduction under stress

Field screening for drought tolerance and high yield potential in

cowpea germplasmC. Fatokun, O. Boukar, S. Muranaka

Fatokun et al. 2012_Plant Gen. Res. 10:171-176


Field screening for drought tolerance and high yield potential in

cowpea germplasm


HT Phenotyping cowpea mini core germplasm collection for adaptation

to drought and low P N. Belko, O. Boukar, C. Fatokun

et al. Screen-house trials for assessing variability in drought-avoidance shoot traits among the cowpea mini core collection in IITA-Kano: Oct 2014 – March 2015 370 lines + 10 checks under non-limiting water condition in 3

replications Gravimetric measurement of whole plant canopy transpiration,

leaf temperature, leaf cholorophyll content, leaf area development and shoot-root biomasses


Extent of phenotypic variations in plant TR, SPAD-CMR, CTD and LA in

cowpea


Field trials for assessing differences in growth and yield performance under well-watered and water-stressed conditions in Minjibir station: March-July 2015 348 lines + 12 checks under 2 water regimes in 3 replications Plant phenology (days to flowering and maturity), yield

components (fodder, pod and grain), visual scoring for (i) Striga emergence, (ii) insect leaf damages and (iii) leaf senescence under drought, and SPAD-CMR and NDVI data on selected genotypes

On-going post-harvest activities and data processing

Field screening for tolerance to drought in cowpea mini core

germplasm collection


2600 2800 3000 3200 3400

600

800

1000

1200

1400

1600

1800

Non-stressed grain yield (kg/ha)

Dro

ught

-stre

ssed

gra

in y

ield

(kg/

ha)

58-53

58-57

IAR8/7-4-5-3Iron-Clay

IT00K-901-6

IT83D-442

IT89KD-288

IT90K-284-2

IT93K-503-1

IT95K-1090-2

IT95K-1095-4

IT96D-610IT97K-207-15

IT97K-556-6IT97K-819-132

IT98K-128-2IT98K-205-8

IT98K-317-2

IT98K-428-3

IT98K-498-1

IT98K-698-2

IT99K-124-5

KVX-396

KVX403

KVX-421-25

KVX-525

MougneN’diambour

Petite-n-grn

Suvita 2

Repeat of the field yield based evaluation under WW & WS in Minjibir: Sept-Dec 2015 Integration of partitioning / grain filling parameters and drought tolerance indices

Beebe et al. 2013_Field Crops Res. 148:24–33

Belko et al. 2014_Crop Science 54:1-9


Field and image based phenotying of root traits for adaptation to drought and low phosphorusField and lab trials for evaluating genotypic differences root

system architecture and anatomy at the ARBC Willcox-AZ with PSU partners: July-Sept 2015

Fifty lines planted in single row plot under WW conditions in 5 reps

5 plants per plot excavated and visually scored for root traits (Shovelomics: angle, number, density, diameter) and root samples taken for cross section anatomy analysis

5 seedlings per line for analysis of root hairs density and length using DIRT & ImageJ


Research plan for genetic improvement (high genetic gain): 1. Identification of plant traits conferring tolerance to drought and adaptation to low P 2. Dissecting traits interactions (trade-off, synergism) and plasticity/stability 3. Parameterization - Modeling the effect of traits across env. and stress scenariosOpportunities and future: • Penn State University and ARBC/G Howard Buffet Foundation

and ICRISAT for high throughput phenotyping of relevant root and shoot traits for adaptation to drought and low fertility

• Institute of Meteorology and Climate Research (IMK-IFU-KIT) Germany, WASCAL Burkina Faso, and Depart. Crop Sci in North Carolina State Univ for modeling to develop guidelines for farming options in response to climate variability

Challenges and resource required: Field and lab research facilities (irrigation, drought and low P screening sites, striga pression, lysimetric system, rain-out shelter, screen-houses, lab space with specific equipment etc.) and human resources (technicians and students).

Opportunities and Way forward Challenges and resource needs


MERCI DE VOTRE ATTENTION

Government & Nonprofit

Phenotyping for tolerance to abiotic stresses