SeminarSeminaron on

Breeding For Tolerance To High Breeding For Tolerance To High Temperature StressTemperature Stress

Guided by:-Guided by:-Dr. G. K. Koutu, Principal ScientistDr. G. K. Koutu, Principal ScientistDr. S.K. Singh, Assistant Professor Dr. S.K. Singh, Assistant Professor

2014-152014-15

Lokesh GourDEPARTMENT OF PLANT BREEDING & GENETICS DEPARTMENT OF PLANT BREEDING & GENETICS

JAWAHARLAL NEHRU KRISHI VISHWA VIDYALAYA JAWAHARLAL NEHRU KRISHI VISHWA VIDYALAYA JABALPUR (M.P.)JABALPUR (M.P.)

IntroductionIntroduction Heat stress/ high temperature stress is defined as increased

temperature level sufficient to cause irreversible damage to plant

growth and development

Between 2000 and 2050, even with no climate change, the price of

rice, maize, soybean, and wheat would rise, the climate change

results in additional price increases which may be drastic effect

(Nelson 2009)

There is a need to search for genotype that can tolerate heat stress

condition and better understanding of genetics and mechanism of

heat tolerance will enable the development of suitable varieties for

such conditions

Contd...Contd... Two types of heat stress mechanism –

• Heat Avoidance : Plant avoid excessive heating of their leaves

by decreasing their absorption of solar radiation

• Heat tolerance : Ability of plant to grow and produce economic

yield under high temperature (Wahid et al. 2007)

All India annual mean temperature anomalies for the period 1901-2009 (based on 1961-1990 average)

• Solid blue curve show sub-decadal time scale variations smoothed with a binomial filter

Morpho-anatomical & Phenological responses

Physiological responses

Molecular responses

Morphological symptoms

Accumulation of compatible osmolytes

Oxidative stress & antioxidants

Anatomical changes Photosynthesis Stress proteins

Phenological changes Cell membrane thermostability

Plant responses to high temperature

Prolongedcause

High Temperatur

e

Occasionalcause

Injury of high temperature on plant body

Heat Shock Sun Scald

Sun Scald

Alocasia macrorrhizos

Tomato Corn

Heat shock

Aglaonema

Crop plants Threshold temp. (◦C)

Growth stage References

Wheat 26 Post-anthesis Stone and Nicol´as (1994)

Corn 38 Grain filling Thompson (1986)

Cotton 45 Reproductive Rehman et al. (2004)

Pearlmillet 35 Seedling Ashraf and Hafeez (2004)

Tomato 30 Emergence Camejo et al. (2005)

Brassica 29 Flowering Morrison and Stewart (2002)

Cool season pulses 25 Flowering Siddique et al. (1999)

Groundnet 34 Pollen production

Vara Prasad et al. (2000)

Cowpea 41 Flowering Patel and Hall (1990)

Rice 34 Grain yield Morita et al. (2004)

Threshold high temperatures for some crop plants

The germination percentage was zero when temperature ranged between 45 and 48 0C.

(Singh and Dhaliwal 1982)

Wide fluctuations in day-night temperature reduces yield of rainfed pulse

Impact of High Temperature in pulses

Impact of High Temperature in rice High temperature may lead to flower abnormality so the pollen viability affected

Long-term increases in night time temperatures decrease rice yields by 10% for every 1 °C (Peng et al.,2004)Albinism of panicles and spikelet's

Albinism- The absence of Chlorophyll producing a white panicles or Spikelets

Shimizu and Kuno, 1975

Stamens stop growth

Two pistils develop

Tissue enlargement in part of ovary

Flower Abnormalities of rice

Impact of High Temperature in wheat Temperature >30ºc – damage pollen & every degree rise in temp reduce yieldBread making quality is decreased at further temperature increases above 35° C in a genotype dependent manner (Blumenthal et al., 1993, Stone and Nicolas, 1995)Terminal heat stress in the post anthesis - reduce grain growth, size and yield because of translocation

Scanning electron microscopy of grains of wheat cultivars (Dias et al. 2008)

Flower development of tolerant (left) and sensitive tomato genotypes (right):-

Pollen viability of tolerant (left two) and sensitive tomato genotypes (right two) :-

Impact of High Temperature in tomato

Under high temperature, anthers showed deformation, dark coloration of the anther tip and elongated pistils. Those flowers had a low percentage of pollen viability (<10%)

Contd…Contd…

a. Rubisco protein2D gel electrophoresis picture of leaf protein of NH219:- The conspicuous protein spot (spot3) found between 50kDa and 60kDa and PI of 6.2 was identified as the Rubisco large chain precursor (EC 4.1.1.39)

Mechanism for High temperature tolerance

b. Heat Shock ProteinClasses of HSPs : HSP 60, HSP 70, HSP 90 , HSP 100

LMW HSPs (Swindell et al. 2007)

Found in Cytosol, Mitochondria, ER, Chloroplast

source : Mirza et al( International Journal of Molecular Science)

Genetic improvement for High temperature tolerance

Conventional Breeding Strategies

Molecular & Biochemical Breeding Strategies

Mutation Breeding Strategies

Conventional breeding strategies

Grow under hot target production

environment

Identify individuals with greater potential

Vegetative StageHeat Tolerance Index (HTI)

Reproductive StagePollen viability, pollen tube growth

and grain filling

Selection criteria for identification or screening of individuals

Workflow of stem cutting tuberisation assay used to screen for heat tolerance on UK varieties potato

Effect of heat stress on tuberisation potential that goes lower

Screening of heat tolerant potato

JG 11JG14(ICCV 92944) KAK 2

JG 14-Screening for heat tolerance at ICRISAT during summer 2008

JG 14 (ICCV 92944) released as Yezin 6 in Myanmar

Screening of heat tolerant Chickpea

M1 seed

M1 Plant

M2 seed

It shows dominant variation

Mutation breeding strategiesTreatment of seeds and vegetative propagules produces chimeras

Molecular & Biochemical breeding strategies

QTL mapping

QTL

Susceptibletolerant

Susceptible tolerant

F2

P2

F1

P1 xTolerantSusceptible

Selection based on presence of marker

Marker Assisted Selection (MAS)

large populations consisting of thousands of plants

Identification of QTL for stay green trait in wheat

Induction of heat-inducible genes

Osmoprotectants,Signaling molecules (e.g., GH),Oxidants (e.g., H2O2)

Induction of temperature tolerance

Foliar application,Pre-sowing seed treatment,Preconditioning of plants,

Low concentrations of inorganic salts

Treatment Crop References High-temperature preconditioning Tomato Morales et al., 2003

Pre-sowing hardening Pearl millet Tikhomirova, 1985

Application of cacl2 - Kolupaev et al., 2005

Seeds pre-treated with Glycinebetaine and polyamines Barley Wahid and Shabbir, 2005

Exogenous application of spermidine tomato Murkowski, 2001

Genetic transformation

Traits selectable for high temperature toleranceNo. Trait Characteristics References(Morphological traits amenable to direct selection)

1 Okra leaf type Higher leaf (N) content Pettigrew (2004)

Higher CO2 exchange rate (CER)

Higher photoelectron transport rate

Reduced individual leaf area Wells et.al., (1986)

2. Lower fruiting height Greater heat tolerance Feaster and Turcotte (1985)

3. Thicker leaves Higher N content Hall (2001)

Higher photosynthetic capacity Wright et.al.,(1993)

4. Earliness Reproductive heat tolerance Ahmed et.al.,(1993), Ehlers and Hall (1996)

5. Stay-green effect Heat tolerance Reynolds et.al.,(1997)

6. Pollen selection Pollen selection through heat treatment Rodriguez-Garay and Barrow (1988)

(Physiological traits for both direct and/or indirect selection)

1. Cell membrane thermostability (CMT)

Measures the resistance of protoplasmic proteins to denaturations

Saadalla et.al.(1990a), Blum & Ebercon (1981)

Heat and drought tolerance Ashraf et.al.(1994)

2. Chlorophyll contents & chlorophyll acb ratio

Dry matter and yield Al-Khatib & Paulsen (1984), Saranga et.al. (2004)

3. Carbon isotopes discrimination differences

Heat tolerance Lu et.al. (1996)

No. Trait Characteristics References(Screening of traits through infrared/ remote sensing for direct selection)

1 Leaf conductance Heat tolerance Lu et.al. (1994)

2. Crop water stress index (CWSI) Transpiration rate

Vapor pressure deficit (VPD)

Plant temperature Wells et.al., (1986)

Net radiation

Canopy temperature Burke et.al.(1990)

Aerodynamics resistance Burke et.al.(1990), Jackson et.al.(1981)

3. Chlorophyll fluorescence Efficiency of PSII Butler (1978)

Indication of damage to PSII Hall (2004)

4. Canopy temperature depression (CTD)

Leaf conductance Reynolds et.al.(1998)

Air temperature Amani et.al.(1996)

Soil water status and RH Amani et.al.(1996)

Heat escape Cornish et.al.(1991)

5. Thermal stress index (TSI) Quantify thermal stress Burke et.al.(1990)

Measures enzymatic functions at high temperature

Contd…

QTLs detected for high temperature tolerance

Plant QTLs Phenotype/ Function of genes

Arabidopsis EDI Flowering time

Arabidopsis PHYA Hypocotyl elongation

Rice Hd1 Flowering time(CONSTANS)

Rice Hd6 Flowering time

Maize Dwarf 8 Flowering time

Rice TGMS Temperature sensitive male sterility

Wheat Xbarc 186

Genes transformed for high temperature tolerance

Transgenic Plants

Transgenes Function of transgenes Source of genes

N. tabacum MT-sHSP Molecular chaperone function in vitro L. esculentum

N. tabacum Dnak 1 High temp. tolerance Aphanothece halophytica

N. tabacum BADH (betain aldehyde dehydrogenase)

Over production of GB osmolyte that enhance heat tolerance

Spinacia oleracea

A. thaliana Cod A (choline oxidase A)

Glycine betaine synthesis during imbibition and seedling germination

A. globiformis

A. thaliana APX1 H2O2 detoxification P. sativum

A. thaliana HvAPX1 H2O2 detoxification H. vulgare

Zea mays & O. sativa

Hsp 100, Hsp 101

HSP synthesis for heat tolerance A. thaliana

N. tabacum Fad 7 Increase the level of unsaturated fatty acids N. tabacum& O. sativa

Daucus carota Hsp 17.7 sHSP synthesis for heat tolerance Daucus carota

High temperature tolerant varieties of crop plants

• Wheat : JW-3020, JW-3173, JW-3211, JW-3288 (Initial & terminal heat tolerance) JW-3336, JW-1202, JW-1203 (terminal heat tolerance) Chirya-3

• Chick pea : JG-14 (Yezin 6 ), Annigeri, ILC 482 and ICCV 10 (Srinivasan et. al., 1996)

• Rice : NH219, Nagina 22 (remain at 40°C temp.), Dular, IR-64

• Papaya : CO7

Centres which works for development of high temperature tolerant crop varieties

IARI, New Delhi

JNKVV , Jabalpur

PAU , Ludhiana

ARI, Pune

NRCPB, New Delhi

ICRISAT, Hyderabad

IIPR, Kanpur

ConclusionConclusion Studies have found tremendous variation within and between species,

thus this will help to breed heat tolerance for future environment. Some of attempts to develop heat-tolerant genotypes are successful. (Ehlers and Hall, 1998; Camejo et al., 2005 )

Important to examine crop responses to a range of possible changes, especially in the nature, frequencies and sequences of extreme climatic changes

Designing/development accurate screening procedures

Heat stress resulted in reduction of other 11 traits except plant height in both N22 and NH219. The extent of reduction was more in N22 than in NH219. Both pollen viability and spikelet fertility were not reduced significantly in N22 and NH219 but reduced by 20% in IR64 (Poli et. al., 2013)

Problem regarding to high temperature tolerance development

References

oFarooq M, Bramley H, Palta JA and Siddique KHM. 2011. Critical Reviews in Plant Sciences, (30):1–17

oHasanuzzaman M, Nahar K, Alam Md M, Roychowdhury R and Fujita M. 2013. Physiological, Biochemical, and Molecular Mechanisms of Heat Stress Tolerance in Plants, Int. J. Mol. Sci., (14): 9643-9684

oKumar U, Joshi AK, Kumari M, Paliwal R, Kumar S, RÖder MS. 2010. Identification of QTLs for stay green trait in wheat (Triticum aestivum L.) in the ‘Chirya 3’ x ‘Sonalika’ population, Euphytica (174):437–445

oWahid A, Gelani S, Ashraf M, Foolad MR. 2007. Heat tolerance in plants: An overview. Environmental and Experimental Botany (61): 199–223