topics term paper assignments¹€อกสาร... · 1 reducing water use, increasing yields...

1

123752 Horticultural Crops Breeding for Pest

Resistance and Environmental Tolerance

123752: Suchila, KKU, Thailand

Assoc. Prof. Suchila Techawongstien (D.Agr.) Plant Breeding Research Center for Sustainable

Agriculture Faculty of Agriculture, Khon Kaen University

1

Course description

Relationship between plant genetics, diseases, insects and environmental stress; Techniques of crop improvement for


resistance to diseases, insects and environmental stress; and

Management of crop resistant variety

2

Topics

Chapter I: Introduction (Terminology, Importance of resistant variety and Principles and methods of breeding for resistance): slide 5‐23

Chapter II: Inheritance of resistance/tolerance traits (Gene action and gene interaction, Sources of resistance, Heritability and breeding method, and assessment of resistance/tolerance): slide


24 ‐158

Chapter V: Plant and environment relationship (Stress environment, Responses of plant to environmental stress and Plant breeding and yield stability): slide 159‐192

Chapter VI: Management of resistance/tolerance varieties: slide 193‐199 3

Term paper assignments

Search and review the update papers Present and discuss the update

papers Suggested topics: bacteria, fungi,


Suggested topics: bacteria, fungi, virus, nematode, insect-pests and environmental resistance/tolerance of economic plants (or the major crops of the student’s research)

4

Lecture Chapter I: Introduction

1.1 Terminology

1 2 Importance of resistant variety

Chapter I: Introduction


1.2 Importance of resistant variety

1.3 Principles and methods of

breeding for resistance

5

1.1 Terminology

1) Parasite

2) Pathogen

3) Disease

7) Tolerance

8) Disease escape

9) Immunity

) ff l h

1.1 Terminology


3) Disease

4) Resistance

5) Susceptible

6) Hypersensitivity

10) Differential host

11) Physiological

race

12) Pathogenicity

6

1

1.1 Terminology (cont.)

13) Vertical resistance14) Horizontal resistance

18) Preference19) Non‐ preference20) Environmental stress

Chapter I: Introduction


15) Disease assessment 16) Persistent vector17) Non‐ persistent vector

21) Phenotypic responses22) Genotypic responses 23) Stability

7

Importance of resistant variety

In 1845-46: Irish Famine-potato late blight (Phytophthora infestans)

40% of potato tuber in Belgium/Ireland was rotten by late blight

1. 2 Importance


More than 1.5 million people died (Population of Ireland declined from 8.2 to 6.2).

https://www.google.co.th/search?q=potato+late+blight+pictures8

At Bengal: Epidemic of Brown Spot (Helminthosporium oryzae) in rice

75-90% of rice yield in Bengal was decreased

Importance of resistant variety (cont.)

1. 2 Importance


More than 2 million people died

Europe: Epidemic of corn caused >20% of maize yield

9

Hurricane Katrina

Tsunami: JapanImpacts of climate changes

http://en wikipedia org/wiki/An Inconvenient Truth

1. 2 Importance


200620052004 2011-4

Tsunami: Thailand Cyclone Nargis

Flooding: Thailand

https://www.google.co.th/search

http://en.wikipedia.org/wiki/An_Inconvenient_Truth

Earthquake Thailand (2014)

10

New lifestyles and new technologies to avert more global warming: what’s the hurry?

It’s getting hotter

1. 2 Importance


June, 2003: 15,000 excess deathsin France alone from heat wave

Source: V. Gutschick, NMSU (Emeritus)11

Rainfall distribution is changing…and becoming more extreme

…which may lead to more frequent droughts, and megadroughts, and wildfires

Source: V. Gutschick, NMSU (Emeritus)

1. 2 Importance


Source: V. Gutschick, NMSU (Emeritus) 12

1

Why hurry?Sea level rise…in the long term

What’s needed already…will have to be multiplied manyfold

Ice caps are melting faster than originally predicted, from “unplugging”

1.2 Importance

123752: Suchila, KKU, ThailandSource: V. Gutschick, NMSU (Emeritus)13

Increased demand 50% by 2030 (IEA)

Energy

Climate Climate ChangeChange

1. Increasing population

2. Increasing levels of urbanisation

The Perfect Storm?The Perfect Storm?1. 2 Importance


Water Increased demand 30% by 2030

(IFPRI)

FoodIncreased demand 50% by 2030

(FAO)

ChangeChangeurbanisation

3. The rightful goal to alleviate poverty

4. Climate Change

Source :John Beddington. 2009. Science in Government: Challenges for the Science in Government: Challenges for the 2121stst Century. Century. Campaign for Science and Engineering, London 10 December 2009.14

Recent effects of global warmingon agriculture and horticultural crops in Japan

• survey on the effect of global warming on agricultural industry in Japan

1.2 Importance


http://www.agnet.org/library.php?func=view&style=type&id=20120104150721 15

White immature rice Cracked rice

1. 2 Importance

123752: Suchila, KKU, ThailandHinohikari Nikomaru (breeded in 2005)

http://www.agnet.org/library.php?func=view&style=type&id=2012010415072116

Rind puffing of Satsuma mandarin

Ch i th di t ib ti

1. 2 Importance


Sunburned fruit Change in the distribution of suitable lands for apple

cultivation

http://www.agnet.org/library.php?func=view&style=type&id=20120104150721 17

1. 2 Importance


Improvement of fruit quality by control of water status using plastic mulching and drip irrigation (fertigation) in citrus orchard


18

1

Reducing water use,increasing yields

drip irrigation From 39% in 2009 to 52% in 2011• Australia, tomatoes using up to 70% less water

• USA tomato yield

1. 2 Importance

Resistant/tolerant varieties are


• USA , tomato yield increase of over 25%.

• India, trials on gherkins save water 40%

• fungicide application rates reduced to 50%.

Source: Kamol Lertrat (2012)

necessary.

19

1.3 Principles and methods of breeding for resistance

Method for controlling disease and insect pests

o Quarantine

o Cultural controlไรตัวห้ํา

Source: รศ.ดร. นุชรีย์ ศิริ

1.3 Principles and methods


o Chemical control

-Hyperparasites and predators

-Resistant varieties

o Integrated control

แมลงช้างปีกใส

20

Host‐parasite Co‐evolution: is defined as the reciprocal adaptive

genetic change of two antagonists through reciprocal selective pressures.

Selection dynamics:



o Negative frequency dependent selectiono Over-dominant selection: Rr > rr, RRo Directional selection

http://en.wikipedia.org/wiki/Host%E2%80%93parasite_coevolution

21

Priorities in breeding objective for resistance to disease/insect:o Great amount of damage to the cropo Controlled at low cost by other means, does

Challenge facing the plant breeder



not merito Screening techniques is available (plant Patho/Ento) + Breeder

o Source of resistance (plant Patho/Ento) + Breeder

22

oTo find out source of resistance:local adapted lines, breeder stock, exotic cultivars, wild and related species and created variation

Challenge facing the plant breeder (cont.)



created variationoMethods to introduce resistance into the existing breeding materialso Agronomic/quality accepted resistant variety, and do not neglect resistance to other pests.

23

2.1) Gene action and gene interaction

2.1.3 Genetics of host‐pathogen reactions

2.1.1 Concept of pathogen and host

2.1.2 Mechanisms of defense in plants against pests

Chapter II: Inheritance of resistance/tolerance


2.2.1 World genetic resources

. .3 Ge et cs o ost pat oge eact o s

2.2) Sources of Resistance

2.2.2 Genetics resource management 24

1

Part II: Inheritance of resistance/tolerance (cont.)

2.3.1 Resistance breeding strategies

2.3) Heritability and breeding method

2.3.2 Breeding for specific resistance


2.4.2 to environmental stress disease

2.4) Assessment of resistance/tolerance

2.4.1 to plant pests (bacteria, fungi and virus)

25

Pathogen Host

Pathogenicity:

Avirulence vs virulence

o Pre-existing defense Mechanisms

2.1) Gene action and gene interaction:

2.1.1 Concept of pathogen and host


Avirulence vs virulence

Non‐aggressive vs aggressive

Races vs pathotypes

o Infection induces defense mechanisms

https://www.google.co.th/search?q=potato+late+blight+pictures

26

Pre‐existing defense

Mechanisms

o Cuticles

o Stomata

Pl t h i

Infection induces

defense mechanisms

o Periderm: plant cell accumulate lignin, turn to necrosis and inhibit disease growth

Mechanism of Resistance

2.1 Gene action and gene interaction


o Plant hairs

o Phyto‐chemical:

‐tomatine anti Fusarium oxysporum

‐unsaturated lactones (glucosides in tulip anti F. oxysporum

o Formation of plant structure: gels and tylose inhibit disease epidemic

o Cell wall modifications: to protect plant cell from disease infection

o Hypersensitivities reaction

27

• Immunity

• Disease escape

2.1.2 Mechanisms of defense in plants against pests



• Defense

• Resistance• Tolerance

28

Resistance: no establishment of

Avoidance or escape: a mechanism that reduces the probability of contact between pathogen or insect pests and the plant

2.1.2 Mechanisms of defense in plants against pests (cont.)



Tolerance: the host develops, continues to grow, and produces well despite the pathogen’s presence

Resistance: no establishment ofthe pathogen in/on the host, or a limited establishment

29

Proposed by H.H. Flor (1950)-study on flax rust “Gene for Gene Hypothesis” Virulence of a pathogen and the resistance of the host have a genetic

2.1.3 Genetic of host-pathogen reactions



resistance of the host have a genetic basis. For each gene that confers resistance in the host, there is a corresponding gene in the pathogen.

30

1

Genetic Variation in Plant

1) Environmental Variation

2) Heredity variation:

Gene Action and Interaction

1) Gene action:

‐ Dominance

‐ Recessive

2) Gene Interaction:



‐ Quantitative Variation

‐Qualitative Variation

2) Gene Interaction:

‐ Complementary action

‐Modifying action

‐ Inhibiting action

‐Masking action (epistasis)

‐ Duplicate action

‐ Additive action31

1) Fungi:1.1 Obligate parasite

1.2 Facultative parasite

Variations: Mutation, genetic recombination

3) Bacteria:3.1 Gall forming bac.

3.2 soft rot bac.

3.3 Physiological or

Genetic Variation in Pathogen



genetic recombinationy g

metabolism altering bac.

Variations: binary fission, genetic recombination, mutation

2) Virus:

Variations: Mutation, genome recombination

32

Type and Mechanism of Resistance

1) Base on heredity of resistance

1.1 Monogenic R.

1.2 Oligogenic R.

1 3 Polygenic R

3) Base on mechanism of R.

3.1 Active R. (Responsive R.)

3.2 Passive R.

4) Base on development stage of plant resistance



1.3 Polygenic R.

2) Base on epidemic of disease (Van de Plank)

2.1 Vertical R.

2.2 Horizontal R.

5) Base on population distribution

5.1 Quantitative R.

5.2 Qualitative R.

p

4.1 Seedling R.

4.2 Adult or mature R.

33

Types of Resistance

o Vertical resistance

o Horizontal resistance



http://en.wikipedia.org/wiki/Heritability

34

• Hypersensitivity

• Specific resistance

• Major gene or oligogenic

Vertical Resistance

Resistance

Susceptibility



oligogenic

• Non‐durable or Qualitative resistance

1 2 3 4 5 6Races

Vertical Resistance to Races 2, 5, and 6

www.uky.edu/...breeding/.../Plant%20Breeding%20S07/bre...

35

• Partial resistance

• Non‐specific resistance

Mi i t

Horizontal Resistance

Resistance



• Minor gene resistance

• Polygenic inheritance

• Quantitative resistance

Horizontal Resistance to all Races

1 2 3 4 5 6Races

www.uky.edu/...breeding/.../Plant%20Breeding%20S07/bre...

36

1

Genes in pathogen

R (resistant, dominant)

r (susceptible, recessive)

A dominant AR (‐) Ar (+)

Gene‐for‐gene reactions

Virulence(a) or Avirulence



a recessive aR (+) Ar (+)

(A) genes In thepathogen

37

Locks and Keys concept(Simplified by J.A. Browning)

Pathogen Host

R1R2 R1r2 r1R2 r1r2

A1A2 - - - +

For resistance:Host has a set of locks (R1R2) as dominant alleles



A1a2 - - + +

a1A2 - + - +

a1a2 + + + +

To establish disease: Pathogen has a set of keys as a virulence allele (a1, a2) to open the locks,or host much have no lock (r1, r2) for that keys

38

Genetics of Host Plant RelationshipGene for gene concept (Flor, 1956)

Genotype

Variety Plant Disease Symptom

Redwood NNPP AnAnApAp R

Genotype of flax and rust



Redwood NNPP ananApAp R

Polk NNpp AnAnapap R

Polk NNpp ananApAp S

Winowa nnpp All genot. S

Plant genot.: NNPP=R, Disease genot.: a=virulence; A=avirulence39

• Over 1750 genebanks exist world wide.

• About 7.4 million germplasm accessions

conserved in ex situ collections,

• More than 70% of the genetic diversity of

some 200-300 crops is already

2.2 Source of Resistance

2.2.1 World Genetic Resources


Establishment of the Svalbard Global Seed

Vault, Norway a last resort safety back-

up repository of genetic resource

safeguard humanity.

conserved in genebanks (SBSTTA, 2010)

• Source: WIEWS 2009; Country

Reports; USDA-GRIN 2009, SOW2

40

USDA, TGRC

The Centre for Genetic Resources,

the Netherlands (CGN)

Active Genetic Resources for Pepper and Tomato

2.2 Source of resistance


41

S. chilenseS. pennellii S. peruvianum

Genetic Resources for Tomato Improvement 2.2 Source of resistance


© TGRCS. habrochaites

S. pimpinellifoliumS. l. var. cerasiforme

S. lycopersicum S. chmielewskii

Source: AVRDC 42

1

Resistance genes against insectsHessian Fly resistance genes H9, H13

Russian Wheat Aphid resistance genes Dn2, Dn4

• Most cultivars from the Pacific North-west carry only resistance gene H3

• RAPDs markers for H9: CCCAGTCACT & H13: TTGCTGGGCG

GrainGenes

(Source: Dweikat et al. 1997 TAG 94: 419-423)

D 2 7DS li k d t i t llit X 111 (3 M)

Grain Genes



Green bug resistance gene Gb5

7A.7S-L7/5

Xpsr129

EcoR I

77SS

77AA

• Gb5: Large interstitial translocation from T. speltoides chromosome 7S-Linto chromosome 7A.

(Source: J. Dubcovsky et al. Crop Sci. 38: 1655-1660)

• Dn4:1DS between RFLP markers Xabc156 (11 cM) and Xksud14 (Source: Me et al 1998 genome 41: 303-306)Gran Genes

GrainGenes

• Dn2:7DS linked to microsatellite Xgwm111 (3 cM)

(Source: Liu et al. 2001 TAG 102: 504-510)

43

TVRC, KU:

2.2.2 Examples of genetic resources in Thailand



Difficult to accessPBRCSA, KKU

Pepper 800 acc.

Tomato 1000 acc.

TVRC, KU:

Pepper 2827

acc. Tomato

acc.?

44

Active genetic resources for hot pepper and tomato

2000

8163

http://www.ars‐grin.gov/npgs/orders.html

Pepper Tomato websiteResources

www avrdc org



1011 1309 http://www.cgn.wageningen‐ur.nl/pgr

Banco de Germoplasma de

Hortalias (BGH; Brazil)www.bgh.ufv.br/

‐ http://tgrc.ucdavis.edu

www.avrdc.org

45

Germplasm*

elite line

Base/improve

Public/ Private

(Seed Co.)/ Grower

Food/health

food

Genetic

resources:

AVRDC

USDA

TGRC TVRC KKU

Seed co.

Col

lect

ion

Evaluation

Gene pool

Germplasm management at KKU



population

Pure/inbred lines

F1-hybrids

Pharmaceutical

products

(Animal) feed

additive

etc.

Selection

Selection

Svalbard Global Seed Vault

46

Collection

Evaluation



Evaluation

Distribution

47

Examples of resistant and other characteristics sources of pepper and tomato

Phyto-nutrient Capsiate, pigment Lycopene,

ß-carotene

Disease Anthracnose Bacterial wilt

Trait Pepper Tomato



Disease

resistance***

Anthracnose,

Gemini virus,

Begomo virus

Bacterial wilt,

TYLCV,

Begomo virus

Others

(Agronomic traits)

Male sterile Raisin tomato

48

1

Evaluation and characterization

Evaluation

100 acc./year

KKU accession number KKU-P 11008Duplicate number -Temporary number -Variant -Species Capsicum annuum L.Subtaxa -Pedigree / Cultivar Name Jinda-YaoCountry Thailand/KKU



Location acc./year

KKU 40

U. Ratchapat CM. 30

U. Ratchapat MSK. 30

(total 300 acc.)

CountryCharacterized year and Rainy Season, 2006Remarks -

Desctiptor Name Value Desctiptor Name Value

Cotyledonous leaf shape - Stem color Green

Cotyledonous leaf color Green Root distribution Low

Hypocotyl pubescence -

Lyfe cycle Biennial Leaf colour Green

Stem color Green with purple stripes Leaf shape LanceolatePlant height (cm.) - Mature leaf length (cm.) 8.5Plant canopy width (cm.) - Mature leaf width (cm.) 3Plant growth habit Intermediate

Days to flowering 40 Anther colour PurpleNumber of flowers per axil One Male sterility AbsentFlower position Erect Calyx annular constriction AbsentCorolla colour White

Fruit colour at intermediate stage Green Neck at base of fruit AbsentFruit colour at mature stage Red Fruit shape at blossom end PointedFruit shape Elongate Fruit surface SmoothFruit length (cm.) 7.5 Fruit pungency High (~ 152,995.80)Fruit width (cm.) 1.4 No. of locules -Fruit shape at pedicel attachment Obtuse

Seed colour Straw 100-seed weight [g] 0.502Seed surface Smooth Number of seeds per fruit -

No. of fruit 300 Dry weight (g) -Fresh weight (g) 530 Remark -

Last update : March' 2007

Seedling Data

Vegetative Data

Inflorescence Data

Fruit Data

Seed Data

Yield Data

49

Germplasm distribution

Field day/Varietal trials

at KKU



Hot pepper in rainy season Tomato in dry season

50

Stock Maintenance:

Cold room conditions

• Medium-term conditions : under refrigeration at 5-10°C.

• All seed samples sealed in a suitable air-tight container in

%

•Drying at 10-25 ° C and 10-15 %RH

• Drying at 5-20 ° C and 15-25% RH



Viability monitoringstorage environment RH 15±3%.

•Germination should exceed 85% & 75 % some

vegetables

•within 12 months after receipt of the sample Regeneration

•Viability drops below 85 % of the

initial viability.

51

Data Base ManagementDatabase QueriesAccessions... Geographic Data on

Wild Species Accessions... Core collectionGenesList of Gene Names and Symbols...View Naming Rules

Example of seed request



gImages ColleaguesOther ResourcesRecent AcquisitionsTop 20 most requested Accessions (2011). Recent database/website updates:(Google maps now available for wild species)(All Wild species localities available)

Solanum lycopersicoides introgression linesSolanum habrochaites (L. hirsutum) introgression lines2nd generation pennellii ILs2005 collections from Chile2001 collections from ChileSeed Request InformationHow to Request SeedContact Information 52

Heritability :is a concept that summarizes how much of the variation in a trait is due to variation in genetic factors. This term is used in reference to the resemblance between parents and their offspring. In this context, high heritability implies a strong resemblance between parents and offspring with regard to a specific trait while low heritability implies a low level

2.3 Heritability and Breeding Methods


regard to a specific trait, while low heritability implies a low level of resemblance.

Figure 1: Heritability estimation.Low (panel a) and high (panel b) heritability can be estimated from the regression (h2) of offspring phenotypic values vs. the average of parental phenotypic values.© 2008 Nature Education All rights reserved

53

Heritability Broad-sense heritability, defined as h2

b = VG/(VG +VE)

Narrow-sense heritability, defined as h2 = VA/(VA + VD + VE)

2.3 Heritability and breeding methods


defined as h n VA/(VA VD VE)

Figure 3. Strength of selection (S) and response to selection (R) in an artificial selection experiment,h2=R/S

Figure 4. Sir Francis Galton's (1889) data showing the relationship between offspring height (928 individuals) as a function of mean parent height (205 sets of parents).

http://en.wikipedia.org/wiki/Heritability54

1

Specificity in the parasite:

2.3.1 Resistance Breeding Strategies

Physiological race: Pathogen genotypes share a group of cultivars

Differential hosts (cultivars): Using differential cultivars toidentify physiological races



share a group of cultivars to which they are virulent

identify physiological races or vice versa.

Cultivars Races

R1 R2 R3 R4

C1 - + - +

C2 - - + +55

Tomato Late Blight Differentials in AVRDC Taiwan

Host Differentials

Pathogen race Ph+,

Ph-1

WV700

(Ph-2)

CLN2037

(Ph-3)

L3708

(Ph-3 + Ph-4)

LA1033

T1 S R R R R

T1,2 S S R R R

T1,3 S R S R R

T1 2 3 S S S R R



T1,2,3 S S S R R

T1,3,5 S R S R S

T1,2,3,4 S S S S R

T1,2,3,5 S S S R S

T1,3,4,5 S R S S S

T1,2,3,4,5 S S S S S

Chen et al., 2008 R= host gene confers resistance to that pathogen race S= susceptible reaction. Pathogen race overcomes resistance

56

Individual Major Genes

2.3.2 Breeding Methods for Specific Resistance

Application of gene pyramiding



Using multi-lines

57

Individual Major Genes

Breed cultivars with major genes that control the

l t t

2.3.2 Breeding Methods for Specific Resistance (cont.)



prevalent pest races

Select progeny from a segregating population or

Transfer major genes from other sources

58

Application of gene pyramiding

Put all knownmajor genes into one line

Analysis of genome

2.3.2 Breeding Methods for Specific Resistance (cont.)



Singh A et al. AoB PLANTS 2012;2012:pls029

Published by Oxford University Press on behalf of the Annals of Botany Company.

Analysis of genomeintrogression associated with resistance genes/QTL. (A) xa13 on chromosome 8 and Xa21, Pi54and qSBR11‐1 on chromosome 11 (B) in ‘Pusa1608’ families.

59

Ty-1

Ty-3

Chromosome 6 Chromosome 11 Chromosome 3

Mapped TY Resistance Genes in Tomato at AVRDC

Ty-5

Chromosome 4



Ty-2

Ty-4

Source: Peter Hanson, AVRDC 60

1

Using multi-lines

Develop many individual lines each with individual major resistance genes (isolines (or

2.3.2 Breeding methods for Specific Resistance (cont.)



near isogenic lines))

Mix the seed of these lines together to get multilines

Protection against a broad spectrum of races

61

2.4) Assessment of Resistance and Tolerance

2.4.1 Concept of evaluation for resistance/tolerance

2.4.2 Assessment for plant pests (bacteria, fungi and virus) resistance


2.4.3 Assessment for environmental tolerance

fungi and virus) resistance

62

under:

o Field condition

o Hot spot condition

A tifi i l/l b t

Resistance levels of disease assessment

o Disease incidence

o Disease severity

2.4.1 Concept of evaluation for resistance or tolerance

Selection/assessment for resistance

2.4 Assessment of resistance and tolerance


o Artificial/laboratory condition

o Disease severity

63

Gene controlling for agronomic traitsand resistance to biotic and abiotic stresses are expected to be located at different regions (Blum, 1980)…

2.4.1 Concept of evaluation for resistance or tolerance



Therefore, assessment the responses of different varieties should be done under separate and suitable conditions, and particular generations.

64

Screening diagram Assessment for resistant to diseases-insect pests and environmental stresses (Adapted from Blum, 1980)



When?: Generations, seasons

Where?: Environmental locations or conditions

How? Inoculations and evaluations

65

2.4.2 Assessment for plant pests resistance

Assessment for bacteria responses

Assessment for fungi responses

Assessment for virus responses



66

1

Breeding tomato varieties for bacterial wilt resistance

Assessment for bacteria responses



67

Bacterial wilt

Ralstonia solanacearum

Serious disease and major constraint in



j

the tomato production

Reduce field stand 30-100 %

68

Tomato Bacterial wilt

Properties

-Soil borne disease

-wide host range

-No effect to pesticide



Characteristics of bacterial wilt resistance

-Location specific

-Strain/temperature dependent

-Not immune: internal bacterial population correlated to

symptom

Wang et al, 2000a; Wang et al, 2008

69

The International set of resistance

source in tomato to bacterial wilt

worldwide evaluated 33 var.

with 1 susceptible check

3 sources of resistance;

Hawaii, Philippine and North Carolina? (Scott et al., 2005)



Hawaiian resistance source:

•a broad-based and the most stable and durable

resistance

•HW7996 the best source of partial resistance to race 3

Wang et al., 1998; Carmeille et al., 2006

70



71

Symptom of wilting caused by

Ralstonia solanacearum


123752: Suchila, KKU, Thailand72

1

Some screening techniques at KKU



73

Materials and methods



74

Table 1. β-carotene content,

fruit yield and some

characteristics of 34 tomato

cultivars grown under field

and plastic-net house

conditions during October

Thirty tomato cultivars from many resources, two resistant and two susceptible

were screened for bacterial wilt resistance.


2008-Febuary 2009 at the

experiment field, Khon Kaen

University, Thailand.

75

Table 2. Responses of fruit yield and bacterial wilt responses in 32 tomato lines/cultivars under the concrete block

with bacterial wilt inoculums during dry season (October 2008-Febuary 2009) and responses to bacterial wilt

disease of 12 tomato cultivars under 2 seasons, dry (October 2008-Febuary 2009) and rainy season (May-

September, 2009) at the experimental field, Khon Kaen University,Thailand.

Bacterial wilt responses and fruit yield were evaluated.


76

Preparation of bacterium inoculums

R. Solanacearum : 3 Isolates ()



Inoculum concentration = 0.2

(107-10

8cfu/g. soil mixed population)

77

Seedling preparation

Seedling: 25-30 days

after sowing



RCBD 3 reps./variety

78

1

Inoculation Methods

1. Root inoculation

2. Micropipette (micropipette technique)



3. Scalpel leaf clip method or leaf clipping

4. Soil drenching method

79

Scalpel leaf clip method or leaf clipping



Micropipette Soil drenching method80

Pathogen morphology and Type of Symptoms

OOZES COLONY


STEM CROSS SECTION

81

Evaluation

Rating score

• 1 : No symptoms

• 2 : One leaf wilted



• 3 : Two to three leaves wilted

• 4 : Four or more leaves wilted

• 5 : Whole wilted (dead plant)82

Wilting intensity computing

% Wilt intensity : %I

I = [ ∑ (Ni x Vi) / (N x V )]x 100



∑Where Ni mean : number of plant wilt respective disease rating

Vi : disease rating (1,2,3,4 or 5)

V : the highest disease rating

N : the number of plant observed

Winstead and Kelman (1952)

Chanh (1989)

83

RM

R

M

SS

20 40 60 80

H

S

100

Highly

resistant


R S S

Bacterial wilt phenotype

84

1

Varietie

s

% dead plant % IntensityWilt intensity observation

(Phenotype)

Week 1 Week 2 Week 3 Week 4 Week 1 Week 2 Week 3 Week 4 Week 1 Week 2 Week 3 Week 4

V1

V2

V3

V4

V5

Data recorded



V5

V6

V7

V8

V9

V10

85

Genotypic analysis

Materials and methods

Dellaportaextraction method

Dellaportaextraction method


200 plants of F2 200 plants of F2

6% poly-acrylamide gel6% poly-acrylamide gelVisualized with the gel

document photographerVisualized with the gel

document photographer

M P1 P2 Homozygouse S

Homozygouse R

Heterozygous

Homozygouse SHomozygouse RHeterozygous

F2

86

DNA Marker linked to Bacterial wilt resistance on chromosome 6 and 12



500 bp

87

M P1 P2

Homozygouse SHeterozygous

F2

Statistical analysis

Segregation ratio by χ2 test =(O‐E)2/E



Homozygouse R

Homozygouse S

Homozygouse R Heterozygous

88

Data recorded

1 1 R1 Hawaii7996 RR RR2 2 L1 PI114968 84 AI SS SS3 3 F1 L X R1 RR RR4 4 F2 L X R1 SS5 5 F3 L X R1 RS RS6 6 F4 L X R1 SS RR7 7 F5 L X R1 RR RS8 8 F6 L X R1 SS SS9 9 F7 L X R1 RS RS

10 10 F8 L X R1 RS SS

No.DNA

No.CCA code

PedigRee name SLM6-17 SLM12-2

Data were analyzed by chi-square

test to as certain the goodness-of-

fit between the expected ratio

following by Mendelian ratio



10 10 F8 L X R1 RS SS11 11 F9 L X R1 RS RR12 12 F10 L X R1 RS RS

Genotype Expected frequency No. of plant Number of plant

χ² P Resistant Susceptible

Hawaii 7996 F2 3:1 200 F2 15:1 200 F2 13:3 200

following by Mendelian ratio

89

Breeding hot pepper for anthracnose resistance

Assessment for fungi responses



90

1

Host : Susceptible cultivars ( Variety and Fruit Stage )

Factors affecting anthracnose disease



Pathogen

:

91

Year Author

species of

capsicum spp.

Resistant source Country reported

Cc Cg Ca

1999 AVRDC C. baccatum PBC81, PBC80 Taiwan

C. chinense PBC932

2003 Yoon C. baccatum PBC81, PBC80 Korae

2004 Yoon etal., 2004 C. baccatum PBC81, PBC80 Korae

2004 Voorrips et al. C. chinense PBC932 Korae

2007 Kim et al. C. baccatum PBC81, PBC80 Korae

C. annuum Daepong-cho

Source of anthracnose resistance


2005 Pakdeevaraporn et al. C. chinense PBC932Thai land

C. baccatum PBC81, PBC80

2005 Park C. baccatum PI594137 Korea

2006 Lin et al. C. chinense PBC932

2008 Prasath C. baccatum PBC81

2005 Pae et al C. baccatum PBC1430

PBC1439

PBC1478

PBC880

C. annuumTCO6903 (R95)

92

Some resistant sources in hot pepper varieties

Progressive lines derived from PBC 932 and PBC 80


Original resistance sources

93

FactorsFruit development



94

C gloesporioides

Pathogen



C. capsiciC. capsiciC. acutatum

C. gloesporioides

Than., 2008

Different species of Colletotrichum grown on PDA

Cc930 Cc1141 Cc388 Ca524 Ca153

95

Pathogen

Cc‐cap26 Ca_MUJ5

- 500 spore/ml

-2 ul, RH 98%, Dark

Microinjection Method

Inoculums preparation for inoculation and evaluation


- Evaluation 5 DAI

Following Park et al (1999) technique96

1

Green mature and ripe mature fruit stage for inoculation


Mature fruit: about 35 and 45 day after anthesis

97

Evaluation of disease reactions



98

score Resistant level Symptom details

0 HR, highly resistant No infection

1 R, resistant 1-2% of fruit area shows necrotic lesion or a larger water-soaked lesion surrounding the infection site

3 MR d l i 2% f h f i h i l i li b

1) Percentage of necrotic lesion on fruit: six rating scores

Two techniques of disease evaluations



3 MR, moderately resistant >2% of the fruit area shows necrotic lesion, acervuli may be present, or water-soaked lesion up to 5% of the fruit surface

5 MS, moderately susceptible

>5-15% of the fruit area shows necrotic lesion , acervuli present , or water-soaked lesion up to 25% of the fruit surface

7 S, susceptible >15-25% of the fruit area shows necrotic lesion with acervuli

9 HS, highly susceptible >25% of the fruit area shows necrosis, lesion often encircling the fruit, abundant acervuli

Montri et al., 2009 99

2) Dimension of necrotic lesion on fruit (1 mm ‐36 mm): two rating scores R S



1) Resistance: necrotic lesion < 4 mm.

2) Susceptible: necrotic lesion > 4 mm.

AVRDC

100

No. Rep box variety lesion green red

1 1 1 1

2 1 1 2

3 1 1 3

4 1 1 4

5 1 1 5

6 1 1 6

7 2 1 1

8 2 1 2

Data collections

Data sheet

2.4 Assessment of resistance and olerance


8 2 1 2

9 2 1 3

10 2 1 4

11 2 1 5

12 2 1 6

13 3 1 1

14 3 1 2

15 3 1 3

16 3 1 4

17 3 1 5

18 3 1 6

Statistic analysis by RCBD, Test significant at p=value 0.05

and 0.01

101

Data analysis

Disease

reaction1/

Disease

reaction1/

101

102

103

104

105

variety

C. acutatum_MUJ5

Ca - green mature Ca - ripe mature

disease incidence disease incidence



106

107

108

109

110

111

susceptible checkResistant check Conclusion for Disease reactions

102

1

Molecular marker aids selection

Distribution of marker

for anthracnose resistance



103

DNA extraction and gel electrophoresis

2.4 Assessment of Resistance and Tolerance


1) Harvest the younger leaves

2) Extraction DNA

3) PCR and running gel (Acrylamide gel)

4) Scoring DNA banding104

genotype Primer name Sequencings Linked

Forward primer Reverse primerC.capsici HpmsE143 CCATTCAGCTAGGGTTCAGTCCA CGACCAAATCGAATCTTCGTGA 9

Anth9-CAPS Unpublished Unpublished 9

C.acutatum HpmsE032 ATGCGCAAAGGGAGAAAATTCA CGAACTAACCGTTCATGGTGGA 12

Primers

2.4 Assessment of Resistance and Tolerance


Anth12-CAPS Unpublished Unpublished 12

SCAR-Indel 100 bp GGTATCTTATTTCATAGGGACCAGGCA TTTGCGGTAGTGACAACAACTTTACAGCCA -

105

Scoring of DNA banding pattern



Lane 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

DNA score 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 3 3 3 3 3

Lane 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43

DNA score 3 1 3 1 3 1 3 1 3 1 3 3 3 3 3 1 3 3

106

Evaluation of Yellow Tomato Leaf Cure Virus (TYLCV)

Assessment for virus responses



Source: AVRDC, 2012

107



108

1



109



Source: AVRDC, 2012 110

Mapped TY Resistance Genes in Tomato



111

Tomato BGVsThe World Vegetable CenterThe World Vegetable Center

AVRDCAVRDCHost range of TYLCV2.4 Assessment of resistance and tolerance


112

Pepper BGVs

Host range of TYLCVThe World Vegetable CenterThe World Vegetable Center

AVRDCAVRDC



113

Pepper BGVs

Host range of TYLCV

The World Vegetable CenterThe World Vegetable Center

AVRDCAVRDC



GV

114

1

Cucurbit BGVs

SquashSquash

Host range of TYLCV


123752: Suchila, KKU, ThailandBottle gourdSquash plantsChayote The World Vegetable CenterThe World Vegetable Center

AVRDCAVRDC 115

Cucurbit BGVs

Host range of TYLCV

The World Vegetable CenterThe World Vegetable Center

AVRDCAVRDC



Ridge gourdCucumber 116

Legume BGVs


AVRDCAVRDC



Mungbean Yard long bean

Soybean

117

Okra BGVs


AVRDCAVRDC



118

Eggplant BGVs


AVRDCAVRDC



TYLCV disease vector and transmission



120

1

Genetic resource from AVRDC

35 accessions of pepper varieties



121

Materials and Methods

V1 V2 V3 V4 V5 V6 V7 V8

V9 V10 V11 V12 V13 V14 V15 V16

V17 V18 V19 V20 V21 V22 V23 V24

LP

LP

LP

LP

Experimental design with 56 tested varieties and local variety



V25 V26 V27 V28 V29 V30 V31 V32

V33 V34 V35 V36 V37 V38 V39 V40

V41 V42 V43 V44 V45 V46 V47 V48

V49 V50 V51 V52 V53 V54 V55 V56

LP

LP LP

LP

LP

LP

LP: Local check

V1-V56: Tested varieties

122

Material and methods: Field evaluation for TYLCV



123

Evaluations of TYLCV: plant symptoms



124

Evaluations of TYLCV: leave symptoms



125



126

1

Percentage of disease reactions

% Disease

symptoms= Σ

(disease score x plant no.)

Total plant no. x maximum disease score

x 100



127

LP 1 2 3 4 5 6 7 8 9 10 11 12 LP LP number 1 2 3 4 5 6 7 8 9 10 11 12 LP

1 105 121

2 127 117

1 126 122

2 133 129

1 114 116

2 128 103

1 124 136

1

2

3

4

Symptom rating (visual) Replication: I Date (day/month):

Block sub block row number

Plant No.

Test plants Test plants

Table for data recorded



2 130 131

1 134 132

2 111 119

1 110 135

2 118 112

1 306 120

2 113 109

1 107 104

2 101 115

1 108 102

2 123 125

1

4

5

6

7

8

9

128

The Responses of Hot pepper to Drought Stress

2.4.3 Assessment for environmental tolerance



129

Chilii pepper production in Thailand



low yield and low qualitylow yield and low quality

80% under rain-fed area

130

400

500

600

700

800

900

Nakorn Rachasima

Chaiyaphum

Loei

Norteast of Thailand

ainfal

l (mm.

)

Fluctuation of rainfall (mm.) in chiili pepper commercial

production area of Northeast of Thailand


0

100

200

300

Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Ra

(a) (b) (c)

Note: (a) vegetative stage, (b) flowering stage and (C) fruiting stage 131

How to improve high pungency, high yield and

produced in short-term drought stress condition?



Breeding Management

132

1

Relation of water in soil, plant and atmosphereAtmospheric water

Transpiration : stomata opening, RH

or PD



Soil water

Water in Plant Absorption : root penitration, moisture

tension of soil

TRANSLOCATION

133

Soil moisture measurement• Gravitational method

soil moisture percentage = (wet soil weight - dry soil weight) x 100/ dry soil weight

• Tensiometric method


http://www.calafrica-sa.co.za/


m m

http://www.calafrica-sa.co.za/http://www.calafrica-sa.co.za/soil_moisture_measurement.htm 134

• Gypsum block

Soil moisture measurement measurement2.4 Assessment of resistance and tolerance


• Neutron probe Hydro probe

http://aces.nmsu.edu/pubs/_h/H640/welcome.html

http://www.agw.kit.edu/english/field_instruments.phphttp://www.pavetesting.com/hydroprobe.php

http://cals.arizona.edu/pubs/water/az1220/

135

Plant water status measurement

Relative water contentRWC = (Fresh weight - Dry weight)/

(Fully turgid weight - DW) X 100

Leaf water Potential



Leaf water PotentialPressure chamber (Kg/cm2)

1 Ib/In2 = 6.895 x 103 Pa

1 Kg = 2.2046 Ib

1 Ib = ½.2046 Kg

1 In = 2.54 cm

½.2046

Kg/(2.542)

= 6.895 x 103 Pa

1 Kg/cm2 = 0.09807 MPa136

Water Potential in Plants

Determine water‐deficit stress in plants.

Determine drought tolerance in plants,

The irrigation needs of different crops

How the water status of a plant affects the quality and yield of plants



How the water status of a plant affects the quality and yield of plants.

Pure Water (no solutes) at normal pressure and elevation where, Ψw = 0

Ψw is increased by an increase in pressure potential (ΨP)

Ψw is decreased by addition of solutes which lowers the solute potential

(ΨS )

137

•Stress types: water (drought and flood), temperature (high temperature

and energy balance, chilling and freezing), irradiance, others

•Level of the stress:

Severe, moderate and mild

•Period of the stress:

Treatment application techniques



Prolong (gradual), short term or transient (sudden)

•Application technigues:

On farm vs Lab. Research:

Drought: lab (water application vs PEG)

Temperature: shading vs Phytotron room or green house

Salinity: saline soil vs Lab

138

1

Experimental design- conducted under greenhouse condition- RCB design with 4 replications- varieties were divided into two groups;

1st gr. were treated with drought stress2nd gr. were control treatment (optimum water applied)



Drought treatment

1-10 DAF100%

11-20 DAF75%

21-30 DAF50%



75% 50%

31-40 DAF25%

-no watered-After severe wilting overnight (re-watered)140

Measurement of water status by Pressure Chamber



• Quick method, commonly used

• Equipment can be used in the field; can

be heavy and bulky

• Dangerous pressure levels are applied in

the chamber Source: Bowers and Sterling, Department of Entomology, Plant Pathology, and Weed Science, NMSU, 2004

141

The positive pressure reading from the plant tissue tested in the

previous slide was 7 bars, a stressed plant.

To estimate the water potential, we must first convert the positive

pressure from bars into MegaPascals (MPa).

Water potential measurement



10 Bars = 1 MPa

7 Bars = 0.7 MPaYP(air) + YP(xylem) = 0

pressure from bars into MegaPascals (MPa).

Ten bars is equal to one MegaPascal, so 7 bars equals 0.7

MegaPascals.

142

60.00

70.00

80.00

90.00

100.00

LWP

(M

Pa

)R

WC

(%

)

KKU-P11039 KKU-P13049 KKU-P11015

Control

Stress

Control

Stress

**

****

** *****

****

-1.00

-0.50

-

LWP

(M

Pa

)

Control

Stress

** ****

**- RWC and LWP of the treated plant were decreased at 21 30 d 31 40 DAF ti l ( d t th

Water status

-1.50

-1.00

-0.50

-

**


-1.50

60.00

70.00

80.00

90.00

100.00

RW

C (

%)

Control

Stress

** ****

**

** **

** *

-1.50

-1.00

-0.50

-

60.00

70.00

80.00

90.00

100.00

LWP

(MP

a)

Control

Stress

Control

Stress

** **

*****

KKU-P12010 TakanotsumeKKU-P11031 KKU-P28006

KKU-P31118

KKU-P33030

21-30 and 31-40 DAF, respectively (compared to the control plant)

143

Plant response to drought stress

Physiological response• stomaltal closure reduce photosynthesis• Leaf development: small, mature and fall.• Produce hormone: ABA, Ethylene ,

Morphological response



Morphological response• Increase leaf thickness and low leaf no. • Low shoot: root ratio• Chlorophyll damage• Leaf roller and wilt

144

1

Cultivars

Yield Growth

Fresh Weight (g) Dry Weight (g) Fruit Length (cm.) Fruit width (cm.) Plant Height (cm.) Stem diameter (cm.)

C S C S C S C S C S C SLow

KKU-P11039 639.96 368.24** 127.99 66.28** 3.74 3.73ns 5.62 5.49* 121.25 120.08ns 1.60 1.34**

KKU-P13049 241.04 178.27** 69.90 51.70** 0.38 0.37ns 3.22 3.18ns 151.85 136.78* 1.99 1.65**

KKU-P11015 503.12 268.00** 120.75 58.96** 1.54 1.52ns 7.30 7.09** 167.83 161.25ns 1.74 1.53**

Medium

Growth and yield responses

- Plant height and Stem diameter were decreased (5-20% and 10-30% of control) in the treated plant dependent on



KKU-P12010 385.94 317.05ns 119.64 98.28ns 0.51 0.50ns 3.62 3.56ns 128.88 138.55* 1.44 1.24**

KKU-P11031 400.17 325.71** 128.05 104.23** 0.58 0.57ns 4.32 4.18* 146.50 133.25* 1.66 1.66**

KKU-P28006 644.41 363.73** 161.10 87.29** 1.72 1.65ns 7.77 7.53** 155.45 139.50** 1.72 1.48**

Takanotsume 165.56 118.15** 41.39 28.36** 2.21 2.20ns 8.38 6.20** 125.31 103.78** 1.22 1.13ns

High

KKU-P31118 318.05 253.71ns 89.05 71.04ns 0.55 0.53ns 2.84 2.80* 144.84 140.25ns 1.44 1.29**

KKU-P33030 356.87 297.94** 96.35 80.44** 0.68 0.67ns 2.77 2.44** 120.55 111.9ns 1.68 1.13**

in the treated plant dependent on varieties- Fruit fresh weight and dry weight were decreased (20-50% of control) in the treated plant dependent on varieties

145

Water Use Efficiency ; WUE

Photosynthetic rate/transpiration of leaf



Total dry mass/total transpiration

Total yield/(water use x HI)

146

EvapWUE

RWCWSDDWWSat

FWWSatWSD

DWWSat

DWFWRWC

mpw

2

2kgorginlossOH

gormginuptakeCONet

5YieldCroporMatterDry

.inUsedWater

4100

3100.

.

2100.

1

EQUATIONS AND ABBREVIATIONS

Abbreviation

w = Water Potential RWC = Relative Water Content ra = Atmospheric Resistance = Osmotic Potential WSD = Water Saturated Deficit rs = Stomatal Resistance p = Turgor Potential WUE = Water Used Efficiency rm = Mesophyll Resistance m = Matrix Potential D = Diffusivity PS = Photosynthesis Trans. = Transpiration Evap. = Evapotranspiration



rmrsra

rsra

OH

COWUE

DOH

Drmrsra

OHOHTrans

DCO

Drmrsra

COCOPS

OH

extint

OH

extint

2

2

2

22

2

22

64.0

76

7resistance

)][]([.

6resistance

64.0

)][]([

2

2

147

Example of drought stress experiment


148

Fig. 1. (A–C) Monthly mean air temperature, relative humidity and light intensity under a plastic net house

and (D) monthly mean rainfall under field condition at the experimental farm at Khon Kaen University

during the rainy season (May to Oct. 2009).

E i t l diti d i t i d


Environmental conditions during stress periods

149

Fig. 2. The values of leaf water potential response at 10-d intervals of the control (C ) and drought-

stressed plants (T ) in nine hot pepper cultivars.**Significant difference between control and drought

treatments within cultivar at P # 0.01 level by t test.


Different species showed different water status under drought stress

150

1

Table 2. Leaf water potential (LWP), relative water content (RWC), leaf area (LA), and specific leaf area

(SLA) between the control plant (C) and drought-stressed plant (T) of nine hot pepper cultivars at 31 to

40 d after flowering.

Different species showed different


Table 3. Plant height, stem diameter,

and shoot-to-root ratio between the

control plants (C) and drought

stressed plants (T) of nine hot pepper

cultivars at second harvest.

pgrowth development under drought stress, compared to their control treatment

151

Table 4. Yield and fruit quality between the control plants (C) and drought-stressed plants (T) of nine

hot pepper cultivars.

Table 5 Capsaicin (Cap) dihydrocapsaicin (Di) capsaicinoid (CAPS) and capsaicinoid yield (CAPS yield) Different species showed different


Table 5. Capsaicin (Cap), dihydrocapsaicin (Di), capsaicinoid (CAPS), and capsaicinoid yield (CAPS yield)

between the control plants (C) and drought-stressed plants (T) of nine hot pepper cultivars.

pYield and pungency under drought stress, compared to their control treatment

152

Cultivar PAL (μmole mg-1

protein) C4H (AU min-1

g-1

) CS (μmole mg-1

protein) POD (AU min-1

g-1

)

C1/

D1/,2/

% increase3/

C1/

D1/,2/

% increase3/

C1/

D1/,2/

% increase3/

C1/

D1/,2/

% increase3/

Yuyi 33.99d 74.02e** 217.8 1.092b 1.167c* 106.9 67.6f 119.6g* 176.9 16.50a 28.13ab* 170.5

Huay-Siiton 92.59bc 135.59b* 146.4 1.181a 1.298a* 110.0 104.9ef 316.7de** 301.9 14.25ab 22.26bc* 156.2

K

Table 2. Phenylalanine ammonialyase (PAL), cinnamic-4-hydroxylase (C4H), capsaicin synthase (CS) and peroxidase (POD)

activities under a control (C) and a drought stress (D) treatments

Bio-molecular Traits for Drought Stress

Different species showed different



Keenoo-

Pama

67.68c 109.16bc* 161.3 1.084b 1.215b* 112.2 546.8c 628.5bc 114.9 1.26f 2.46g* 195.2

BGH 1719 148.91b 165.23ab 110.9 1.177a 1.214b* 103.2 752.4ab 578.3c 79.9 5.06d 31.31a** 618.8

Perennial 173.54a 188.32a 108.5 1.093b 1.191b* 109.0 936.4a 982.7a 104.9 3.13de 20.63c** 659.1

Mean 97.34 134.46 154.6 1.125 1.217 108.2 481.6 525.2 155.1 8.04 20.96 360.0

enzymes activities under drought stress, compared to their control treatment

153

Bio-molecular Traits Under Drought Stress

Figure 1. Sum of Cap and Di (CAPs) which

were redrawn from Phimchan et al9

and

activities of phenylalanine ammonia-lyase

(PAL), cinnamic-4-hydroxylase (C4H),

capsaicin synthase (CS) and peroxidase

(POD) in hot pepper fruit between non-

drought and drought stressed plants.

L 1 M 1 M 2 H 1 H 20

20000

40000

60000

80000

100000

120000

140000

160000

180000

Non-drought stressed plantDrought stressed plant

Capsaicinoids (CAPs)

CA

Ps

(SH

U)

Cultivars

150

200

250

Non-drought stressed plantDrought stressed plant

Phenylalanine ammonia-lyase (PAL)

e m

g-1

prot

ein)

Cinnamic-4-hydroxylase (C4H)

U m

in-1

g-1 )

1.0

1.5

2.0



L 1 M 1 M 2 H 1 H 20

50

100

PA

L (u

mol

e

Cultivars Cultivars

L 1 M 1 M 2 H 1 H 2

C4H

(A

U

0.0

.5

Capsaicin synthase (CS)

L 1 M 1 M 2 H 1 H 20

200

400

600

800

1000

1200

CS

(um

ole

mg-

1 pr

otei

n)

CultivarsL 1 M 1 M 2 H 1 H 2

0

10

20

30

40

Peroxidase (POD)

PO

D (

AU

min

-1g-

1 )

Cultivars

Only phenylalanine ammonia-lyase (PAL) showed positive association

to capsaicinoids contents under drought stress

Source: Pimchan et al, 2014 154

Experiments: 3 chilli pepper cultivars, 3 replications

and 3 plants per replication 3 levels of drought stress treatment

Exercise for drought stress



Data recorded: LWP, RWC, plant height, plantcanopy, Stem diameter and SPAD chlorophyllreading

155

Cultivar Rep.LWP RWC

T1 T2 T3 T1 T2 T3

Bhut jolokia

1

2

3

Table 1 Leaf water potential (LWP) and relative water content (RWC) of 3 hotpepper cultivars treated by 3 levels of drought stress



Akanee pirot

1

2

3

Habanero

1

2

3

156

1

Cultivar Plant no.Plant height (cm.) Plant canopy (cm.)

T1 T2 T3 T1 T2 T3

Bhut jolokia

1

2

3

Table 2 Plant height and plant canopy of 3 hot pepper cultivars treated by3 levels of drought stress



Akanee pirot

1

2

3

Habanero

1

2

3

157

Cultivar Plant no.Stem diameter (cm.)

SPAD chlorophyll reading

(SCMR)

T1 T2 T3 T1 T2 T3

Bhut jolokia

1

2

3

Table 3 Stem diameter and SPAD chlorophyll reading of 3 hot pepper cultivarstreated by 3 levels of drought stress



3

Akanee pirot

1

2

3

Habanero

1

2

3

158

Chapter V: Plant and environmental stress

Chapter V: Plant and Abiotic Stress


159

Chapter V: Plant and environment stress

5.1 The stress environment (Dr. Suchila)

5 3 Pl t b di d i ld t bilit

5.2 Responses of plant to environmental stress (Dr. Suchila)



5.3 Plant breeding and yield stability (Dr. Suchila)

5.5 Selection for yield and environment (Dr. Sanun)

5.6 Physiology and genetic implication (Dr. Sanun)

5.4 Models and applications (Dr. Sanun)

160

5.1) The Stress Environment

• The 1972 crisis (World imbalance of population and food supply) demonstrated how closely the world borders on severe food shortages The food production limitations

5.1 The stress environment


shortages. The food production limitations other than man‐made stresses, such as air pollution, and pest losses are: (a) availability of productive soil, and (b) a climate supportive to crop growth (Christiansen, 1982)

161

Land availability• ~10% of the world’s arable may be categorized as free of stress

• ~20% of the land is under some kind of mineral stress



• ~26% is affected by drought stress

• ~15% is affected by freezing stress

Source: Dudal (1976, quoted by Blum, 1986)

162

1

Climatic restriction

It has been estimated that 60‐80% of variability in crop production is a result of whether fluctuation (temperature and water availability) Well known and senior breeder in



availability). Well known and senior breeder in USA concluded that modern technology has little influence on the susceptibility of non‐irrigated crops to whether (Christiansen, 1982).

163

Types of abiotic environmental stress

Temperature: heat, cold

Water: drought, water logging



p

Salinity & Mineral

164

Stress environments:

Water stress

• Very little of the world’s water is used by plants. Nace (1969, quoted by Christiansen, 1982) estimated that only 2.5% of the world supply is non‐oceanic or fresh, and that only 0.007% of all water is used by



y yplants.

• In rain‐fed agriculture the problem of short‐term (10‐20 day) drought is common and exacts an intangible and real toll on productivity. This incidence at critical

periods of cropping cycles can be disastrous.165

Stress Environment: Heat stress

• Most of the world’s agriculture is restricted by temperature to the confines of 50° South to 50° North latitudes (low temperature). High

temperature is a limiting factor in much of lowland tropics.

• The critical thermal regimes affect plant phenology, developmental phases, growth rates, yield components and final yield. Growth and development processes follow distinct temperature‐response



development processes follow distinct temperature response curves, displaying a peak or a plateau at what are defined as an optimum temperature. Any temperature outside the optimum is non‐optimum, and may be defined as stress temperature. Various plant processes (different crops and growth stages) may have different temperature optima, or different threshold stress temperature (related to heat‐sums or degree‐days).

166

According to threshold temperatures: two annual crops groups:

Cool season vegetable

cabbage

cauliflower

Warm season vegetable

o Eggplant

o Corn



sweet pea

melon

sweet pepper

Allium

o pumpkin

o Water melon

o Cucumber

o Yard long bean

167

Stress environment: salinity stress

• Soil properties that determine its mineral effects on plants are mineral content, organic matter content, CEC, permeability etc.

• The ultimate definition of a given soil‐nutritional problem, at the breeder’s level is best reached by growing and analyzing



the given crop in the target environment.

• The complexity of the formation of saline soils is affected by rainfall, mineral weathering, fossil salts etc. It is important for the breeder to recognize two major and different saline environments (saline soil, and non‐saline soil irrigated with saline or brackish water).

168

1

• Avoidance: The environmental factor is excluded from the plant tissue (J. Lewitt, )

5.2) Responses of Plant to Abiotic Stress

Physiological standpoint:



• Tolerance: The environmental factor penetrates the tissue but tissue survives

• Resistance imply to Tolerance

169

Criterion for the development and use of

screening tests:

Generic variation should occur in germplasm

Screening for environmental stress

5.2 Responses of plant to abiotic stress


heritability (h2) for the trait should be greater than h2 for yield

Trait should be correlated with yield

Screening test should be easy, rapid and economical to apply

170

Importance of resistant or tolerant variety

• The genetic improvement of stress resistance is an economically viable solution. On the other hand, stress‐resistant crop varietiesshould not necessary associated with

5.2 Responses of plant to abiotic stress


ysituations of poor or marginal crop performance, or their importance should not be limited to conditions of low‐input and high‐risk farming (value added crop, nitch market;Blum, 1986).

171

5.3) Plant Breeding and Yield Stability



172

Drought Stress for Breeder standpoints

“Drought stress”:short term (duration) drought stress

5.3 Plant breeding and yield stability

5.3.1 Plant breeding for Environmental stress: “Drought stress”


Drought stress is implied a total yield loss.

short-term (duration) drought stress

Drought stress occurs at any subpotential yield level. The improvement of yield at any of these levels is the main purpose of breeder.

173

“Drought is responsible for severe food shortages and famine in developing countries.”

Breeding for drought stress resistance

Drought stress is induced by inability of the



Drought stress is induced by inability of the crop to meet its evapotranspiration demand. Therefore the reduction of the ETactual/ETmaximum

is interested. Potential evapotranspiration = PEActual evapotranspiration = ET

174

1

The goal of breeder: to obtain economic yield

Breeding for drought stress resistance

Yield = T x WUE x HI



Where T= total seasonal crop transpiration

WUE = water use efficiency

HI = Crop harvest index

(Source: J.B. Passioura and C.T. de Witt)

175

Plant traits affecting drought response

Phenology: early or late

Plant development and size

Non‐senescence

Stem reserve utilization

Chapter V: Plant and Abiotic Stress


Plant surface: wax

Root: length & dept

Photosynthetic systems

Water use efficiency

176

Mechanisms of drought resistance

• Escape: early maturity characteristic under optimum condition at the beginning season

• Avoidance: Decreasing water loss by extract soil moisture efficiency

Chapter 5: Plant and Abiotic Stress5.3 Plant breeding and yield stability


• Tolerance: Some species more toleranat of post anthesis drought, and can produce appreciable yield under stress

• Recovery: The environmental factor is excluded some species are able to recover after short duration drought stress

177

5.3.1 Plant breeding for environmental stress: “Heat (high temperature) stress”

Different objective between the breeder inTropical vs temperate area

Screen under hot target environment (hot spot):



Using controlled green house of growth chamber or phytotron room

g ( p )worldwide research stations of international seed Co.

178

5.3.1 Plant breeding for environmental stress: “Salinity Stress”

Soil salinity: ~ 95% million hectares worldwide

Soil salinity: the accumulation of dissolved salts in the soil

Different responses among different crops



Tolerant Moderately tolerant

Moderately sensitive

sensitive

Cotton Sugar beet Peanut Strawberry

Barley Soybean Corn Potato

Sorghum Rice Tomato

Sweet potato Onion179

Management of resistant/tolerant varieties

The concept and role of genotype x environment interaction in plant breeding

5.3.2) Yield Stability



Management of resistant/tolerant

varieties

180

1

The concept and role of genotype x environment (G x E) interaction in plant breeding

Plant breeder are interested in developing high yielding cultivars with sustained or stable high performance over seasons and years; that is “Yield stability”



years; that is Yield stability .

G x E occurs when two or more genotypes are compared across different environments, and their relative performance are found to differ.

181

Stability is the ability of plant that can maintain yield under the environment variables.



Different responses of chilli cultivars under two different location environment

182

1. Estimation of plant growth or characters in the different environments for many years

2. Study of the Regression coefficient between the output of each crop varieties with yield of all cultivars grown at that l ti (Fi l &

Models and application



all cultivars grown at that location (Finlay & Wilkinson and Eberhart & Russel)

3. Measuring of the combining ability of varieties, that growing in different environments.

4. Determined by GGE biplot

183

GGE-Biplot

GGE‐biplot is an effective tool for:

Mega‐environment analysis (e.g. “which‐won‐where”

pattern), whereby specific genotypes can be recommended

to specific mega‐environments (Yan, 2003; Yan, 2006)



to specific mega environments (Yan, 2003; Yan, 2006)

Genotype evaluation (the mean performance and stability)

Environmental evaluation (the power to discriminate

among genotypes in target environments)

184

GGE-Biplot



In total phenotypic variation, E explains most of the variation and G and G x E are usually small (Yan, 2002)

185

Classification of G x E interaction

No G x E interaction: one genotype performs better than the others across all environments

A non cross over G x E interaction: one genotype outperforms the others across all environments, but that genotype may exceed the other genotype 20 unit in one



g yp y g ypenvironment (env.) But 60 unit in the other env.

A crossover G x E interaction: (Most important to plant breeder)

a genotype A is more productive in one env., while the other genotype more productive in the other env.

Combined G x E interaction: one genotype increase the other genotype decrease 186

1

Finlay & Wilkinson and Eberhart & Russel



187

Stability analysis



188

Descriptions of environments where trials were conducted

during 2009–2011

Different altitudes temperatures



Different altitudes, temperatures, RH%, rainfalls, solar radiations and soil properties

(6 environments)

189

Combined analysis of variance for yield and capsaicinoid traits of six

chili cultivars evaluated in six environments during 2009–2011



Interactions between cultivars and environments (C x E) was found in all traits studied.

190

Stability analyses for yield and capsaicinoids of six chili cultivars

grown at six environments during 2009-2011

Dallay Khorsaney performed combined G x E inteaction



KKU-P-21041 performed stable fruit yield across all environments

191

GGE-Biplot

GGE biplot for dry fruit yield (A) and capsaicinoids content (B) of six cultivars across six environments. Environment

identification, KK1: Khon Kaen rainy season 2009; CM1: Chiang Mai rainy season 2009; LB: Lobesa rainy season

2010; KB: Kabesa rainy season 2010; KK2: Khon Kaen dry season 2010-2011; CM2: Chiang Mai dry season

2010 -2011.



GxE interactions complicate the process of selecting of genotypes with superior performance.192

1

Gerplasm

Chapter VI: Management of resistance /tolerance varieties


Breeding Variety release

193

Remarks:

Breeder should note that resistance or tolerance

is not durable.

Chapter VI: Management of R & T var.


“The resistant/tolerant varieties need to be improved time to time”i.e. genes pyramiding or multi-lines improvement.

194

Breeder seed

Foundationseed

breeder and institute

breeder and institute

Seed production of resistant/tolerant varieties(similar protocol to the normal seed production)



Certified seed

Registered seed

grower or government or private sectors

seed registered officer

195

Variety registration & release

International procedure

(see CSSA guild lines)

Search and discuss about:



Thailand procedure(see www.doa.go.th/pvp/)

196

Registration procedureInternational procedure(see CSSA guild lines)

• Cultivar name• Scientific name• Designation during

Thailand procedure(see www.doa.go.th/pvp/

• Cultivar name• Scientific name• Name of organizations



g gdevelopment

• Name of organizations or institutions

• Brief description of the cultivars

• etc.

gor institutions

• Brief description of the cultivars

• Information for cultivar development

• etc.197

www.doa.go.th/pvp/


Registered for PVP in Thailand

Department of Agriculture, Ministry of Agriculture and cooperatives 198

1

Patents: USA vs Thai



http://www.google.com/patents/US20090019600

199

Acquaah, G. 2007. Principles of Plant Genetics and Breeding. Blackwell Publishing.

Blum, A. 1988. Plant breeding for stress environments. CRC Press, Inc. Florida.

www.sciencecampaign.org.uk/.../2009/beddingtonpresentat...

Key References



http://www.google.com/Adaptation and yield stability.htm

http://www.google.com/Patent WO2009098685A2 ‐ Disease resistant pepper plants

http://plantandsoil.unl.edu/croptechnology2005/pages/animationOut.cgi?anim_name=RootUptake.swf

http://www.google.com/patents/US8642845200

topics term paper assignments¹€อกสาร... · 1 reducing water use, increasing yields...

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