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 123752: Suchila, KKU, Thailand 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 123752: Suchila, KKU, Thailand 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, 123752: Suchila, KKU, Thailand 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 123752: Suchila, KKU, Thailand 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 lh 1.1 Terminology 123752: Suchila, KKU, Thailand 3) Disease 4) Resistance 5) Susceptible 6) Hypersensitivity 10) Differentialhost 11) Physiological race 12) Pathogenicity 6
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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
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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,
123752: Suchila, KKU, Thailand
Suggested topics: bacteria, fungi, virus, nematode, insect-pests and environmental resistance/tolerance of economic plants (or the major crops of the student’s research)
New lifestyles and new technologies to avert more global warming: what’s the hurry?
It’s getting hotter
1. 2 Importance
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
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
1.3 Principles and methods
123752: Suchila, KKU, Thailand
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.)
1.3 Principles and methods
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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.)
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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
2.2 Source of resistance
123752: Suchila, KKU, 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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
population
Pure/inbred lines
F1-hybrids
Pharmaceutical
products
(Animal) feed
additive
etc.
Selection
Selection
Svalbard Global Seed Vault
46
Collection
Evaluation
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
2.2 Source of resistance
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
regard to a specific trait, while low heritability implies a low level of resemblance.
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
123752: Suchila, KKU, Thailand
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
2.3 Heritability and breeding methods
123752: Suchila, KKU, Thailand
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
2.3 Heritability and breeding methods
123752: Suchila, KKU, Thailand
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
2.3 Heritability and breeding methods
123752: Suchila, KKU, Thailand
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.)
2.3 Heritability and breeding methods
123752: Suchila, KKU, Thailand
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.)
2.3 Heritability and breeding methods
123752: Suchila, KKU, Thailand
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
2.3 Heritability and breeding methods
123752: Suchila, KKU, Thailand
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.)
2.3 Heritability and breeding methods
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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)
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
66
1
Breeding tomato varieties for bacterial wilt resistance
Assessment for bacteria responses
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
67
Bacterial wilt
Ralstonia solanacearum
Serious disease and major constraint in
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
j
the tomato production
Reduce field stand 30-100 %
68
Tomato Bacterial wilt
Properties
-Soil borne disease
-wide host range
-No effect to pesticide
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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)
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
71
Symptom of wilting caused by
Ralstonia solanacearum
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand72
1
Some screening techniques at KKU
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
73
Materials and methods
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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.
123752: Suchila, KKU, Thailand
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.
123752: Suchila, KKU, Thailand
76
Preparation of bacterium inoculums
R. Solanacearum : 3 Isolates ()
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
Inoculum concentration = 0.2
(107-10
8cfu/g. soil mixed population)
77
Seedling preparation
Seedling: 25-30 days
after sowing
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
RCBD 3 reps./variety
78
1
Inoculation Methods
1. Root inoculation
2. Micropipette (micropipette technique)
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
3. Scalpel leaf clip method or leaf clipping
4. Soil drenching method
79
Scalpel leaf clip method or leaf clipping
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
Micropipette Soil drenching method80
Pathogen morphology and Type of Symptoms
OOZES COLONY
123752: Suchila, KKU, Thailand
STEM CROSS SECTION
81
Evaluation
Rating score
• 1 : No symptoms
• 2 : One leaf wilted
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
• 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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
∑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
123752: Suchila, KKU, Thailand
R S S
Bacterial wilt phenotype
84
1
Varietie
s
% dead plant % IntensityWilt intensity observation
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
500 bp
87
M P1 P2
Homozygouse SHeterozygous
F2
Statistical analysis
Segregation ratio by χ2 test =(O‐E)2/E
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
90
1
Host : Susceptible cultivars ( Variety and Fruit Stage )
Factors affecting anthracnose disease
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
Original resistance sources
93
FactorsFruit development
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
94
C gloesporioides
Pathogen
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
- Evaluation 5 DAI
Following Park et al (1999) technique96
1
Green mature and ripe mature fruit stage for inoculation
123752: Suchila, KKU, Thailand
Mature fruit: about 35 and 45 day after anthesis
97
Evaluation of disease reactions
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
103
DNA extraction and gel electrophoresis
2.4 Assessment of Resistance and Tolerance
123752: Suchila, KKU, Thailand
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
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
2.4 Assessment of resistance and tolerance
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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
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
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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.
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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
123752: Suchila, KKU, Thailand
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 2 Plant height and plant canopy of 3 hot pepper cultivars treated by3 levels of drought stress
2.4 Assessment of resistance and tolerance
123752: Suchila, KKU, Thailand
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
2.4 Assessment of resistance and tolerance
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3
Akanee pirot
1
2
3
Habanero
1
2
3
158
Chapter V: Plant and environmental stress
Chapter V: Plant and Abiotic Stress
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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)
Chapter V: Plant and environment stress
123752: Suchila, KKU, Thailand
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
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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
5.1 The stress environment
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• ~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
5.1 The stress environment
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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
5.1 The stress environment
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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
5.1 The stress environment
123752: Suchila, KKU, Thailand
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
5.1 The stress environment
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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
5.1 The stress environment
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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
5.1 The stress environment
123752: Suchila, KKU, Thailand
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:
Chapter V: Plant and environment stress
123752: Suchila, KKU, Thailand
• 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
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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
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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
Chapter V: Plant and environment stress
123752: Suchila, KKU, Thailand
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”
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
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
123752: Suchila, KKU, Thailand
• 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):
5.3 Plant breeding and yield stability
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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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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”
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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.
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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)
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
187
Stability analysis
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
188
Descriptions of environments where trials were conducted
during 2009–2011
Different altitudes temperatures
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
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.
5.3 Plant breeding and yield stability
123752: Suchila, KKU, Thailand
GxE interactions complicate the process of selecting of genotypes with superior performance.192
1
Gerplasm
Chapter VI: Management of resistance /tolerance varieties
123752: Suchila, KKU, Thailand
Breeding Variety release
193
Remarks:
Breeder should note that resistance or tolerance
is not durable.
Chapter VI: Management of R & T var.
123752: Suchila, KKU, Thailand
“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)