Breeding approaches for pest resistance in rice crop

1. “Breeding for Major Insect Pests Resistance
in Rice”
BY
S.PRIYANKA

2. BROWN PLANTHOPPER
Nilaparvata lugens
GREEN LEAFHOPPER
Nephotettix virescens
YELLOW STEM BORER
Scirpophaga incertulas
RICE LEAFFOLDER
Cnaphalocrocis medinalis
THRIPS
Stenchaetothripsbiformis
GALL MIDGE
Orseolia oryzae
WBPH
Sogatella furcifera
EAR HEAD BUG
Leptocorisa oratorius
MAJOR PESTS OF RICE

3. Estimated rice yield losses caused by insect pests on a world basis
(Cramer, 1967)
• Region Yield loss (%)
• Asia 31.5
• People's Republic of China 15.0
• Africa 4.4
• South America 3.5
• North and Central America 3.4
• Europe 2.0
SEVERITY OF YIELD LOSSES DUE TO PEST:
About 2.6 million persons were affected and l2,000 died from hunger.
losses occured ranging from 31.5%in Asia to 2% in Europe.
Annual rice loss of the world due to pest accounts for more than 5% of
the total output of rice (Wang et al., 2009).
Brown planthopper outbreak in l733 was reported as one of the most
damaging insect outbreaks in the history of rice production in Japan.

4.  14% estimated yield loss of all important crops on global bases.
Methods of insect control:
1) Biological method (predators,parasites,natural pesiticides..)
2) Chemical method(chemical insecticides)
Biological method is cheap and does not have any adverse effect on the
ecosystem
To cope with the increasing demand for rice a key element is the development and
implementationof effective rice insect management strategies
R.F.CHANDLERPEST MANAGEMENT

5. Genetic resistance- ability of some genotypes to give higher yields of
good quality than susceptible varieties at the same initial level of insect
attack under similar environmental conditions.
Genetic resistance is the cheapest and the best method of
insect control in crop plants.
One of the component in IPM
GENETIC RESISTANCE??????
•Cost of cultivation
• Reduces populationof predators
and parasites.
• Environmental pollution
• Development of pesticides
resistant biotypes.
Adverse effects of
chemical method

6. Genetic Resistance
A. Based on number of genes:
• Monogenic resistance: Controlled by single gene
• Oligogenic resistance: Controlled by few genes
• Polygenicresistance:Controlled by many genes
B. Based on biotype reaction:
• Vertical resistance:
Effective against specific biotypes (specific resistance)
• Horizontal resistance:
Effective against all the known biotypes (Non specific
resistance)

7. Mechanisms of Insect Resistance:
1) Non preference
2) Antibiosis
3) Tolerance
4) Avoidance or escape.
Painter, 1951

8. NON-PREERENCE
(Non acceptance and Antixenosis)
• Make the host undesirable for unattractive to insects for food, shelter, or
reproduction.
• Various plant character which are associated with non preference include
colour, light penetration, hairiness, leaf angle, odour and taste.
HOST CROP INSECT PEST NON-PREFERENCE PREFERENCE
Rice Rice stem borer Lignified stem Nonlignified stem
Brown planthopper Low asparagines High asparagines
Red Pericarp White Pericarp
Purple stigma Yellow stigma

9. CROP HOST INSECT PEST CAUSE OF ANTIBIOTICS
Rice Rice stem borer High silica content
Adverse effect of host plant on the development and reproduction of insect
pests which feed on resistantplant.
Retard the growth and rate of reproduction of insect pest sometimes lead to
death of an insect.
Involve morphological, physiological and biochemical features of the host
plant.
ANTIBIOSIS (True form of resistance)

10. • Ability of a variety to produce greater yield than susceptible variety at the
same level of insect attack.
• Greater recovery of damaged parts than susceptible ones.
• Avoidance refers to escape of a variety from insect attack either due to
earliness or its cultivation in the season where insect population is very
low.
• Early maturing varieties.
TOLERANCE
AVIODANCE

11. • .
• .
Sources of Insect Resistance
• Utilizationof resistance present in a cultivated
variety for crop improvementCultivated variety
• If desired resistance is not available in a
cultivated variety it should be searched in the
national and or world germplasm collections.
Germplasm of the
crop species
A related wild
species
• Transgenics.
• Eg: Bt
Unrelatedorganisms
•Resistance found in wild relatives.

12. SOURCE OF INSECT RESISTANCE
SCREENING:
• Insect resistantcultivars for use as donors in the breeding programs have
been developed for more than 30 rice insect species throughout the
world.
• In Asia and South America major emphasis has been on the leafhoppers,
planthoppers and stem borers.
Rice Germplasm collectionsin IRRI

13. BREEDING METHODS FOR BIOTIC STRESS
Conventional methods
INTRODUCTION
SELECTION
HYBRIDIZATION
- Backcross breeding
MULTILINE BREEDING
Modern methods
MUTATION BREEDING
TILLING
MOLECULAR BREEDING AND GENOMICS
TRANSGENIC APPROACH

14. Insect Resistance Breeding (Katiyar ,1999)
Production of novel geneticvariation
Selectionof improvedvariants
Screening of rice germplasmto identify noval donars
Use of donarsin hybridizationwith elite cultivarsto create new
combination of genes.
Using standardizedphenotypic testing procedurein green house
and field.

15. Biotechnological approaches ( Katiyar ,1999)
Biotechnological Approches
For Resistance
DNA markers
and
fingerprinting
Transformation
Wide
hybridization
Novel geneticvariation
and selection

16. S.
no
ResistanceTraits Biotechnological Approaches
1 Brown plant hopper DNA marker technology
2 Gall midge resistance Transformation technology, DNA marker technology,DNA
finger printing of insect.
3 Green leaf hopper DNA marker technology
4 Yellow stem borrer Transformation technology, DNA marker technology
5 Leaf folder , hispa Transformation technology
6 Striped stem borrer Transformation technology
Biotechnological Approach For Insect Resistance
(Toenniessen)

17. Molecular markers in rice breeding ( Katiyar, 1999)
•RAPD
•RFLP
• PCR based
markers:
 SCAR
 STS
 Mini Satellites
 AFLP
Helped to locate resistant factors for :
•Brown planthopper
•Green leafhopper
•White backed planthopper
•Gall midge
ADVANTAGES:
Used in identificationof number of
genes invovled andmagnitude o
their contribution.
 screening of resistant genes in
absence of pest.
Horizontal resistance
Gene pyramiding

18. BROWN PLANT HOPPER(Nilaparvata lugens)
•Causes yield loss up to 60 per cent (Panda and Khush, 1995).
•China - 2.7 millions tons ( direct damage)
• Vietnam- 0.4 milliontons (grassy stunt and ragged stunt) (Brar et al.,
2005).
INDIA
Tamilnadu-1.8milliontons.
Severe outbreak occurred in Kerala state at the end of 1973 and early in
1974 (Koya 1974; Nalinakumariand Mammen, 1975).
Severe yield loss
Andhra Pradesh- high yieldingvarieties like SambamashuriandSwarna
were developed and being cultivatedbut these varieties lack resistance
to BPH(Mathur et al., 1999 and Krishnaiah et al., 1999)

19. Evolution of BPH resistance varieties.
Sources of resistance to BPH were first identified in 1967 (Pathak et al.,
1969).
 Donors used in breeding BPH resistantvarieties (Heong and Hardy, 2009)
 Mudgo
 ASD 7
 Rathu
 Heenathi
 Babawee
 ARC 10550
 Swarnalata
Identification of new donorsto breed new BPH resistantvarieties can show
resistanceto newly evolved BPH biotypes
Identification of BPH donors

20. Gene action:(Additive) (Aiyswariya, 2004)
L X T analysis:
5 lines (MDU5,ASD 6, ADT43,ADT36 &IR 50)
8 testers (ASD 7, PTB 33,W123, IR 72,IR 36, ACM9803, ACM9818, Mudgo 1):
Sl.no characters GCA
variance
SCA
variance
Additive
variance
Dominance
variance
ratio
1 Daysto 50%
flowering
4.43 1.92 8.86 1.92 4.61
2 Plant height 16.34 27.58 32.68 27.58 1.18
3 No. of productive
tillers
0.55 2.61 1.10 2.61 0.42
4 No. of
grains/paniicle
1.48 7.76 2.96 7.76 0.38
5 100 grain weight 0.01 0.01 0.01 0.01 1.0
6 BPH resistance 1.56 0.62 3.12 0.62 5.04
7 Grain yield/plant 1.34 0.65 2.74 0.65 4.22

21. International Rice ResearchInstitute (IRRI) and Japan.
Mass-screening techniques such as bulked seedling test and mass-rearing
methods of BPH (Athwal et al. 1971, Kaneda & Kisimoto 1979, Choi et al.
1979)accelerated the work on varietal resistance againstBPH
Resistantvarieties were bred and cultivated in the Philippines, Japan and
other Asian countries since 1970s..
Continuous cultivation leads to breakdown due to the evolvement of new
biotypes
Need for molecularstudies

22.  Breakdown of monogenic resistance by new BPH biotypes has been a
serious threat .
 To overcome monogenic resistance, pyramiding of BPH resistance genes
and quantitative trait loci (QTLs), through marker-assisted method, is
needed.
 Genes and QTLs have identified and mapped on rice chromosomes.
Molecular studies in BPH

23. Genes chromosome references
Bph3 6S Jairin et al.,2007
Bph12(t) 4S Yang et al,2002;Rahman et al .2009
Bph13(t) 2L Liu et al,2001
Bph13(t) 3S Renganayakiet al.2002
Bph14(t) 3L Yang et al,2004
Bph15(t) 4S Yang et al,2004;Rahman et al .2009
Bph16(t) 12 Hirabayashi etal .2004,Myint et al .2012
Bph17(t) 4S Sun et al.2005, Rahman et al .2009
Bph18(t) 12L Jena et al.2006, Myint et al .2012
Bph19(t) 3S Chen et al,2006
Bph20(t) 4 Rahman et al .2009
Bph21(t) 12 Rahman et al .2009
Bph22(t) Ram et al.2010, Rahman et al .2009
Bph23(t) Ram et al.2010, Rahman et al .2009
Bph24(t) Deen et al.2010, Rahman et al .2009
BPH25 6S Myint et al .2012
BPH26 12L Myint et al .2012
Mapped additional major brown plant hopper resistant genes:

24. Brown planthopper resistance QTLs mapped on rice
chromosomes
QTL s Chromosome Reference
Qbp1 3L Huang et al 2001
Qbp1 3 Ren et al 2004
Qbp2 4 Ren et al 2004
Qbp1 3L Yang et al. 2004
Qbp2 4S Yang et al. 2004
Qbp3 3 Sun et al. 2005
Qbp4 4 Sun et al. 2005
Qbp4 4S Liu et al. 2009
Qbp6 6S Sun et al. 2005
Qbp7 7 Liu et al. 2009
Qbp9 Liu et al. 2009
Qbp10 10 Sun et al. 2005

25. White backed plant hopper: Sogatella furcifera
• Attained a place of major pest of paddy in Gujarat.
• The outbreak during 1991, 1992, 1993 and 1996 ruined the crop
completely (Korat et al., 1999)
• Genetic analysis of resistantvarieties has identified eight genes for WBPH
resistance.(Padmavathi, 2007)
• Four of these genes are dominant
WBPH 1, WBPH 2, WBPH 3 , WBPH 5 , WBPH 6 , WBPH 7, WBPH 8
• WBPH 4 is recessive (Angeles et al., 1981; Saini et al., 1982;Wu and Khush,
1985)

26. Six genes for resistance (Wbph1, Wbph2, Wbph3, wbph4, Wbph5, wbph6)
have been identified (Khush 1984).
Tan et al (2004)- Wbph7 (t) and Wbph8(t)
Kadrivel et al (1999)mapped QTLs for resistance to whitebacked
planthopper.
Yamasaki et al 1999; Sogawata et al 2001-QTLs for ovicidal response
Gene Source Chromosome
Wbph1 N22 7
Wbph2 ARC10239 6
Wbph3 ADR52
Wbph4 Podiwi A8
Wbph5 N’ Diang Marie
Wbph6
Wbph7

27. • Varietal resistance is considered as the most promising and practical
approach in the integrated control of this pest (Khan and Saxena,1986).
• Efficiency improved by using molecular markers for detecting genes
governing WBPH resistance.
TRANSGENICS (Planthoppers):
• Snowdrop lectin [Galanthus nivalis agglutinin (GNA)] obtained by
Agrobacterium-mediated genetic transformation showed high resistance to
WBPH (D. Nagadhara, 2004).
• Transgenics also showed substantialresistance againstthe brown plant
hopper and green leaf hopper

28. Green leafhopper (Nephotettix virescens)
•Severe outbreaks in India occurred in 1968 and 1969 and in
Philippines 1971.
•Epidemics of rice tungro virus .
• Native varieties-Peta and Sigadis were resistant to
N.virescens.
•Because of their frequent use as parents, a high proportion
of breeding material from IRRI and in many national
programs is resistant to GLH.

29. Gene Source Chromosome
Glh1 Pankhari 203 5
Glh2 ASD7 11
Glh3 IR8 6
Glh4 Ptb8 3
Glh5 ASD8, O. rufipogon 8
Glh6 TAPL796 5
Glh7 MaddaniKaruppan
Glh8 DV85
Glh9
Glh10
Glh11
Glh12
Glh13
Glh14 ARC11554 4
Genetics of Green Leafhopper

30. RESISTANCE FOR RICE HOPPERS (D.S.Brar,2009)
INSECT PESTS DONORS FOR RESISTANCE
Brown planthopper Mudgo, ASD7, Ptb33, Rathu Heenati,
Babawee, ARC10550,Swarnalata,
Oryza officinalis, O. australiensis, O.
minuta
Green leafhopper Peta, Pankhari 203, Sigadis, Ptb8,
ARC10313, ASD7, ASD8, DV85,
Asmaita
Whitebacked planthopper N22, ARC10239, ADR52, Podiwi-A8
Zigzag leafhopper Rathu Heenati, Ptb21, Ptb33

31. PLANT HOPPER GENES IDENTIFIED
Brown planthopper Bph1, bph2, Bph3, bph4, bph5, bph6, bph7,
bph8, Bph9, Bph10, bph11, bph12, bph13,
Bph14, bph15, Bph16, Bph17, Bph18(t),
bph19(t), bph20, bph21
Green leafhopper Glh1, Glh2, Glh3, glh4, Glh5, Glh6, Glh7,
Glh8, Glh9, glh10
Whitebacked planthopper Wbph1, Wbph2, Wbph3, wbph4, Wbph5,
Wbph6, Wbph7(t), Wbhp8(t)
Zigzag leafhopper Zlh1, Zlh2, Zlh3
Genes for plant hoppers identified in rice (2009)

32. Yellow Stem Borer: Scirpophaga incertulas
• Yieldlosses due to YSB accounts around 20–70 %
(Catling , 1987; Chelliah , 1989).
• Both biotic and abiotic factors are believed to be
responsible for pest population dynamics (Singh , 2009).
• Climatic factors such as temperature, rainfall, and
relative humidity greatly influence the outbreak of the
insect population (Chen , 1968; Heong , 2007;Siswanto
2008).

33. Resistant sources:
• TKM 6
• Chianan 2
• Taichung 16
• Ptb 10
• Su-Yai 20
• WC1263
• Oryza officinalis and O. ridleyi- Highly resistant
Morphological and Anatomical characteristics:
Tall varieties with long, wide leaves and large stems are more
susceptible.
More layers of lignified tissue and sclerenchymatous tissue,
and presence of silica cells are more resistant.
IR20 - the first borer-
resistant, improved-
plant-typevariety, was
developedby crossing
TKM6 with Peta/TN1.
Moderate resistant

34. • Traditional breeding for stem borer resistance in is not successful.
• More than 30,000 rice accessions for resistance to different stem borers .
• Good level of resistance againstthe yellow stem borer has been rare in
the rice germplasms (Deka and Barthakur, 2010).
Control of stem borer through common insecticide is major constraint.
NEED FOR TRANSGENICS IN YSB
No rice variety with sufficient level of resistancehas been developed.

35. GENETIC TRANSFORMATION
Transformation of indica rice has been carried out using :
• Direct gene transfer using protoplast (Daatta S K)
• Particle bombardement (christou P)
• Agrobacterium mediated gene delivery (Aldemitaa R)
cry1 Ab and cry 1Ac – effective control of yellow stem borer.
Molecular studies:
Genetic studies of YSB resistance revealed polygenic nature of trait
.(chaudary et al ., 1984).
Screening with molecular markers are preferable.

36. Control of yellow stem borer in rice with respect to naturally
occurring insecticidal proteins.
• The protease like inhibitor from mature jackfruit (Artocarpus
heterophyllus)Inhibits the activity of midgut protease of yellow stem
borer. (Shamim,2011).
• Significant reduction in larval weight and mortality was observed, when
fresh rice culms with protease inhibitor was feeded to the yellow stem
borer larvae.
Insecticidal proteins

37. Gall midge
• The extent and severity of gall midge infestation has
significantly increased since 1970’s.
• The cultivation of high tillering varieties, intensive
managementand low parasitization are conducive to
the rapid multiplication of this pest.
• In India, serious pest in Madhya Pradesh, Manipur,
Orissa, Telangana, Andhra Pradesh, Bihar, Karnataka
and Kerala.
(Suvendhu et al., 2014)

38. Disadvantages in current control measures
• The gall midge attacks rice crop from the nursery to the end of
tillering stage – Hence all stage are vulnerable.
• At the onset of monsoons, it becomes active and completes one to
two generations on grasses before it moves to rice crop.
• In some cases, the pest survives on weeds and also in rice stubbles
left after the first crop.
• At present tolerant or resistant varieties are not 100% resistant.
BASE LINE – ERADICATION BY PESTICIDES IS TEMPORARY,
Hence breeding new resistance varieties is only hope
(Brar et al. 2009)

39. • National Rice Improvement Programs in Bangladesh, India,
Indonesia, Sri Lanka and Thailand
- currently screening germplasm for resistance to the gall midge
Indian varieties:
• Eswarakora
• HR42
• HR63
• Ptb 18
• Ptb 21
• Siam 29
• Thai variety- Leuang 152

40. Breeding for gall midge resistance - case study 1

41. Breeding for gall midge resistance - case study 2

42. Procedure – case study 1 &2
Test entries
Gall midge resistant
genotypes was collected
Phenotypic screening
The test entries screened
against gall midge biotype
in greenhouse
Genotypic screening & characterization
Gm6 gene- flanking SSR marker
Gene tagging
And mapping
Crossing
GM resistant and hybrid
Conformation
The crossed seeds were grown
and checked for the presence of
resistant gene

43. Rice Thrips: Stenchaetothrips biformis
commonspecies :
• Stenchaetothripsbiformis (Bagnall) (Thripidae)
• Haplothrips aculeatus (Fabricius) Phlaeothripidae
S. biformis - major rice pest (In Bangladesh, China, India, Indonesia, Japan,
and Sri Lanka)
Varieties :
• IR62
• IET1444
Traditional varieties:
• Ptb 21
• Ptb 33 f
Highly resistantvariety should be
derivedby utilizing wild species.

44. Wild species with resistance to rice thrips
Species (IRRI accessionno.) origin Reference
O.eichingeri
101171 tanzania Veluswamy et al (1981)
101418 Uganda Nugaliyadde and Heinrichs (1984a)
101422 Uganda Nugaliyadde and Heinrichs (1984a)
101424 Uganda Nugaliyadde and Heinrichs (1984a)
O.glaberrima
103437 Senegal Veluswamy et al (1981)
103438 Senegal Veluswamy et al (1981)
103443 Senegal Veluswamy et al (1981)
O.minuta
101079 Philippines Nugaliyadde and Heinrichs (1984a)
101083 Philippines Nugaliyadde and Heinrichs (1984a)
101097 Philippines Nugaliyadde and Heinrichs (1984a)
O.nivara
100897 Burma Nugaliyadde and Heinrichs (1984a)
101510 india Veluswamy et al (1981)
103836 bangaladeh Veluswamy et al (1981)
O.nivara/O.sativa
103791 venezula Veluswamy et al (1981)

45. Wild species with resistance to rice thrips
Wild species location reference
O.officinalis
100181 Burma Nugaliyaddeand Heinrichs (1984a)
100973 philippines Nugaliyaddeand Heinrichs (1984a)
101073 philippines Nugaliyaddeand Heinrichs (1984a)
101117 philippines Nugaliyaddeand Heinrichs (1984a)
101155 malaysia Nugaliyaddeand Heinrichs (1984a)
O.perennis
103849 India Veluswamyet al (1981)
O.rufipongan/O.nivara
101993 srilanka Veluswamyet al (1981)
O.sativa
103826 Veluswamyet al (1981)
103831 Veluswamyet al (1981)

46. Earhead bug: Leptocorisa oratorius
All cultivated rice varieties are susceptible to grain-sucking pests
Sathi
Panicle remains enclosed in the leaf sheath and offers a sort of mechanical
resistance to insect sucking.
Demerit: Threshing
In India:
•Varieties Mundagakuttyfrom Tamil Nadu
• Soma from Bihar.
Varietal resistance(Insectpests of Rice)

47. RICE LEAFFOLDER: Cnaphalocrocis medinalis
Recorded to cause 63% to 80% yield loss in rice
- (Rajendaran et al, 1986).
Host Plant Resistance:
• Trichome density on the abaxial surface (TKM6) Ramachandran and
Khan (1991).
Food searching capabilities:
Plant height, productive tiller number, leaf length, leaf area and leaf
thickness.
Longer and broaderleaves recordedmore
damage

48. Varietal resistance:
Donars offering multiple resistance to leaffolder:
• TKM6
• WC1263
• Ptb 33
• GEB24
• Muthumanikam
SCREENING:
IRRI-out of 18000 rice accessions,115 were found to be moderately resistant.
Wild species resistant to
leafolder:
Oryza brachyantha,
O. Nivara
O. rufipogon
O. perennis
Traditional varieties:
Ptb 21, Ptb 33, TKMI,
TKM2, TKM6,
Muthumanikam,and
WC1263,

49. QTL’ s for rice leaffolder resistance
• :Traits Chromoso
me
Interval LOD Score Phenotypic
Variance
Additive
Effect
qRLF-1 1 RM3412-RM6716 3.08 15.68% -9.1
qRLF-2 2 RM207-RM48 2.64 8% -6.4
qRLF-3 3 RM1022-RM7 3.29 16.4% 9.5
qRLF-4 4 RM3276-RM255 2.81 13.5% 8
qRLF-5 8 RM72-RM331 4.01 21.1% 10

50. Transgenics:
Cry1Ac and CpTI expressing transgenic lines
Offered resistant :
• In feeding behaviour
• Mortality rate was more than 90%.
-(Han et al., 2007).
Potato Proteinase Inhibitor II
out of 10 transgenic lines of TNG 67(Japonica) and aromatic
indica varieties (HBC 19 & Pusa basmathi)
5 lines exhibited:
high mortality rate (87.5% & 92.5%)
lowest larval feeding rate
-(REKHA et al., 2008)

51. Constraints in Insect Resistance Breeding
Long term process (10-15 years).
Breeding for resistanceto one pest leads to the susceptibility
to another pest.
Genes are associated with some undesirable characters.
Interspecific gene transfer(wild species)
Expensive method
Screening for insect resistance is the most critical and difficult
step in plant breeding programme.
Evolution of new virulent biotypes.