Академический Документы
Профессиональный Документы
Культура Документы
discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/259874835
CITATIONS READS
24 760
5 authors, including:
SEE PROFILE
Some of the authors of this publication are also working on these related projects:
Successfully completed NAIP funded project "Allele Mining and Expression Profiling for R-gene and
Avr gene in Rice Blast Pathosystem for Development of Race Non Specific Disease Resistance" View
project
All content following this page was uploaded by Shamshad Alam on 27 January 2014.
Pratyoosh Shukla
Abstract Molecular screening and genetic diversity specific STS marker, 195R-1, for Pi9 gene produced
of major rice blast resistance (R) genes were deter- positive bands in only two germplasm, Kalchatti and
mined in 32 accessions of rice germplasm from North Bachi thima. The Uniform Blast Nursery (UBN)
East and Eastern India with ten gene based single analysis showed that out of 32, six germplasm was
nucleotide polymorphisms and sequence tagged sites resistant, ten moderately resistant and 16 germplasm
(STS) markers, namely z56592, zt56591, k39512, were susceptible. Presence of Piz-t, Pita/Pita-2 and
k3957, candidate gene marker, Pita3, YL155/YL87, Pi9 gene ensured a resistant reaction in outdoor blast
YL183/YL87, Pb28, 195R-1 which showed close-set nursery whereas germplasm carrying Pib was suscep-
linkage to nine major rice blast resistance (R) genes, tible when present alone. Presence of multiple genes,
Piz, Piz-t, Pik, Pik-p, Pik-h, Pita/Pita-2, Pib and Pi9 however, contributed to slow blasting resistance in the
and one susceptible pita gene. Among the 32 acces- field. These results are useful in identification and
sions, 13 were positive for Piz gene and six for Piz- incorporation of resistant genes from the germplasm
t gene. Six accessions were positive for Pik gene, into elite cultivars through marker assisted selection in
seven for Pik-p and 16 for Pik-h gene. One accession, rice breeding programs.
Atte thima, was positive for three of Pik multiple
genes. Out of 32, only two germplasm, Dudhraj and Keywords Indian rice germplasm R genes
Nepali dhan, were detected with both Pita3 and screening Blast resistance SNP markers
YL155/YL87 marker for Pita/Pita-2 gene. The Pib Marker assisted selection
gene appeared to be omnipresent and was detected in
31 of 32 germplasm with marker Pb28. The gene Abbreviations
R Resistance
Avr Avirulent
SNPs Single nucleotide polymorphisms
J. Imam S. Alam N. P. Mandal M. Variar STS Sequence tagged sites
Central Rainfed Upland Rice Research Station (CRRI),
UBN Uniform blast nursery
Hazaribagh 825301, Jharkhand, India
e-mail: mukund.variar@gmail.com MR Moderately resistant
S Susceptible
J. Imam P. Shukla (&) MAS Marker assisted selection
Enzyme Technology and Protein Bioinformatics
NE North East
Laboratory, Department of Microbiology, Maharshi
Dayanand University, Rohtak 124001, Haryana, India SES Standard evaluation system
e-mail: pratyoosh.shukla@gmail.com DAS Days of sowing
123
Euphytica
123
Euphytica
identified/cloned are effective against the different for 24 weeks for genomic DNA extraction. Rice
lineages prevalent in the region. Eastern India is leaves were collected from plants and were imme-
considered to be a rich pocket of rice genetic resources diately frozen in liquid nitrogen and stored at
in the world owing to the extremely diverse rice -80 C for DNA extraction. Genomic DNA was
growing conditions as compared to other parts of the isolated using Plant Mini Kit (Qiagen, Valencia,
country. Selection made unknowingly by various CA, USA) from 100 mg of leaf tissue according to
ethnic groups inhabiting at different altitudes and manufacturers instruction. The extracted genomic
climatic situations, practicing different forms of DNA was estimated on 1 % agarose gels for
cultivation might have contributed to some extent quality. The quantity of extracted DNA was
towards the diversity of rice crop in this region. It is measured by Nano Drop system, 2000c (Thermo,
roughly estimated that during the past more than USA) and were diluted to 100 ng/ll with TE buffer
30,000 rice cultivars were grown in North East and and stored at -20 C.
Eastern parts of India (Ngachan et al. 2011). Mahender
et al. 2012 demonstrated the presence of six to seven Markers specific for rice blast R genes
genes in rice accessions from the North Eastern state
of Manipur which was related to high level of The germplasm accessions were screened for the
resistance in the accessions. presence of nine major blast resistance (R) genes, Piz,
Molecular genetic markers are now widely used to Piz-t, Pik, Pik-p, Pik-h, Pita/Pita-2, Pib, Pi9 and one
characterize gene bank collections that contain susceptible pita gene using a set of ten SNP and gene
untapped resources of distinct alleles which will based STS markers (Table 1). PCR based allele-
remain hidden unless efforts are initiated to screen specific SNP and gene based STS markers were
them for their potential use and function. Abundance obtained for identification of blast resistance genes in
of SNP polymorphisms has made it an attractive tool the selected germplasm. All the markers were synthe-
for allele mining and marker assisted selection. SNPs sized by Operon Eurofins (USA).
can be detected using allele specific PCR primers and
typed by the presence or absence of PCR amplified DNA marker analysis
products on standard agarose gels. Presence of differ-
ent blast resistance genes in a collection of 32 rice An allele specific PCR marker assays the genotype by
germplasm from North East and Eastern India was examining the presence or absence of a PCR ampli-
examined in the present investigation using PCR fication product. The PCR analyses were conducted
based SNP markers. and templates for PCR reaction set up for 20 ll as
follows: 19 PCR buffer, 0.2 mM of dNTPs mix,
1.5 mM of MgCl2, 0.2 lM each primer, 50 ng of
Materials and methods genomic DNA template, 1 unit of taq DNA polymer-
ase (Fermentas Life Sciences, Burlington, Canada)
Plant materials and DNA isolation and MilliQ water to final volume of 20 ll. The PCR
amplification program consisted of (i) 5 min initial
Thirty-two germplasm including local land races denaturation at 94 C (ii) 35 cycles of 45 s denatur-
and primitive cultivars originating from North East ation at 94 C, 45 s primer annealing at different Tm
and Eastern India and a cultivated check Vandana (Table 1), and 2 min extension at 72 C, and a final
was selected from the genetic stock maintained at extension for 5 min at 72 C. PCR products were
Central Rainfed Upland Rice Research Station, separated in 23 % agarose gels in 19 TBE buffer,
Hazaribag, based on their partial/complete resis- stained in ethidium bromide and visualized under UV
tance to blast resistance in outdoor blast nurseries. transilluminator. All PCR reactions for each sample
The seeds were germinated in the dark on moist- were repeated at least once to confirm the results. The
ened filter papers for 2 days at 30 C in a plant amplified fragments were scored as presence (1) or
growth chamber, planted in plastic pots and shifted absence (0) of amplicon linked to each gene DNA
to greenhouse for growth at 27 C with 16 h light fragment.
123
Euphytica
Qu et al. (2006)
The NE and Eastern India rice germplasm, selected for
this study based on earlier reports of their complete/
References
1,042
2,000
292
257
112
148
1,500
861
1,043
388
Table 1 Details of single nucleotide polymorphisms (SNP) and gene based STS markers tightly linked to the major rice blast resistant genes
59
55
60
56 Scores 03 were considered resistant (R), 45 as
moderately resistant (MR) and 69 as susceptible (S).
gcattctccaacccttttgcatgcat
acattggtagtagtgcaatgtca
ctgcgccaagcaataaagtc
ctaccaacaagttcatcaaa
ctaccaacaagttcatcaaa
gactcggtcgaccaattcgcc
agcaggttataagctagctat
agcaggttataagctaggcc
ttgctgagccattgttaaaca
atggtcctttatctttattg
Primer sequence
195R-1
Marker
K3957
Pita3
Pb28
PCR results for the Piz and Piz-t rice blast resistance
genes were determined by visualization of amplicons
number
of 292 and 257 bp, using SNP primer, z56592 for Piz
6
6
2
6
11
11
11
12
12
(Fig. 1) and zt56591 for Piz-t (Fig. 2). Piz gene was
scored on 13 accessions and Piz-t gene was detected in
Pita/Pita-
Piz-t
Pita
Pib
Pi9
Pik
Piz
123
Table 2 SNPs and STS markers associated with ten blast resistance genes in North East and Eastern Indian rice germplasm and their reaction to blast at UBN, Hazaribag, WS
2012
Euphytica
S. No. Varieties HRC State/ Piz Piz-t Pik Pik-p Pik-h Pita/ Pita/Pita-2 Pita Pi9 Pib Highest
Location Pita-2 scorea
Z56592 Zt56591 K39512 K3957 Gene based Pita3 YL155/ YL183/ 195R-1 Pb28 (09)
marker YL87 YL87
123
Euphytica
Highest
scorea
(09)
5
5
4
Pb28
Pib
1
1
1
195R-1
Pi9
0
0
0
0 means the absence of amplicons linked to the rice blast resistant genes 1 means the presence of amplicons linked to the rice blast resistant genes
YL183/
YL87
Pita
1
1
1
Pita/Pita-2
YL155/
0
0
0
K3957
Pik-p
0
0
0
K39512
0
1
0
Zt56591
Piz-t
0
0
0
Tripura
State/
1,511
1,002
808
123
Euphytica
123
Euphytica
The Pib-specific PCR primer set, Pb28 marker was Reaction of the set of 32 germplasm to blast under
developed to produce a 388 bp amplicon based on its UBN was evaluated at CRURRS, Hazaribag. Disease
123
Euphytica
Fig. 11 Frequency distribution of blast disease score of 32 North East and Eastern India rice germplasm
reaction after 35 days of sowing ranged from 0 to 6 in resistance in the uniform blast nursery. When the
the SES scale when the susceptible spreader CO39 resistance pattern of the germplasm is examined vis a
was completely killed in the nursery (score 9). Six vis the presence of amplicon products of the major
germplasm were resistant (score 03), ten moderately genes it was noted that though resistance is generally
resistant (score 4) and sixteen susceptible ([5) proportional to the frequency of the gene(s), certain
(Fig. 11). All the six resistant germplasm originated genes (Pi9, Pita-2, Piz-t) were more effective than
from Sikkim and were positive for the markers for Piz- others in thwarting infection. They were also less
t, Pita-2 or Pi9. Presence of Pita-2 or Pi9, invariably frequently detected by marker amplification. Con-
contributed to resistance in the nursery. None of the versely, the less effective genes were more predomi-
other 26 moderately resistant/susceptible germplasm nant but mostly ineffective against the different
harboured markers for these genes. Piz-t appeared to pathotypes/lineages that are normally present in the
be associated with moderate resistance in two germ- nursery, albeit with different frequencies. Multi-loca-
plasm in addition to the two germplasm which tion evaluation of a set of isogenic lines carrying 24
recorded resistant reaction. Though the presence of major blast resistance genes had earlier indicated that
multiple resistance genes generally appeared to have Pi9, Piz-t and Pita-2 were resistant at most locations,
higher level of resistance, there were also instances indicating their broader spectrum of resistance to
where the presence of several genes did not ensure pathotypes prevalent in Eastern India (Variar et al.
resistance. Atte thima, for example, harboured five 2009). Among the multi-genes near the waxy gene
resistance genes, but was susceptible. Pib, Pita, Pik- locus on chromosome 6 (Ballini et al. 2008) Pi9 was
p and Pik-h were the most abundant genes detected in more effective than Pi2 (Piz-t) in the present investi-
the germplasm but they were also the least effective gation. Piz, Piz-t and Pi40(t) were commonly detected
against the population of the blast fungus present in the in germplasm originating from North East India earlier
nursery at Hazaribag. It is however, noted that all the but Pi9 was not detected among them (Mahender et al.
tested germplasm possessed low levels of resistance as 2012). Since the present study was taken up with a
evidenced by their relative scores vis a vis the select set of rice germplasm covering a wider geo-
susceptible spreader, CO39 which possessed none of graphical region (Chattisgarh, Jharkhand, Sikkim and
the genes evaluated in this study (data not shown). Assam), the chances improved for detection of rare
Genetic frequency of the nine major rice blast alleles like Pi9 which was identified in two germplasm
resistance genes, Piz, Piz-t, Pik, Pik-p, Pik-h, Pita/ Kalchati and Bachi thima. While Pi9 gene conferred
Pita-2, pita, Pi9 and Pib, ranged from 6 to 97 % in the complete resistance in Kalchati and Bachi thima, Piz-t,
select set of germplasm that exhibited different level of along with other genes, contributed to complete
123
Euphytica
resistance in three germplasm (Nepali dhan, Chirakey- virulent on the isogenic lines carrying Pib and Pik
C and Phaudel) and moderate resistance in others. (Unpublished data).
Thirteen accessions (41 %) showed positive bands for Genetic variability among the rice accessions and
the Piz gene with z56592 SNP marker but its presence within the pathogen often leads to inconsistent marker
independently did not contribute to complete resis- and phenotype analysis. MAS have the advantage in
tance in any of the germplasm evaluated in this study. identifying R genes, but its power lies in the robustness
Both Piz and Piz-t genes have been used for conferring of the markers used. The identification of rice blast
blast resistance to Japanese cultivars, because their resistance gene Pib and analysis of rice blast resistance
importance was emphasized by Hayashi et al. (2004) in gene Piz suggests that DNA markers derived from the
rice breeding in Japan. The Piz-t and Piz genes, co- gene is a valuable tool for blast gene identification and
segregated with z56592 and zt56591 markers, were screening among the rice germplasm (RoyChowdhury
flanked on one side by z4794 and on the other side by et al. 2012a, b). In this study, the PCR based markers
z60510 and z5765 markers (Hayashi et al. 2006). employed for screening of different blast resistance
Two blast resistance genes, Pita and Pita-2, have genes are well established and effective. The consis-
been located at the Pita locus near the centromere of tent results showed with the selected SNPs and STS
chromosome 12 and are tightly linked to each other markers for respective genes was highly reliable and
(Wang et al. 2002). These two genes were interacting make them the marker of choice for molecular
in terms of their resistance specificity. Pita-2 has a screening of rice blast resistant genes among the rice
broader resistance spectrum than Pita (Rybka et al. germplasm. For the molecular identification of blast
1997; Bryan et al. 2000). In the present study, only resistance genes, isogenic lines for the selected genes
two accessions Dudhraj and Nepli dhan were positive were taken as positive control and CO39 as negative
of Pita/Pita-2 genes using the SNP marker Pita3 and control.
dominant marker YL155/YL87. These two acces- Many rice varieties have been developed as com-
sions showed resistance pattern in UBN. The pletely resistant to M. oryzae strains, but soon
isogenic lines of Pita and Pita-2 gene, evaluated breakdown of rice blast resistant genes occurred
earlier, were resistant to blast in multi environment because of the emergence of stronger virulent isolates
testing and this confirmed that the germplasm having of rice blast fungus (Bonman et al. 1986; Mackill and
Pita/Pita-2 gene is likely be effective in rice Bonman 1992). Genotyping of the germplasm with
breeding program. The 32 accessions were also allele specific markers helped to identify nine major
screened for pita susceptible gene with dominant blast resistance genes Piz, Piz-t, Pik, Pik-p, Pik-h,
marker YL183/YL87 and found that 29 accessions Pita/Pita-2, Pib and Pi9 and one recessive pita gene in
gave amplicons of 1,043 bp. The Pita/Pita-2 markers 32 North East and Eastern Indian rice germplasm. The
used in this study were based on DNA polymorphism UBN analysis revealed the blast disease pattern of
of nucleotides between the resistant indica Pita allele these germplasm and showed that germplasm con-
and the susceptible japonica pita allele (Jia et al. taining Pita/Pita-2, Pi9 are also effective against the
2002, 2003, 2004a, b). Identification of Pita/Pita-2 prevalent pathotypes of M oryzae. Interestingly,
genes and its validation with differential isolates of several germplasm had multiple disease resistance
M. oryzae reveals that the Indian rice germplasm are genes. Virulence analyses using specific isolates
diverse and potential source of blast resistant lines would help unravel the response of the resistance
which can be exploited in rice blast breeding genes against specific lineages. Results of this work
programs (Shikari et al. 2013). showed that all of the 32 accessions of North East and
Members of the Pik multi-gene family and Pib were Eastern India rice germplasm possessed more than one
the most frequently detected genes in the present study blast resistance gene. Being adapted to the locations
but neither the germplasm possessing them nor the where these germplasm originated, and having co-
isogenic lines in the previous evaluations (Variar et al. evolved with the local population of the blast fungus,
2009) exhibited resistance. Virulence analyses using use of these germplasm could have a competitive edge
50 M. oryzae isolates collected from North East and over other exotic resistance donors currently being
Eastern India also indicated that most isolates are used in the breeding programs.
123
Euphytica
Acknowledgments The authors acknowledge the National Hayashi N, Inoue H, Kato T, Funao T, Shirota M, Shimizu T,
Agriculture Innovative Project- Component 4, Indian Council of Kanamori H, Yamane H, Hayano-Saito Y, Matsumoto T,
Agricultural Research (ICAR), on Allele mining blast resistance Yano M, Takatsuji H (2010) Durable panicle blast-resis-
genes for financial support for this research. This work was tance gene Pb1 encodes an atypical CC-NBS-LRR protein
performed at the Central Rainfed Upland Rice Research Station, and was generated by acquiring a promoter through local
Hazaribag, Jharkhand, India. genome duplication. Plant J 64:498510
Hittalmani S, Parco A, Mew TW, Zeigler RS, Huang N (2000)
Fine mapping and DNA marker-assisted pyramiding of the
References three major genes for blast resistance in rice. Theor Appl
Genet 100:11211128
Ashikawa I, Hayashi N, Yamane H, Kanamori H, Wu J, Mat- Huang H, Huang L, Feng G, Wang S, Wang Y, Liu J, Jiang N,
sumoto T, Ono K, Yano M (2008) Two adjacent nucleo- Yan W, Xu L, Sun P, Liu Z, Pan S, Liu X, Xiao Y, Liu E,
tide-binding site-leucine-rich repeat class genes are Dai L, Wang G (2010) Molecular mapping of the new blast
required to confer Pikm-specific rice blast resistance. resistance genes Pi47 and Pi48 in the durably resistant
Genetics 180:22672276 local rice cultivar Xiangzi 3150. Phytopathology
Ballini E, Morel JB, Droc G, Price A, Courtois B, Notteghem JL, 101:620626
Tharreau D (2008) A genome-wide meta-analysis of rice Inukai T, Nelson RJ, Zeigler RS, Sarkarung S, Mackill DJ,
blast resistance genes and quantitative trait loci provides Bobman JM, Takamure I, Kinoshita T (1994) Allelism of
new insights into partial and complete resistance. Mol blast resistance genes in near-isogenic lines of rice. Phy-
PlantMicrobe Interact 21:859868 topathology 84:12781283
Bonman JM, Vergel de Dios TI, Khim MM (1986) Physiologic Jeung JU, Kim BR, Cho YC, Han SS, Moon HP, Lee YT, Jena
specialization of Pyricularia oryzae in the Philippines. KK (2007) A novel gene, Pi40(t), linked to the DNA
Plant Disease 70:767769 markers derived from NBS-LRR motifs confers broad
Bryan GT, Wu KS, Farrall L, Jia YL, Hershey HP, McAdams spectrum of blast resistance in rice. Theor Appl Genet
SA, Faulk KN, Donaldson GK, Tarchini R, Valent B 115:11631177
(2000) A single amino acid difference distinguishes resis- Jia Y, Wang Z, Singh P (2002) Development of dominant rice
tant and susceptible alleles of the rice blast resistance gene blast Pi-ta resistance gene markers. Crop Sci
Pi-ta. Plant Cell 12:20332045 42:21452149
Chen X, Shang J, Chen D, Lei C, Zou Y, Zhai W, Liu G, Xu J, Jia Y, Bryan GT, Farrall L, Valent B (2003) Natural variation at
Ling Z, Cao G, Ma B, Wang Y, Zhao X, Li S, Zhu L (2006) the Pi-ta rice blast resistance locus. Phytopathology
A B-lectin receptor kinase gene conferring rice blast 93:14521459
resistance. Plant J 46:794804 Jia Y, Redus M, Wang Z, Rutger JN (2004a) Development of a
Chen J, Shi Y, Liu W, Chai R, Fu Y, Zhuang J, Wu J (2011) A SNLP marker from the Pi-ta blast resistance gene by tri-
Pid3 allele from rice cultivar Gumei2 confers resistance to primer PCR. Euphytica 138:97105
Magnaporthe oryzae. J Genet Genomics 38:209216 Jia Y, Wang Z, Fjellstrom RG, Moldenhauer KAK, Azam MA,
Das A, Soubam D, Singh PK, Thakur S, Singh NK, Sharma TR Correll J, Lee FN, Xia Y, Rutger JN (2004b) Rice Pi-ta
(2012) A novel blast resistance gene, Pi54rh cloned from gene confers resistance to the major pathotypes of the rice
wild species of rice, Oryza rhizomatis confers broad blast fungus in the US. Phytopathology 94:296301
spectrum resistance to Magnaporthe oryzae. Funct Integr Kang S, Lebrun MH, Farrall L, Valent B (2001) Gain of viru-
Genomics 12:215228 lence caused by insertion of a Pot3 transposon in a Mag-
Deng Y, Zhu X, Shen Y, He Z (2006) Genetic characterization naporthe grisea avirulence gene. Mol PlantMicrobe
and Wne mapping of the blast resistance locus Pigm(t) Interact 14:671674
tightly linked to Pi2 and Pi9 in a broad-spectrum resistant Kiyosawa S, Mackill DJ, Bonman JM, Tanaka Y, Ling ZZ
Chinese variety. Theor Appl Genet 113:705713 (1986) An attempt of classification of worlds rice varieties
Flor HH (1971) Current status of gene-for-gene concept. Annu based on reaction pattern to blast fungus strains. Bull Natl
Rev Phytopathol 9:275296 Inst Agrobiol Resour 2:1339
Fukuoka S, Saka N, Koga H, Ono K, Shimizu T, Ebana K, Lee SK, Song MY, Seo YS, Kim HK, Ko S, Cao PJ, Suh JP, Yi
Hayashi N, Takahashi A, Hirochika H, Okuno K, Yano M G, Roh JH, Lee S, An G, Hahn TR, Wang GL, Ronald P,
(2009) Loss of function of a proline-containing protein Jeon JS (2009) Rice Pi5-mediated resistance to Magna-
confers durable disease resistance in rice. Science porthe oryzae requires the presence of two coiled-coil-
325:9981001 nucleotide-binding-leucine-rich repeat genes. Genetics
Hayashi K, Yoshida H (2009) Refunctionalization of the ancient 181:16271638
rice blast disease resistance gene Pit by the recruitment of a Lin F, Chen S, Que Z, Wang L, Liu X, Pan Q (2007) The blast
retrotransposon as a promoter. Plant J 57:413425 resistance gene Pi37 encodes a nucleotide binding site-
Hayashi K, Hashimoto N, Daigen M, Ashikawa I (2004) leucine-rich repeat proteinand is a member of a resistance
Development of PCR-based SNP markers for rice blast gene cluster on rice chromosome 1. Genetics
resistance genes at the Piz locus. Theor Appl Genet 177:18711880
108:212220 Liu X, Lin F, Wang L, Pan Q (2007) The in silico map-based
Hayashi K, Yoshida H, Ashikawa I (2006) Development of cloning of Pi36, a rice coiled-coil nucleotide-binding site
PCR-based allele specific and InDel marker sets for nine leucine-rich repeat gene that confers race-specific resis-
rice blast resistance genes. Theor Appl Genet 113:251260 tance to the blast fungus. Genetics 176:25412549
123
Euphytica
Liu J, Wang X, Mitchell T, Hu Y, Liu X, Dai L, Wang GL Shikari AB, Khanna A, Krishnan SG, Singh UD, Rathour R,
(2010) Recent progress and understanding of the molecular Tonapi V, Sharma TR, Nagarajan M, Prabhu KV, Singh
mechanisms of the rice-Magnaporthe oryzae interaction. AK (2013) Molecular analysis and phenotypic validation
Mol Plant Pathol 11:419427 of blast resistance genes Pita and Pita2 in landraces of rice
Liu Y, Liu B, Bordeos A, Leung H, Zhu X, Wang G, Yang J, (Oryza sativa L.). Indian J Genet 73:131141
Leach JE (2013) Fine-mapping and molecular marker Takahashi A, Hayashi N, Miyao A, Hirochika H (2010) Unique
development for Pi56(t), a NBS-LRR gene conferring features of the rice blast resistance Pish locus revealed by
broad-spectrum resistance to Magnaporthe oryzae in rice. large scale retrotransposon-tagging. BMC Plant Biol 10:175
Theor Appl Genet 126:985998 Tang J, Zhu X, Wang Y, Liu L, Xu B, Li F, Fang J, Chu C (2011)
Mackill DJ, Bonman JM (1992) Inheritance of blast resistance in Semi-dominant mutations in the CC-NB-LRR-type R gene,
near-isogenic lines of rice. Phytopathology 82:746749 NLS1, lead to constitutive activation of defense responses
Mahender A, Swain DM, Gitishree D, Subudhi HN, Rao GJN in rice. Plant J 66:9961007
(2012) Molecular analysis of native Manipur rice accessions Tanksley SD, McCouch SR (1997) Seeds banks and molecular
for resistance against blast. Afr J of Biotech 11:13211329 maps: unlocking genetic potential from the wild. Science
McCouch SR, Nelson RJ, Tohme J, Zeigler RS (1994) Mapping 277:10631066
of blast resistance genes in rice. In: Zeigler RS, Leong SA, Valent B (1990) Rice blast as a model system for plant pathol-
Teng PS (eds) Rice blast disease. CAB International, ogy. Phytopathology 80:3336
Wallingford, pp 167186 Valent B, Chumley FG (1994) A virulence genes and mecha-
Ngachan SV, Mohanty AK, Pattanayak A (2011) Status Paper nism of genetic instability in the rice blast fungus. In:
on Rice in North East IndiaRice in North East India. Rice Zeigler RS, Leong SA, Teng PS (eds) Rice blast disease.
Knowledge Management Portal pp 82. http://www.rkmp. CAB International, Wallingford, pp 111134
co.in. Accessed 28 June 2011 Valent B, Farrall L, Chumley FG (1991) Magnaporthe grisea
Okuyama Y, Kanzaki H, Abe A, Yoshida K, Tamiru M, Saitoh genes for pathogenicity and virulence identified through a
H, Fujibe T, Matsumura H, Shenton M, Galam DC, Undan series of backcrosses. Genetics 127:87101
J, Ito A, Sone T, Terauchi R (2011) A multi-faceted Variar M, Vera Cruz CM, Carrillo MG, Bhatt JC, Sangar RBS
genomics approach allows the isolation of rice Pia blast (2009) Rice blast in India and strategies to develop durably
resistance gene consisting of two adjacent NBS-LRR resistant cultivars. In: Xiaofan W, Valent B (eds) Advances
protein genes. Plant J 66:467479 in genetics, genomics and control of rice blast disease.
Qu S, Liu G, Zhou B, Bellizzi M, Zeng L, Dai L, Han B, Wang Springer, New York, pp 359374
GL (2006) The broad spectrum blast resistance gene Pi9 Wang ZX, Yano M, Yamanouchi U, Iwamoto M, Monna L,
encodes a nucleotide-binding site leucine-rich repeat pro- Hayasaka H, Katayose Y, Sasaki T (1999) The Pib gene for
tein and is a member of a multigene family in rice. Genetics rice blast resistance belongs to the nucleotide-binding and
172:19011914 leucine-rich repeat class of plant disease resistance genes.
RoyChowdhury M, Jia Y, Jackson A, Jia MH, Fjellstrom R, Plant J 19:5564
Cartwright R (2012a) Analysis of rice blast resistance gene Wang C, Hirano K, Kawasaki S (2002) Cloning of Pita-2 in the
Pi-z using pathogenicity assays and DNA markers. Eu- centromeric region of chr 12 with HEGS: high efficiency
phytica 184:3547 genome scanning. Third International Rice Blast Confer-
RoyChowdhury M, Jia Y, Jia MH, Fjellstrom B, Cartwright R ence, p 25
(2012b) Identification of the rice blast resistance gene Pi- Wang Z, Jia Y, Rutger JN, Xia Y (2007) Rapid survey for
b in the national small grains collection. Phytopathology presence of a blast resistance gene Pi-ta in rice cultivars
102:700706 using the dominant DNA markers derived from portions of
Rybka K, Miyamoto M, Ando I, Saito A, Kawasaki S (1997) the Pi-ta gene. Plant Breed 126:3642
High resolution mapping of the Indica-derived rice blast Wang X, Fjellstrom RG, Jia Y, Yan W, Jia MH, Scheffer BE,
resistance genes II. Pi-ta2 and Pi-ta and a consideration of Wu D, Shu Q, McClung AM (2010) Characterization of Pi-
their origin. Mol Plant Microbe Interact 10:517524 ta Blast resistance gene in an international rice core col-
Shang J, Tao Y, Chen X, Zou Y, Lei C, Wang J, Li X, Zhao X, lection. Plant Breed 129:491501
Zhang M, Lu Z, Xu J, Cheng Z, Wan J, Zhu L (2009) Xia JQ, Correll JC, Lee FN, Marchetti MA, Rhoads DD (1993)
Identification of a new rice blast resistance gene, Pid3, by DNA fingerprinting to examine microgeographic variation in
genome-wide comparison of paired nucleotide-binding the Magnaporhe grisea (Pyricularia grisea) population in
site-leucine-rich repeat genes and their pseudogene alleles two rice fields in Arkansas. Phytopathology 83:10291035
between the two sequenced rice genomes. Genetics Xiao WM, Yang QY, Wang H, Guo T, Liu YZ, Zhu XY, Chen
182:13031311 ZQ (2010) Identification and fine mapping of a resistance
Sharma TR, Madhav MS, Singh BK, Shanker P, Jana TK, Dalal gene to Magnaporthe oryzae in a space-induced rice
V, Pandit A, Singh A, Gaikwad K, Upreti HC, Singh NK mutant. Mol Breed 28:303312
(2005) High-resolution mapping, cloning and molecular Yaegashi H (1994) Use of resistant varieties and disease control
characterization of the Pi-kh gene of rice, which confers for paddy rice. Agric Hortic 69:149154
resistance to Magnaporthe grisea. Mol Genet Genomics Yang JY, Chen S, Zeng LX, Li YL, Chen Z, Li CY, Zhu XY
274:569578 (2009a) Race specificity of major rice blast resistance
Sharma TR, Rai AK, Gupta SK, Vijayan J, Devanna BN, Ray S genes to Magnaporthe grisea isolates collected from indica
(2012) Rice blast management through host-plant resis- rice in Guangdong. China Rice Sci. doi:CNKI.SUN.SDKE.
tance: retrospect and prospects. Agric Res 1:3752 0.2008.04.011
123
Euphytica
Yang QZ, Lin F, Feng SJ, Wang L, Pan QH (2009b) Recent blast resistance gene which emerged after rice domestica-
progress on molecular mapping and cloning of blast tion. New Phytol 189:321334
resistance genes in rice (Oryza sativa L.). Sci Agric Sin Zhou B, Qu S, Liu G, Dolan M, Sakai H, Lu G, Bellizzi M,
42:16011615 Wang GL (2006) The eight amino-acid differences within
Yuan B, Zhai C, Wang W, Zeng X, Xu X, Hu H, Lin F, Wang L, three leucine-rich repeats between Pi2 and Piz-t resis-
Pan Q (2011) The Pik-p resistance to Magnaporthe oryzae tance proteins determine the resistance specificity to
in rice is mediated by a pair of closely linked CC-NBS- Magnaporthe grisea. Mol PlantMicrobe Interact 19:
LRR genes. Theor Appl Genet 122:10171028 12161228
Zhai C, Lin F, Dong Z, He X, Yuan B, Zeng X, Wang L, Pan Q
(2011) The isolation and characterization of Pik, a rice
123