Rice Genetics IV - IRRI books - International Rice Research Institute

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analysis suggests that the mutation in GR-740-12-8 is dominant. Another gammaray-inducedmutant, GR-978-18-18, shows strong resistance to 10 races of Xoo fromthe Philippines and to two isolates of the blast pathogen. This mutant is slightly shorterthan the wild-type IR64. Another mutant, GR-282-12-15, showed a gain in resistanceto a single isolate of Xoo and two blast isolates. Segregation patterns in the M 3 generationsuggest that both mutations in GR-978-18-18 and in GR-282-12-15 are recessive.Overall, a majority of these gain-in-resistance mutants are morphologically abnormal,ranging from semidwarf to severe stunting. In Arabidopsis, some gain-inresistancemutants were also morphologically abnormal (Clough et al 2000). As ofnow, we have not identified mutants showing susceptibility to multiple pathogens,indicating a need to expand the screening to detect these presumably rare mutations.The most common class of disease-response mutants is those with lesion mimics.Yin et al (2000) demonstrated that some but not all lesion mimic mutants (in anothergenetic background of indica rice IR68) exhibit enhanced resistance to blast and bacterialblight. To further delineate the pathways involved in disease and defense responses,we selected 21 lesion mimic mutants from the IR64 mutant collection. Wefound that these mutants showed a varying degree of enhanced resistance to differentbacterial blight strains and that resistance was not race-specific. Two mutants, D6-1851 and D6-242, with reduced resistance to Xoo and M. grisea were monitored forexpression of a pathogenesis-related (PR) protein gene PBZ1 (Fig. 2). In the wildtypeand single mutants, the expression of PBZ1 was strongest in the second youngestleaf (L2) but decreased in the older leaves (L3 and L4). In the double mutant, whichshowed enhanced resistance, high expression of PBZ1 was maintained in the olderleaves.Double mutants have been created by crossing independently inherited lesionmimic mutants to evaluate the epistatic relationships among these mutations. Defenseresponse (DR) genes PR-1, peroxidase 22.3 (POX22.3), and peroxidase POC1(POC1) were used to probe mRNA from four leaf stages of the single and doublemutants derived from D6-1851 × D6-242. POX22.3 expression was high in youngleaves and diminished as lesions developed. PR-1 and POC1 expression was enhancedrelative to that observed in IR64 in the second and third leaves. In general, DR geneexpression was positively correlated with enhanced disease resistance to bacterialblight. These results suggest that the expression of these DR genes is developmentallycontrolled and regulated by the lesion mimic genes and that mis-regulation ofcertain defense-related genes was correlated with the resistant phenotypes. Severalsubtractive cDNA libraries induced by pathogens have been constructed and a panelof candidate genes is being used to correlate gene expression with broad-spectrumdisease resistance (J. Leach, published data).Sheath blight caused by Rhizoctonia solani is a difficult disease to evaluate becauseof the variability associated with the disease response and the strong environmentalinfluence on pathogen-host interactions. However, there is an urgent need tosearch for endogenous genes that can enhance sheath blight resistance as no geneticresistance has yet been found through conventional screening of rice germplasm. Ourstrategy is to first identify mutants with a gain or loss in resistance to blast and bacte-244 Leung et al
ALesion length (cm)252015105IR64D6-1851D6-242DM09 11 13 15Days after inoculationBLeaf ageIR64 D6-1851 D6-242 DML1 L2 L3 L4 L1 L2 L3 L4 L1 L2 L3 L4 L1 L2 L3 L4PBZ128SrRNAFig. 2. (A) Reduced susceptibility to bacterial blight in single and double mutants (DM) carryinglesion mimic mutations. (B) Expression of PBZ1 in leaf of different ages (L1 is youngest andL4 is oldest). The double mutant maintains a high level of expression in older leaves, whichcorresponds to enhanced resistance to bacterial blight.rial blight, and then evaluate these mutants with sheath blight under field conditions.So far, several lesion mimic mutants have been evaluated for reaction to sheath blightbut no enhanced resistance has been found. In addition, we have begun screening fortolerance of tungro viruses using viruliferous green leafhopper carrying both the spherical(RTSV) and bacilliform (RTBV) viruses. From a screening of about 2,000 lines,six putative mutants with different degrees of tolerance of RTSV were identified, andthese are being confirmed for both the phenotypes and genotypes.Abiotic stressesWe have initiated mutant screenings under drought, submergence, salinity, and zincdeficiency conditions. In preliminary screening experiments, quantitative variationwas observed in zinc deficiency tolerance but no variability for salinity stress (100mM NaCl) was found. From a screening of approximately 3,000 lines for submergencetolerance under field conditions, G. Gregorio and D. Adorada (personal communication)identified six putative mutants that give 75% recovery after 14-d submergencetreatment at the seedling stage. These mutants are being reevaluated toconfirm inheritance of submergence tolerance and yield performance.Deletion mutants for functional genomics: . . . 245
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Retrotransposons of rice as a tool
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First, the Rice Genetics Cooperativ
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OverviewRice genetics from Mendel t
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Rice genetics from Mendelto functio
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nese cultivar Nipponbare. The chrom
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Gene symbolization in riceIn the ab
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Table 2. Some examples of mapping g
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and YAC libraries, respectively. Th
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gene from wild species is the intro
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ReferencesAbbasi FM, Brar DS, Carpe
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Nagao S. 1951. Genic analysis and l
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Yoshimura S, Yamanouchi U, Katayose
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important traits for which segregat
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trolled by a single recessive gene,
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AromaAromatic rice varieties often
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Submergence tolerance is an interes
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Kinoshita T, Maekawa M. 1986. Genet
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NotesAuthors’ addresses: J.N. Rut
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The Rockefeller Foundation has a lo
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Evolution and implementation of the
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Scientific progress and outputsIn t
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Another salient example is that of
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BOX NO. 1recent international works
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For the purposes of this discussion
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Because of the great diversity (sci
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eeders where final products to enha
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Chen S, Lin XH, Xu CG, Zhang Q. 200
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Toenniessen GH. 1998. Rice biotechn
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Molecular markers,genetic diversity
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Evolution and domestication of rice
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ABCDO. barthiiO. rufipogonO. longis
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Wild(O. rufipogon)Geographical diff
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South Asia (particularly on the wes
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al 1992, Sun et al 1996a,b,c), thou
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wild and cultivated types (Est10, W
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Cai HW, Morishima H. 2000a. Diversi
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Sato YI, Tang SX, Yang LU, Tang LH.
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The solid base laid by classical ri
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The synthesis shown in Figure 1 is
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Arabidopsis, long heralded as a “
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Monocots and eudicots: Is there sti
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Zhang H, Jia J, Gale MD, Devos KM.
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ility of rice productivity (Lu 1999
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Table 1 continuedIntrageneric class
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Molecular markers and evolutionary
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A100100521010078100991009894100O. s
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Two distinct types of sequences wer
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arthii and O. longistaminata, and t
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testing Ka/Ks between the homeologo
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Roschevicz RI. 1931. A contribution
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Miniature inverted repeattransposab
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eau and Wessler 1992, 1994), rice (
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MITEs as a source of allelic divers
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ATIR Olo-D* Olo-CTIRBCas Exp Wan Sn
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Le QH, Wright S, Yu Z, Bureau T. 20
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Microsatellite markers in rice:abun
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make the best SSR markers for rice.
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Predicted frequency100,00080,000Cla
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Relationship between SSRs and genes
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are indicated with RM locus designa
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SSRs have been used to define intro
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provide a bridge for moving rapidly
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Ishii T, McCouch SR. 2000. Microsat
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NotesAuthors’ addresses: S.R. McC
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traits of economic importance (Mack
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Table 2. Tagging and mapping of som
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even at the juvenile stage. Several
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Chen et al (2000) transferred the b
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genome sequence of rice, this will
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Kurata N, Nagamura Y, Yamamoto K, H
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Witcombe JR, Hash CT. 2000. Resista
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QTL mapping in rice:a few critical
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M-QTLsM-QTLs are defined as single
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(genetic/statistical models and cor
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Table 5. The percentage of three ty
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Table 7. Some environment-specific
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ferently to the environments. Simil
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(case 3), one may infer that this g
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Simultaneous QTL introgression and
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Doebley J, Stec A, Gustus C. 1995.
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Visscher PM, Haley CS, Thompson R.
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unavailable until recently with the
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Table 1 continued.Trait QTL a Flank
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the remaining parts of the linkage
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Table 3. QTLs detected for yield an
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Table 6. Correlations of various pa
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The functions of these sequences ne
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Structural and functional genomicsT
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The International Rice GenomeSequen
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the RGP, a PAC (P1-derived artifici
Page 205 and 206: Progress of sequencing in the RGPWe
Page 207 and 208: ice genome sequencing to organize a
Page 209 and 210: Strategies and techniquesfor finish
Page 211 and 212: uses the quality values generated b
Page 213 and 214: with the advanced techniques requir
Page 215 and 216: times contain sufficient data to ma
Page 217 and 218: amplification of regions that previ
Page 219 and 220: 4. Structure problems—subcloning
Page 221 and 222: estriction maps or optical maps are
Page 223 and 224: nal assembly may point to areas tha
Page 225 and 226: McMurray AA, Sulston JE, Quail MA.
Page 227 and 228: Sequence-tagged connector/DNA finge
Page 229 and 230: 1999). The development of multiple-
Page 231 and 232: Transposable elements in the rice H
Page 233 and 234: ers, maps, mapping populations, and
Page 235 and 236: Miropeats-OSJNBa0065H03 1.2 cMThres
Page 237: NotesAuthors’ addresses: R.A. Win
Page 240 and 241: Sasaki 1997). Such analysis is ofte
Page 242 and 243: ABNipponbare × KasalathF 2QTL anal
Page 244 and 245: were classified into two groups bas
Page 246 and 247: effective at determining the genoty
Page 248 and 249: addition, some accessions of wild r
Page 250 and 251: Sasaki T, Burr T. 2000. Internation
Page 252 and 253: Mutational analysis has long been t
Page 254 and 255: marked by a deletion/chromosomal re
Page 258 and 259: For drought-response screening, our
Page 260 and 261: Pi-7(t)Xa21Xa10Xa3Xa4Pi-1(t)Pi-k, P
Page 262 and 263: Although mutants with discrete gene
Page 265 and 266: Generation of T-DNA insertionaltagg
Page 267 and 268: Aprobe AEEprobe BpGA1633RBgus Tn p3
Page 269 and 270: Number of loci36%224%170%010 20 30F
Page 271 and 272: Table 2. GUS assay in roots of tran
Page 273 and 274: 1999, Krysan et al 1999, Sato et al
Page 275 and 276: Transposons and functionalgenomics
Page 277 and 278: cesses. To study the interactions a
Page 279 and 280: Constructs were made with the aim o
Page 281 and 282: RBcis-effect of the adjacent strong
Page 283 and 284: Ac knockout mutagenesisThe Ac lines
Page 285 and 286: Table 2 summarizes the transformati
Page 287 and 288: ReferencesBaldwin D, Crane V, Rice
Page 289: NotesAuthors’ addresses: R. Greco
Page 292 and 293: These include T-DNA (Azpiroz-Leehan
Page 294 and 295: polyproteins and two identical 138-
Page 296 and 297: mutant. The mutation in the ent-kau
Page 298 and 299: primers (G1, G2) and two Tos17-spec
Page 300 and 301: of insertion mutants by sequencing
Page 302 and 303: Grandbastien M-A, Spielman A, Caboc
Page 304 and 305: NotesAuthors’ addresses: H. Hiroc
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human endeavor, greatly contributed
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Equally important as the genomic in
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Oryzabase is one of the most recent
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Database structureA characteristic
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types of users. An annotation datab
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ReferencesAntonio BA, Fang Z, Sanch
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Gene isolation and functionEnhancin
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Enhancing deployment of genes forbl
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mochi and Tsuyuake (Wu et al 1996).
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tive indica rice varieties supports
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transcription factor, resulting in
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facilitate breeding durable resista
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(Inukai et al 1994, Yu et al 1996,
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Rossman AY, Howard RJ, Valent B. 19
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Molecular signaling in diseaseresis
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AOsRac1IED II III IV 214aaOsRac1 KC
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Japonica rice variety Kinmaze, whic
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Table 1. Characteristics of lesion-
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an effective inducer of host resist
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Takahashi A, Kawasaki T, Henmi K, S
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et al 2000). Sequence analysis of t
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612-amino acid protein, including t
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Genetic dissection of the Xa21-medi
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ReferencesAnderson PA, Lawrence GJ,
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Isolation of candidate genesfor tol
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Materials and methodsPlant material
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Microarray hybridization and data a
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Table 3. Most abundant transcripts
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Microarray technologyThe rapid accu
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view of procedures, pitfalls, and n
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Log(2) ratio1.51.0OC104E01—WCP1OC
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Table 5. Strongly regulated transcr
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ReferencesAdams MD, Soares MB, Kerl
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Acknowledgments: We thank Chris Bor
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y cells separating during tissue de
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(Erwee and Goodwin 1983). The sympl
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tion between the abundance of CDK t
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in a complex of 105 kDa. These resu
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Drew MC, Jackson MB, Giffard S. 197
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Umeda M, Umeda-Hara C, Yamaguchi M,
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asexual reproduction through apomix
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MPManual pollination by breeders×
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Search for apomixis in riceThere ar
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Issues related to achieving synthet
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come and may use imprinting to achi
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Embryo-inducing geneInactive ablati
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M 1 3 3 4 5 6 7 8 9 10 11 12 13 14
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embryos, several genes characterist
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Our search for a rice homologue of
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sequenced the two rice DMC1 genes a
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Gustine DL, Sherwood RT, Huff DR. 1
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Peel MD, Carman JG, Leblanc O. 1997
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TransformationEngineering for virus
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Engineering for virusresistance in
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proteins, (2) the expression of dys
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Antisense and cosuppression RNA. Ex
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Table 1. Transgenic rice with patho
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the disease symptoms. Efforts from
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an ambisense coding strategy (Fig.
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first produced and shown to have vi
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McLean GD, Evans G. 1997. Some pote
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Waterhouse PM, Upadhyaya NM. 1998.
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duction in response to dehydration
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(KIKEKLPG) in the carboxyl terminus
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These three plasmids were used to t
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We now show the effects of the toba
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Since one major goal of plant scien
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Table 4. Copy number of transgenes
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Chan M-T, Chang H-H, Ho S-L, Tang W
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NotesAuthors’ address: Department
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initially fixed by phosphoenolpyruv
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High-level expression of maize C 4-
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The effect of illumination on PPDK
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ate in 2% O 2 can be limited by Pi
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Transgene integration,organization,
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duced by organogenesis, somatic emb
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ing transgene organization. FISH an
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ments and the integration of exogen
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Level 2: activation of local repair
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strongly suggested that plant facto
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other cases it was not. We observed
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Gorbunova V, Levy AA. 1997. Non-hom
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Gene silencing and itsreactivation
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Carbofuran (trade name, Furadan) is
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Akb4xHpaIIJKA52 (R 0)52-6 (R 1)52-9
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ABCDhptuidAmUbi1kb10.08.05.04.13.0F
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Fig. 4. X-gluc staining of germinat
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eflect different causes for silenci
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Table 4. Suppressors of silencing f
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Garrick D, Fiering S, Martin DI, Wh
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Xu Y, Buchholz WG, DeRose RT, Hall
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Rice molecular breeding workshopThi
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the working group can develop a net
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Rice genetics cooperativeA meeting
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