Isolates Of Magnaporthe Grisea Research Articles

GC-MS analysis of metabolites from blast pathogen infected from foxtail millet

Foxtail millet blast caused by the fungal pathogen Magnaporthe grisea (Anamorph: Pyricularia grisea) is an important and most destructive disease. M. grisea is a ubiquitous plant pathogen with a wide host range. To understand the host-pathogen interaction system, a knowledge about the role of toxins in disease development is paramount. This may helpful in development of a new resistant cultivar screening procedures. The crude toxin extracted from Magnaporthe grisea (TNFxM1) isolate was determined through Gas chromatography–mass spectrometry (GC-MS) technique. The main constituents were Glycerol and 1, 6 Dihydroxynaphthalene.

Inheritance and allelic relationship among gene(s) for blast resistance in pearl millet [Pennisetum glaucum (L.) R. Br.

AbstractSix blast‐resistant pearl millet genotypes, ICMB 93333, ICMB 97222, ICMR 06444, ICMR 06222, ICMR 11003 and IP 21187‐P1, were crossed with two susceptible genotypes, ICMB 95444 and ICMB 89111 to generate F1s, F2s and backcrosses, BC1P1 (susceptible parent × F1) and BC1P2 (resistant parent × F1) for inheritance study. The resistant genotypes were crossed among themselves in half diallel to generate F1s and F2s for test of allelism. The F1, F2 and backcross generations, and their parents were screened in a glasshouse against Magnaporthe grisea isolates Pg 45 and Pg 53. The reaction of the F1s, segregation pattern of F2s and BC1P1 derived from crosses involving two susceptible parents and six resistant parents revealed the presence of single dominant gene governing resistance in the resistant genotypes. No segregation for blast reaction was observed in the F2s derived from the crosses of resistant × resistant parents. The resistance reaction of these F2s indicated that single dominant gene conferring resistance in the six genotypes is allelic, that is same gene imparts blast resistance in these genotypes to M. grisea isolates.

Deciphering Genome Content and Evolutionary Relationships of Isolates from the Fungus Magnaporthe oryzae Attacking Different Host Plants.

Deciphering the genetic bases of pathogen adaptation to its host is a key question in ecology and evolution. To understand how the fungus Magnaporthe oryzae adapts to different plants, we sequenced eight M. oryzae isolates differing in host specificity (rice, foxtail millet, wheat, and goosegrass), and one Magnaporthe grisea isolate specific of crabgrass. Analysis of Magnaporthe genomes revealed small variation in genome sizes (39–43 Mb) and gene content (12,283–14,781 genes) between isolates. The whole set of Magnaporthe genes comprised 14,966 shared families, 63% of which included genes present in all the nine M. oryzae genomes. The evolutionary relationships among Magnaporthe isolates were inferred using 6,878 single-copy orthologs. The resulting genealogy was mostly bifurcating among the different host-specific lineages, but was reticulate inside the rice lineage. We detected traces of introgression from a nonrice genome in the rice reference 70-15 genome. Among M. oryzae isolates and host-specific lineages, the genome composition in terms of frequencies of genes putatively involved in pathogenicity (effectors, secondary metabolism, cazome) was conserved. However, 529 shared families were found only in nonrice lineages, whereas the rice lineage possessed 86 specific families absent from the nonrice genomes. Our results confirmed that the host specificity of M. oryzae isolates was associated with a divergence between lineages without major gene flow and that, despite the strong conservation of gene families between lineages, adaptation to different hosts, especially to rice, was associated with the presence of a small number of specific gene families. All information was gathered in a public database (http://genome.jouy.inra.fr/gemo).

Fertility status of Magnaporthe grisea populations from finger millet

Two hundred twenty nine isolates of Magnaporthe grisea were collected from finger millet cultivars at 14 locations in north India (Uttarakhand) and two locations in south India (Tamilnadu&Karnataka). Each isolate was tested for the mating types (Male fertile and female sterile) using standard fertile testers. Per cent isolates that neither mated nor produced perithecia was high among isolates collected from south India than those from Uttarakhand. Male fertile and hermaphrodite isolates from cooler region showed greater efficiency to produce perithecia. Results revealed that distribution of mating type varied with geographical region. Male fertile and female sterile isolates segregated in 1:1 ratio among field isolates collected from both north and south India. Field isolates from north India showed equal probability of getting male fertile, female sterile and hermaphrodite isolates, which suggested that probability for sexual reproduction in the population of M. grisea existed in nature in the central Himalayas of India.

Relationship Between Resistance Gene Analogue and Blast Resistance in Rice

DNA fragments of 43 rice varieties were amplified with 11 pairs of primers designed based on resistance gene analogue (RGA) of plants, and the blast resistance of the varieties was identified by inoculation with 33 isolates of Magnaporthe grisea collected from Yunnan Province, China. Clustering results revealed a significant correlation between the blast resistance and DNA bands with a correlation coefficient of 0.6117 ( α=0.01), indicating that the resistance analysis based on RGA-PCR clustering analysis coincided with that based on inoculation. The correlation coefficients, ranging from 0.1701 to 0.535, however, depended on the primers. Five pairs of primers, S1/AS3, S1 INV/S2 INV, XLRR For/XLRR Rev, Pto-Kin1 IN/Pto-Kin2 IN, and NLRR For/NLRR Rev might be applied for blast resistance identification in consideration of their band numbers and polymorphisms, and their correlation coefficients with blast resistance were 0.5305, 0.4898, 0.4059, 0.3719 and 0.3524, respectively. Besides, indica and japonica rice except two highly susceptible varieties, CO39 and Lijiangxintuanheigu, could be well classified by the 11 pairs of primers.

Genetic Mapping of Magnaporthe grisea Avirulence Gene Corresponding to Leaf and Panicle Blast Resistant QTLs in Jao Hom Nin Rice Cultivar

AbstractThe avirulence characteristic of Magnaporthe grisea isolate TH16 corresponding to Jao Hom Nin (JHN) rice cultivar was studied by mapping population of 140 random ascospore progenies derived from the cross between B1‐2 and TH16 isolates. Segregation analyses of the avirulence characteristic performing on JHN rice at the seedling and flowering stages were performed in this mapping population. We used the reference map of Guy11/2539 to choose microsatellite DNA markers for mapping the avirulence gene. The genetic map of this population was constructed from 39‐microsatellite markers. The genetic map was spanned by covering seven chromosomes with an average distance of 11.9 cM per marker. In mapping population the distribution of pathogenic and non‐pathogenic progenies on JHN rice were found to be fitted to 1 : 1 ratio for two of the rice stages, seedling and flowering stages. The Quantitative Trait Loci (QTL) analysis for avirulence genes corresponding to two rice stages were located at the same region on chromosome 2 between markers Pyms305 and Pyms435. The LOD score and percentage of phenotypic variance explained (PVE) on two rice stages were 5.01/16.69 and 6.73/20.26, respectively. These loci were designated as Avr‐JHN(lb) and Avr‐JHN(pb) corresponding to leaf and panicle blast characteristics. The findings of this study can be the initial step for positional cloning and identifying any function of avirulence genes corresponding to leaf and panicle blast characteristics.

Development of simple sequence repeat markers for Japanese isolates of Magnaporthe grisea

We developed nine simple sequence repeat (SSR) markers useful for differentiating Japanese isolates of Magnaporthe grisea through a bioinformatic approach. Repeat sequences in the genome of M. grisea were identified by a Tandem Repeat Finding program. Length polymorphisms at 28 loci were examined, nine of which were selected on the basis of detected polymorphisms. These nine SSR markers showed a Nei's gene diversity ranging from 0.23 to 0.91 among 48 field isolates of two natural populations. These SSR markers are well suited for M. grisea epidemiology and population genetics.

Race Specificity of Major Rice Blast Resistance Genes to Magnaporthe grisea Isolates Collected from indica Rice in Guangdong, China

Race-specific resistance and field resistance of 30 rice blast resistance monogenic lines derived from different resources were evaluated. The spectra of resistance to 163 Magnaporthe grisea isolates collected from indica rice in Guangdong Province, China ranged from 0.6% to 89.6%. Most of the monogenic lines showed a narrow resistance spectrum and high susceptibility in rice blast area, whereas the lines with Pik h and Pi1(t) had the broad resistance spectra of 89.6% and 82.2% respectively, showing a high and stable blast resistance in fields. According to the cluster analysis of specific resistance to 163 blast isolates tested, the 30 monogenic lines were divided into 15 groups, and based on the principal factor analysis, nine kinds of race-specific resistance were identified. Pik, Piz 5 , Pi9 and Pish can be used as candidate resistance genes for rice breeding since their specific resistance differed from those of the backbone parents in Guangdong, China. Gene pyramiding of Pik h [or Pi1(t)], Pi9 (or Piz 5 ) and Pish (or Pita 2 ) will be effective to obtain broad-spectrum blast resistance in rice breeding program in Guangdong, China. The strategies for studying and application of rice blast resistance genes were discussed.

Genetic Differentiation of Rice and Non‐rice Populations of Magnaporthe grisea from North‐western Himalayas using Native Protein and Isozyme Polymorphisms

AbstractNative protein and isozyme markers were used to analyse the population structure of rice, finger millet (Eleusine coracana), jungle rice (Echinochloa colonum), goosegrass (Eleusine indica) and crabgrass (Digitaria sanguinalis) infecting isolates of Magnaporthe grisea from the north‐western Himalayan region of India. Both protein and isozyme analyses revealed a high level of genetic diversity among different host‐limited populations of the pathogen including those infecting rice. Cluster analysis of protein and isozyme data permitted the grouping of isolates in accordance with their host specificity. A close association was observed between specific esterase and shikimate dehydrogenase (SDH) isozyme electromorphs and the host specificity of the isolates, indicating that these electromorphs might be linked to the genes conditioning host specialization. Cluster analysis indicated the presence of two lineages in M. grisea species complex: first lineage with moderate bootstrap support was constituted by the isolates from rice, finger millet, goosegrass and jungle rice and a second statistically robust lineage was represented by the isolates from crabgrass. Subpopulations of the pathogen attacking rice and weeds in the same field were genetically distinct and the isolates from different hosts never exhibited a same multilocus haplotype. The overall results indicated that M. grisea populations infecting different hosts were genetically isolated and there was no gene flow among rice and non‐rice isolates of the pathogen.

The Eight Amino-Acid Differences Within Three Leucine-Rich Repeats Between Pi2 and Piz-t Resistance Proteins Determine the Resistance Specificity to Magnaporthe grisea

The rice blast resistance (R) genes Pi2 and Piz-t confer broad-spectrum resistance against different sets of Magnaporthe grisea isolates. We first identified the Pi2 gene using a map-based cloning strategy. The Pi2 gene is a member of a gene cluster comprising nine gene members (named Nbs1-Pi2 to Nbs9-Pi2) and encodes a protein with a nucleotide-binding site and leucine-rich repeat (LRR) domain. Fine genetic mapping, molecular characterization of the Pi2 susceptible mutants, and complementation tests indicated that Nbs4-Pi2 is the Pi2 gene. The Piz-t gene, a Pi2 allele in the rice cultivar Toride 1, was isolated based on the Pi2 sequence information. Complementation tests confirmed that the family member Nbs4-Piz-t is Piz-t. Sequence comparison revealed that only eight amino-acid changes, which are confined within three consecutive LRR, differentiate Piz-t from Pi2. Of the eight variants, only one locates within the xxLxLxx motif. A reciprocal exchange of the single amino acid between Pi2 and Piz-t did not convert the resistance specificity to each other but, rather, abolished the function of both resistance proteins. These results indicate that the single amino acid in the xxLxLxx motif may be critical for maintaining the recognition surface of Pi2 and Piz-t to their respective avirulence proteins.

Isolation, Characterization and Preliminary Genetic Analysis of Laboratory Tricyclazole‐resistant Mutants of the Rice Blast Fungus, Magnaporthe grisea

AbstractThe minimum inhibitory concentration of tricyclazole for hyphal melanization (MIC‐H) was adopted to detect the sensitivity of 129 Magnaporthe grisea isolates collected in China in 2000. Results showed that the mean MIC‐H was 0.2 μg/ml and no isolate with a MIC‐H ≥1 μg/ml was detected. Therefore, 1 μg/ml was chosen as a discriminatory dose to identify resistant mutants generated by ultraviolet (UV) radiation. Only three low‐level resistant (R) mutants derived from the sensitive (S) isolate TH16 were obtained. In addition, fitness decrease was observed for all mutants, with lower sporulation ability and pathogenicity to rice than that of the wild strain TH16. Four crosses between S × R and S × S were tested to determine the inheritance mode of resistance during the process of sexual recombination by analysing the sensitivity of hybrid F1 progeny to tricyclazole. Progeny of crosses between a tricyclazole‐sensitive strain and tricyclazole‐resistant mutants segregated in a 1 : 1 (R : S) ratio and no segregation was found in the cross of S × S, indicating that each mutant contained a single gene for resistance. No nucleotide differences leading to amino acid changes in the coding sequences for 1,3,6,8‐tetrahydroxynaphthalene reductase (4HNR) and 1,3,8‐trihydroxynaphthalene reductase (3HNR) were found between resistant mutants and sensitive strains. Therefore, it is preliminarily concluded that tricyclazole resistance in M. grisea was conferred by a one‐locus mutation in a single Mendelian gene other than those encoding for 4HNR or 3HNR.

Identification and fine mapping of AvrPi15, a novel avirulence gene of Magnaporthe grisea

Avirulence of Magnaporthe grisea isolate CHL346 on rice cultivar GA25 was studied with 242 ascospore progenies derived from the cross CHL346 x CHL42. Segregation analysis of the avirulence in the progeny population was in agreement with the existence of a single avirulence (Avr) gene, designated as AvrPi15. For mapping the Avr gene, we developed a total of 121 microsatellite DNA markers [simple sequence repeat (SSR)], which evenly distributed in the whole-genome of M. grisea through bioinformatics analysis (BIA) using the publicly available sequence. Linkage analysis of the AvrPi15 gene with these SSR markers showed that six markers on chromosome 6, MS6-1, MS6-2, MS6-3, MS6-7, MS6-8 and MS6-10, were linked to the AvrPi15 locus. To further define the chromosomal location of the AvrPi15 locus, two additional markers, MS6-17 and STS6-6, which were developed based on the sequences of telomeric region 11 (TEL11), were subjected to linkage analysis. The results showed that MS6-17 and STS6-6 were associated with the locus by 3.3 and 0.8 cM, respectively. To finely map the Avr gene, two additional candidate avirulence gene (CAG) markers, CAG6-1 and CAG6-2, were developed based on the gene annotation of the sequence of TEL 11. Linkage analysis of the Avr gene with these two markers revealed that both of them completely cosegregated with the AvrPi15 locus. Finally, this locus was physically mapped into approximately 7.2-kb interval of the TEL11 by BIA using these sequence-ready markers. This is the key step toward positional cloning of the AvrPi15 gene.

Recovery and characterization of asexual recombinants ofMagnaporthe grisea

Eleven nitrate non-utilizing (nit) mutants were recovered from six field isolates ofMagnaporthe grisea that had different degrees of sensitivity to the blasticide kitazin P (iprobenfos, IBP). Allnit mutants were resistant to chlorate and there were no significant differences in hyphal growth rate, conidial production, sensitivity to IBP, and pathogenicity betweennit mutants and their parent isolates.nit phenotypes and IBP-resistance were used as two independent genetic markers to study asexual recombination inM. grisea. Asexual recombinants were recovered from the heterokaryotic mycelium of three inter-strain pairings, namely, DY2-3(nitl-LR) + A7-3 (nitM-S); A7-3 (nitM-S) + F4-2 (nitl-MR); and F4-2 (nitl-MR) + DY2-4 (nitA-LR), at a frequency of 7.31%, 14.00% and 8.63%, respectively. The growth rate, conidial production and pathogenicity of asexual recombinants were similar to those of parental strains. We concluded that asexual recombination resulting from hyphal fusion might contribute to variability inM. grisea.

한국·중국 벼 도열병균 생리형 분석 및 품종 저항성 특성 비교

한 중 벼 도열병균의 특성 검정과 저항성원의 공동 평가를 통하여 병원균의 특성을 구명하고 주요 저항성원에 대한 정보와 한국 도열병균의 분화 현상이 중국으로부터 유래하였는지를 분석하였다. 각국의 대표균주 각각 100균주씩을 한국, 중국 판별품종 및 단인자 저항성 품종에 접종한 결과, 한국 병원균은 중국 단인자 저항성 품종에서 29개의 반응형을 보였으며, 한국균주 46%가 중국 NIL(LTH/일본품종 저항성 유전자)을 모두 침해하여, 우리나라 벼 육종 시 저항성원으로 적합지 않은 것으로 나타났다. 또한 한국판별품종에 대한 중국균주의 반응결과, KJ-201, KJ-301, KJ-403, KJ-105, KJ-101 레이스가 각각 11%, 10%, 9% , 8%, 7%의 비율로 나타났다. 그러나 RFLP-MGR586 profile은 한국 균주가 중국균주보다 band 수와 양상이 다양하여 변이가 심한 것으로 나타났으며, IRRI에서 육성한 11개의 단인자 저항성 품종에 대하여 한-중균주의 유전자 침해비율은 대체로 비슷한 경향이었다. Pi-z, Pi-3, Pi-5(t)는 한국 균주가 각각 56.6%, 58.1%, 45.4%침해하는 반면, 중국 균주는 각각 85.2%, 75.9%, 75.9%를 침해하여 중국 균주의 침해 비율이 높았고 Pi-b 유전자의 경우, 한국 균주의 82.6%가 침해하는 반면 중국 균주는 59.3%가 침해하여 한국 균주의 침해 비율이 높았다. One hundred isolates of Magnaporthe grisea from Korea and China were characterized for pathogenicity using eight Korean differential varieties(KDV), six Chinese differential varieties(CDV) and six near isogenic lines(NILs) developed in China. The restriction length polymorphism of M. grisea isolates from each country also was analyzed using MGR586 as a probe. One hundred Korean isolates classified into 17 races on KDV were grouped into 29 pathotypes on Chinese near isogenic lines(NILs). Virulence of 46% of Korean isolates against all the six Chinese NILs indicated that the current six Chinese NILs alone was not enough to be used as differential varieties in Korea. Especially, susceptibility of the BL1 carrying resistance gene Pi-b to 70% of tested Korean isolates suggested that BL1(Pi-b) may not be a useful resistance source to Korean blast. Based on the virulence assays of M. grisea populations from each country were divided into two groups. About 50% of Chinese isolates showed similarity to the 30% of the Korean isolates. Especially, the isolates from northern part of China, where Japonica rice cultivars were grown, showed high similarity to the Korean isolates, while isolates from southern part of China, where Indica rice were mainly grown, showed low similarity to Korean isolates. The genome RFLPs of Korean isolates were quite different from those of southern part of China using MGR586 as a probe. These data indicated that the physiological and genetical characteristics of M. grisea population might be determined by strong interaction with cultivated rice.

Differential expression of rice Nramp genes in response to pathogen infection, defense signal molecules and metal ions

The Nramp genes encode a class of metal transporters that universally exist in both eukaryotes and prokaryotes. In animals such as the mouse, the Nramp1 gene is important for natural host resistance to the infection of intracellular pathogens. To investigate the potential role of Nramp genes in plant defense response, we have conducted expression analysis of three OsNramp genes in rice seedlings and cell suspension cultures. In 2-week-old rice seedlings, the expression of OsNramp1 appeared to be suppressed by Burkholderia glumae during the susceptible interaction, unaffected by a virulent Magnaporthe grisea isolate, but was significantly induced by an avirulent M. grisea isolate during the resistant interaction. The expression of OsNramp2 and OsNramp3 was also significantly induced by B. glumae and M. grisea, but their induction by the M. grisea isolate was weak during the susceptible interaction. In rice leaf tissues, OsNramp1 transcription was delayed and suppressed by jasmonic acid (JA) and abscisic acid (ABA), respectively, while OsNramp2 was induced by JA, ABA and ethephon. In rice suspension cells, OsNramp3 was activated by the M. grisea cell wall elicitor and ABA. The elicitor-stimulated OsNramp3 expression was Ca 2+ dependent and could be accentuated by cycloheximide (CHX). Both OsNramp1 and OsNramp2 had relatively high basal expression in suspension cells, and their expression was not significantly affected by the defense signal molecules. However, OsNramp1 transcript was increased drastically with EDTA, while OsNramp1 and OsNramp2 were slightly suppressed by 10 mM ferrous iron. In contrast, OsNramp3 was induced by 10 mM ferrous iron and zinc. These results indicate that rice Nramp genes are differentially expressed in response to pathogen infection, defense signals and metal ions, and are potentially involved in plant defense response.

Resistance spectra of wheat cultivars and virulence diversity of Magnaporthe grisea isolates in Brazil

Seventy-two monoconidial isolates of Magnaporthe grisea were obtained from the States of Mato Grosso do Sul and Paraná. The isolates were inoculated on seedlings of 20 wheat (Triticum aestivum) cultivars under greenhouse conditions. The virulence diversity of M. grisea was assessed based on compatible and incompatible reactions of leaf blast on wheat cultivars. Fifty-four distinct virulence patterns were identified on test cultivars among the isolates collected from the two wheat growing States. Sixteen of these isolates corresponding to 22.2% showed similar virulence pattern. None of the wheat cultivars was resistant to all isolates of M. grisea, but the cultivars differed in degree of resistance as measured by the relative spectrum of resistance (RSR) and disease index (DI). Among the cultivars the RSR ranged from 0 to 53.3% and DI from 0.4662 to 0.9662 (0 to 1 scale). The wheat cultivar BR18 exhibited a broad resistance spectrum in relation to the rest of the tested cultivars to the isolates of M. grisea, and can be used in wheat resistance breeding.

Identification of avirulence genes in the rice blast fungus corresponding to three resistance genes in Japanese differentials

The pathogenicity of progeny from crosses among three Chinese isolates of Magnaporthe grisea collected from rice was tested on three Japanese differentials (Ishikarishiroke, Aichiasahi, K 59) having the blast resistance genes Pii, Pia, and Pit, respectively. Monogenic control was demonstrated for avirulence to the differentials. To identify resistance genes corresponding to the avirulence genes, the resistance and susceptibility in F3 lines of the cultivars in response to the parents of the crosses were analyzed genetically. The three avirulence genes identified, designated Avr-Pii, Avr-Pia, and Avr-Pit, appear to correspond to resistance genes Pii, Pia, and Pit, respectively. The monogenic control of avirulence in the fungus and monogenic dominant resistance in rice cultivars supports a gene-for-gene relation in the Pii-, Pia-, or Pit-dependent resistance to the rice blast fungus in rice cultivars.

Occan, a novel transposon in the Fot1 family, is ubiquitously found in several Magnaporthe grisea isolates.

We investigated a DNA fragment and its flanking region deleted in the spontaneous Pi-a virulent mutant of Magnaporthe grisea Ina168. A new transposon-like sequence was identified from a region adjacent to the deleted fragment and was named Occan. Occan contained a 2,259-bp ORF interrupted by one 63-bp intron and had both a TA dinucleotide and 77 bp of perfect inverted repeats at both termini, without direct repeats. These features indicated that Occan is a member of the Fot1 family. RT-PCR analysis confirmed the expression of the putative transposase and the presence of an intron. Southern analysis of pulse-field gel electrophoresis-separated chromosomes indicated that Occan was dispersed in all chromosomes of the rice pathogen, Ina168. Copy numbers of Occan were also preserved in a host-specific manner amongst M. grisea isolates. In particular, rice pathogens contained a large number of the element inserted into their genome. Phylogenetic analysis with other known members of the Fot1 family revealed that Occan was dissimilar to any other known elements and it is thus proposed that Occan be separated to a new subfamily.

Relationship between Avirulence Genes of the Same Family in Rice Blast Fungus Magnaporthe grisea

A genetic cross between rice-field isolates of Magnaporthe grisea produced progeny segregating for avirulence/ virulence on six rice cultivars among nine race differentials, while on three other cultivars, Shin 2 (Pik-s), Aichi Asahi (Pia) and Ishikari Shiroke (Pii), parental and progeny isolates were all virulent. Based on segregation ratios in 115 progeny isolates, avirulence on Kanto 51 (Pik), Yashiro-mochi (Pita), Fukunishiki (Piz) and Toride 1 (Piz-t) is under monogenic control. On Tsuyuake (Pik-m) and Pi No. 4 (Pita-2), however, a disproportionate ratio in the segregation was observed, suggesting that avirulence on these two cultivars is controlled by two or more genes. Assuming that the avirulence gene AvrPik-m consists of at least two genes, AvrPik-m1 and AvrPik-m2, each of which functions in the whole gene AvrPik-m, and that one of AvrPik-m1 and AvrPik-m2 is AvrPik, we could account for the disproportion in the avirulence/virulence segregation of the progeny. This hypothesis would also be consistently applied for avirulence gene AvrPita-2. There seem to be two types of the avirulence genes : AvrPik-m, that is comprised of the tightly linked genes, AvrPik-ml (=AvrPik) and AvrPik-m2, and AvrPita-2, that is comprised of the loosely linked genes AvrPita-2A (=AvrPita) and AvrPita-2B. As one possible explanation of the rice resistant reaction to blast, multiple specificity was suggested for the first time for the blast fungus. On the contrary, the avirulence genes AvrPiz and AvrPiz-t were inherited independently, despite the corresponding genes for resistance (Piz and Piz-t) being located at the same locus. The cross of rice blast isolates (races 447 and 337) produced only 25 kinds of races in the progeny, although theoretically about 64 kinds of races should be produced if six avirulence genes segregated independently. Because no progeny are with AvrPik (or AvrPita) and without AvrPik-m (or AvrPita-2), the number of races theoretically should be 36 at most. A number of strains, such as races 377 and 737, with a single avirulence gene were obtained from this cross. These strains may be valuable for analysis of resistance genes in rice plant.

RAPD and Pathotype Analyses of Magnaporthe grisea Populations from the north‐western Himalayan Region of India

AbstractUsing random amplified polymorphic DNA (RAPD) analysis, 250 isolates of Magnaporthe grisea collected from the north‐western Himalayan region were separated into 25 DNA fingerprint groups or lineages. Of these 25 groups, 13 were exclusive to isolates obtained from Himachal Pradesh (HP), five from Uttaranchal and one from Jammu and Kashmir (J & K), India. Seven remaining groups were composed of isolates from different locations and 26 isolates could not be classified. Although RAPD analysis revealed high genetic variability among M. grisea populations from HP and J & K, genetic variation was low in the isolates collected from Uttaranchal. DNA fingerprint groups specific to a particular geographical region were also obtained. Pathogen population consisting of 119 isolates from north‐western Himalayan region has been grouped into 52 pathotypes on the basis of disease reaction on international differential rice lines. Highest frequency of virulence was trapped on the rice line Caloro (Pi−ks) followed by NP125 (Pi−? and K‐60 (Pi−kp) while it was lowest on Tadukan (Pi−ta and/or Pi−ta2) and BL‐1(Pi−b and Pi−sh). Virulence was rare on Fukunishiki (Pi−zs). Rice line Tetep (Pi−kh+?) was found to be highly effective in north‐western Himalayan region as none of the isolate could infect this line. These blast resistant rice lines can be used in resistance breeding for the effective management of rice blast in this region of India.