Genetic Analysis of Host Species Specificity of Magnaporthe oryzae Isolates from Rice and Wheat

ABSTRACT A Triticum isolate (pathogenic on wheat) of Magnaporthe oryzae was crossed with an Oryza isolate (pathogenic on rice) to elucidate mechanisms of their parasitic specificity on wheat. When the pathogenicity of their F (1) cultures (hybrids between a Triticum isolate and an Oryza isolate) was tested on wheat, avirulent and virulent cultures segregated in a 7:1 ratio. This result suggests that three loci are involved in avirulence of the Oryza isolate on wheat. One of the three loci conditioned papilla formation, whereas the others conditioned the hypersensitive reaction. Allelism tests revealed that the locus conditioning papilla formation is Pwt2 while one of the two loci conditioning the hypersensitive reaction is Pwt1. The other locus conditioning the hypersensitive reaction was different from any other known loci and, therefore, was designated as Pwt5.     

Characterization of interactions between barley and various host-specific subgroups of Magnaporthe oryzae and M. grisea

The Magnaporthe oryzae–M. grisea species complex is composed of several host-specific subgroups, but does not contain a barley-specific subgroup. To characterize the relationship between barley and these subgroups, we inoculated 24 barley cultivars separately with each of 18 isolates from various hosts. The interactions between these cultivars and isolates included various reactions from nonhost-like immune responses to typical host responses. Evenly closely related isolates of the blast fungi caused such contrasting reactions. The immune responses of barley cultivars against a Setaria isolate, Si-1J, were examined in detail. An infection assay with near-isogenic fungal strains suggested that PWT1, which was first identified as a major gene conditioning the avirulence of Si-1J on wheat, was involved in the avirulence on two-thirds of the barley cultivars. At the cytological level, the immune responses were associated with both papilla formation and hypersensitive reaction (HR). Of these two, however, HR played a more critical role than papilla formation. Studying the interactions of barley with M. oryzae and M. grisea may reveal various steps in the process of host specialization of a parasite species and the concomitant evolution of host resistance.     

Inheritance of resistance to bacterial blight in 21 cultivars of rice

ABSTRACT Genetic analysis for resistance to bacterial blight (Xanthomonas oryzae pv. oryzae) of 21 rice (Oryza sativa L.) cultivars was carried out. These cultivars were divided into two groups based on their reactions to Philippine races of bacterial blight. Cultivars of group 1 were resistant to race 1 and those of group 2 were susceptible to race 1 but resistant to race 2. All the cultivars were crossed with TN1, which is susceptible to all the Philippine races of X. oryzae pv. oryzae. F(1) and F(2) populations of hybrids of group 1 cultivars were evaluated using race 1 and F(1) and F(2) populations of hybrids of group 2 cultivars were evaluated using race 2. All the cultivars showed monogenic inheritance of resistance. Allelic relationships of the genes were investigated by crossing these cultivars with different testers having single genes for resistance. Three cultivars have Xa4, another three have xa5, one has xa8, two have Xa3, eight have Xa10, and one has Xa4 as well as Xa10. Three cultivars have new, as yet undescribed, genes. Nep Bha Bong To has a new recessive gene for moderate resistance to races 1, 2, and 3 and resistance to race 5. This gene is designated xa26(t). Arai Raj has a dominant gene for resistance to race 2 which segregates independently of Xa10. This gene is designated as Xa27(t). Lota Sail has a recessive gene for resistance to race 2 which segregates independently of Xa10. This gene is designated as xa28(t).     

A Gene-for-Gene Relationship Underlying the Species-Specific Parasitism of Avena/Triticum Isolates of Magnaporthe grisea on Wheat Cultivars

ABSTRACT To elucidate genetic mechanisms of the species-specific parasitism of Magnaporthe grisea, a Triticum isolate (pathogenic on wheat) was crossed with an Avena isolate (pathogenic on oat), and resulting F(1) progeny were subjected to segregation analyses on wheat cvs. Norin 4 and Chinese Spring. We found two fungal loci, Pwt3 and Pwt4, which are involved in the specific parasitism on wheat. Pwt3 operated on both cultivars while Pwt4 operated only on 'Norin 4'. Using the cultivar specificity of Pwt4, its corresponding resistance gene was successfully identified in 'Norin 4' and designated as Rmg1 (Rwt4). The presence of the corresponding resistance gene indicated that Pwt4 is an avirulence locus. Pwt3 was assumed to be an avirulence locus because of its temperature sensitivity. We suggest that gene-for-gene interactions underlie the species-specific parasitism of M. grisea.     

The identification of a gene for race-specific resistance to Peronospora parasitica (downy mildew) in Brassica napus var. oleifera (oilseed rape)

The oilseed rape cultivar Cresor was resistant to 14 isolates of Peronospora parasitica derived from crops of Brassica napus in the UK. Segregation for resistance to one isolate among F₂ plants and F₃ progeny of crosses between Cresor and the susceptible cultivars Victor and Jet Neuf indicated that resistance was controlled by a single gene. There was evidence that genetic background and environment could influence the phenotypic expression of this resistance. Two sexual progeny isolates derived from a homothallic isolate of P. parasitica avirulent on Cresor were completely virulent on this cultivar. This suggested that the parental isolate was heterozygous at a matching locus or loci for avirulence and demonstrated the race-specific nature of the resistance.     

谷瘟病菌无毒基因AVR1-CO39序列变异及遗传多样性分析

 谷瘟病是谷子上毁灭性病害之一,为了探讨不同地区谷瘟病菌群体的遗传多样性,对我国11个省(自治区)171株谷瘟病菌的无毒基因AVR1-CO39进行扩增测序,并利用ClustalX2.0和DnaSP5.0软件对测序结果进行分析。结果表明,171株谷瘟病菌单孢菌株AVR1-CO39的CDS编码区共有40个多态性位点,依据序列之间的核苷酸差异划分为37个单倍型,H1型为绝对优势单倍型。我国11个省份谷瘟病菌群体的AVR1-CO39具有较高的遗传多态性,由于存在较为频繁的基因交流,种群之间没有明显的遗传分化。种群内部的遗传分化是谷瘟病菌遗传分化的主要方式,错配分布检测结果显示进化过程中可能出现群体扩张,并且谷瘟病菌的聚类与地理来源没有显著的关系。研究结果表明,谷瘟病菌无毒基因AVR1-CO39具有较高的变异性,以H1为核心单倍型在不断地变异衍生出新的等位基因类型,并且这种变异衍生趋势并不受地理隔离的影响。研究结果可为谷子抗病品种选育,揭示谷瘟病菌无毒基因与谷子抗病基因之间的互作机制提供理论支持。     

Pi34-AVRPi34: a new gene-for-gene interaction for partial resistance in rice to blast caused by Magnaporthe grisea

The japonica rice (Oryza sativa) cultivar Chubu 32 has a high level of partial resistance to blast, which is mainly controlled by a dominant resistance gene located on chromosome 11. The partial resistance to the rice blast fungus (Magnaporthe grisea) in Chubu 32 has isolate specificity; isolate IBOS8-1-1 is more aggressive on Chubu 32 than are other isolates. We hypothesized that the gene-for-gene relationship fits this case of a partial resistance gene in Chubu 32 against the avirulence gene in the pathogen. The partial resistance gene in Chubu 32 was mapped between DNA markers C1172 (and three other co-segregated markers) and E2021 and was designated Pi34. In the 32 F₃ lines from the cross between a chromosome segment substitution line (Pi34⁻) from Koshihikari/Kasalath and Chubu 32, the lines with high levels of partial resistance to the M. grisea isolate Y93-245c-2 corresponded to the presence of Pi34 estimated by graphic genotyping. This indicated that Pi34 has partial resistance to isolate Y93-245c-2 in compatible interactions. The 69 blast isolates from the F₁ progeny produced by the cross between Y93-245c-2 and IBOS8-1-1 were tested for aggressiveness on Chubu 32 and rice cultivar Koshihikari (Pi34⁻). The progeny segregated at a 1 : 1 ratio for strong to weak aggressiveness on Chubu 32. The results suggested that Y93-245c-2 has one gene encoding avirulence to Pi34 (AVRPi34), and IBOS8-1-1 is extremely aggressive on Chubu 32 because of the absence of AVRPi34. This is the first report of a gene-for-gene relationship between a fungal disease resistance gene associated with severity of disease and pathogen aggressiveness.     

Genetic analysis of host–pathogen incompatibility between Lolium isolates of Pyricularia oryzae and wheat

Lolium isolate TP2 of Pyricularia oryzae, causal agent of gray leaf spot of perennial ryegrass (Lolium perenne), is virulent on perennial ryegrass, but avirulent on wheat cultivars. Genetic analysis of wheat F₂ populations revealed that the resistance of wheat cultivars Chinese Spring, Shin-chunaga, and Norin 4 to TP2 was conditioned by two genes, R1 and R2. R1 was highly effective, while R2 was less effective. The strong resistance gene R1, designated Rmg6, was mapped on chromosome 1D using microsatellite markers. For revealing genetic mechanisms of avirulence, TP2 was crossed with Triticum isolate Br48. Segregation analysis of their F₁ progenies revealed that the avirulence of TP2 on the three wheat cultivars was conditioned by two unlinked genes, one (A1) highly effective and the other (A2) less effective. These results suggest that the incompatibility between TP2 and the common wheat cultivars is conditioned by two gene pairs; the Rmg6–A1 interaction results in strong resistance, and the R2–A2 interaction results in moderate resistance.     

The genetic control and expression of specificity in Bremia lactucae (lettuce downy mildew)

The inheritance of specific virulence in Bremia lactucae was studied in crosses involving 12 heterothallic isolates of the fungus. In one cross, virulence to eight of the II specific resistance factors examined segregated in the F₁ generation. Although there were exceptions, most of the data were consistent with the hypothesis that pathogenicity was controlled by independent single loci with avirulence dominant to virulence. Linkage between loci determining virulence on R2 and R11 was confirmed. Loci controlling virulence on R5 and R8 also appeared to be linked but these may be identical R-factors. Contrary to a previous suggestion, the locus determining virulence to R10 was independent of that for R5/R8. The expression of virulence to three R-factors (Rl, R4 and R5/R8) was influenced by independent second loci. The presence of a dominant allele at the second locus inhibited avirulence. The expression of avirulence on R6 seemed to be influenced by modifier genes and environment in some isolates. Although the cultivars Mildura, Bourguignonne, Sucrine and Captain were originally thought to contain a single resistance factor, these data suggest that Sucrine carries R5/R8 in addition to R10 whilst Mildura may carry Rl in addition to R3. The stock of Bourguignonne appeared to be a mixture of resistance genotypes. The data add additional support to the suggestion that Capitan (Rll) may carry two resistance factors.     

A new blast resistance gene identified in the Indian native rice cultivar Aus373 through allelism and linkage tests

Blast, caused by Pyricularia grisea , is a major constraint on rice production. To widen genetic diversity for disease resistance, the Indian native rice cultivar Aus373 was screened by F₂ segregation analyses to investigate the genetic basis of its high resistance. Aus373 was crossed with a series of Japanese differential cultivars (JDCs) and the Chinese susceptible cultivar Lijiangxintuanheigu (LTH). The resistance ratios of subsequent F₂ progenies were used to determine the number of blast-resistance loci present as well as allelic relationships with known loci. Resistance of Aus373 was governed by dominant alleles at two loci, one at the Pi-k locus and the second apparently at a new locus linked to an isozyme gene Amp-1 with a recombination fraction of 37.9 ± 3.0% on chromosome 2. This putative new locus and allele were designated Pi16 (t).     

Genetics of specific resistance in pea (Pisum sativum) cultivars to seven races of Pseudomonas syringae pv. pisi

Seven races of Pseudomonas syringae pv. pisi were distinguished using eight differential cultivars of pea (Pisum sativum). Segregation among F₂ populations of crosses between differential cultivars sequentially inoculated with races of P.s. pv. pisi provided evidence for four and possibly six putative resistance(R)/avirulence(A) gene pairs. R1, R2 and R3 are dominant resistance alleles at single loci, R4 is a dominant allele at a single locus which exhibits variable expression possibly dependent on genetic background. There is evidence that R3 and R4 are at linked loci. Homology tests provided proof of the occurrence of the alleles R2, R3 and R4 in more than one cultivar. Two other alleles, R5 and R6, were postulated to explain the observed segregation ratios in certain crosses.
It can be inferred that P.s. pv. pisi races 2, 3 and 4 each carry a different single a virulence gene, race 6 carries no apparent avirulence genes, and race 7 carries at least A2, A3 and A4. Race 1 carries Al, A3, A4 and possibly A6; race 5 carries A2, A4 and possibly A5 and A6.     

华东地区水稻品种条纹叶枯病抗性鉴定

为了丰富水稻条纹叶枯病的抗性资源和加快条纹叶枯病抗性品种培育,对69份来自华东及华南地区的籼粳不同亚种材料,在浙江、江苏两地进行苗期人工接虫和大田自然诱发筛选条纹叶枯病抗性资源.11份材料表现出较好的条纹叶枯病抗性,抗性水平与目前华东地区广泛利用的抗源镇稻88相当,且抗性表现为显性遗传.分子标记鉴定发现,其中3份材料与镇稻88抗性位点一致,而其余材料与镇稻88不同.这些抗性材料育成的后代抗性好、产量高,是培育江浙沪等晚粳稻区条纹叶枯病抗性品种的优质亲本.     

Natural Variation at the Pi-ta Rice Blast Resistance Locus

ABSTRACT The resistance gene Pi-ta protects rice crops against the fungal pathogen Magnaporthe grisea expressing the avirulence gene AVR-Pita in a gene-for-gene manner. Pi-ta, originally introgressed into japonica rice from indica origin, was previously isolated by positional cloning. In this study, we report the nucleotide sequence of a 5,113-base pair region containing a japonica susceptibility pi-ta allele, which has overall 99.6% nucleotide identity to the indica Pi-ta allele conferring resistance. The intron region shows the levels of sequence diversity that typically differentiate genes from indica and japonica rices, but the other gene regions show less diversity. Sequences of the Pi-ta allele from resistant cultivars Katy and Drew from the southern United States are identical to the resistance Pi-ta sequence. Sequences from susceptible cultivars El Paso 144 and Cica 9 from Latin America define a third susceptibility haplotype. This brings the total number of Pi-ta haplotypes identified to four, including the resistance allele and three susceptibility alleles. The Pi-ta locus shows low levels of DNA polymorphism compared with other analyzed R genes. Understanding the natural diversity at the Pi-ta locus is important for designing specific markers for incorporation of this R gene into rice-breeding programs.     

Allelism of rice blast resistance genes in two Chinese rice cultivars, and identification of two new resistance genes

Blast, caused by Pyricularia grisea , is a major constraint on rice production. To broaden genetic diversity for resistance to this disease, two rice cultivars, GA20 and GA25 from Yunnan Province, China, were analysed for the genetic basis of their high resistance to blast. GA20 was crossed with 10 Japanese differential cultivars, and GA25 was crossed with nine of them and with the susceptible Chinese cultivar Lijiangxintuanheigu (LTH). The resistance of GA20 was governed by two dominant genes allelic to genes at the Pi-k and Pi-ta loci. The allele at the Pi-k locus was new, based on a reaction pattern different from known alleles at this locus. It could not be shown whether or not the allele at the Pi-ta locus is new, because races with virulence for Pi-ta were not tested. GA25 has one resistance gene, which is not allelic to genes at the loci Pi-a , Pi-k , Pi-z , Pi-ta , Pi-b , and Pi-t , but is linked to the Pi-i gene on chromosome 9 with a recombination frequency of 15.1 ± 2.8%.
The new allele at the Pi-k locus in GA20 is designated as Pi-k^g (t), and the new resistance gene in GA25 as Pi15(t) .     

Nitrogen-induced changes in colony density and spore production of Erysiphe graminis f.sp. hordei on seedlings of six spring barley cultivars

The influence of increasing nitrogen supply (30, 60, 120 and 240 mg N per pot) on susceptibility was studied on seedlings of six cultivars of spring barley inoculated with virulent isolates of powdery mildew. The colony density (CD) measured as colonies per cm² was significantly increased with increasing application of nitrogen on all cultivars, and a significant interaction was found between N and cultivar. The different reactions of the cultivars could not be ascribed to lack of N uptake. In general, increasing N application enhanced the sporulation capacity of colonies (CSC) irrespective of increased CD and the cumulative production of spores per cm² leaf (CSCM) increased strongly with N application in all cultivars. No interaction between N and cultivar was found for the latter component. The increase in CSCM closely corresponded with the increase in N content and fresh weight of uninoculated leaves. No interaction between N treatment and powdery mildew isolates was found for infection efficiency and spore production per colony, when tested on one cultivar. The N-induced changes in infection and sporulation can explain the main part of the increasing effect of N fertilization on powdery mildew development in the field. The results indicate that it may be possible to breed for or select barley cultivars with low N impact on powdery mildew development.     

Genetic differentiation of Magnaporthe oryzae populations from scouting plots and commercial rice fields in Korea

A previous study of the diversity and population structure of the rice blast fungus, Magnaporthe oryzae, over a 20-year period in Korea, found novel fingerprint haplotypes each year, and the authors hypothesized that populations might experience annual bottlenecks. Based on this model, we predicted that M. oryzae populations would have little or no genetic differentiation among geographic regions because rice blast is commonly found throughout Korea each year and M. oryzae would have to disperse from small populations surviving annually between rice crops. To test this hypothesis, we sampled M. oryzae from rice fields in eight provinces in Korea in a single year (1999). In four provinces, we sampled from a set of rice cultivars commonly grown in commercial fields (group I); because of low disease incidence in four other provinces, we could not sample from commercial fields and instead sampled from scouting plots of different cultivars set up for detecting new pathotypes of M. oryzae (group II). All isolates were genotyped with DNA fingerprint probes MGR586 and MAGGY, a telomere-linked gene family member TLH1, the PWL2 host specificity gene and mating type. Fingerprint haplotypes clustered into two distinct lineages corresponding to the two sets of cultivars (groups I and II), with haplotype similarities of 71% between lineages and >76% within lineages. Isolates from the same cultivar within group I were genetically differentiated among locations, and isolates within the same location were differentiated among cultivars. Differentiation for TLH1 and PWL2 was significant (P < 0.03), but not as strong as for fingerprint markers. Similar analyses were not possible among group II isolates because too few isolates were available from any one cultivar. All isolates were in the same mating type, Mat1-1, ruling out sexual reproduction as a source of novel haplotypes. When the 1999 samples were compared with the historical samples from the previous study, haplotypes of group I formed a separate cluster, while those of group II clustered with haplotypes from the historical sample. Altogether, geographic subdivision, monomorphism of mating type, and correlation of haplotypes to sets of cultivars are not consistent with the hypothesis of repeated turnover of haplotypes. Instead, the previous correlations of haplotypes to year might have been caused by inadequate sampling of haplotypes each year, highlighting the need for studies of population genetics to be conducted with systematic samples collected to address specific questions.     

Genetic Constitution and Pathogenicity of Lolium Isolates of Magnaporthe oryzae in Comparison with Host Species-Specific Pathotypes of the Blast Fungus

ABSTRACT Fungal isolates from gray leaf spot on perennial ryegrass (prg isolates) were characterized by DNA analyses, mating tests, and pathogenicity assays. All of the prg isolates were interfertile with Triticum isolates and clustered into the crop isolate group (CC group) on a dendrogram constructed from rDNA-internal transcribed spacer 2 sequences. Since the CC group corresponded to a newly proposed species, Magnaporthe oryzae, all of the prg isolates were designated M. oryzae. However, DNA fingerprinting with MGR586, MGR583, and Pot2 showed that the prg isolates are divided into two distinct populations, i.e., TALF isolates and WK isolates. The TALF isolates were virulent only on Lolium species, whereas the WK isolates were less specific, suggesting that gray leaf spot can be caused not only by Lolium-specific isolates but also by less specific isolates. We designated the TALF isolates as Lolium pathotype. The TALF isolates showed diverse karyotypes in spite of being uniform in DNA fingerprints, suggesting that theyare unstable in genome organization.     

福建省稻瘟病菌致病性及其无毒基因分析

利用41个已知抗性基因水稻品种测定2003—2006年从福建省闽东、闽南、闽西、闽北和闽中5个主要稻区采集分离的87个稻瘟病单孢菌株的致病性。结果表明,福建省稻瘟病菌群体含有与所有测试抗病基因相应的无毒基因,其中66.67%的稻瘟病菌株表现较强致病力。病菌群体对水稻抗病基因Pi-d2、Pi-k(1)、Pi-km、Pi-kh、Pi-1(1)、Pi-z5(1)、Pi-z5(2)和Pi-1(2)的毒力频率均低于10%,提示这些抗病基因在福建省可作抗源使用。2003—2006年福建省稻瘟病菌群体中分别出现了40、37、36和38个无毒基因,其中有34个无毒基因在各年份均有分布,有30个无毒基因在5个主要稻区均有分布,Avr-a(2)、Avr-3(2)、Avr-ks、Avr-4b、Avr-b、Avr-kp(C)、Avr-km(C)、Avr-ta(C)、Avr-11(C)、Avr-19(t)、Avr-t和Avr-a(1)无毒基因的出现频率均低于30%,提示与之相对应的抗病基因在福建省水稻品种抗稻瘟病育种中应慎用。含有17、14、23、18和16个无毒基因组合的病菌较多,其组合频率分别为13.79%、10.34%、9.20%、8.05%和8.05%。     

Aggressiveness of Mycosphaerella graminicola Isolates from Susceptible and Partially Resistant Wheat Cultivars

ABSTRACT The selective effect of quantitative host resistance on pathogen aggressiveness is poorly understood. Because two previous experiments with a small number of bread wheat cultivars and isolates of Mycosphaerella graminicola had indicated that more susceptible hosts selected for more aggressive isolates, we conducted a larger experiment to test that hypothesis. In each of 2 years, six cultivars differing in their levels of partial resistance were planted in field plots, and isolates were collected from each cultivar early and late in the growing season. The isolates were inoculated as populations bulked by cultivar of origin, field replicate, and collection date on seedlings of the same six cultivars in the greenhouse. The selective impact of a cultivar on aggressiveness was measured as the difference in aggressiveness between early and late isolates from that cultivar. Regression of those differences on disease severity in the field yielded significance values of 0.0531 and 0.0037 for the 2 years, with moderately resistant cultivars selecting for more aggressive isolates. In a related experiment, the protectant fungicide chlorothalonil was applied to plots of two susceptible cultivars to retard epidemic development. When tested in the greenhouse, isolates of M. graminicola from those plots were significantly more aggressive than isolates from the same cultivars unprotected by fungicide.     

Inheritance and molecular mapping of barley genes conferring resistance to wheat stripe rust

ABSTRACT Most barley cultivars are resistant to stripe rust of wheat that is caused by Puccinia striiformis f. sp. tritici. The barley cv. Steptoe is susceptible to all identified races of P. striiformis f. sp. hordei (PSH), the barley stripe rust pathogen, but is resistant to most P. striiformis f. sp. tritici races. To determine inheritance of the Steptoe resistance to P. striiformis f. sp. tritici, a cross was made between Steptoe and Russell, a barley cultivar susceptible to some P. striiformis f. sp. tritici races and all tested P. striiformis f. sp. hordei races. Seedlings of parents and F(1), BC(1), F(2), and F(3) progeny from the barley cross were tested with P. striiformis f. sp. tritici races PST-41 and PST-45 under controlled greenhouse conditions. Genetic analyses of infection type data showed that Steptoe had one dominant gene and one recessive gene (provisionally designated as RpstS1 and rpstS2, respectively) for resistance to races PST-41 and PST-45. Genomic DNA was extracted from the parents and 150 F(2) plants that were tested for rust reaction and grown for seed of F(3) lines. The infection type data and polymorphic markers identified using the resistance gene analog polymorphism (RGAP) technique were analyzed with the Mapmaker computer program to map the resistance genes. The dominant resistance gene in Steptoe for resistance to P. striiformis f. sp. tritici races was mapped on barley chromosome 4H using a linked microsatellite marker, HVM68. A linkage group for the dominant gene was constructed with 12 RGAP markers and the microsatellite marker. The results show that resistance in barley to the wheat stripe rust pathogen is qualitatively inherited. These genes might provide useful resistance against wheat stripe rust when introgressed into wheat from barley.