Small differential interactions for partial resistance in rice cultivars to virulent isolates of the blast pathogen

Summary Six rice genotypes, differing in partial resistance, were exposed to three isolates of the blast pathogen. Of the variance due to host and pathogen genotypes, 39% was due to host genotype effects, 60% was due to isolate effects, and only 1% was due to host genotype × isolate interactions. Although small, this interaction variance was highly significant and mainly due to the IR50 × W6-1 and IR37704 × JMB8401-1 combinations. Although behaving largely as race-non-specific (large main effects only), the partial resistance cannot be classified as race-non-specific. The results suggest that minor genes for partial resistance operate in a gene for gene relationship with minor genes in the pathogen.     

Effect on leaf photosynthetic rate by leaf blast for rice cultivars with different types and levels of resistance

Summary The effect of an inoculation with Pyricularia oryzae (isolate P06-6) on net leaf photosynthetic rate of rice (Oryza sativa) was studied with four cultivars. Measurements were taken on the sixth leaf of the main culm of plants in the early tillering stage. On cultivars CO39, IR50 and IR64 a susceptible infection type developed, but a clear difference in relative infection efficiency of the cultivars was observed. The highest number of lesions developed on leaves of CO39, whereas the lowest number was found on leaves of IR64. For all three cultivars the effect of a single lesion on the reduction in net leaf photosynthetic rate was found to be equal to a reduction in leaf area of three times the area occupied by the visible lesion. On IR68, a cultivar with complete resistance, brown specks of pinpoint size appeared without any effect on net leaf photosynthetic rate.     

Response to selection for high and low partial resistance to leaf blast in F2 populations of three rice crosses

Summary Divergent selection for higher and lower partial resistance to leaf blast was applied in F₂ populations of crosses between the rice cultivars IR36, IR64 and CO39 after exposure to a virulent isolate. IR36 and IR64 are partially resistant while CO39 is highly susceptible. As selection criteria the number of sporulating lesions in leaves of the main culm or the lesion density in the topmost leaf relative to other F₂ plants with the same stage of development were used. A highly significant and meaningful response to selection was obtained in most cases, but the heritability was low. Realized heritabilities varied from 0.14 to 0.25 depending on the cross and were similar for both selection criteria. Selection for improved partial resistance to leaf blast is possible as early as the F₂. The efficiency of selection is probably much higher if replicated tests could be made, and better results are therefore expected if selection among F₃ lines is carried out. The results indicated that the relatively low infection efficiency in IR36 and IR64 is oligo- or polygenically controlled.     

Partial resistance to neck blast influenced by stage of panicle development and rice genotype

Summary Neck nodes of eight rice genotypes were inoculated with a virulent isolate of the blast pathogen at four slightly different, increasingly older stages of panicle development shortly after flowering. Resistance to infection as well as resistance to growth of the pathogen after infection was assessed. Significant differences between genotypes were found for both components of resistance. Resistance to growth of the pathogen sharply increased with aging of the neck node as was indicated by a large reduction of the length of the lesions on the culm, but resistance to infection did not change much. The lesion length was closely related to yield loss. A small delay in time of infection can lead to a large decrease in yield reduction. Meaningful comparison of partial resistance to neck blast between genotypes therefore requires infection at an identical stage of panicle development. In the field, where panicles are in different stages of development, selection of genotypes that do show the symptoms of neck blast together with relatively little yield loss in infected panicles is recommended.     

Durable resistance to rice blast disease-environmental influences

Summary Blast is one of the most serious diseases of rice worldwide. The pathogen,Pyricularia grisea, can infect nearly all parts of the shoot and is commonly found on the leaf blade and the panicle neck node. Host resistance is the most desirable means of managing blast, especially in developing countries. Rice cultivars with durable blast resistance have been recognized in several production systems. The durable resistance of these cultivars is associated with polygenic partial resistance that shows no evidence of race specificity. This partial resistance is expressed as fewer and smaller lesions on the leaf blade but latent period does not appear to be an important component. Partial resistance to leaf blast is positively correlated with partial resistance to panicle blast, although some cultivars have been found showing leaf-blast susceptibility and panicle-blast resistance. A diverse set of environmental factors can influence the expression of partial resistance, including temperature, duration of leaf-wetness, nitrogen fertilization, soil type, and water deficit. Because of the great diversity of rice-growing environments, resistance that proves durable in one system may or may not prove useful in another. In highly blast-conducive environments, other means of disease management must be applied to assist host-plant resistance.     

Molecular mapping of a new gene for resistance to rice blast (Pyricularia grisea Sacc.)

A single dominant blast resistance gene conferring resistance to a Korean rice blast isolate was identified in rice variety `Suweon 365'. We report the chromosomal localization and molecular mapping of this blast resistance gene designated as Pi-18, which confers resistance to Korean isolate `KI-313' of the blast pathogen. To know whether there is a relationship among genes conditioning resistance to location-specific isolates of the blast pathogen and thereby to identify linked markers to resistance gene for isolate KI-313 collected in Korea, RFLP markers previously reported to be linked to major blast resistance genes in different rice germplasm and other markers mapped to nearby regions were surveyed for polymorphism between a resistant (`Suweon 365') and a susceptible (`Chucheongbyeo') parent. Linkage associations of the RFLP markers with the resistance gene were verified using an F2 and F3 segregating population of known blast reaction. RFLP analysis showed that Pi-18 was located near the end of chromosome 11, linked to a single copy clone RZ536 at a distance of 5.4 centiMorgans (cM) and that this gene was different from Pi-1(t). An allelism test revealed that this gene was also different from Pi-k. Currently, a combination of RAPD and microsatellite primers is being employed to find additional markers in this region. Tightly linked DNA markers will facilitate selection for resistant genotypes in breeding programs and provide the basis for map based cloning of this new blast resistance gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic variation in resistance to blast disease (Pyricularia oryzae Cavara) in Japanese rice (Oryza sativa L.), as determined using a differential system

A total of 324 Japanese rice accessions, including landrace, improved, and weedy types were used to 1) investigate genetic variations in blast resistance to standard differential isolates, and 2) across the genome using polymorphism data on 64 SSR markers. From the polymorphism data, the accessions were classified into two clusters. Accessions from irrigated lowland areas were included mainly in cluster I, and upland and Indica types were mainly in cluster II. The accessions were classified into three resistance subgroups, A2, B1 and B2, based on the reaction patterns to blast isolates. The accessions in A2 were postulated to have at least two resistance genes Pish and Pik-s, whereas those in B1 had various combinations of the resistance genes Pish, Pia, Pii, Pi3, Pi5(t), and Pik alleles. The B2 accessions were resistant to almost all isolates, and many accessions of cluster II were included, and had Pish, Pia, Pii, Pi3, Pi5(t), certain Pik, Piz and Pita alleles, and unknown genes. The frequencies of accessions of B1 originating in Hokkaido, and those of B2 originating in the Kanto and Tohoku regions were remarkably higher than in the other regions.     

Inheritance of resistance to rice stripe virus in rice line `BL 1'

Rice stripe is the most serious virus disease in temperate rice-growing countries. The most economical and environmentally safe practice for controlling this disease is virus-resistant cultivars. ‘BL 1’ is an elite germplasm line with the blast resistance gene Pib, and has been used as a differential line for testing the pathogenicity of the blast fungus. We found that certain progenies from BL 1 showed resistance to both blast and rice stripe virus (RSV). The objectives of this study were to evaluate the RSV resistance in the field and under artificial conditions, to assess the reaction to the insect vector(small brown plant hopper, SBPH), and to examine its inheritance and its relationship to blast resistance in BL 1.BL 1 was susceptible to SBPH, but resistant to RSV in field and artificial inoculation tests. The inheritance of RSV resistance in F₃ lines from the cross Nipponbare (NPB)/BL 1 was studied using artificial inoculation with a population of viruliferous SBPH. A serological assay for RSV infection using an enzyme-linked immunosorbent assay (ELISA) was used. RSV resistance in BL 1 was controlled by a single major gene with incomplete dominance. The locus responsible for RSV resistance was genetically independent of the blast resistance gene Pib. The resistance gene for RSV infection in BL 1 was also independent of Stvb-i, a gene widely distributed in resistant Japanese cultivars. Resistance to RSV must be diversified in rice cultivars considering the potential for future emergence of new RSV strains. The new resistance gene identified in BL 1, which has improved plant type and blast resistance, is considered useful for breeding RSV-resistant cultivars in japonica rice. This revised version was published online in August 2006 with corrections to the Cover Date.     

Mapping of leaf and neck blast resistance genes with resistance gene analog, RAPD and RFLP in rice

An F₈ recombinant inbred population was constructed using a commercial indica rice variety Zhong 156 as the female parent and a semidwarf indica variety Gumei 2 with durable resistance to rice blast as the male parent. Zhong 156 is resistant to the fungus race ZC₁₅ at the seedling stage but susceptible to the same race at the flowering stage. Gumei 2 is resistant to ZC₁₅ at both stages. The blast resistance of 148 recombinant inbred lines was evaluated using the blast race ZC₁₅. Genetic analysis indicated that the resistance to leaf blast was controlled by three genes and the presence of resistant alleles at any loci would result in resistance. One of the three genes did not have effects at the flowering stage. Two genes, tentatively assigned as Pi24(t) and Pi25(t), were mapped onto chromosome 12 and 6,respectively, based on RGA (resistance gene analog), RFLP and RAPD markers. Pi24(t) conferred resistance to leaf blast only, and its resistance allele was from Zhong 156. Pi25(t) conferred resistance to both leaf and neck blast, and its resistance allele was from Gumei 2. In a natural infection test in a blast hot-spot, Pi25(t) exhibited high resistance to neck blast, while Pi24(t) showed little effect. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification of microsatellite markers linked to the blast resistance gene <Emphasis Type="Italic">Pi-1(t)</Emphasis> in rice

The present work was conducted to identify microsatellite markers linked to the rice blast resistance gene Pi-1(t) for a marker-assisted selection program. Twenty-four primer pairs corresponding to 19 microsatellite loci were selected from the Gramene database (www. gramene.org) considering their relative proximity to Pi-1(t) gene in the current rice genetic map. Progenitors and DNA bulks of resistant and susceptible families from F₃ segregating populations of a cross between the near-isogenic lines C101LAC (resistant) and C101A51 (susceptible) were used to identify polymorphic microsatellite markers associated to this gene through bulked segregant analysis. Putative molecular markers linked to the blast resistance gene Pi-1(t) were then used on the whole progeny for linkage analysis. Additionally, the diagnostic potential of the microsatellite markers associated to the resistance gene was also evaluated on 17 rice varieties planted in Latin America by amplification of the specific resistant alleles for the gene in each genotype. Comparing with greenhouse phenotypic evaluations for blast resistance, the usefulness of the highly linked microsatellite markers to identify resistant rice genotypes was evaluated. As expected, the phenotypic segregation in the F₃ generation agreed to the expected segregation ratio for a single gene model. Of the 24 microsatellite sequences tested, six resulted polymorphic and linked to the gene. Two markers (RM1233*I and RM224) mapped in the same position (0.0 cM) with the Pi-1(t) gene. Other three markers corresponding to the same genetic locus were located at 18.5 cM above the resistance gene, while another marker was positioned at 23.8 cM below the gene. Microsatellite analysis on elite rice varieties with different genetic background showed that all known sources of blast resistance included in this study carry the specific Pi-1(t) allele. Results are discussed considering the potential utility of the microsatellite markers found, for MAS in rice breeding programs aiming at developing rice varieties with durable blast resistance based on a combination of resistance genes. Centro Internactional de Agricultura Tropical (CIAT) institute where the research was carried out     

Identification of a major blast resistance gene in the rice cultivar 'Tetep'

Analysis of near‐isogenic lines (NILs) indicated the presence of a novel resistance gene in the indica rice cultivar ‘Tetep’ which was highly resistant to the rice blast fungus Magnaporthe grisea.‘Tetep’ was crossed to the widely used susceptible cultivar ‘CO39’ to generate the mapping population. A Mendelian segregation ratio of 3 : 1 for resistant to susceptible F₂ plants further confirmed the presence of a major dominant locus, in ‘Tetep’, conferring resistance to the blast fungal isolate B157, corresponding to the international race IC9. Simple sequence length polymorphism (SSLP) was used for molecular genetic analysis. The analysis revealed that the SSLP marker RM 246 was linked to a novel blast resistance gene designated Pi‐tp(t) in ‘Tetep’.     

Evaluation of a simple technique for characterizing rice root systems in relation to drought resistance

Summary Drought is a constraint to rice yields in much of Asia's rainfed rice crop. Although genotypic variation for root system characteristics is well documented in this semiaquatic species, little improvement of rice for drought resistance has been attempted. No simple method to evaluate root systems exists. In this paper we report the development and testing of a simple device to measure the pulling force required to uproot rice seedlings. The force required to pull rice seedlings from the paddy soil was correlated with root weight, root branching and thick root number. The capability to separate rice cultivars by this technique and relationship of the root pulling force to other drought screening methods are illustrated.     

Induction and selection of mutations for resistance against bacterial leaf blight in rice

Summary Dry seed lots of a rice cultivar, Harebare, susceptible for bacterial leaf blight (BLB), were exposed to thermal neutrons with and without pre-treatment of seed for boron-enrichment, and to gamma-rays. M₁ plants of each of the treatments were grown and their seeds were separately harvested. M₂ populations were raised in rice fields of a farmer in a region where BLB is epidemic every year, and M₂ and control plants which expressed resistant reactions to BLB were selected. M₃ and control lines as plant progenies of the selected M₂ plants were raised in the same rice fields as in M₂ generation in order to investigate their BLB resistance and other agronomically important traits. Variances for disease severity of M₂ populations were significantly larger than those of the control population. whereas their means were not singificantly different from that of the control population, suggesting induction of mutations toward both resistance and susceptibility to BLB. Mean disease severity values of the M₃ lines from selected M₂ plants were significantly smaller than those of the control lines, indicating gains of selection in M₂ for quantitative resistance against BLB. Thermal neutrons, especially with boron-enrichment pre-treatment were effective to induce mutations for resistance against BLB. Some M₃ mutant lines with quantitatively enhanced resistance against BLB were not modified in other agronomic traits from those of the original cultivar. Significance of the induced quantitative resistance in breeding programmes for BLB resistance is discussed.     

Comparison of selection indices to screen lowland rice for drought resistance

Summary Although many selection indices have been used to screen rices (Oryza sativa L.) for drought resistance, there has been little comparison of the relative merits of these indices. Research was conducted to compare drought resistance as estimated from grain yields, canopy-temperature-based stress indices, visual scoring, and uprooting force for 30 rice genotypes grown in the field with a puddled Maahas clay (Typic Tropaquept) and to evaluate traits related to drought resistance from nonstressed plants grown in the field and in aeroponic culture. Water deficit was imposed in the field by withholding irrigation from 45 to 75 days after transplanting compared to a continuously flooded control. Grain yields in the stress treatment were most strongly correlated with visual assessment of drought stress symptoms according to a standard evaluation system (r = 0.66). Canopy-temperature-based indices were also significantly correlated with grain yields of the stress treatment (r from –0.55 to –0.63). No trait of aeroponically grown plants was correlated with traits of stressed plants in the field. We conclude that visual scoring of stressed plants was the best method of screening for drought resistance, but if controlled water deficit cannot be imposed, then drought resistance may be estimated by measuring both uprooting force and grain yield.     

A new gene for resistance in rice to Asian rice gall midge (Orseolia oryzae Wood Mason) biotype 1 population at Raipur, India

The Asian rice gall midge, Orseolia oryzae Wood Mason (Diptera: Cecidomyiidae), is a major pest of rice in several South and South East Asian countries. The maggots feed internally on the growing tips of the tillers and transform them into tubular galls, onion leaf-like structures called ‘silver shoots’ resulting into severe yield loss to the rice crop. We studied the mode of inheritance and allelic relationships of the resistance genes involved in resistant donor Line 9, a sib of a susceptible cultivar ‘Madhuri’. The segregation behaviour of F₁, F₂ and F₃ populations of the cross between Line 9 and susceptible cultivar MW10 confirmed the presence of a single dominant gene for resistance. Tests of allelism with all the known genes giving resistance to this population indicated that Line 9 possessed a new gene which was designated Gm 9 This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of resistance to bacterial blight disease of rice

Summary Inheritance of resistance to the Punjab isolate of Xanthomonas campestris pv. oryzae of bacterial blight disease of rice was studied in seven breeding lines resistant to the disease. The results revealed that resistance in breeding lines PAU 122-73-1-4-1, PAU 164-102-1-2-1-1-1, KJT 24, IR 5657-33-2-1-2 and IR 22082-41-2-2 was controlled by single dominant genes allelic to the dominant gene which confers resistance to the Punjab isolate in Patong 32. Resistance to the Punjab isolate in breeding lines IET 7172 and RP 2151-40-1 was found to be controlled by single recessive resistance genes allelic to one of the recessive resistance genes present in BJ 1. The two genes are independently inherited and are being used to develop bacterial blight resistant varieties.     

Molecular mapping of a novel blast resistance gene Pi38 in rice using SSLP and AFLP markers

Blast, caused by Magnaporthe grisea, is the most devastating disease of rice worldwide. In this study, the main objective was to identify and map a new gene for blast resistance, in an indica rice cultivar ‘Tadukan’ against blast fungal isolate B157, using molecular tools. F₂ segregating population was derived from ‘CO39’ (susceptible) and ‘Tadukan’ (resistant), and molecular mapping of the blast resistance gene was carried out using simple sequence length polymorphism (SSLP) and amplified fragment length polymorphism (AFLP) methods. Two SSLP markers, RM206 and RM21 and three AFLP markers (AF1: E‐aca/M‐ctt; AF2: E‐aca/M‐cat and AF3: E‐acc/M‐cac2) were identified to be linked to the resistance gene. The co‐segregation analysis using SSLP markers implied that the blast resistance gene designated Pi38 resides on rice chromosome 11.     

Development of a SNLP marker from the Pi-ta blast resistance gene by tri-primer PCR

The Pi-ta gene from indica introgressed into japonica rice has been used to control the blast disease caused by the fungal pathogen Magnaporthe grisea (Herbert) Barr. (anamorph Pyricularia oryzae Cav.) worldwide. A single nucleotide length polymorphism (SNLP) was identified at the intron region of the Pi-ta gene to develop a codominant Pi-ta gene marker suitable for genotyping with an automated machine. The DNA primer specific to the resistant Pi-taallele was labeled with blue dye (FAM, 6-carboxyfluorescein) as a forward primer, the DNA primer specific to the susceptible pi-ta allele was labeled with green dye (HEX, 4,7,2′,4′,5′,7′-hexachloro-6-carboxyfluorescein) as another forward primer and the DNA primer identical to both Pi-ta/pi-ta alleles was unlabeled as the reverse primer for polymerase chain reaction (PCR). Using these three primers, a 181-bp blue peak in homozygous resistant and a green peak of 182–183 bp in homozygous susceptible, and both peaks in heterozygous plants were produced by PCR. The utility of marker was verified using a segregating F₂ population, inbred cultivated lines, dominant markers and pathogenicity testing. A codominant Pi-ta marker was thus developed for effective Pi-ta assisted selection for crop improvement. Using highly homologous competitive primers for allele detection by PCR can benefit the study of genome organization of the complex locus. This revised version was published online in August 2006 with corrections to the Cover Date.     

Environmental stability of partial resistance in spring wheat to wheat leaf rust

Summary Five spring wheat cultivars differing in partial resistance (PR) to wheat leaf rust were tested at Wageningen (the Netherlands) on a sandy and a clay site, El Batan (CIMMYT, Mexico) and Ponta Grossa (Brazil) over two years. The cultivars were Skalavatis 56, Little Club (both very susceptible), Westphal 12A, Akabozu and BH 1146 (all three with high levels of PR). The results showed that PR was expressed at all four locations in both years. The level of expression was influenced by the environment but the cultivar ranking was hardly affected. Selection for PR in the field can therefore be carried out over a wide range of environments.     

Residual effects of the Xa-4 resistance gene in three rice cultivars when exposed to a virulent isolate of Xanthomonas campestris pv. oryzae

Summary F₂ plants of five, and F₃ plants of three, crosses between genotypes carrying the race-specific resistance gene Xa-4 and genotypes not carrying this gene were inoculated with two isolates of Xanthomonas campestris pv. oryzae. Half the tillers of each plant received isolate PX061, avirulent on the Xa-4 gene, the other half of the tillers received isolate PX099, virulent for the Xa-4 gene. The F₂ and F₃ populations segregated for a single dominant resistance gene, Xa-4.The parental, F₂ and F₃ genotypes not carrying Xa-4 had mean lesion lengths between 28 and 29 cm for both isolates. The Xa-4 carrying parents showed a mean lesion length of 2.7 cm with the avirulent isolate and of 12.4 cm with the virulent isolate. The Xa-4 carrying F₂ and F₃ genotypes had mean lesion lengths of 5.2 and 20.1 cm for the two isolates, respectively. These observations strongly indicate that the Xa-4 gene, carried by the rice genotypes studied (IR28, Cisadane and BR51-282-8), had a considerable residual effect when exposed to virulent isolate PXO99.