Evaluation of common wheat cultivars for tan spot resistance and chromosomal location of a resistance gene in the cultivar 'Salamouni'

A total of 50 wheat (Triticum aestivum L.) cultivars were evaluated for resistance to tan spot, using Pyrenophora tritici‐repentis race 1 and race 5 isolates. The cultivars ‘Salamouni’, ‘Red Chief’, ‘Dashen’, ‘Empire’ and ‘Armada’ were resistant to isolate ASC1a (race 1), whereas 76% of the cultivars were susceptible. Chi‐squared analysis of the F₂ segregation data of hybrids between 20 monosomic lines of the wheat cultivar ‘Chinese Spring’ and the resistant cultivar ‘Salamouni’ revealed that tan spot resistance in ‘Salamouni’ was controlled by a single recessive gene located on chromosome 3A. This gene is designated tsn4. The resistant cultivars identified in this study are recommended for use in breeding programmes to improve tan spot resistance in common wheat.     

Allelic variation at the storage protein loci of 55 US-grown white wheats

Fifty soft white and hard white wheat cultivars (Triticum aestivum L.), and five club wheat cultivars (T. compactum L.) were partially characterized in terms of their storage protein compositions, i.e. gliadins, and high molecular weight and low molecular weight glutenin subunits (HMW-GS and LMW-GS, respectively). At the Glu-1 loci, HMW-GS composition 1,7 + 9,2+ 12 was found to be predominant, being expressed in 11 cultivars out of 55. The most common alleles at the loci coding for gliadins and LMW-GS were found to be Gli-A1/Glu-A3a (43.6%), Gli-B1/Glu-B3b (36.4%), Gli-D1a/Glu-D3a (38.1%) and Gli-Dli/Glu-D3a (21.8%). Two-dimensional fractionation (acid-poly-acrylamide gel electrophoresis (A-PAGE) × sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)) of reduced and alkylated glutenins revealed that the number and the relative mobility of LMW-GS polypeptides were different from those reported for the corresponding Glu-3 alleles of hard-bread wheat cultivars. This result could account for the different technological properties of soft white wheats compared with hard-bread wheat cultivars, owing to the major impact of LMW-GS on dough quality.     

The use of microsatellite markers to determine the relative proportions of grain produced by cultivars and the frequency of hybridization in bread wheat mixtures

Two polymorphic microsatellite markers were selected to identify 24 bread wheat cultivars commonly grown in France and to estimate the proportions of cultivar and hybrid grains in the harvests of four 4‐cultivar mixtures (CM 1‐4) planted in equal proportions in farmers’ fields. This technology was used not only to determine whether a mixture comprises the declared cultivars, but also whether there is any contamination with other cultivars, to identify the contaminants and to estimate their proportions. At harvest, the cultivar proportions ranged from 15 to 37%. Only in the mixture CM2 (‘Malacca’, ‘Somme’, ‘Renan’ and ‘Soissons’), did the cultivars contribute equally to the harvest. The other cultivar mixtures CM1 (‘Malacca’, ‘Somme’, ‘Renan’ and ‘Camp Remy’), CM3 (‘Malacca’, ‘Texel’, ‘Apache’ and ‘Aligre’) and CM4 (‘Malacca’, ‘Somme’, ‘Apache’ and ‘Virtuose’) showed significantly unequal cultivar proportions with ‘Somme’ dominating ‘Renan’ and ‘Camp Remy’ in CM1, and ‘Apache’ dominating ‘Malacca’ and ‘Aligre’ in CM3 and ‘Malacca’ in CM4. Similar cultivar proportions were measured with gliadin and glutenin markers in the mixtures CM3 and CM4, confirming the results with microsatellites. No contamination was found. Hybrids accounted for between 1.3 and 6.3% of the grains produced in the four cultivar mixtures.     

Genetic relationships between South African wheat cultivars as measured by gliadin banding patterns

The use of gliadins in cultivar identification is a well‐known practice that is used in many countries. The aim of this study was to determine the gliadin banding patterns of 35 commercial South African wheat cultivars and to use these data to determine genetic relationships between the cultivars using sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. The index of genetic similarity was used to calculate pairwise distance matrices, which were converted to a dendogram. Most of the gliadin bands fell within the nomenclature used. Fourteen cultivars had one novel band and six had two novel bands. All the cultivars could be distinguished from one another. Genetic distances between clusters were small. This suggested that there was not enough unique genetic variability to set any cluster apart from the others.     

DROUGHT STRESS: Soil Water Availability Alters the Inter‐ and Intra‐Cultivar Competition of Three Spring Wheat Cultivars Bred in Different Eras

Competition for water generates a classic aspect of the tragedy of the commons, the ‘race for fish’, where crops must allocate more resource to acquisition of the limiting resource than is optimal for crop yield allocation. A pot experiment using a simple additive (target–neighbour) design was conducted to examine the above‐ground and below‐ground growth of three spring wheat (Triticum aestivum L.) cultivars when grown alone and in mixtures at three levels of water availability. The effects of competition and water availability were compared by observing patterns of growth, biomass allocation and below‐ground outcomes. Competitive interactions were investigated among cultivars ‘HST’, ‘GY602’ and ‘LC8275’, target plant of each cultivar grown without neighbouring plants are referred to herein as control plant and one target plant of each cultivar sown surrounded either by same or another cultivar as intra‐ or inter‐cultivar competition. Competitive ability was assessed as the response ratio (lnRR) between the target plant surrounded by six other plants and the target plant in isolation. Our results showed that the cultivar ‘HST’, released over a century ago, produced a higher biomass and grain yield than the more recently released cultivars ‘LC8275’ and ‘GY602’ when grown as isolated plants with sufficient water supply. However, competition for resources from neighbours led to target plant biomass and grain yield being significantly reduced relative to controls in all three cultivars, particularly in ‘HST’. When subjected to intra‐cultivar competition, the two recently released cultivars ‘LC8275’ and ‘GY602’ had higher grain yields and water use efficiency for grain than ‘HST’ in all three water regimes. The landrace ‘HST’ had better and significantly linear relationships between biomass and biomass allocation, root length and specific root length, whereas the recent and modern cultivars had much more water‐related species‐specific changes in root morphology and allocation patterns. These results suggest that crop traits that influence competitive ability, such as biomass allocation to roots and root plasticity in response to drought have changed in modern wheat cultivars because of breeding and selection.     

小麦品种的蛋白电泳鉴定

由于小麦不同品种的种子含有不同的麦醇溶蛋白(gliadin),因此利用聚丙烯酰胺凝胶电泳可以分析比较不同品种的小麦。这种方法不受栽培条件的影响。本文分析了25个不同品种小麦的麦醇溶蛋白 SDS-聚丙烯酰胺凝胶电泳图谱,并对乙醇、脲提取的麦醇溶蛋白电泳图谱进行了比较,结果表明不同品种小麦麦醇溶蛋白电泳图谱差异明显,将疑胶     

Powdery mildew resistance in Czech and Slovak barley cultivars

Fifteen powdery mildew resistance genes and the gene MlaN81 derived from ‘Nepal 81’were found in 76 Czech and Slovak spring and winter barley cultivars when tested for reaction to a set of powdery mildew isolates. Nine cultivars (‘Donum’, ‘Expres’, ‘Jubilant’, ‘Orbit’, ‘Primus’, ‘Progres’, ‘Stabil’, ‘Vladan’ and ‘Zlatan’) are composed of lines with different resistance genes. The Mlat gene is present in nine cultivars and was transferred from the Anatolian landrace ‘A‐516′. The resistances derived from ‘KM‐1192’and ‘CI 7672’were identical and designated Ml(Kr). Five winter barley cultivars possess the Ml(Bw) resistance. The winter barley line ‘KM‐2099’carries the mlo gene. The parental cultivar ‘Palestine 10’was also tested in which the genes Mlk1, MlLa were identified. The German cultivar ‘Salome’, a parent of seven cultivars tested, probably carries the gene MlLa in addition to mlo and Mla7. The gene mlo6 may be present in the cultivar ‘Heris’. Most of the results were confirmed by the pedigrees of the cultivars.     

Genes for Powdery Mildew Resistance in Cultivars of Spring Wheat

Twenty-three cultivars of spring wheat were inoculated with nineteen different powdery mildew isolates; their ruction patterns hive been compared with those of twenty-two cultivars/lines carrying identified powdery mildew resistance genes. Applying the gene-for-gene hypothesis, it is evident that three cultivars have none of the resistance genes used, seven others (including ‘Solo’) may carry Pm4b, only. The resistance pattern of ‘Selpek’ is identical to A/-1 resistant cultivars of winter wheat and may be explained by the presence of Pm5. The resistance pattern of Pm5 (Mt-i) cultivars is very different from a number of ‘Kolibri’-related cultivars of spring wheat. Since either all or nothing of that specific pattern has been transferred to all cross progenies of ‘Kolibri’, a single gene is assumed to oe responsible for it, preliminarily designated as Ml-k. The cultivar ‘Mephisto’ carries the ‘Normandie’ resistance (Pwl 2, 9). In five cultivars to spring wheat the combined effects of at least two of the above-mentioned sources have been found. Despite the fact that ‘Normandie’ and ‘Sappo’ are not closely related. ‘Sappo’ shows the complete ‘Normandie’ resistance pattern plus that of Pm4b. The same is true for ‘Planet’ and ‘Walter’.     

Identification of Rye Chromosome 1R Translocations and Substitutions in Hexaploid Wheats Using Storage Proteins as Genetic Markers

Grain protein compositions of 106 advanced generation backcross lines from crosses involving ‘Amigo’ (1AL.1RS), ‘Aurora’, ‘Kavkaz’, ‘Skorospelka-35’ and ‘Sunbird’ (all 1BL.1RS) and ‘Gabo’ 1DL.1RS parents and 152 cultivars with unknown pedigree were analysed by one-dimensional SDS-PAGE. Eighty seven backcross lines and 16 cultivars carried one or other of these translocations, 2 cultivars had a 1R (1B) substitution, whereas 5 backcross lines were found to be heterogeneous for the 1BL.1RS translocation. The translocation lines were easily identified by the presence of secalins (Sec-1) controlled by rye chromosome arm IRS and a simultaneous loss of the gliadin (Gli-1) and/or triticin (Tri-1) protein bands controlled by the replaced wheat chromosome arm (1AS, 1BS or 1DS). Certain gliadins, showing no allelic variation among the genotypes analysed, were identified as markers for chromosome arms 1AS (M_r= 34 kd) and IBS (M_r= 42,33 kd). The whole chromosome substitutions 1R (1B) were recognized by scoring for the presence of Sec-1 and HMW secalin bands, Sec-3 (controlled by rye chromosome arm 1RL) and the absence of Gli-B1 and HMW glutenin subunits, Glu-B1 (controlled by wheat chromosome arm 1BL). The results have shown that protein electrophoresis provides a rapid and reliable technique for screening genotypes for these translocations and substitutions in a breeding programme.     

冀西北冬麦北移适宜播期和品种的研究

为了进一步明确冀西北冬麦北移的适宜播期和品种,选用3个冬小麦品种（‘京冬1号’、‘苏引6号’和‘02铁品5’）,通过分期播种,研究不同播期对小麦越冬和产量的影响。结果表明,以2008年9月23日为播种期的小麦越冬性和产量最高,播期主要通过影响有效穗数多少决定产量高低,播种越晚影响越明显;‘02铁品5’与‘苏引6号’产量极显著高于‘京冬1号’。通过对2005-2010年的冬前积温进行分析,认为冀西北冬小麦适宜播期为9月20日至9月25日左右,同时,应根据气候及品种特性进行适当调整,即较暖地区可适当延后,冷凉地区要适当提前;生育期短、冬性弱的品种要适当晚播,生育期长和冬性强的品种可适当早播;播期推迟时,要适当加大播种密度,以弥补越冬性差造成的群体不足。     

Genes for wheat stem rust resistance postulated in German cultivars and their efficacy in seedling and adult‐plant field tests

Stem rust of wheat (caused by Puccinia graminis f.sp. tritici) gained high international attention in the last two decades, but does not occur regularly in Germany. Motivated by a regional epidemic in 2013, we analysed 15 spring and 82 winter wheat cultivars registered in Germany for their resistance to stem rust at the seedling stage and tested 79 of these winter wheat cultivars at the adult‐plant stage. A total of five seedling stem rust resistance genes were postulated: Sr38 occurred most frequently (n = 29), followed by Sr31 (n = 11) and Sr24 (n = 8). Sr7a and Sr8a occurred only in two spring wheat genotypes each. Four cultivars had effective seedling resistance to all races evaluated that could only be explained by postulating additional resistance genes (‘Hyland’, ‘Pilgrim PZO’, ‘Tybalt’) or unidentified gene(s) (‘Memory’). The three winter wheat cultivars (‘Hyland’ ‘Memory’ and ‘Pilgrim PZO’) were also highly resistant at the adult‐plant stage; ‘Tybalt’ was not tested. Resistance genes Sr24 and Sr31 highly protected winter wheat cultivars from stem rust at the adult‐plant stage in the field. Disease responses of cultivars carrying Sr38 varied. Mean field stem rust severity of cultivars without postulated seedling resistance genes ranged from 2.71% to 41.51%, nine of which were significantly less diseased than the most susceptible cultivar. This suggests adult‐plant resistance to stem rust may be present in German wheat cultivars.     

谷蛋白和醇溶蛋白的电泳分析及小麦品种鉴定

应用不同浓度梯度的聚丙烯酰胺凝胺SDS电泳比较分析了18个小麦品种的、用不同提取方法制备的醇溶蛋白和谷蛋白.结果表明,经1M脲素提取掉多数醇溶蛋白、清蛋白及其它醇溶成分后的小麦样品,用加有2%SDS和5%β—巯基乙醇的裂解缓冲液提取的高分子量谷蛋白和醇溶蛋白,在8—12%梯度的SDS胶中显示出良好的分辨率和极大的多态性,适于小麦品种鉴定.用裂鲜缓冲液直接提取半粒小麦的全蛋白可用于逐粒品种鉴别.     

Identification of powdery-mildew-resistance genes in common wheat (Triticum aestivum L. em. Thell.). V. Old German cultivars and cultivars released in the former GDR

A total of 59 old wheat cultivars grown in Germany prior to 1960 were tested for mildew response using a collection of 12 differential isolates of Erysiphe graminis DC f. sp. tritici Marchal (Blumeria graminis (DC) Speer f. sp. tritici). Nineteen cultivars did not possess any major resistance gene and 25 were characterized by susceptible or intermediate responses. Fifteen cultivars revealed isolate-specific response patterns that could not be attributed to known major resistance genes or gene combinations. Many of the old German cultivars inherited a mildew-resistance gene from the Canadian cultivar ‘Garnet’ which is tentatively designated M1-Ga. Cultivars ‘Bretonischer Bartweizen’ (designated M1-Br) and ‘Adlungs Alemannen’ (designated M1-Ad) appeared to carry unknown resistance genes. Among 18 winter wheat cultivars released in the former GDR. eight showed susceptibility to all isolates used. Cv. “Borenos” carries resistance gene Pm3c. Five cultivars possess gene Pm4b. two cultivars gene pm5 and one cultivar a combination of genes Pm2 and Pm4b. Cultivar ‘Zentos’ was resistant to almost all isolates used. Its resistance might be conditioned by different unknown major resistance genes.     

Effect of Cultivar, Incubation Temperature, and Stage of Microspore Development on Anther Culture in Wheat and Triticale

Effects of incubation temperature and developmental stage of microspores on polyhaploid production in three wheat cultivars‘Pavon 76′, ‘Kitt’, and ‘Chris’ and one triticale cultivar, ‘T81′, were studied using a one-step medium. Calli failing to differentiate on the one-step medium were placed on a medium containing 1 mg/l indole-3-acctic acid (IAA) and 2 mg/1 6-furfurylaminopunne (KIN). Anthers containing either early- or late-uninucleate microspores were incubated in dark at 26, 28 or 32°C lor 3 days prior to transfer to 26°C. Averaged over temperatures and microspore stages, frequency of calli and green plantlets were 8.9 % and 3.4 %, respectively, for wheat cultivar‘Pavon 76′, 8.4 % and 1.6 % for cultivar ‘Kitt’, 4.5 % and 0.25 % for cultivar ‘Chris’, and 2.9 % and 0.12 % for the triticale cultivar‘T81′. However, cultivar-by-developmental-stage interaction was significant for frequency of callus induction. Temperature had no significant effects on callus induction and plantlet regeneration. Anthers containing early-unmucleate microspores produced no polyhaploids.     

An STS marker distinguishing the rye-derived powdery mildew resistance alleles at the Pm8/Pm17 locus of common wheat

A sequence‐tagged site marker has been developed from restriction fragment length polymorphism marker probe IAG95 for the rye‐derived powdery mildew resistance Pm8/Pm17 locus of common wheat. This polymerase chain reaction marker enables the amplification of DNA fragments with different sizes from T1AL.1RS and T1BL.1RS wheat‐rye translocation cultivars with chromatin from ‘Insave’ and ‘Petkus’ rye, respectively, and therefore will be very useful in distinguishing Pm8‐carrying cultivars from Pm17‐carrying cultivars. Results obtained with that marker were compared with resistance tests performed on detached primary leaves of 29 wheat lines from two populations derived from doubled haploid production. The molecular assay corresponded well with the resistance tests in all the lines, and therefore will be helpful for the identification of Pm17 in lines in which other Pm genes or quantitative trait loci are present.     

Evaluation of cold hardiness in two sets of near-isogenic lines of wheat (Triticum aestivum) with polymorphic vernalization alleles

In wheat, variation at the orthologus Vrn‐1 loci, located on each of the three genomes, A, B and D, is responsible for vernalization response. A dominant Vrn‐1a allele on any of the three wheat genomes results in spring habit and the presence of recessive Vrn‐1b alleles on all three genomes results in winter habit. Two sets of near‐isogenic lines (NILs) were evaluated for DNA polymorphisms at their Vrn‐A1, B1 and D1 loci and for cold hardiness. Two winter wheat cultivars, ‘Daws’ and ‘Wanser’ were used as recurrent parents and ‘Triple Dirk’ NILs were used as donor parents for orthologous Vrn‐1 alleles. The NILs were analysed using molecular markers specific for each allele. Only 26 of 32 ‘Daws’ NILs and 23 of 32 ‘Wanser’ NILs had a plant growth habit that corresponded to the marker genotype for the markers used. Freezing tests were conducted in growth chambers programmed to cool to ?21.5°C. Relative area under the death progress curve (AUDPC), with a maximum value of 100 was used as a measure of death due to freezing. The average relative AUDPC of the spring habit ‘Daws’Vrn‐A1a NILs was 86.15; significantly greater than the corresponding winter habit ‘Daws’Vrn‐A1b NILs (42.98). In contrast, all the ‘Daws’Vrn‐A1bVrn‐B1aVrn‐D1b and Vrn‐A1bVrn‐B1bVrn‐D1a NILs (spring habit) had relative AUDPC values equal to those of their ‘Daws’ sister genotypes with Vrn‐A1bVrn‐B1bVrn‐D1b NILs (winter habit). The average AUDPC of spring and winter habit ‘Wanser’ NILs differed at all three Vrn‐A1, Vrn‐B1 and Vrn‐D1 locus comparisons. We conclude that ‘Daws’ and ‘Wanser’ have different background genetic interactions with the Vrn‐1 loci influencing cold hardiness. The marker for Vrn‐A1 is diagnostic for growth habit and cold hardiness but there is no relationship between the Vrn‐B1 and Vrn‐D1 markers and the cold tolerance of the NILs used in this study.     

Alkaloid accumulation in capsules of the selfed and cross-pollinated poppy

In harmony with international regulations the creation of poppy cultivars with high alkaloid content as well as free of alkaloids is in progress. In the course of selection procedures, five poppy cultivars of high chemical diversity, ‘Kheops’, ‘AT, ‘Tebona’, ‘Kék Gemona’ and ‘Przemko’, were selfed and crossed in two successive years (2000‐01). After full ripening, both the biomass production and the alkaloid content of capsules of selfed and crossed plants were determined. When the castrated alkaloid‐free cultivar ‘Przemko’ (accumulating only 0.01 mg/100 g morphine in capsules) was pollinated with cultivars rich in alkaloids, the morphine content of its capsules increased to 0.9‐7.5 mg/100 g‐values. Based on the well‐known biosynthetic background of poppy alkaloids, this can be explained by promotion of (S)‐norcoclaurine synthesis, which seems to be suppressed in alkaloid‐free plants; but tissues of the developing hybrid seeds, which contain a combined gene pool of both alkaloid‐free cultivar and that of the pollen donor plants, release this suppression. The chemical metaxenia also manifested itself in other combinations. In particular, the appearance of narcotine (0.1‐1.1 mg/100 g) in the narcotine‐free cultivars ‘Kheops’, ‘A1’ and ‘Przemko’ can be explained biochemically. If the narcotine‐type plants ‘Kék Gemona’ (accumulating 11.3 mg/g narcotine) were used for pollination, the developing seed tissue, which contains the gene pool of narcotine biosynthesis, might contribute to the suppression of the activity of 1,2‐dehydroreticuline reductase, which leads to the accumulation of narcotine at the cost of morphinanes.     

Genetic basis of seedling-resistance to leaf rust in bread wheat 'Thatcher'

The bread wheat cultivar ‘Thatcher’ is documented to carry the gene Lr22b for adult‐plant resistance to leaf rust. Seedling‐resistance to leaf rust caused by Puccinia triticina in the bread wheat cultivar ‘Thatcher’, the background parent of the near‐isogenic lines for leaf rust resistance genes in wheat, is rare and no published information could be found on its genetic basis. The F₂ and F₃ analysis of the cross ‘Agra Local’ (susceptible) × ‘Thatcher’ showed that an apparently incompletely dominant gene conditioned seedling‐resistance in ‘Thatcher’ to the three ‘Thatcher’‐avirulent Indian leaf rust pathotypes – 0R8, 0R8‐1 and 0R9. Test of allelism revealed that this gene (temporarily designated LrKr1) was derived from ‘Kanred’, one of the parents of ‘Thatcher’. Absence of any susceptible F₂ segregants in a ‘Thatcher’ × ‘Marquis’ cross confirmed that an additional gene (temporarily designated LrMq1) derived from ‘Marquis’, another parent of ‘Thatcher’, was effective against pathotype 0R9 alone. These two genes as well as a second gene in ‘Kanred’ (temporarily designated LrKr2), which was effective against all the three pathotypes, but has not been inherited by ‘Thatcher’, seem to be novel, undocumented leaf rust resistance genes.     

Seasonal Changes in Carbohydrate and Protein Content of Seeded Bermudagrasses and Their Effect on Spring Green‐Up

The widespread use of warm‐season turfgrasses in transition zones of Europe, such as northern Italy, has been hampered by the long dormancy periods. To encourage the conversion from cool‐ to warm‐season grasses, research is needed to identify cultivars that exhibit early spring green‐up. A 2‐year study was conducted at the agricultural experimental farm of Padova University from November 2006 to October 2008 to compare water‐soluble carbohydrate and protein content in stolons of four bermudagrass [Cynodon dactylon (L.) Pers.] cultivars and determine their effect on spring green‐up. Samples of ‘La Paloma’, ‘NuMex Sahara’, ‘Princess 77’ and ‘Yukon’ were collected monthly, and water‐soluble carbohydrates (WSC) and crude protein (CP) content of stolons were measured. Dry weight values of WSC and CP for each cultivar were regressed against days needed to reach 80 % green cover in spring (D80). ‘Yukon’ exhibited the highest rhizome dry weight and WSC content during the winter months and was the fastest to reach 80 % green cover. Conversely, ‘Princess 77’ was the slowest cultivar to green‐up in both years. Regression analysis revealed a stronger relationship between D80 and WSC than between CP content and D80.     

Comparison of two fertility restoration systems against photoperiod-sensitive cytoplasmic male sterility in wheat

A ‘two‐line system’ using photoperiod‐sensitive cytoplasmic male sterility (PCMS) caused by Aegilops crassa cytoplasm under a long‐day photoperiod ( 15 h) has been proposed as a new means of producing hybrid varieties in common wheat. The PCMS line is maintained by self‐pollination under short‐day conditions, and hybrid seeds can be produced through outcrossing of the PCMS line with a pollinator under long‐day conditions. Two kinds of fertility restoration systems against the PCMS are known. One is involved with a set of multiple fertility‐restoring (Rf) genes in the wheat cultivar ‘Norin 61’ located on (at least) chromosomes 4A, 1D, 3D and 5D. The other is controlled by a single dominant major Rf gene, Rfd1, located on the long arm of chromosome 7B in the wheat cultivar ‘Chinese Spring’. To examine the degree of fertility restoration by these two systems, nine PCMS lines were crossed with ‘Norin 61’ and ‘Chinese Spring’ as the restorer lines, and the F₁ hybrids were investigated. The degree of fertility restoration was estimated by comparing the seed set rates in the F₁ hybrids having the Ae. crassa cytoplasm and those with normal cytoplasm. The results revealed that the fertility restoration ability of a set of multiple Rf genes in ‘Norin 61’ was higher than that of the Rfd1 gene in ‘Chinese Spring’.