Biochemical markers for cabbage seedpod weevil (<Emphasis Type="Italic">Ceutorhynchus obstrictus</Emphasis> (Marsham)) resistance in canola (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

In the last decade, the cabbage seedpod weevil (Ceutorhynchus obstrictus (Marsham)) has become a major insect pest of canola (Brassica napus L.) in Canada reducing seed yields up to 35%. Therefore, the benefits of developing weevil resistant germplasm to canola breeders and the industry would reduce input costs, pesticide use, environmental degradation and increase yield. Yellow mustard (Sinapis alba L.) is resistant to C. obstrictus (CSPW), although the exact mechanism is not known (McCaffrey et al. 1999). A unique canola population was generated at the University of Guelph from a cross between B. napus and S. alba through embryo rescue and backcrossed to canola several times prior to double haploid (DH) production. Approximately one-half of this DH population had canola quality glucosinolate concentration (<16 μmol/g) and was used for further breeding. The hypothesis was that some DH progeny from this cross inherited resistance to CSPW from S. alba. Weevil infestation levels were assessed for the B. napus × S. alba BC2 and BC3 DH populations in the field over 7 years in Alberta where weevil pressure is strong to establish the resistant or susceptible status of these lines. The basic objectives for this study were to confirm field resistance in the B. napus × S. alba germplasm in Ontario and to identify any biochemical markers associated with resistance/susceptibility. Canola doubled haploid lines derived from BC2 or BC3 families were field screened for resistance (R) followed by chemical analysis of glucosinolates to detect biochemical polymorphisms correlated with CSPW resistance using High Performance Liquid Chromatography (HPLC). Two polymorphic peaks were found, one each, from extracts of upper cauline leaves and Stage 3 pod seed, with retention times of ~23 and 19 min, respectively. These HPLC peaks consistently correlated with larval infestation data and the peak differences between R and S DH lines were significant. Therefore, these two peaks can be considered as biochemical markers in this breeding germplasm and may play a role in rapid and early detection of CSPW resistance.     

Differential preference of <Emphasis Type="Italic">Capsicum</Emphasis> spp. cultivars by <Emphasis Type="Italic">Aphis gossypii</Emphasis> is conferred by variation in volatile semiochemistry

The aim of this study was to demonstrate that Capsicum spp. cultivars are differentially preferred by the cotton aphid, Aphis gossypii, and to investigate the role of volatile semiochemicals in conferring differences in host preferences. Two preference assays were conducted in 2008 under greenhouse conditions. Fourteen different commercially available cultivars were grown in cages protected by an anti-aphid net, and were infested 60 days after planting, through the release of ten adult female A. gossypii per plant. The results showed that after a five-day infestation period, statistically significant differences in the mean number of A. gossypii between cultivars were observed, with Sweet Pepper Hybrid Green Belt (SPHGB) being one of the cultivars with the lowest number of A. gossypii per plant. To test the hypothesis that the preference of cultivars was associated with release of volatile, Capsicum spp-derived semiochemicals, olfactometer behavior bioassays were conducted with A. gossypii, using volatile organic compounds (VOCs) collected from non-preferred SPHGB and preferred SPAB cultivars. A. gossypii was significantly repelled only by the VOCs of infested SPHGB. Furthermore, coupled gas chromatography-mass spectrometry (GC-MS) analysis of VOCs released by plants prior to, and after, A. gossypii infestation, revealed that the non-preferred SPHGB cultivar released nine additional compounds after infestation, including 6-methyl-5-hepten-2-one, a known plant defense semiochemical involved in plant—aphid interactions. These data suggest that non-preferred cultivars releasing this semiochemical have the potential to be used in breeding programs aimed at producing A. gossypii-resistant Capsicum spp. cultivars.     

Interspecific hybridization of fig (<Emphasis Type="Italic">Ficus carica</Emphasis> L.) and <Emphasis Type="Italic">Ficus erecta</Emphasis> Thunb., a source of Ceratocystis canker resistance

Ceratocystis canker, which is caused by the fungus Ceratocystis fimbriata Ellis et Halsted, is one of the most severe diseases of the common fig (Ficus carica L.). In contrast, the wild fig species F. erecta Thunb. is resistant to this fungus. We performed interspecific hybridization between the common fig (seed parent) and F. erecta (pollen parent) through artificial pollination. Even though hybrid seeds showed high germination rates, the seedling survival rates were low. All of the seedlings contained the expected simple sequence repeat (SSR) alleles from both common fig and F. erecta at each of the three loci tested, thus confirming the parent–offspring relationships of the interspecific hybrids. The leaf morphological characters of hybrid seedlings were intermediate between those of the parents. Cuttings of cultivars of common fig, F. erecta, and hybrid seedlings were inoculated with C. fimbriata by direct wounding of the shoot. All of the common fig cultivars tested withered and died within 10 weeks. Leaves and shoots of the hybrids and F. erecta were healthy 100 days after inoculation. Our results suggest that interspecific hybridization between the common fig and the wild species F. erecta is a breakthrough in the breeding of a new fig rootstock source with resistance to Ceratocystis canker.     

Ploidy level studies on the <Emphasis Type="Italic">Dioscorea cayenensis</Emphasis>/<Emphasis Type="Italic">Dioscorea rotundata</Emphasis> complex core set

The Guinea yams, Dioscorea cayenensis Lam. and D. rotundata Poir. (D. cayenensis–D. rotundata complex), represent a highly important crop, widely distributed in the humid and semi-humid tropics. The ploidy levels of 170 accessions of the core set of Guinea yams from West African countries was determined using flow cytometry with propidium iodide staining. One hundred and eight of the genotypes were found to be tetraploid, 47 were hexaploid and five were octoploid. One mixoploid individual containing tetraploid and hexaploid nuclei was also detected. A deeper analysis considering each separate taxon revealed that while for D. rotundata the majority of individuals were tetraploid, for D. cayenensis this ploidy level was not detected in any of the accessions. Also, no association between ploidy level and place of cultivation was found for the evaluated germplasm. The obtained data is highly valuable for breeding programs of Guinea yam, especially for the optimization of future hybridization experiments directed to the genetic improvement of this economically important crop.     

Search in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm for sources of broad-spectrum resistance to four <Emphasis Type="Italic">Tospovirus</Emphasis> species

The genus Tospovirus was considered as monotypic with Tomato spotted wilt virus (TSWV) being the only assigned species. However, extensive studies with worldwide isolates revealed that this genus comprises a number of species with distinct virulence profiles. The Neotropical South America is one center of Tospovirus diversity with many endemic species. Groundnut ringspot virus (GRSV), TSWV, Tomato chlorotic spot virus (TCSV), and Chrysanthemum stem necrosis virus (CSNV) are the predominant tomato-infecting species in Brazil. Sources of resistance were found in Solanum (section Lycopersicon) mainly against TSWV isolates from distinct continents, but there is an overall lack of information about resistance to other viral species. One-hundred and five Solanum (section Lycopersicon: Solanaceae) accessions were initially evaluated for their reaction against a GRSV isolate by analysis of symptom expression and systemic virus accumulation using DAS-ELISA. A subgroup comprising the most resistant accessions was re-evaluated in a second assay with TSWV, TCSV, and GRSV isolates and in a third assay with a CSNV isolate. Seven S. peruvianum accessions displayed a broad-spectrum resistance to all viral species with all plants being free of symptoms and systemic infection. Sources of resistance were also found in tomato cultivars with the Sw-5 gene and also in accessions of S. pimpinellifolium, S. chilense, S. arcanum, S. habrochaites, S. corneliomuelleri, and S. lycopersicum. The introgression/incorporation of these genetic factors into cultivated tomato varieties might allow the development of genetic materials with broad-spectrum resistance, as well as with improved levels of phenotypic expression.     

Genetic effects and genetic relationships among <Emphasis Type="Italic">shrunken</Emphasis> (<Emphasis Type="Italic">sh2</Emphasis>) sweet corn lines and F1 hybrids

The objectives of this study were to quantify the components of genetic variance and the genetic effects, and to examine the genetic relationship of inbred lines extracted from various shrunken2 (sh2) breeding populations. Ten diverse inbred lines developed from sh2 genetic background, were crossed in half diallel. Parents and their F1 hybrids were evaluated at three environments. The parents were genotyped using 20 polymorphic simple sequence repeats (SSR). Agronomic and quality traits were analysed by a mixed linear model according to additive-dominance genetic model. Genetic effects were estimated using an adjusted unbiased prediction method. Additive variance was more important than dominance variance in the expression of traits related to ear aspects (husk ratio and percentage of ear filled) and eating quality (flavour and total soluble solids). For agronomic traits, however, dominance variance was more important than additive variance. The additive genetic correlation between flavour and tenderness was strong (r = 0.84, P < 0.01). Flavour, tenderness and kernel colour additive genetic effects were not correlated with yield related traits. Genetic distance (GD), estimated from SSR profiles on the basis of Jaccard’s similarity coefficient varied from 0.10 to 0.77 with an average of 0.56. Cluster analysis classified parents according to their pedigree relationships. In most studied traits, F1 performance was not associated with GD.     

Allelic variation for the rust resistance gene <Emphasis Type="Italic">Lr34</Emphasis>/<Emphasis Type="Italic">Yr18</Emphasis> in Canadian wheat cultivars

The wheat (Triticum aestivum L.) gene Lr34/Yr18 conditions resistance to leaf rust, stripe rust, and stem rust, along with other diseases such as powdery mildew. This makes it one of the most important genes in wheat. In Canada, Lr34 has provided effective leaf rust resistance since it was first incorporated into the cultivar Glenlea, registered in 1972. Recently, molecular markers were discovered that are either closely linked to this locus, or contained within the gene. Canadian wheat cultivars released from 1900 to 2007, breeding lines and related parental lines, were tested for sequence based markers caSNP12, caIND11, caIND10, caSNP4, microsatellite markers wms1220, cam11, csLVMS1, swm10, csLV34, and insertion site based polymorphism marker caISBP1. Thirty different molecular marker haplotypes were found among the 375 lines tested; 5 haplotypes had the resistance allele for Lr34, and 25 haplotypes had a susceptibility allele at this locus. The numbers of lines in each haplotype group varied from 1 to 140. The largest group was represented by the leaf rust susceptible cultivar “Thatcher” and many lines derived from “Thatcher”. The 5 haplotypes that had the resistance allele for Lr34 were identical for the markers tested within the coding region of the gene but differed in the linked markers wms1220, caISBP1, cam11, and csLV34. The presence of the resistance or susceptibility allele at the Lr34 locus was tracked through the ancestries of the Canadian wheat classes, revealing that the resistance allele was present in many cultivars released since the 1970s, but not generally in the older cultivars.     

Resistance screening of <Emphasis Type="Italic">Coffea</Emphasis> spp. accessions for <Emphasis Type="Italic">Pratylenchus coffeae</Emphasis> and <Emphasis Type="Italic">Radopholus arabocoffeae</Emphasis> in Vietnam

Coffee varieties with resistance for the plant-parasitic nematodes Pratylenchus coffeae and Radopholus arabocoffeae are limited in Vietnam. A selection of imported varieties and high yield varieties of Arabica coffee in Vietnam were evaluated for resistance to both plant-parasitic nematode species in Northern Vietnam. The same experiments were carried out with hybrid arabica coffee, three selected clones of Coffea canephora and one clone of Coffea excelsa in the Western Highland of Vietnam. The screened coffee accessions from Ethiopia (KH1, KH13, KH20, KH21, KH29, and KH31) were susceptible and good host for P. coffeae. Also accessions 90P4 (Portugal) and Oro azteca (Mexico) had a reproduction factor Rf > 1. Pluma Hidalgo (Mexico), 90/6 (Vietnam), 90P3 (Portugal), 90P2 (Vietnam), Variedad (Mexico), 90T (Portugal), and Garnica (Mexico) were poor hosts (Rf < 1) but not tolerant to P. coffeae, expressed by a reduction of root weight compared to untreated control plants. Most of the coffee accessions tested in Northern Vietnam were intolerant to R. arabocoffeae, except 90T which showed no reduction of root weight, even at high initial nematode densities (4,000/pot). Good hosts for R. arabocoffeae were Variedad, KH1, KH21, KH29, KH20, KH31, and KH13 with Rf > 1. Pluma Hidalgo, 90/6, 90P3, 90P2, 90T, Oro azteca, and Garnica were poor hosts (Rf < 1). In the Western Highland experiment, all arabica coffee accessions were susceptible for P. coffeae with Rf ranging from 1.41 to 1.59. Tolerance to P. coffeae was found in C. liberica var. Dewevrei, Hong34 and Nhuantren. Coffea excelsa, Hong34, Nhuantren, and H1C19 were tolerant to R. arabocoffeae at the highest inoculation density (4,000 nematodes/pot). The most susceptible accessions were Nhuantren and K55. Resistance (Rf < 1) to R. arabocoffeae was found in C. liberica var. Dewevrei and Hong34. This article reports on the first screening for resistance and tolerance to P. coffeae and R. arabocoffeae in coffee accessions in Vietnam and shows promising results for enhanced coffee-breeding.     

Increased resistance to <Emphasis Type="Italic">Ustilago zeae</Emphasis> and <Emphasis Type="Italic">Fusarium verticilliodes</Emphasis> in maize inbred lines bred for <Emphasis Type="Italic">Fusarium graminearum</Emphasis> resistance

Preliminary field observations in our maize breeding nurseries indicated that breeding for improved resistance to gibberella ear rot (Fusarium graminearum) in maize may indirectly select for resistance to another ear disease, common smut (Ustilago zeae). To investigate this, we compared the disease severity ratings obtained on 189 maize inbreds, eight of which included our inbreds developed with selection for gibberella ear rot resistance after field inoculation and breeding for 8–10 years. No correlation was found between disease severities for the 189 inbreds but the eight gibberella-resistant lines were consistently more resistant to smut. To further examine this relationship and to determine if these eight inbreds would be useful for developing inbreds with either common smut or fusarium ear rot (F. verticilliodes) resistance, we conducted a Griffing’s diallel analysis on six inbreds of maize, four with high levels of gibberella ear rot resistance representing all of the pedigree groups in our eight gibberella lines, and two with very low levels. Our most gibberella ear rot resistant inbreds, CO433 and CO441, had the lowest disease ratings for all three diseases, the consistently largest general combining ability effects and several significant specific combining ability effects. It was concluded that some inbreds bred specifically for gibberella ear rot would also be useful in breeding for resistance to common smut and fusarium ear rot.     

Genetic and histological characterization of a novel recessive genic male sterile line of <Emphasis Type="Italic">Brassica napus</Emphasis> derived from a cross with <Emphasis Type="Italic">Capsella bursa</Emphasis>-<Emphasis Type="Italic">pastoris</Emphasis>

In a previously made cross Brassica napus cv. Oro (2n = 38) × Capsella bursa-pastoris (2n = 4x = 32), one F₁ hybrid with 2n = 38 was totally male sterile. The hybrid contained no complete chromosomes from C. bursa-pastoris, but some specific AFLP (amplified fragment length polymorphism) bands of C. bursa-pastoris were detected. The hybrid was morphologically quite similar to ‘Oro’ except for smaller flowers with rudimentary stamens but normal pistils, and showed good seed-set after pollination by ‘Oro’ and other B. napus cultivars. The fertility segregation ratios (3:1, 1:1) in its progenies indicated that the male sterility was controlled by a single recessive gene. In the pollen mother cells of the male sterile hybrid, chromosome pairing and segregation were normal. Histological sectioning of its anthers showed that the tapetum was multiple layers and was hypertrophic from the stage of sporogenic cells, and that the tetrads were compressed by the vacuolated and disaggregated tapetum and no mature pollen grains were formed in anther sacs, thus resulting in male sterility. The possible mechanisms for the production of the male sterile hybrid and its potential in breeding are discussed.     

Meiotic behavior as a selection tool in the breeding of <Emphasis Type="Italic">Brachiaria humidicola</Emphasis> (Poaceae)

Brachiaria humidicola is a tropical grass that grows in seasonally swampy grasslands in Africa. In Brazil, two apomictic cultivars (2n = 54) of this species are widely used as pastures in poorly drained soils. The recent discovery of a sexual polyploid accession (2n = 36) in the germplasm collection at the Embrapa Beef Cattle Research Center allowed intraspecific hybridization with the objective of broadening the genetic variability and selection of superior genotypes in this species. Hybridization, however, depends on accessions with the same ploidy level. Cytological analyses of 55 accessions revealed that 19 apomictic accessions also presented 2n = 36 chromosomes. Chromosome pairing in hexavalent association at diakinesis and metaphase I suggested that the basic chromosome number for this species is x = 6. Cytological analysis revealed abnormalities in variable frequencies in the meiosis of these hexaploid (2n = 6x = 36) accessions. The most common were those related to irregular chromosome segregation which led to unbalanced gamete formation, but chromosome stickiness was also recorded. These results clearly demonstrate the value of cytogenetics in the choice of genitors and for superior hybrids to be obtained in the breeding of this species. For that both the ploidy level and the frequency of abnormalities need to be considered, besides other favorable agronomic characteristics.     

RDA derived <Emphasis Type="Italic">Oryza minuta</Emphasis>-specific clones to probe genomic conservation across <Emphasis Type="Italic">Oryza</Emphasis> and introgression into rice (<Emphasis Type="Italic">O. sativa</Emphasis> L.)

Molecular markers have been successfully used in rice breeding however available markers based on Oryza sativa sequences are not efficient to monitor alien introgression from distant genomes of Oryza. We developed O. minuta (2n = 48, BBCC)-specific clones comprising of 105 clones (266–715 bp) from the initial library composed of 1,920 clones against O. sativa by representational difference analysis (RDA), a subtractive cloning method and validated through Southern blot hybridization. Chromosomal location of O. minuta-specific clones was identified by hybridization with the genomic DNA of eight monosomic alien additional lines (MAALs). The 37 clones were located either on chromosomes 6, 7, or 12. Different hybridization patterns between O. minuta-specific clones and wild species such as O. punctata, O. officinalis, O. rhizomatis, O. australiensis, and O. ridleyi were observed indicating conservation of the O. minuta fragments across Oryza spp. A highly repetitive clone, OmSC45 hybridized with O. minuta and O. australiensis (EE), and was found in 6,500 and 9,000 copies, respectively, suggesting an independent and exponential amplification of the fragment in both species during the evolution of Oryza. Hybridization of 105 O. minuta specific clones with BB- and CC-genome wild Oryza species resulted in the identification of 4 BB-genome-specific and 14 CC-genome-specific clones. OmSC45 was identified as a fragment of RIRE1, an LTR-retrotransposon. Furthermore this clone was introgressed from O. minuta into the advanced breeding lines of O. sativa.     

Application of the TRAP technique to lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L<Emphasis Type="Italic">.</Emphasis>) genotyping

Summary To demonstrate the applicability of the target region amplification polymorphism (TRAP) marker technique to lettuce genotyping, we fingerprinted 53 lettuce (Lactuca sativa L.) cultivars and six wild accessions (three from each of the two wild species, L. saligna L. and L. serriola L.). Seven hundred and sixty-nine fragments from 50 to 900 bp in length were amplified in 10 PCR reactions using 10 fixed primers in combination with four fluorescent labeled arbitrary primers. Three hundred and eighty-eight of these fragments were polymorphic among the 59 Lactuca entries and 107 fragments were polymorphic among the 53 lettuce cultivars and the six wild accessions; 251 fragments were present only in the wild species. These markers not only discriminated all cultivars, but also revealed the evolutionary relationship among the three species: L. sativa, the cultivated species, is more closely related to L. serriola than to L. saligna. Cluster analysis grouped the cultivars by horticultural types with a few exceptions. These results are consistent with previous findings using RFLP, AFLP, and SAMPL markers. The TRAP markers revealed significant differences in genetic variability among horticultural types, measured by the average genetic similarity among the cultivars of the same type. Within the sample set, the leaf type and butterhead types possessed relatively high genetic variability, the iceberg types had moderate variability and the romaine types had the lowest variability. The genetic behavior of TRAP markers was assessed with a mapping population of 45 recombinant inbred lines (RILs) derived from an interspecific cross between L. serriola and L. sativa. Almost all the markers segregated in the expected 1:1 Mendelian ratio and are being incorporated into the existing lettuce linkage maps. Our results indicate that the TRAP markers can provide a powerful technique for fingerprinting lettuce cultivars. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

Fingerprinting temperate <Emphasis Type="Italic">japonica</Emphasis> and tropical <Emphasis Type="Italic">indica</Emphasis> rice genotypes by comparative analysis of DNA markers

This paper describes the relative efficiency of three marker systems, RAPD, ISSR, and AFLP, in terms of fingerprinting 14 rice genotypes consisting of seven temperatejaponica rice cultivars, three indica near-isogenic lines, three indica introgression lines, and one breeding line of japonica type adapted to high-altitude areas of the tropics with cold tolerance genes. Fourteen RAPD, 21 ISSR, and 8 AFLP primers could produce 970 loci, with the highest average number of loci (92.5) generated by AFLP. Although polymorphic bands in the genotypes were detected by all marker assays, the AFLP assay discriminated the genotypes effectively with a robust discriminating power (0.99), followed by ISSR (0.76) and RAPD (0.61). While significant polymorphism was detected among the genotypes of japonica and indica through analysis of molecular variance (AMOVA), relatively low polymorphism was detected within the genotypes of japonica rice cultivars. The correlation coefficients of similarity were significant for the three marker systems used, but only the AFLP assay effectively differentiated all tested rice lines. Fingerprinting of backcross-derived resistant progenies using ISSR and AFLP markers easily detected progenies having a maximum rate of recovery for the recurrent parent genome and suggested that our fingerprinting approach adopting the ‘undefined-element-amplifying’ DNA marker system is suitable for incorporating useful alleles from the indica donor genome into the genome of temperate japonica rice cultivars with the least impact of deleterious linkage drag.     

SSR markers for marker assisted selection of root-knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) resistant plants in cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L)

Cotton (Gossypium hirsutum L) cultivars highly resistant to the southern root-knot nematode (RKN) [Meloidogyne incognita (Kofoid and White) Chitwood] are not available. Resistant germplasm lines are available; however, the difficulty of selecting true breeding lines has hindered applied breeding and no highly resistant cultivars are available to growers. Recently, molecular markers on chromosomes 11 and 14 have been associated with RKN resistance, thus opening the way for marker assisted selection (MAS) in applied breeding. Our study aimed to determine the utility of these markers for MAS. Cross one was RKN resistant germplasm M240 RNR × the susceptible cultivar, FM966 and is representative of the initial cross a breeder would make to develop a RKN resistant cultivar. Cross two consists of Clevewilt 6 × Mexico Wild (PI563649), which are the two lines originally used to develop the first highly RKN resistant germplasm. Mexico Wild is photoperiodic. We phenotyped the F₂ of cross one for gall index and number of RKN eggs per plant and genotyped each plant for CIR 316 (chromosome 11) and BNL 3661 (chromosome 14). From this, we verified that MAS was effective, and the QTL on chromosome 14 was primarily associated with a dominant RKN resistance gene affecting reproduction. In the first F₂ population of cross two, we used MAS to identify 11 plants homozygous for the markers on chromosomes 11 and 14, and which also flowered in long days. Progeny of these 11 plants were phenotyped for RKN gall index and egg number and confirmed as RKN highly resistant plants. Generally about 7–10 generations of RKN phenotyping and progeny testing were required to develop the original RKN highly resistant germplasms. Our results show that commercial breeders should be able to use the markers in MAS to rapidly develop RKN resistant cultivars.     

Molecular markers linked to <Emphasis Type="Italic">Bn</Emphasis>;<Emphasis Type="Italic">rf</Emphasis>: a recessive epistatic inhibitor gene of recessive genic male sterility in <Emphasis Type="Italic">Brassica napus </Emphasis>L.

7–7365AB is a recessive genic male sterile (RGMS) two-type line, which can be applied in a three-line system with the interim-maintainer, 7–7365C. Fertility of this system is controlled by two duplicate dominant epistatic genes (Bn;Ms3 and Bn;Ms4) and one recessive epistatic inhibitor gene (Bn;rf). Therefore an individual with the genotype of Bn;ms3ms3ms4ms4Rf_ exhibits male sterility, whereas, plant with Bn;ms3ms3ms4ms4rfrf shows fertility because homozygosity at the Bn;rf locus (Bn;rfrf) can inhibit the expression of two recessive male sterile genes in homozygous Bn;ms3ms3ms4ms4 plant. A cross of 7–7365A (Bn;ms3ms3ms4ms4RfRf) and 7–7365C (Bn;ms3ms3ms4ms4rfrf) can generate a complete male sterile population served as a mother line with restorer in alternative strips for the multiplication of hybrid seeds. In the present study, molecular mapping of the Bn;Rf gene was performed in a BC₁ population from the cross between 7–7365A and 7–7365C. Bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) technique was used to identify molecular markers linked to the gene of interest. From a survey of 768 primer combinations, seven AFLP markers were identified. The closest marker, XM5, was co-segregated with the Bn;Rf locus and successfully converted into a sequence characterized amplified region (SCAR) marker, designated as XSC5. Two flanking markers, XM3 and XM2, were 0.6 cM and 2.6 cM away from the target gene, respectively. XM1 was subsequently mapped on linkage group N7 using a doubled-haploid (DH) mapping population derived from the cross Tapidor × Ningyou7, available at IMSORB, UK. To further confirm the location of the Bn;Rf gene, additional simple sequence repeat (SSR) markers in linkage group N7 from the reference maps were screened in the BC₁ population. Two SSR markers, CB10594 and BRMS018, showed polymorphisms in our mapping population. The molecular markers found in the present study will facilitate the selection of interim-maintainer.     

Combining different resistance components enhances resistance to <Emphasis Type="Italic">Fusarium</Emphasis> head blight in spring wheat

Fusarium head blight (FHB) is a devastating disease in wheat throughout the world. FHB resistance consists of two components: resistance to initial infection (type I) and resistance to spread within infected spikes (type II). Current wheat breeding programs for FHB focus on type II resistance, which limits pathogen spread but may not be sufficiently durable. To combine type I with existing type II resistance, 113 F₉-derived recombinant inbred lines (RILs) were developed from a cross between three wheat genotypes Frontana, W9207, and Alsen. The RILs were evaluated for resistance to initial infection, FHB spread within spike, kernel damage, and deoxynivalenol (DON) content in two independent greenhouse experiments in 2006 and 2007. Among the 113 RILs, 20% lines showed ≤10% initial disease severity (IDS) and ≤11 to 30% final disease severity (FDS), and 19% had DON content ≤5 μg/g. Approximately 11% of the RILs showed tendency of higher resistance (as exhibited by lower IDS, FDS, and DON content) than the resistant parents. The 42 of the FHB-resistant RILs were analyzed with seven simple sequence repeat (SSR) markers or microsatellites known to be linked to FHB resistance. Approximately half of the RILs had molecular markers linked to both types of FHB resistance indicated the presence of type I and II resistance alleles in the RILs. The resistant RILs identified in this study should be useful for the future improvement of FHB resistance in spring wheat.     

Identification of quantitative trait loci involved in resistance to <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">syringae</Emphasis> in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

Pseudomonas syringae is the main pathogen responsible for bacterial blight disease in pea and can cause yield losses of 70%. P. syringae pv. pisi is prevalent in most countries but the importance of P. syringae pv. syringae (Psy) is increasing. Several sources of resistance to Psy have been identified but genetics of the resistance is unknown. In this study the inheritance of resistance to Psy was studied in the pea recombinant inbred line population P665 × ‘Messire’. Results suggest a polygenic control of the resistance and two quantitative trait loci (QTL) associated with resistance, Psy1 and Psy2, were identified. The QTL explained individually 22.2 and 8.6% of the phenotypic variation, respectively. In addition 21 SSR markers were included in the P665 × ‘Messire’ map, of which six had not been mapped on the pea genome in previous studies.     

Analysis of rice blast resistance gene <Emphasis Type="Italic">Pi</Emphasis>-<Emphasis Type="Italic">z</Emphasis> in rice germplasm using pathogenicity assays and DNA markers

The Pi-z gene in rice confers resistance to a wide range of races of the rice blast fungus, Magnaporthe oryzae. The objective of this study was to characterize Pi-z in 111 rice germplasm accessions using DNA markers and pathogenicity assays. The existence of Pi-z in rice germplasm was detected by using four simple sequence repeat (SSR) markers (RM527, AP4791, AP5659-1, AP5659-5) closely linked to Pi-z, and was verified using pathogenicity assays with an avirulent strain (IE1k) and two virulent races (IB33 and IB49). Among 111 germplasm accessions evaluated, 73 were found to contain the Pi-z gene using both SSR markers and pathogenicity assays. The remaining 38 germplasm accessions were found to be inconsistent in their responses to the blast races IB33, IEIk and IB49 with expected SSR marker alleles, suggesting the presence of unexpected SSR alleles and additional R gene(s). These characterized germplasm can be used for genetic studies and marker-assisted breeding for improving blast resistance in rice.