Functional markers delimiting a <Emphasis Type="Italic">Medicago</Emphasis> orthologue of pea symbiotic gene <Emphasis Type="Italic">NOD3</Emphasis>

Symbiotic gene mutated in the pea (Pisum sativum L.) line RisfixC is a determinant of the number of symbiotic root nodules. In parallel to a sharp increase in nodule number, its mutational inactivation brings about the insensitivity of nodulation to the ambient nitrate level (Nts trait). Using the established localization to the SYM2-NOD3 region of the pea linkage group I, functional PCR markers were developed for the orthologous region on the chromosome 5 of the model species Medicago truncatula. Owing to the conservation of the binding regions of the designed primers, pea orthologues were successfully amplified with 60% of the primer pairs tested. When applied to a mapping pea population from the cross of the line RisfixC x Afghanistan L1268 (sym2), the new markers allowed to localize the supernodulation mutation within 2.5 cM confidence interval in the pea genome. The placement of the functional markers on the M. truncatula chromosome 5 confined the orthologous gene location to eight overlapping BACs spanning approximately 710 kbp (positions 37,755,678–38,467,472). The narrowed list of the annotated Medicago genes in combination with the published data on their symbiotic and nitrate regulation can be used for the candidate gene identification, together with the requirements imposed by the known function in nodule number initiation and nitrate sensing. In addition, the new markers are applicable for tracking the RisfixC allele in breeding programmes aimed at the improvement of symbiotic performance.     

Relevance of unilateral and bilateral sexual polyploidization in relation to intergenomic recombination and introgression in <Emphasis Type="Italic">Lilium</Emphasis> species hybrids

Sexual polyploids were induced in diploid (2n = 2x = 24) interspecific F1 hybrids of Longiflorum × Asiatic (LA) and Oriental × Asiatic (OA) Lilium hybrids by backcrossing to Asiatic (AA) parents as well as by sib-mating of the F1 LA hybrids. A majority of the BC1 progenies were triploid and the progenies from sib-mating were tetraploid or near tetraploids. Genomic in situ hybridization (GISH) technique was applied to assess the intergenomic recombination in the BC1 populations of LA and OA hybrids obtained after unilateral sexual polyploidization. A total of 63 LA (LA × AA and AA × LA) and 53 OA hybrids were analysed. LA hybrids were originated through the functioning of 2n gametes either as 2n eggs or 2n pollen while those of OA hybrids originated through functional 2n pollen of diploid OA genotype. In both type of crosses, a majority of the progenies had originated through First Division Restitution (FDR) mechanism of functional 2n gamete either with or without a cross over. However, there were nine LA- and four OA-genotypes where Indeterminate Meiotic Restitution (IMR) was the mechanism of 2n gamete formation. Based on GISH, total amount of introgression of Longiflorum and Oriental genome into Asiatic genome was determined. Most of the BC progenies exhibited recombination and the amount of recombination was higher in LA hybrids as compared to OA hybrids. Intergenomic recombination was also determined cytologically in the 16 plants of sib-mated LA hybrids where both parents had contributed 2n gametes. Based on these results the nature of interspecific lily hybrids obtained from uni- and bilateral sexual polyploidization leading to allotriploid and allotetraploid formation is discussed in the context of introgression and intergenomic recombination.     

Development of microsatellite markers in cultivated and wild species of sections <Emphasis Type="Italic">Cepa</Emphasis> and <Emphasis Type="Italic">Phyllodolon</Emphasis> in <Emphasis Type="Italic">Allium</Emphasis>

The potential of microsatellite markers for use in genetic studies has been evaluated in Allium cultivated species (Allium cepa, A. fistulosum) and its allied species (A. altaicum, A. galanthum, A. roylei, A. vavilovii). A total of 77 polymerase chain reaction (PCR) primer pairs were employed, 76 of which amplified a single product or several products in either of the species. The 29 AMS primer pairs derived from A. cepa and 46 microsatellites primer pairs from A. fistulosum revealed a lot of polymorphic amplicons between seven Allium species. Some of the microsatellite markers were effective not only for identifying an intraspecific F₁ hybrid between shallot and bulb onion but also for applying to segregation analyses in its F₂ population. All of the microsatellite markers can be used for interspecific taxonomic analyses among two cultivated and four wild species of sections Cepa and Phyllodolon in Allium. Generally, our data support the results obtained from recently performed analyses using molecular and morphological markers. However, the phylogeny of A. roylei, a threatened species with several favorable genes, was still ambiguous due to its different positions in each dendrogram generated from the two primer sets originated from A. cepa and A. fistulosum.     

Obtainment of an intergeneric hybrid between <Emphasis Type="Italic">Forsythia</Emphasis> and <Emphasis Type="Italic">Abeliophyllum</Emphasis>

Forsythia suspensa and F. ‘Courtaneur’ were used as female parents to cross with Abeliophyllum distichum in 2011 and an intergeneric hybrid of F. suspensa × A. distichum was obtained, though with very low seed set. The morphological characteristics, flower fragrance and volatile organic compounds of flowers were analysed. The intergeneric hybrid had intermediate morphological characteristics of both parents and flower fragrance and was confirmed as a true intergeneric hybrid by amplified fragment length polymorphism (AFLP) markers. Compared with its mother parent (F. suspensa), flowers of the intergeneric hybrid are pale yellow with delicate fragrance. Volatile organic compounds of flowers were retrieved by purge-and-trap techniques, and determined by gas chromatography and mass spectrometry (GC–MS). The main volatile organic components of F. suspensa were isoprenoids, while the main volatile organic components of A. distichum and the hybrid of F. suspensa × A. distichum were aliphatics. To determine the time and the site of intergeneric hybridizing barriers occured, the pollen tubes’ behavior after pollination was observed under fluorescence microscopy. It was found that significant pre-fertilization incompatibility existed in intergeneric crossing combinations [F. ‘Courtaneur’ (Pin) × A. distichum (Thrum) and F. suspensa (Pin) × A. distichum (Thrum)], and only a few pollen tubes of A. distichum penetrated into the ovaries of Forsythia. In our research, an intergeneric hybrid between Forsythia and Abeliophyllum was obtained for the first time, which will provide a solid foundation for expanding the flower color range of Forsythia and breeding fragrant-flowered cultivars.     

Identification of resistant sources against <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> in <Emphasis Type="Italic">Brassica</Emphasis> species with emphasis on <Emphasis Type="Italic">B. oleracea</Emphasis>

Stem rot caused by Sclerotinia sclerotiorum is one of the most devastating diseases of rapeseed (Brassica napus L.) which causes huge loss in rapeseed production. Genetic sources with high level of resistance has not been found in rapeseed. In this study, 68 accessions in six Brassica species, including 47 accessions of B. oleracea, were evaluated for leaf and stem resistance to S. sclerotiorum. Large variation of resistance was found in Brassica, with maximum differences of 5- and 57-folds in leaf and stem resistance respectively. B. oleracea, especially its wild types such as B. rupestris, B. incana, B. insularis, and B. villosa showed high level of resistance. Our data suggest that wild types of B. oleracea possess tremendous potential for improving S. sclerotiorum resistance of rapeseed.     

Screening global <Emphasis Type="Italic">Medicago</Emphasis> germplasm for weevil (<Emphasis Type="Italic">Hypera postica</Emphasis> Gyll.) tolerance and estimation of genetic variability using molecular markers

The genus Medicago encompasses many important forage species for both temperate and tropical regions. M. sativa L., commonly known as lucerne, is one of the most important forage species grown worldwide, but its production suffers seriously from weevil (Hypera postica Gyll.) infestation. The aim of this work was to identify species/accessions with tolerance to weevil and their molecular analysis using simple sequence repeat (SSR) markers. After screening 197 global germplasm encompassing 50 Medicago species for weevil tolerance, 22 lines representing 13 species were identified where leaf damage was ≤15% (P ≤ 0.05). In total, 37 accessions of the 22 lines, five Indian lucerne cultivars with leaf damage ≥75% and 10 accessions of the 13 Medicago species with low to high infestation (>25%) were molecularly assessed using 11 SSR markers (5 newly developed) to delineate closest to lucerne lines for breeding. In total, 33 bands were scored. The SAHN clustering using UPGM algorithm resulted into two main clusters supported with high boot strap values and with genetic similarity ranging from 0.33 to 0.96. Two accessions of M. tenoreana were observed closest to Indian lucerne cultivars. The rich variability revealed can be used as potential resource for transferring genes across species. Although the inter-specific hybridization is difficult preposition in genus Medicago largely due to post fertilization barrier, the identified species/accessions can be utilized on priority in breeding programs especially employing biotechnological tools like culturing of fertilized pods, ovule-embryo culture and electroporation.     

Genetics of resistance to <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> in <Emphasis Type="Italic">Cucumis sativus</Emphasis> var. <Emphasis Type="Italic">hardwickii</Emphasis> R. Alef

The genetics of resistance to Cucumber mosaic virus (CMV) in Cucumis sativus var. hardwickii R. Alef, the wild progenitor of cultivated cucumber was assessed by challenge inoculation and by natural infection of CMV. Among the 31 genotypes of C. sativus var. hardwickii collected from 21 locations in India the lowest mean percent disease intensity (PDI) was recorded in IC-277048 (6.33%) while the highest PDI was observed in IC-331631 (75.33%). All the four cultivated varieties (DC-1, DC-2, CHC-1 and CHC-2) showed very high PDI and susceptible disease reaction. Based on mean PDI, 8 genotypes were categorized as resistant, 13 as moderately resistant, 9 as moderately susceptible and one as susceptible. A chi-square test of frequency distribution based on mean PDI in F₂ progenies of six resistant × susceptible crosses revealed monogenic recessive Mendelian ratio 1(R):3(S) to be the best fit. This monogenic recessive model was further confirmed by 1(R):1(S) ratio as the best fit for back cross with resistant parent and no fit for either 3:1 or 1:1 in the back cross with the susceptible parent. The results revealed that CMV resistance in C. sativus var. hardwickii was controlled by a single recessive gene. Considering the cross compatibility between C. sativus var. hardwickii and cultivated cucumber, the resistance trait can be easily transferred to cultivated species through simple backcross breeding.     

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.     

Estimation of phenotypic divergence in a collection of <Emphasis Type="Italic">Capsicum</Emphasis> species for yield-related traits

Phenotypic divergence among the 24 accessions belonging to a collection of six species of Capsicum from different geoclimatic regions available in field gene bank of CIMAP, Lucknow, India, was quantified by multivariate analysis for 12 quantitative and qualitative traits. Based on their values, all 24 accessions were grouped into six clusters such that the genetic stocks within cluster had smaller D ² values among themselves than those belonging to different clusters. The accessions of Capsicum annuum were distributed in different clusters in morphotyping representing different species, suggesting that taxonomic characteristics are not always related to agronomic traits. No parallelism/association was found between geographical and phenotypic diversity. Accessions 15 and 23 (clusters V and VI, respectively) had distinct identity. The three characteristics that played the greatest role in differentiation were fruit diameter, number of fruits per plant, and leaf diameter, which can be utilized as conventional/morphological markers for the improvement of chilli yield and obtaining good segregants in chilli breeding programs.     

Resistance to <Emphasis Type="Italic">Cucumber green mottle mosaic virus</Emphasis> in <Emphasis Type="Italic">Cucumis sativus</Emphasis>

Cucumber green mottle mosaic virus (CGMMV) is a severe threat for cucumber production worldwide. At present, there are no cultivars available in the market which show an effective resistance or tolerance to CGMMV infection, only wild Cucumis species were reported as resistant. Germplasm accessions of Cucumis sativus, as well as C. anguria and C. metuliferus, were mechanically infected with the European and Asian strains of CGMMV and screened for resistance, by scoring symptom severity, and conventional RT-PCR. The viral loads of both CGMMV strains were determined in a selected number of genotypes using quantitative RT-PCR. Severe symptoms were found following inoculation in C. metuliferus and in 44 C. sativus accessions, including C. sativus var. hardwickii. Ten C. sativus accessions, including C. sativus var. sikkimensis, showed intermediate symptoms and only 2 C. sativus accessions showed mild symptoms. C. anguria was resistant to both strains of CGMMV because no symptoms were expressed and the virus was not detected in systemic leaves. High amounts of virus were found in plants showing severe symptoms, whereas low viral amounts found in those with mild symptoms. In addition, the viral amounts detected in plants which showed intermediate symptoms at 23 and 33 dpi, were significantly higher in plants inoculated with the Asian CGMMV strain than those with the European strain. This difference was statistically significant. Also, the amounts of virus detected over time in plants did not change significantly. Finally, the two newly identified partially resistant C. sativus accessions may well be candidates for breeding programs and reduce the losses produced by CGMMV with resistant commercial cultivars.     

Production and characterization of inter-sectional hybrids between <Emphasis Type="Italic">Kalanchoe spathulata</Emphasis> and <Emphasis Type="Italic">K. laxiflora</Emphasis> ( = <Emphasis Type="Italic">Bryophyllum crenatum</Emphasis>)

The genus Kalanchoe is currently divided into section Kalanchoe and section Bryophyllum, and there has been no successful report on the production of inter-sectional hybrids. Therefore, reciprocal crosses were made between Kalanchoe spathulata (sect. Kalanchoe) and K. laxiflora (sect. Bryophyllum) in order to obtain basic information on the reproductive barriers between these two sections. The seeds were aseptically germinated in vitro and the plants were grown in greenhouse till flowering. When K. spathulata was used as a maternal donor, 39 out of 80 plants showed intermediate characteristics between K. spathulata and K. laxiflora. In contrast, no plants were obtained in the reverse crosses. Hybridity of these plants was confirmed by flow cytometric analysis, chromosome numbers and RAPD analysis. Bulbil formation on the leaf margin as one of the conspicuous characteristics of K. laxiflora was not observed in the hybrids. Some of the hybrid lines showed some pollen fertility, but failed to yield viable seeds by self-pollination or backcross-pollination. Successful production of the inter-sectional hybrid between the two species suggests that they are not so distantly related as considered previously.     

Role of <Emphasis Type="Italic">Berberis</Emphasis> spp. as alternate hosts in generating new races of <Emphasis Type="Italic">Puccinia graminis</Emphasis> and <Emphasis Type="Italic">P. striiformis</Emphasis>

The common barberry and several other Berberis spp. serve as the alternate hosts to two important rust pathogens of small grains and grasses, Puccinia graminis and P. striiformis. Barberry eradication has been practiced for centuries as a means to control stem rust. Diverse virulence variations have been observed in populations of P. graminis f. sp. tritici that were associated with susceptible barberries in North America. Barberry likely has played a role in generating new races of P. striiformis f. sp. tritici in some regions in the world. Several North American stem rust races, namely races 56, 15B and QCC, initially originated from barberry, were subsequently responsible for generating large-scale epidemics. Thus, sexual cycles on Berberis spp. may generate virulence combinations that could have serious consequences to cereal crop production.     

Dominant gene for common bean resistance to common bacterial blight caused by <Emphasis Type="Italic">Xanthomonas</Emphasis><Emphasis Type="Italic">axonopodis</Emphasis> pv. <Emphasis Type="Italic">phaseoli</Emphasis>

The common bacterial blight pathogen [Xanthomonas axonopodis pv. phaseoli (Xap)] is a limiting factor for common bean (Phaseolus vulgaris L.) production worldwide and resistance to the pathogen in most commercial cultivars is inadequate. Variability in virulence of the bacterial pathogen has been observed in strains isolated from Puerto Rico and Central America. A few common bean lines show a differential reaction when inoculated with different Xap strains, indicating the presence of pathogenic races. In order to study the inheritance of resistance to common bacterial blight in common bean, a breeding line that showed a differential foliar reaction to Xap strains was selected and was crossed with a susceptible parent. The inheritance of resistance to one of the selected Xap races was determined by analysis of segregation patterns in the F₁, F₂, F₃ and F₄ generations from the cross between the resistant parent PR0313-58 and the susceptible parent ‘Rosada Nativa’. The F₁, F₂ and F₃ generations were tested under greenhouse conditions. Resistant and susceptible F_3:4 sister lines were tested in the field. The statistical analysis of all generations followed the model for a dominant resistance gene. The resistant phenotype was found to co-segregate with the SCAR SAP6 marker, located on LG 10. These results fit the hypothesis that resistance is controlled by a single dominant gene. The symbol proposed for the resistance gene is Xap-1 and for the bacterial race, XapV1.     

Hybrids of <Emphasis Type="Italic">Passiflora</Emphasis>: <Emphasis Type="Italic">P. gardneri</Emphasis> versus <Emphasis Type="Italic">P. gibertii,</Emphasis> confirmation of paternity,morphological and cytogenetic characterization

Two new varieties of interspecific hybrids of Passiflora have been developed from the cross between P. gardneri versus P. gibertii, both registered under the Passiflora Society International. Twelve putative hybrids were analyzed. Hybridization was confirmed using RAPD and SSR markers. Primer UBC11 (5′-CCGGCCTTAC-3′) generated informative bands. Primer SSR Pe75 has amplified species-specific fragments and a heterozygote status was observed with two parent bands 300 and 350 bp. The molecular markers generated have been analyzed for the presence or absence of specific informative bands. Based on the morphological characterization, we have identified two hybrid varieties: P. ‘Gabriela’ and P. ‘Bella’. P. ‘Gabriela’ produced flowers in bluish tones, bluish petals on the adaxial and abaxial faces, light blue sepals on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. P. ‘Bella’ produced flowers in lilac tones, intense lilac petals on the adaxial and abaxial faces, dark lilac sepals with whitish edges on the adaxial and light green on the abaxial faces, corona with the base of filaments in intense lilac color and white apex. The cytogenetic analysis verified that the hybrids have the same chromosomal number as the parents (2n = 18); the formation of bivalents between the homeologous chromosomes (n = 9) was observad, leading to regular meiosis, which allows the sexual reproduction and use of these hybrids in breeding programs.     

Identification of genetic sources with attenuated <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis>-induced symptoms in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm

The whitefly-transmitted Tomato chlorosis virus (ToCV) (genus Crinivirus) is associated with yield and quality losses in field and greenhouse-grown tomatoes (Solanum lycopersicum) in South America. Therefore, the search for sources of ToCV resistance/tolerance is a major breeding priority for this region. A germplasm of 33 Solanum (Lycopersicon) accessions (comprising cultivated and wild species) was evaluated for ToCV reaction in multi-year assays conducted under natural and experimental whitefly vector exposure in Uruguay and Brazil. Reaction to ToCV was assessed employing a symptom severity scale and systemic virus infection was evaluated via RT-PCR and/or molecular hybridization assays. A subgroup of accessions was also evaluated for whitefly reaction in two free-choice bioassays carried out in Uruguay (with Trialeurodes vaporariorum) and Brazil (with Bemisia tabaci Middle-East-Asia-Minor1—MEAM1?=?biotype B). The most stable sources of ToCV tolerance were identified in Solanum habrochaites PI 127827 (mild symptoms and low viral titers) and S. lycopersicum ‘LT05’ (mild symptoms but with high viral titers). These two accessions were efficiently colonized by both whitefly species, thus excluding the potential involvement of vector-resistance mechanisms. Other promising breeding sources were Solanum peruvianum (sensu lato) ‘CGO 6711’ (mild symptoms and low virus titers), Solanum chilense LA1967 (mild symptoms, but with high levels of B. tabaci MEAM1 oviposition) and Solanum pennellii LA0716 (intermediate symptoms and low level of B. tabaci MEAM1 oviposition). Additional studies are necessary to elucidate the genetic basis of the tolerance/resistance identified in this set of Solanum (Lycopersicon) accessions.     

Genetic effects of introgression genomic components from Sea Island cotton (<Emphasis Type="Italic">Gossypium barbadense</Emphasis> L.) on fiber related traits in upland cotton (<Emphasis Type="Italic">G</Emphasis>. <Emphasis Type="Italic">hirsutum</Emphasis> L.)

The germplasm with exotic genomic components especially from Sea Island cotton (Gossypium barbadense L. Gb) is the dominant genetic resources to enhance fiber quality of upland cotton (G. hirsutum L., Gh). Due to low efficiency of phenotypic evaluation and selection on fiber quality, genetic dissection of favorable alleles using molecular markers is essential. Genetic dissection on putative Gb introgressions related to fiber traits were conducted by SSR markers with mapping populations derived from a cross between Luyuan343 (LY343), a superior fiber quality introgression line (IL) with genomic components from Gb, and an elite Upland cotton cv. Lumianyan#22 (LMY22). Among 82 polymorphic loci screened out from 4050 SSRs, 42 were identified as putative introgression alleles. A total of 29 fiber-related QTLs (23 for fiber quality and six for lint percentage) were detected and most of which clustered on the putative Gb introgression chromosomal segments of Chr.2, Chr.16, Chr.23 and Chr.25. As expected, a majority of favorable alleles of fiber quality QTLs (12/17, not considering the QTLs for fiber fineness) came from the IL parent and most of which (11/12) were conferred by the introgression genomic components while three of the six (3/6) favorable alleles for lint percentage came from the Gh parent. Validation of these QTLs using an F₈ breeding population from the same cross made previously indicated that 13 out of 29 QTLs showed considerable stability. The results suggest that fiber quality improvement using the introgression components could be facilitated by marker-assisted selection in cotton breeding program.     

A simple,rapid and reliable bioassay for evaluating seedling vigor under submergence in <Emphasis Type="Italic">indica</Emphasis> and <Emphasis Type="Italic">japonica</Emphasis> rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Submergence is a major stress causing yield losses particularly in the direct-seeded rice cultivation system and necessitates the development of a simple, rapid and reliable bioassay for a large scale screening of rice germplasms with tolerance against submergence stress. We developed two new bioassay methods that were based primarily on the seedling vigor evaluated by the ability of fast shoot elongation under submerged conditions, and compared their effectiveness with two other available methods. All four bioassay methods using cultivars of 7 indica and 6 japonica types revealed significant and consistent cultivar differences in seedling vigor under submergence and/or submergence tolerance. Japonica cultivars were more vigorous than indica cultivars, with Nipponbare being the most vigorous. The simplest test tube method showed the highest correlations to all other methods. Our results suggest that seedling vigor serves as a submergence avoidance mechanism and confers tolerance on rice seedlings to flooding during early crop establishment. A possible relationship is discussed between seedling vigor based on fast shoot elongation and submergence tolerance defined by recovery from submergence stress.     

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.     

Allelic composition and associated quality traits of the <Emphasis Type="Italic">Glu-1</Emphasis> and <Emphasis Type="Italic">Glu-3</Emphasis> loci in selected modern Ethiopian durum wheat varieties

Gluten protein determines the processing quality of both durum wheat and bread wheat. The glutenin subunits compositions and associated quality traits of 20 Ethiopian durum wheat varieties were systematically analyzed using SDS-PAGE and Payne numbers. A total of 16 glutenin patterns were identified. At the Glu-A1 locus, all varieties scored the null allele. The predominant glutenin alleles at the Glu-B1 locus were Glu-B1b (7+8) and Glu-B1e (20). In Glu-3, the most abundant glutenin subunits were Glu-A3a and Glu-B3c. Based on the Payne scores, the varieties Yerer, Ginchi, Candate, and Foka were identified to have allelic composition suitable for pasta making. The cluster analysis using agglomerative hierarchical clustering (AHC) method classified the varieties into four similarity classes. Based on the findings of this experiment, suggestions were made for allelic composition improvement through introgression of superior alleles from known Glu-1 and Glu-3 sources.     

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.