Genetic analysis of high amylose content in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) using a triploid endosperm model

Traditionally, high amylose starch (HAS) from maize (Zea mays L.) has been mainly used as an ingredient in gum candies and as an adhesive for corrugated cardboard. Two recent advances have increased interest in the use of HAS. The first one has been the development of starch-based biodegradable thermo plastics. Second, high amylose maize is a source of resistant starch (RS), a type of starch that resists digestion. As a food additive, consumers can benefit from added RS since it will lower the glycemic index and the risk of colon cancer in accordance with recent research in food science. Normal maize has about 25% amylose starch. A maize inbred line, GEMS-0067 (Reg. no GP-550, PI 643420) possesses high amylose modifier gene(s) that, together with the recessive amylose extender (ae) gene, raises the starch amylose percentage to at least 70%. The objective of this study was to determine the gene effects, non-allelic interactions and heritability of high amylose content in maize using Bogyo’s triploid model. Nine populations were derived from a cross between H99ae, a maize inbred line with 55% amylose starch, and GEMS-0067. Data were collected from two locations in Missouri (MO) and South Dakota (SD) over 2 years (2005 and 2006). Incomplete dominance explained some of the inheritance of HAS. Maternal effects were also detected. The triploid models for MO and SD were separately established based on the corresponding data in 2005 and 2006. The additive and type 1 dominance effects in MO, and the additive, type 1 dominance, type 2 dominance, and additive × additive in SD were significantly different from zero meaning that those effects played an important role in amylose synthesis. Both broad-sense and narrow-sense heritabilities were high indicating that high amylose content could be effectively selected for in a segregating population.     

Genotype × environment interactions in populations possessing <Emphasis Type="Italic">Ga1</Emphasis>-<Emphasis Type="Italic">s</Emphasis> and <Emphasis Type="Italic">ga1</Emphasis> alleles for cross incompatibility in maize

Use of cross incompatibility in corn (Zea mays L.) by the Ga1-s allele may reduce cross-fertilization in specialty and conventional organic corn with pollen from genetically-modified (GM) corn. For effective use, information about environment, and genotype × environment effects on cross-fertilization by ga1 as well as heritability of cross incompatibility in maize is necessary. Our objective was to obtain this information. Four population pairs differing in their genotype at ga1 were evaluated for cross incompatibility with ga1 pollen in different environments. Populations were derived by crossing the recurrent parents B116, PHG35, ARZM16035:S19, and (CHZM05015:Mo17)Mo17 to Ga1-s donor parent Mo508W/Mo506W. Two replicates of each treatment were grown in the center of 952 m² fields planted with purple corn as an adventitious source of ga1/ga1 pollen. Open pollination was allowed and the amount of cross-fertilization estimated by averaging the percentage of purple seeds. Environment and genotype × environment effects were not significant. Contrasts to evaluate differences in cross-fertilization between Ga1-s and ga1 populations revealed that mean percentages of cross-fertilization in Ga1-s populations of B116, ARZM16035:S19, and (CHZM05015:Mo17)Mo17 were significantly lower than in ga1 populations. The estimated broad-sense heritability on an entry-mean basis for cross incompatibility was 0.81. Results suggest differences in genotype at ga1 played a major role in cross-fertilization of populations differing in their genotype at the ga1 locus. Incompatibility may be selected effectively over different environments and the Ga1-s system may be of value to reduce cross-fertilization with GM corn pollen.     

Complementary DNA (cDNA) cloning and functional verification of resistance to head smut disease (<Emphasis Type="Italic">Sphacelotheca reiliana</Emphasis>) of an NBS–LRR gene <Emphasis Type="Italic">ZmNL</Emphasis> in maize (<Emphasis Type="Italic">Zea mays</Emphasis>)

The nucleotide-binding site (NBS)-leucine-rich repeat (LRR) gene family comprises the largest number of known disease resistance (R) genes and is one of the largest gene families in plants. In the present study, the full-length cDNA of ZmNL (GenBank Accession Number KF765443) was isolated using Rapid Amplification of cDNA Ends. The nucleotide sequence of ZmNL contains an open reading frame of 3156 bp that encodes the ZmNL protein, which is comprised of 1051 amino acid residues. This putative protein has high homology to other known resistance proteins (84% to Triticum aestivum LR10) and belongs to the CC–NBS–LRR type R gene family. The ZmNL gene was introduced into the maize inbred line of Huangzao4 which was highly susceptible to head smut under the control of the maize ubiquitin promoter by Agrobacterium-mediated transformation. The head smut disease incidence of 3 T₂ transgenic lines was significantly reduced (by 18.38–29.40%) compared with the wild type, which indicated that the overexpression of ZmNL gene in maize enhanced the resistance to the fungus Sporisorium reilianum (Kühn) Clint of these plants.     

Combining ability amongst CIMMYT’s early maturing maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm under stress and non-stress conditions and identification of testers

CIMMYT (International Maize and Wheat Research Institute) Zimbabwe’s early maturing maize program, which aims to supply seed to approximately 4 million hectares of maize area in eastern and southern Africa, lacks adequate information on heterotic relationships among early maturing germplasm and has no early maturing testers for hybrid development. Open-pollinated varieties (OPVs) and hybrids are the products targeted for this region. Among the hybrids, three-way and double-cross hybrids are desired. Thus the use of single crosses as testers would be an appropriate choice for such a breeding program as one could potentially identify three-way combinations during the early generation test. A twelve-parent diallel was formed and crosses evaluated to identify heterotic groups and single-cross testers. Crosses were evaluated under four different environments in Zimbabwe, two optimal, one low nitrogen stress and one drought stress. P5 (an early maturing line from heterotic group A) and CML 395 (a late maturing inbred line from heterotic group B) were used as reference parents to establish heterotic groupings of germplasm used in the diallel. The single cross (P7/P8) was identified as a potential group A tester because of: (a) co-classification of inbred lines into heterotic group A, (b) good yields-9.8 t/ha (optimal), 3.4 t/ha (low nitrogen) and 2.1 t/ha (drought); and (c) good GCA effects for grain yield (0.49 t/ha) of line P7 while line P8 contributed to reduced height and anthesis-silking interval.     

<Emphasis Type="Italic">Platanus</Emphasis> × <Emphasis Type="Italic">acerifolia</Emphasis> genotypes surviving to inoculation with <Emphasis Type="Italic">Ceratocystis platani</Emphasis> (the agent of canker stain): first screening and molecular characterization

Canker stain, caused by the fungus Ceratocystis platani, is a destructive disease in Platanus spp. It has been recently proved that resistant accessions can be produced and grown in Europe. However, additional resistant genotypes are still needed in order to avoid the onset of virulent pathogen strains favoured by the selection pressures exerted by genetically homogeneous resistant plane tree plantings. In this study we present the results of two parallel experiments performed on 975 accessions of P. × acerifolia seedlings and P. × acerifolia clones derived by cutting propagation from mature trees grown in the urban environments. The selection process was based on inoculation with C. platani and yielded 13 accessions that showed different types of resistant reactions and survived in a stable manner thoughout the period of observation. Selected accessions were characterised by sequencing the rDNA-ITS region and by developing PCR procedures capable of detecting P. orientalis and P. occidentalis LEAFY homoeologues. These molecular analyses enabled us to confirm the identification of the species, its hybrid origin and to assess an evident genetic variability among the accessions, which therefore have to be considered as different genotypes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Potential new genes for resistance to <Emphasis Type="Italic">Mycosphaerella graminicola</Emphasis> identified in <Emphasis Type="Italic">Triticum aestivum</Emphasis> × <Emphasis Type="Italic">Lophopyrum elongatum</Emphasis> disomic substitution lines

Lophopyrum species carry many desirable agronomic traits, including disease resistance, which can be transferred to wheat by interspecific hybridization. To identify potentially new genes for disease and insect resistance carried by individual Lophopyrum chromosomes, 19 of 21 possible wheat cultivar Chinese Spring × Lophopyrum elongatum disomic substitution lines were tested for resistance to barley yellow dwarf virus (BYDV), cereal yellow dwarf virus (CYDV), the Hessian fly Mayetiola destructor, and the fungal pathogens Blumeria graminis and Mycosphaerella graminicola (asexual stage: Septoria tritici). Low resistance to BYDV occurred in some of the disomic substitution lines, but viral titers were significantly higher than those of two Lophopyrum species tested. This suggested that genes on more than one Lophopyrum chromosome are required for complete resistance to this virus. A potentially new gene for resistance to CYDV was detected on wheatgrass chromosome 3E. All of the substitution lines were susceptible to Mayetiola destructor and one strain of B. graminis. Disomic substitution lines containing wheatgrass chromosomes 1E and 6E were significantly more resistant to M. graminicola compared to Chinese Spring. Although neither chromosome by itself conferred resistance as high as that in the wheatgrass parent, they do appear to contain potentially new genes for resistance against this pathogen that could be useful for future plant-improvement programs.     

Quantitative trait loci analysis of citrus leprosis resistance in an interspecific backcross family of (<Emphasis Type="Italic">Citrus reticulata</Emphasis> Blanco × <Emphasis Type="Italic">C. sinensis</Emphasis> L. Osbeck) × <Emphasis Type="Italic">C. sinensis</Emphasis> L. Osb

Leprosis, caused by citrus leprosis virus (CiLV) and transmitted by the tenuipalpid mite Brevipalpus phoenicis, is one of the most important viruses of citrus in the Americas. Sweet oranges (Citrus sinensis L. Osb.) are highly susceptible to CiLV, while mandarins (C. reticulata Blanco) and some of their hybrids have higher tolerance or resistance to this disease. The mechanisms involved in the resistance and its inheritance are still largely unknown. To study the quantitative trait loci (QTL; quantitative trait loci) associated with the resistance to CiLV, progeny analyses were established with 143 hybrid individuals of ‘Pêra’ sweet orange (C. sinensis L. Osb.) and ‘Murcott’ tangor (C. reticulata Blanco × C. sinensis L. Osb.) from controlled crossings. Disease assessment of the hybrid individuals was conducted by infesting the plants with viruliferous mites in the field. The experiment consisted of a randomized completely block design with ten replicates. The evaluated phenotypic traits were incidence and severity of the disease on leaves and branches, for a period of 3 years. The MapQTL™ v.4.0 software was used for the identification and location of possible QTL associated with resistance to CiLV on a genetic map obtained from 260 AFLP and 5 RAPD markers. Only consistent QTLs from different phenotypic traits and years of evaluation, with the critical LOD scores to determine the presence or absence of each QTL calculated through the random permutation test, were considered. A QTL was observed and had a significant effect on the phenotypic variation, ranging from 79.4 to 84% depending on which trait (incidence or severity) was assessed. This suggests that few genes are involved in the genetic resistance of citrus to CiLV.     

Fine mapping of the <Emphasis Type="Italic">fg</Emphasis> gene using the interspecific cross of <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> × <Emphasis Type="Italic">Gossypium barbadense</Emphasis>

Frego (fg) bract is an important agronomic trait in tetraploid cotton, which has been widely introduced into several cotton varities or lines in the past several years. In order to help us further understand the underlying molecular mechanism of frego bract development, a map-base cloning strategy was used to localize the fg locus. An F₂ population which comprised of 290 fg individuals derived from a cross of the multiple-marker line T582 (G. hirsutum, carrying the fg gene) with Hai7124 (G. barbadense) was constructed. Genetic linkage analysis was carried out to map of the fg locus with SSR and EST-SSR markers in tetraploid cotton. Genetic linkage analysis showed that the fg locus was flanked by the marker NAU3016 and NAU3172 on the long arm of chromosome 3, with the genetic distance of 0.3 cM and 4.7 cM, respectively. The information of fg locus provided the basic information for the final isolation of this important gene in tetraploid cotton, these marker information could be used in marker-assisted selection in cotton.     

Complex segregation analysis of day-neutrality in domestic strawberry (<Emphasis Type="Italic">Fragaria ×ananassa</Emphasis> Duch.)

Thirty bi-parental progenies were generated using 45 genotypes from the University of California (UC) strawberry breeding population as parents. Both parent genotypes and their offspring were classified for photoperiod insensitivity, or day-neutrality, for flowering using a late-season flowering score and the number of inflorescences per plant recorded during late summer. Complex Segregation Analysis of these traits indicated that their distributions were best explained by a genetic model that postulates a single major locus with partial dominance for day-neutrality in combination with a background of polygenic and environmental variation. The frequency of the allele conferring day-neutrality was estimated as p = 0.59–0.62 in this experimental population. Genotypic values for the inferred major locus were estimated as a = 1.12 and d = −0.81 for the flowering score, and a = 4.93 and d = 2.41 for inflorescence number. Further resolution of inheritance patterns were obtained by comparing the phenotypic variance for each trait with estimates obtained by insertion of these genotypic class values and allele frequencies into standard quantitative genetic models, and by the comparing variance components estimated using a mixed model analysis with and without inferred genotypic classes as a fixed effect. These comparisons suggest that the major gene determines 80.5% and 73.9% of the additive genetic variance for flowering score and inflorescence number respectively. One complicating feature of the results obtained here is that a non-Mendelian model of segregation fit statistically better than a fixed Mendelian model. The genetic parameters estimated using this non-Mendelian model were essentially identical to those obtained with fixed segregation; hybrid and octoploid ancestry, selection affecting flowering response, and limited number of generations in the analysis are discussed as possible explanations of this result.     

Performance of various grapefruit (<Emphasis Type="Italic">Citrus paradisi</Emphasis> Macf.) and pummelo (<Emphasis Type="Italic">C. maxima</Emphasis> Merr.) cultivars under the dry tropic conditions of Mexico

Grapefruit growers in the tropics require information about existing and new citrus cultivars with high productivity potential. The objective of this study was to determine the growth, yield, and fruit quality performance of seven pigmented and four white grapefruit cultivars under the dry tropic conditions of Colima, Mexico. The trees were budded on sour orange (Citrus aurantium L.) rootstock and planted at a distance of 8 × 4 m. ‘Oroblanco’ and ‘Marsh Gardner’ white-fleshed grapefruit cultivars and ‘Chandler’, a pink-fleshed pummelo, were the largest trees with the greatest height (5.0–5.6 m), canopy diameter (6.2–6.3 m), trunk diameter (21.9–23.3 cm), and canopy volume (109–123 m³). Lower height (4.3–4.8 m) and canopy volume (73–96 m³), but with similar canopy diameter to the previously mentioned cultivars, were recorded for the remaining pigmented cultivars. ‘Chandler’ pummelo and four pigmented grapefruit cultivars (‘Shambar’, ‘Río Red’, ‘Ray Ruby’, and ‘Redblush #3’) had yearly productions of 34.8, 34.9, 34.1, 32.7, and 30.6 ton ha⁻¹, respectively. The most productive white grapefruit cultivar was ‘Marsh Gardner’ (30.5 ton ha⁻¹). Grapefruit cultivars having the largest fruit size showed a higher inverse relationship between fruit weight and yield than those with small fruit. Most genotypes had higher values of fruit weight, juice content, and maturity index than those required by the local market. The most promising grapefruit cultivars based on their acceptable growth, yield superior to 30 ton ha⁻¹, and acceptable fruit color were ‘Río Red’, ‘Shambar’, ‘Ray Ruby’, and ‘Redblush #3’.     

Additional evidence for oligogenic inheritance of durable host resistance to coffee berry disease (<Emphasis Type="Italic">Colletotrichum kahawae</Emphasis>) in arabica coffee (<Emphasis Type="Italic">Coffea arabica</Emphasis> L.)

Breeding for host resistance to coffee berry disease (CBD) in arabica coffee (Coffea arabica) was initiated some 35–40 years ago in Kenya, Ethiopia and Tanzania in response to severe CBD epidemics. The release of CBD resistant cultivars to the coffee growers has been in progress since 1985. The resistance of cultivars like Ruiru 11 (Kenya) and Ababuna (and other cvs in Ethiopia) appears to be of a durable nature, since confirmed cases of a breakdown of host resistance under field conditions have not been reported over the past 20 years. Host resistance to the hemibiotrophic fungus Colletotrichum kahawae is of a quantitative nature, but nevertheless can be practically complete in some genotypes of arabica coffee. There is still no consensus on the genetics of CBD resistance, some claiming convincing evidence for oligogenes (1–3 major genes) and others for polygenes determining CBD resistance. Results from genetic studies with germplasm from the centre of genetic diversity for C. arabica in Ethiopia are presented here. These together with the recent identification of molecular markers associated with and the mapping of one major gene, provides additional evidence for oligogenic inheritance of CBD resistance. The development of cultivars combining yield and quality with durable host resistance to CBD has contributed greatly to increased sustainability of arabica coffee production in Africa. It has also considerable relevance to arabica coffee in Latin America and Asia, where CBD is still a quarantine disease but with a risk of becoming endemic one day, just as has happened earlier with coffee leaf rust (Hemileia vastatrix).     

A new gene <Emphasis Type="Italic">Bph33</Emphasis>(<Emphasis Type="Italic">t</Emphasis>) conferring resistance to brown planthopper (BPH), <Emphasis Type="Italic">Nilaparvata lugens</Emphasis> (Stål) in rice line RP2068-18-3-5

The rice brown planthopper (BPH) Nilaparvata lugens (Stål) is one of the major pests of rice across Asia. Host-plant resistance is the most ecologically acceptable means to manage this pest. A rice breeding line RP2068-18-3-5 (RP2068) derived from the land race Velluthacheera is reported to be resistant to BPH populations across India. We identified a new R gene [Bph33(t)] in this line using advanced generation RILs derived from TN1 × RP2068 cross through phenotyping at two locations and linkage analysis with 99 polymorphic SSR markers. QTL analysis through IciMapping identified at least two major QTL on chromosome 1 influencing seedling damage score in seed box screening, honey dew excretion by adults and nymphal survival. Since no BPH R gene has been reported on chromosome 1, we designate this locus as a new gene Bph33(t) which accounted for over 20% of phenotypic variance. Scanning the region for candidate gene suggested two likely candidates a leucine rich repeat (LRR) gene and a heat shock protein (HSP) coding gene. Expression profiling of the two genes in the two contrasting parents and RILs showed induction of the HSP gene (LOC_Os01g42190.1) at 6 h after infestation while LRR gene did not show such induction. It is likely that the HSP represented Bph33(t).     

Induction of <Emphasis Type="Italic">Fusarium</Emphasis> wilt (<Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">pisi</Emphasis>) resistance in garden pea using induced mutagenesis and in vitro selection techniques

Wilt caused by Fusarium oxysporum f. sp. pisi is a serious production constraint for peas worldwide. An attempt was made to isolate wilt-resistant mutants in two susceptible pea genotypes, Arkel and Azad P-1, employing induced mutagenesis and in vitro selection techniques. Two thousand seeds of each genotype were mutagenized either with ethyl methane sulfonate (EMS, 0.2% and 0.3%) or gamma rays (5-22.5 kR) in ⁶⁰Co gamma cell for three consecutive years. Screening of different mutagenized populations under wilt-sick plots resulted in the isolation of 25 mutants exhibiting complete or enhanced wilt resistance compared to parental genotypes. Five of these wilt-resistant mutants also outperformed the susceptible background genotypes in terms of yield and other horticultural traits. Efforts were also made to isolate wilt-resistant regenerants from callus cultures exhibiting insensitivity to culture filtrate (CF) of F. oxysporum f. sp. pisi. A total of 250 regenerants (R ₀) were obtained from CF-insensitive calli selected from medium supplemented with 20% culture filtrate. When evaluated in artificially inoculated sick plots, only five R ₂ lines obtained from the regenerants exhibited enhanced wilt resistance compared to parental cultivars. However, the selected lines did not exhibit resistance levels equivalent to those shown by wilt-resistant lines isolated through in vivo mutagenesis. To conclude, induced mutagenesis through irradiation and EMS treatments exhibited superiority over in vitro selection for inducing wilt resistance in peas.     

Nuclear genome size and chromosome analysis in <Emphasis Type="Italic">Chenopodium quinoa</Emphasis> and <Emphasis Type="Italic">C. berlandieri</Emphasis> subsp. <Emphasis Type="Italic">nuttalliae</Emphasis>

Cytogenetic characterization by karyotyping and determination of DNA content by flow cytometry of seven cultivated varieties of Chenopodium was performed. Chenopodium quinoa cultivar Barandales and C. berlandieri subsp. nuttalliae cultigens Huauzontle, Quelite and Chia roja showed 2n = 4x = 36, x = 9. Statistically insignificant genome size differences for studied varieties ranged from 2.96 pg/2C (1 Cx = 724 Mbp) in C. quinoa to 3.04 pg/2C (1 Cx = 743 Mbp) in Huauzontle. Karyotype analyses revealed the presence of nine groups of four metacentric chromosomes, including two pairs of chromosomes with satellites. The first pair of satellites was located on the largest pair of chromosomes and the second on a different pair of chromosomes in all accessions analyzed. Variation among varieties was evident in chromosome size, genome length (GL) and the position of satellites. Chia roja exhibited greatest GL (58.82 μm) and biggest chromosomes (2.04 μm). Huauzontle showed the smallest GL (45.02 μm) and shortest chromosomes (1.60 μm). Comparison of GL in studied taxa was statistically significant and allowed to define three groups according to the use given to these plants. These data indicate that they are small, very stable genomes in terms of DNA content, and they support the allotetraploid origin(s) of C. berlandieri subsp. nuttalliae and C. quinoa.     

Genetic parameters for growth traits and stem-straightness in <Emphasis Type="Italic">Eucalyptus urophylla</Emphasis> × <Emphasis Type="Italic">E. camaldulensis</Emphasis> hybrids from a reciprocal mating design

An inter-specific hybrid breeding program involving Eucalyptus urophylla (U) and Eucalyptus camaldulensis (C) was implemented in order to provide genotypes better adapted to southern China with improved growth rate, stem-straightness and wind-resistance. A trial involving 36 reciprocal crosses from six parents each of C and U that had been preselected for superior growth and stem-straightness was established at a site in Luokeng in Guangdong province. Ten, pure-species families using the hybrid parents as open-pollinated female parents were included as controls. Survival and growth traits at ages 2 and 8.3 years and stem-straightness at age 2 years were assessed. Inter-specific hybrids generally performed better than the pure species in terms of survival, growth traits and stem-straightness. Female U by male C crosses generally outperformed those involving male U and female C. Further indication of the significant reciprocal effect was supported by negative and low correlations between paired groups of full-sibs that differed only in the direction of the cross. Components of female additive genetic variance and narrow-sense heritability calculated from this estimate (\(\sigma_{\rm Af}^{2}\) and \(h_{\rm f}^{2}\)), respectively) were generally higher than those of the male \(\left( {\sigma_{\rm Am,}^{2} h_{\rm m}^{2} } \right)\), providing evidence for maternal effects. The narrow-sense heritability (h²) estimates based on general hybridizing ability for growth traits and stem-straightness were generally low at both ages, and of low precision at 8.3 years. The ratio of hybrid additive-to-dominance variance \(\left( {{{\sigma_{\rm A}^{2} } \mathord{\left/ {\vphantom {{\sigma_{\rm A}^{2} } {\sigma_{\rm D}^{2} }}} \right. \kern-0pt} {\sigma_{\rm D}^{2} }}} \right)\) was of little practical consequence at age 2 years and had further decreased by age 8.3 years. Trait-trait genetic correlations amongst hybrids were generally positive and moderate to high. Hybrid vigour, gauged by comparison with the performance of the pure species progeny was significant, though correlations between pure species and hybrid progeny-based estimates of parental performance were weak, indicating that making parental selections in pure species trials may not be a successful strategy.     

Estimation of outcrossing rates in intraspecific (<Emphasis Type="Italic">Oryza sativa</Emphasis>) and interspecific (<Emphasis Type="Italic">Oryza sativa</Emphasis> × <Emphasis Type="Italic">Oryza glaberrima</Emphasis>) rice under field conditions using agro-morphological markers

Rice is mainly a self-pollinating crop, but some outcrossing has been reported. Outcrossing with an undesirable donor would lead to the creation of segregants or off-types, which would adversely affect genetic purity and uniformity of the crop. Outcrossing rates in rice under field conditions were investigated using cultivar WAB96-1-1 as a pollen donor and WAB56-104, NERICA 2, NERICA 4 and NERICA 7 as pollen recipients. Levels of outcrossing were investigated up to 30 m from the pollen donor. Dominant morphological markers of red kernel colour and pubescent leaves of the donor were used to identify hybrids. A total of 721 134 plants were investigated. There was an average outcrossing rate of 0.7 ± 0.51%, with a potential outcrossing rate of 2.45 ± 0.86%. Outcrossing rates decreased with increase in distance. It ranged from 2.45% at 0.2 m from the donor to 0.05% at 25 m from the donor. Differences were observed between genotypes and seasons. In season 1 the highest average outcrossing rate of 1.2 ± 0.63% was with WAB56-104 and in season 2 it was 1.1 ± 0.69% with NERICA 4. Outcrossing occurred up to 30 m from the donor. This has implications for germplasm management and conservation and the production of high quality seed. Spatial isolation remains the most practical method to prevent undesirable gene flow. The study indicated that red kernel colour and leaf pubescence can be used to effectively assess outcrossing under field conditions in rice.     

Reciprocal differences in DNA sequence and methylation status of the pollen DNA between F<Subscript>1</Subscript> hybrids of <Emphasis Type="Italic">Solanum tuberosum</Emphasis> × <Emphasis Type="Italic">S. demissum</Emphasis>

Reciprocal differences, mostly caused by cytoplasmic effects, are frequently observed in interspecific hybrids. Previously, we found that crosses onto Solanum demissum were much successful with the pollen of interspecific hybrids between S. tuberosum as female and S. demissum as male (TD hybrids) than the reciprocal ones (DT hybrids). To elucidate this reciprocally different crossability, we analyzed the pollen DNA of TD and DT using methylation-sensitive amplified polymorphism (MSAP) analysis. Using 126 primer combinations, MSAP analysis revealed 57 different bands between bulked pollen DNA samples of TD and DT. Individual examination of 16 TD and 9 DT plants disclosed eight bands uniformly different between TD and DT. Their sequencing results revealed two pairs of bands to be identical to each other, resulting in six distinct sequences. As expected, one band shared high homology with chloroplast DNA, and another one with mitochondrial DNA. However, one band that was apparently different at DNA sequence level and maternally transmitted from S. demissum, showed no homology with any known sequence. The remaining three bands were of DNA methylation level differences with no or uncertain homology to known sequences. To our knowledge, this is the first report detecting reciprocal differences in DNA sequence or DNA methylation other than those in cytoplasmic DNA.     

Intergeneric hybridization and relationship of genera within the tribe Anthemideae Cass. (I. <Emphasis Type="Italic">Dendranthema crassum</Emphasis> (kitam.) kitam. × <Emphasis Type="Italic">Crossostephium chinense</Emphasis> (L.) Makino)

An intergeneric cross has been made between Dendranthema crassum (kitam.) kitam. (2n = 90; ♀) and Crossostephium chinense (L.) Makino (2n = 18; ♂). Most of the hybrid embryos aborted at an early developmental stage. Using ovule rescue, it was possible to establish a single intergeneric hybrid plant showing 2n = 54 chromosomes. The leaf length, leaf width and epidermal hair density of the hybrid were all intermediate between those of the parents. However the flower diameter, number of tubular florets, epidermal hair height and epidermal hair length exceeded those of both parents. A genomic in situ hybridization approach was able to distinguish between the parental genomes in the hybrid plant.     

Mapping quantitative trait loci for cottonseed oil,protein and gossypol content in a <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> × <Emphasis Type="Italic">Gossypium barbadense</Emphasis> backcross inbred line population

Cotton is one of the most important oil-producing crops and the cottonseed meal provides important protein nutrients as animal feed. However, information on the genetic basis of cottonseed oil and protein contents is lacking. A backcross inbred line (BIL) population from a cross between Gossypium hirsutum as the recurrent parent and G. barbadense was used to identify quantitative trait loci (QTLs) for cottonseed oil, protein, and gossypol contents. The BIL population of 146 lines together with the two parental lines was tested in the same location for three years in China. Based on a genetic map of 392 SSR markers and a total genetic distance of 2,895.2 cM, 17 QTLs on 12 chromosomes for oil content, 22 QTLs on 12 chromosomes for protein content and three QTLs on two chromosomes for gossypol content were detected. Seed oil content was significantly and negatively correlated with seed protein content, which can be explained by eight QTLs for both oil and protein contents co-localized in the same regions but with opposite additive effects. This research represents the first report using a permanent advanced backcross inbred population of an interspecific hybrid population to identify QTLs for seed quality traits in cotton in three environments.     

Mapping a gene conferring resistance to <Emphasis Type="Italic">Wheat yellow mosaic virus</Emphasis> in European winter wheat cultivar ‘Ibis’ (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Wheat yellow mosaic, caused by Wheat yellow mosaic virus (WYMV), is one of the most devastating soil-borne diseases of winter wheat (Triticum aestivum L.) in Japan. Yellow-striped leaves and stunted spring growth, symptomatic of WYMV infection, result in severe yield loss. A new putative WYMV resistance gene in the European wheat cultivar ‘Ibis’ was mapped in the cluster of microsatellite markers including Xcfd16, Xwmc41, Xcfd168 and Xwmc181 on the long arm of chromosome 2D at the distances of 2.0 cM, 4.0 cM, 7.1 cM and 12.4 cM, respectively. WYMV-resistant cultivars contained a common haplotype of the four markers, whereas moderately susceptible and susceptible cultivars did not. These results should be useful in marker-assisted selection for WYMV resistance in wheat.