Outcrossing Rates in Autumn and Spring-Sown Barley

Natural outcrossing is a potential cause of the admixtures which are often observed, especially in winter barley. Outcrossing was studied in yield trials with hap laid -derived homozygous offspring, lines from tour crosses between winter and spring barley varieties. Outcrosses were recognised by heterozygosity in storage protein loci Hor1 and Hor2 as revealed by electrophoresis. Outcrossing, was, on average, about 5% in lines sown in the autumn and less than 0.5% after spring sowing. Autumn-sown lines from one of the four crosses studied had about 10% outcrosses.     

Variation in outcrossing rate and genetic structure on six cultivars of Vicia faba L. as affected by geographic location and year

The present study was conducted to evaluate the importance of two environmental factors, year and geographic location, on the cultivar outcrossing rate and genetic structure of Vicia faba L. Under natural field conditions. Outcrossing rates were estimated for six cultivars using isozyme markers, the mixed mating model and the multilocus method. The genetic structure was described by Wright's fixation index and the inbreeding equilibrium coefficient. Faba bean outcrossing rate, as measured by multilocus estimates was relatively invariable among cultivars within locations. Our results suggest substantial variation in outcrossing rate associated with geographic location (Mediterranean subtropical and temperate climates enhancing the cross-pollination). Year variation was far less important than geographic variation.     

Wheat pollen dispersal under semiarid field conditions: potential outcrossing with Triticum aestivum and Triticum turgidum

Isolation distance is the main barrier to crop-to-crop gene-flow. A 3-year study assessed the maximum potential outcrossing under field conditions between two wheat cultivars (Triticum aestivum L.) and between wheat and durum wheat (Triticum turgidum L. var. durum). Outcrossing was measured by seed set on emasculated recipient plants placed at four sides with different distances from a 3 m × 3 m T. aestivum (cultivar Chinese Spring) pollen source. Frequencies of seed set at 0 m distance were 45% (37–56%) for T. aestivum cultivars and 18% (5–30%) with T. turgidum. These values agree with hybridization in non-limiting pollen conditions measured by manual crosses in greenhouse. The number of pollen grains and the outcrossing frequencies decreased at increasing distances influenced by the prevailing wind direction. Under semiarid conditions of this assay, viable pollen was found 14 m from the pollen source, with a maximum distance of 8 m at which cross-pollination decreases below 1%. Ambient conditions affect pollen viability, hybridization and pollen dispersal. Data presented in this paper emphasize the major role played by environmental conditions in outcrossing. Data obtained in one area may therefore not coincide with the prevailing situation in different locations and climates.     

Esterase 1 and Hordein C allelic and genotypic frequencies in CC XXXI,a male sterile barley population

Summary Seeds from the 5th, 6th, 12th and 18th generation of enforced outcrossing in CCXXXXI-B, a barley (Hordeum vulgare L.) population containing genetic male sterility, were assayed for genotypic distribution of Est 1 and Hor C alleles. The result suggest that natural selection was effective in altering allelic frequencies at both loci and that genotypic frequencies were often different from what would be expected if allelic frequencies solely determined genotypic frequencies. Departures of actual from expected genotypic frequencies occurred in that shifts in allelic frequencies were detected.Joint contribution of the USDA-ARS and published with the approval of the Director of the Montana Agric. Exp. Sta. Journal Series No. 2281.     

Outcrossing rate of barley genotypes with different floral characteristics in drought‐stressed environments in Jordan

The flowering characteristics of a highly inbreeding species such as barley might influence the extent of outcrossing that can be expected. Increased outcrossing is a prerequisite to exploit heterosis in inbreeding species, which can lead to considerable yield increase as has been demonstrated for barley grown under marginal conditions. Anther extrusion (AE) in barley has been demonstrated to be a genetically controlled, quantitative trait with intermediate to high heritability. Some authors consider AE and open flowering in barley as identical. The present study was conducted to investigate whether barley genotypes with increased AE exhibit increased outcrossing rates under field conditions in comparison with genotypes showing no AE. Field experiments were conducted in three contrasting environments in Jordan in the 2001/02 growing season using six adapted barley genotypes, three with high AE and three with no AE. Outcrossing rates were investigated employing codominant DNA markers (microsatellites). No significant difference in outcrossing rates was detected between anther‐extruding and non‐extruding barley genotypes. Results indicate that high AE is not a sufficient condition to cause increased outcrossing rates and thus cannot be termed open flowering.     

Spontaneous outcrossing in tomato depends on cultivar and environment and varies between individual flowers

Knowledge about the degree of spontaneous outcrossing of diverse genotypes is essential for breeding programmes, maintenance breeding, and seed production. For tomato (Lycopersicon esculentum Mill.), very limited scientific evidence for genotypic differences is available and evidence from Europe is scarce. To close this knowledge gap, six cultivars were investigated in three Central European locations as part of the Organic Outdoor Tomato Project. To determine outcrossing rates, the monogenetic “cut‐leaf” trait, which is dominant over the “potato‐leaf” trait, was used as morphological marker. The observed range of outcrossing was 0.0%–5.2%. Outcrossing was significantly influenced by cultivar and environment. The outcrossing rate of individual flowers varied within cultivars ranging from 0% to 37%. The potential of newly opened flowers to accept foreign pollen varied largely with the cultivar. Genotypic differences could partly be linked to flower morphology traits. The potential for recombination between tomato genotypes is generally very low but can be a source for new variation in on‐farm management.     

Mapping of Malt Endopeptidase, NADH Diaphorase and Esterase Loci on Barley Chromosome 3L

Isoelectric focusing (IEF) and immunoblotting were used to detect genetic variants of malt endopeptidase (MEP1), an enzyme related to malting quality in barley and coded by the CepB locus on barley chromosome 3 (= 3H). A variant was found in a Turkish accession of wild barley (Hordeum vulgare ssp. spontaneum). The self progeny of a hybrid between this accession and the barley cultivar ‘Clipper’ were analyzed for recombination between the CepB locus and other isozyme loci. The estimates of recombination linked the CepB locus to an NADH diaphorase locus (Ndh2), which in turn was linked to the seedling esterase complex (Est1, Est2, and Est4) situated near the distal end of the long arm of chromosome 3. The Ndh2 locus was independent of two other NADH dehydrogenase loci (Ndh3, Ndh5) which were mapped on barley chromosome 5 (= 1H) in relation to the three hordein loci (Hor1, Hor2, and Hor3).     

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.     

Evaluation of floral characteristics of barley in the semi-arid climate of north Syria

Previous studies have revealed that, in areas affected by severe drought, yielding ability and stability of barley (Hordeum vulgare L.) could be improved by increasing the level of heterozygosity. One possible approach to increase the heterozygosity level is to improve the outcrossing rate. In view of that, the genetic variability and phenotypic correlation coefficients involving four floral traits related to outcrossing behaviour were investigated in 274 drought adapted barley lines from the International Center for Agricultural Research in the Dry Areas (ICARDA's) current breeding programme. A significant quantitative variation existed in anther length and width, stigma length, and anther extrusion, which indicate that selection for these traits may be effective. Anther extrusion showed weak but highly significant positive correlations with anther and stigma length. Somewhat higher positive correlations were found between anther and stigma length. The results indicate that selection for high anther extrusion and longer anthers and stigmata may increase outcrossing in cultivated barley and consequently improve yielding ability through enhanced heterozygosity.     

Estimation of Outcrossing Rate in Chickpea (Cicer Arietinum L.) Sown in Autumn

Gene flow via outcrossing from transgenic plants to relatives will be one of the most important concerns to grow of the transgenic chickpea (Cicer arietinum L.) in European Union (EU). This report is therefore focused on spontaneous outcrossing rate in chickpea. A total of 39 kabuli type mutants with white flower and one desi type with pink flower were grown to estimate spontaneous outcrossing rate. Outcrossing rate ranged from 0.0 to 1.25% in mutant materials. Since labelling threshold for transgenic contamination in food and feed in European Union (EU) is 0.9%, outcrossing rate of 1.25% is higher than threshold of 0.9% in EU, and this result suggests that cultivation of transgenic chickpea will be under high risk to be contaminated chickpeas in neighbourhood fields.     

Linkage of Rust Resistance Genes from Wild Barley (Hordeum spontaneum) with Isozyme Markers

Eighty-three third backcross lines which comprise a set of near isogenic lines (NIL's) of the barley cultivar ‘Clipper’ but each carrying a different chromosomal segment from Hordeum spontaneum, marked with a distinct isozyme, were tested for resistance to three races of the barley leaf rust pathogen (Puccmia hordei). Fourteen lines showed resistance to at least one race and three showed resistance to all three races. The resistance in two of these lines was controlled by separate, single partially dominant genes. In one case the resistance gene named Rph1O was on chromosome 3 and linked (r = 0.15 ±0.05) with the isozyme locus Est2. In the second case, the gene (Rph11) was on barley chromosome 6 and linked (r = 0.07±0.02) with the isozyme locus Acp3 and (r = 0.11±0.02) with Dip2.     

Potential and limitations of isozymes for chromosomal localization of resistance genes against barley mild mosaic virus (BaMMV)

Summary To assess the possibilities offered by isozymes to locate resistance genes against barley mild mosaic virus (BaMMV), the isozyme patterns of 19 barley (Hordeum vulgare L.) genotypes carrying genes different from ym4 were determined. Of the 15 isozyme systems tested, only three were polymorphic, namely aconitate hydratase, esterase, phosphogluconate dehydrogenase, providing markers on four of the seven barley chromosomes. Studies of F₂ progenies derived from three crosses between resistant genotypes and susceptible varieties failed to reveal linkage between resistance genes and isozymes. Another goal of the experiment was to study the linkage relationships between ym4 and the esterase locus (Est1-Est2-Est4). Our estimates of the recombination rate between these two loci (3.41 and 8.32%) were in the range of those reported between these esterases and one of the resistance genes of the Chinese variety Mokusekko 3.     

Sources of Resistance to Powdery Mildew in Barley Lines Derived from Hordeum spontaneum Collected in Israel

Fourty two barley lines direved from the F₇ of crosses between barley cultivars and different accessions of Hordeum spontaneum collected in Israel and 30 lines or varieties with known genes for resistance to powdery mildew were included m this study. Eleven European and three Israeli powdery mildew cultures, possessing virulence genes corresponding to known resistance genes, were used to make comparisons between the varieties with known resistance genes and H. spontaneum derived lines. The reaction pattern of 39 H. spontaneum derived lines was clearly different from the reaction pattern o; any of the known genes for mildew resistance included in this study. Only two cases were observed in which the reaction pattern of H. spontaneum derived lines agreed with reaction patterns of known genes for mildew resistance viz. Ml-a9 and Ml-p. Trie Mildew resistance of one line apparently traces back to uncontrolled outcrossing with a Ml-a.6+Ml-g resistant cultivar. Since the majority of the 42 host genotypes tested showed distinctive variation in resistant reaction types against different mildew cultures, this study docs not support the assumption that differences in resistant infection types against distinct mildew cultures are sufficient to indicate the presence of supplementary genes for resistance in a given genotype of the host. The results justify the conclusion that the natural population of H. spontaneum in Israel forms a large gene pool for mildew resistance which is not yet used m cultivated barley.     

Estimation of the outcrossing rate for annual Asian wild rice under field conditions

The Asian wild rice, Oryza rufipogon, has partial outcrossing behavior and shows high levels of genetic variation. To estimate an accurate outcrossing rate of annual form of O. rufipogon, two backcross lines (Lines R1 and R2) between Oryza sativa Nipponbare and O. rufipogon W630 were examined under field conditions. A chromosome survey confirmed that these lines had wild chromosomal segments at more than 92% of marker loci. As for the traits of glume, stamen and pistil, Line R1 showed similar floral morphology as that of O. rufipogon W630, whereas Line R2 had larger glumes. In 2005, 2006 and 2008, a total of 22 backcross plants were planted in the middle of wild rice plots. The successive progenies of each plant were examined using microsatellite markers that could clearly detect self-pollination and outcrossing. The outcrossing rates of Line R1 plants ranged from 4.04% to 25.50% with an average of 10.20%. This indicates that cross-pollination of wild rice is a chance event affected by many environmental factors. The outcrossing rates of Line R2 plants also varied, however, no significant difference was observed between the averages of Lines R1 and R2, suggesting that the glume sizes are not critical for outcrossing ability.     

Outcrossing Rates of Experimental Populations of Cuphea lanceolate

Outcrossing rates of Cuphea lanceolata Ait., a new Dilseed crop, have not been estimated; however, this species is known to be insect-pollinated, self-compatible, and autosterile. We used five allozyme loci and the mixed mating model to estimate the outcrossing rates of experimental populations of Cuphea lanceolata Ait. grown at Corvallis, Oregon in 1987 and 1988. Populations were grown using densities of one and 27 plants m^?2. Multilocus out-crossing rate estimates of populations grown at low and high density ranged from 0.61 to 0.85 and 0.93 to 0.94, respectively. Density caused differences may be caused by differences in pollinator movement. In open plantings, e.g., space-plant nurseries, selfing rates are increased by increased within plant pollination by bumblebees, the primary pollinators of C. lanceolata in temperate zones. The movement of pollinators between plants is enforced in dense plantings. We found C. lanceolata is predominantly al-logamous, but selfing rates as great as 39.0 % were observed among space-plants; thus, the progeny of an open-pollinated individual plant grown in this type of nursery is a mixed half-sib and S₁ family. Maximum outcrossing can be achieved by using dense plantings.     

Evidence for phenotypic assortative mating for flower colour in periwinkle

Two experiments were conducted with periwinkle, Catharanthus roseus, to determine the extent of natural outcrossing. Three white-flowered, fully self-fertile, monogenic recessive mutants, namely, dwarf, wavy leaf margin and curved leaf were used, together with their parental white-flowered variety, ‘Nirmal’, and a normal pink-flowered variety, PS-1. The extent of total outcrossing ranged from 43.4 to 79.3% among mutants. Outcrossing between white × white-flowered plants ranged from 28.3 to 65.3% and was two to four times greater than that between white × pink-flowered plants in the three mutants. The extent of out-crossing between white × pink-flowered plants was similar “02.2-15.0%” in all mutants and also similar to that in the normal white-flowered variety,‘Nirmal’(00.4%), where white × white flower out-crossing could not be estimated. There were no large differences in the number of seeds per fruit, percentage fruit set and germination percentage of seeds obtained from self, white × white and white × pink flower crosses made in the glasshouse. There were also no significant differences in the number of flowers produced by the genotypes used in the study. The observed higher frequency of white × white flower matings compared with white × pink flower matings appeared to be due to the constancy of flower colour exhibited by the butterfly pollinators Pachliopta hector and Catopsilia pyranthae during their flower visits. Observations made on the occurrence of natural self-pollination revealed that automatic self-pollination did not occur in periwinkle.     

Pathogenic variation among isolates of Rhynchosporium secalis from barley grass growing in South Eastern Australia

Summary The pathogenicity of 182 single spore isolates of Rhynchosporium secalis from Hordeum leporinum and 94 isolates from H. vulgare collected from throughout southeastern Australia was tested on 15 barley varieties, each having different combinations of resistance genes. Forty five percent of the barley grass isolates were pathogenic on 5 or more varieties but only 6% of the cultivated barley isolates attacked this range of varieties. On the basis of reaction type 20 different pathogenicity groups were recognised, with barley grass isolates being classified into 19 and the cultivated barley isolates into 5, four of which were the same as the barley grass isolates. Numerical analysis of data on leaf area damage inferred 33 groups, 24 of which were unique to barley grass isolates, two to cultivated barley isolates and 9 common to both groups. There was as much variation in pathogenicity among single spore isolates from the same lesion as between isolates from different lesions collected from the same or different locations.     

Risk assessment of the transfer of imazethapyr herbicide tolerance from Clearfield rice to red rice (Oryza sativa)

Hybridization between Clearfield rice and weedy red rice would have a direct impact on management and long-term strategies of imazethapyr technology for rice weed control. The objective of this research was to determine rates and agronomic consequences for outcrossing between Clearfield rice and red rice. Red rice populations showed extensive variation for plant height, panicle length, tillers/plant, seeds/plant, seed set and grain weight. Outcrossing was detected from all Clearfield rice cultivars (‘CL121’, ‘CL141’, ‘CL161’, and ‘CLXL8’) to red rice and was confirmed by phenotypic and DNA marker analyses. An overall outcrossing frequency of 0.17% was observed in 2002 red rice samples with a range from 0% to 0.46%. Tolerance of 2002 red rice samples to imazethapyr corresponded to levels of acetohydroxyacid synthase (AHAS) activity. A majority (94%) of the progeny from the 2002 samples segregated 3 resistant:1 susceptible for tolerance to imazethapyr, indicating that a single dominant gene from Clearfield rice was associated with tolerance in the hybrid material. The remaining samples did not segregate for tolerance, suggesting that spontaneous mutations for tolerance were present in this material before or after crossing with Clearfield rice. A four-fold increase in outcrossing frequency of 0.68% was observed in 2003 red rice samples with the highest outcrossing frequency for a single location at 3.2%. Results from this study indicate that outcrossing between Clearfield and red rice will occur rapidly at rates that warrant early-season field scouting and a crop rotation scheme to prolong usefulness of the Clearfield technology.     

Gain from marker‐based recurrent selection for increased heterozygosity in barley

Barley (Hordeum vulgare L.) is a highly adaptable cereal crop grown under a wide range of agro‐ecological conditions. Various literature reports demonstrate that heterozygosity may enhance the yielding level and stability of barley particularly in stress‐prone environments. The major aim of this study was to investigate whether recurrent selection (RS) for heterozygosity at highly polymorphic codominant molecular marker loci increases outcrossing in a genetically broad‐based population. Four to six SSR marker loci were used for this purpose. Selection was conducted over four cycles. Progress from selection was evaluated in a terminal experiment comprised of population samples from all four RS cycles. All experiments were conducted under greenhouse conditions. The starting population was composed of 201 gene bank accessions (178 lines, 12 landraces and 11 wild barley populations) sampled in West Asia and North Africa. Selection led to a stepwise increase in heterozygosity from 0.23% to 1.50%. Correspondingly, the estimated multilocus outbreeding rate rose from 1.4% to 3.5%. This positive selection response offers new perspectives for improving the productivity of barley in stress‐prone areas.     

Allotetraploid hybrids produced by protoplast fusion for seedless triploid <Emphasis Type="BoldItalic">Citrus</Emphasis> breeding

In connection with crop improvement strategies for marginal regions it has been proposed to increase the outcrossing rate in barley which would presuppose high and persistent pollen viability to ensure successful cross fertilisation. The present study was designed to investigate the viability of mature pollen from extruded and non-extruded anthers of cultivated and wild barley (Hordeum vulgare ssp vulgare and ssp spontaneum, respectively) in comparison with the closely related but obligatory outcrossing species H. bulbosum. Pollen viability (PV) was assessed employing the p-phenylenediamine-peroxidase-test on pollen derived from spikes or anthers immediately after collection and after treatment at 20, 30 and 40 ^∘C for 1, 2, 4, 8 and 16 h. The latter treatment was interposed after 8 h by a 10 h period of darkness at 12 ^∘C and thus called 26 h treatment, consecutively. Initial PV was high with 98% across all genotypes and even at 40 ^∘C did not fall below 80% after 8 h. After the 26 h treatment, PV of two H. vulgare genotypes originating from semi-arid regions and of H. bulbosum fell below 60% while the other genotypes retained a PV of > 80%. Viability of pollen of extruded anthers in H. vulgare ssp. spontaneum was on average slightly but significantly lower than PV of non-extruded anthers but still remained above 90%, even after the 26 h treatment at 40 ^∘C. Pollen viability of the outcrossing species H. bulbosum ranged on a very similar level as PV of H. vulgare. Results indicate that pollen of H. vulgare retains a sufficiently high level of viability to ensure successful cross-fertilisation over a period of at least 26 h even at high temperatures of up to 40 ^∘C.