Outcrossing and isolation requirement in kenaf (Hibiscus cannabinus L.)

Summary To find the isolation distance in Hibiscus cannabinus L. (kenaf), experiments were conducted for three years with two planting designs. The extent of cross pollination irrespective of the planting designs was inversely related with the distance. At a distance beyond 10.0 m, the contamination was found to be absent. Directions had no influence on the contamination. The amount of contamination varied considerably between the years.     

Natural outcrossing rate in Vernonia galamensis

Vernonia galamensis is a potential new industrial crop growing wild in Ethiopia. The seed oil is rich in vernolic acid, an epoxy fatty acid, which is of interest for oleochemical uses. Basic information on the reproductive system of Vernonia is still very limited. The amount of natural outcrossing was estimated at two locations in Ethiopia (Alemaya and Babile) using flower colour as a marker. Single plants with white flowers, which is a monogenic recessive trait, were planted in plots with normal pink flowers and the outcrossing rate was estimated from the frequency of pink‐flowered plants in the progeny of the white‐flowered plants. Estimates of the natural outcrossing rate ranged between individual plants from 3.5 to 16% at Alemaya and 2.5 to 12% at Babile. Vernonia galamensis can be classified as a mainly self‐pollinated species.     

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.     

Outcrossing in celery (Apium graveolens)

Summary Isozyme marker loci were used to measure outcrossing in artificial and natural populations of celery (Apium graveolens). In three separate experiments involving populations of uniform density and flowering time, outcrossing rate ranged from 47 to 87% and the mean was 71.4%. In a wild population, outcrossing was estimated at 32.4 and 53.1%, using different homozygotes as the fixed female, and the weighted mean was 49.4%. A slight correlation of outcrossing rate with population density was observed. Mean outcrossing rates estimated from departures from panmictic expectations at polymorphic isozyme loci among open-pollinated plant introductions were 67.6% (celeries), 79.8% (celeriacs), and 85.1% (smallages). Based on these results, it was concluded that celery is best described as intermediate with respect to self- and cross-pollination.     

Estimation of outcrossing rate in landraces of tetraploid wheat (Triticum turgidum L.)

A study was undertaken to assess the amount of natural outbreeding in tetraploid wheat landraces of Ethiopian origin. The study consisted of two trials which were planted separately at two locations representing different environmental conditions for two consecutive seasons. The design employed was the randomized complete block with three replications. Two polymorphic loci, glume pubescence and awn condition were utilized as markers. The rate of outcrossing was based on the number of hybrids found in the recessive class of each marker. The overall level of outcrossing was higher in the pure stand trial than in the mixed stand trial. A higher rate of outcrossing occurred in the higher altitude area in the pure stand trial while the opposite was the case in the mixed stand trial. The average rate of outcrossing due to the marker loci was also different, with the long awn displaying more outbreeding tendency. Both locational and seasonal differences in the amount of outcrossing were observed. Generally, the study has revealed that there is a considerable amount of outbreeding in the landraces and that this genetic factor could play a role in preserving variation in the landrace populations. The level of outcrossing detected in this study could have a bearing on the maintenance of the genetic purity of diverse breeding lines being handled together in a breeding nursery and varietal purity in seed multiplication.     

Development of a co-dominant DNA marker tightly linked to gene <Emphasis Type="Italic">tardus</Emphasis> conferring reduced pod shattering in narrow-leafed lupin (<Emphasis Type="Italic">Lupinus angustifolius</Emphasis> L.)

The reduced pod shattering gene tardus is one of the most important domestication genes in narrow-leafed lupin (Lupinus angustifolius L.). In development of a molecular marker linked to the tardus gene, we incorporated the concept of marker validation during the initial candidate marker identification stage. Four dominant microsatellite-anchored fragment length polymorphism (MFLP) markers were identified as candidate markers based on their banding patterns in an F₈ recombination inbred line (RIL) population. One specific marker best correlating with phenotypes in the representative germplasm was selected and converted to a simple PCR-based marker. This established marker, designated as “TaLi”, is located at a distance of 1.4 cM from the tardus gene. DNA sequencing revealed six insertion/deletion sites between the non-shattering marker allele and the shattering marker allele. Validation of marker TaLi on 25 domesticated commercial cultivars and 125 accessions of the lupin core collection found a 94% marker and tardus phenotype match. Marker TaLi is the first simple PCR-based marker that can be widely used for non-shattering pod selection in narrow-leafed lupin breeding program.     

Genetic linkage of QTLs for late blight resistance and foliage maturity type in six related potato progenies

A set of test crosses of diploid potatoes was used to identify QTLs for foliage resistance against Phytophthora infestans and QTLs for foliage maturity type, and to assess their genetic relationship. The most important locus for both traits was found on chromosome 5 near marker GP21: the allele of marker GP21 that is associated with resistance to late blight is also associated with late foliage maturity. An additional QTL with a small effect on foliage maturity type was identified on chromosome 3, and additional QTLs for late blight resistance were found on chromosomes 3 and 10. Another QTL was detected on chromosome 7 when resistance was adjusted for the effect of foliage maturity type. The additional QTLs for resistance against P. infestans on chromosomes 3 and 10 seem to be independent of foliage maturity type and are not affected by epistatic effects of the locus on chromosome 5. The effects of the additional QTLs for resistance are small, but early maturing genotypes that necessarily have the allele for susceptibility for late blight on chromosome 5 may benefit from the resistance that is provided by these QTLs on chromosomes 3 and 10.     

Natural outcrossing in common buckwheat

Common buckwheat is an obligate cross pollinating crop because of its sporophytic self-incompatibility system. Therefore a study to assess the extent of natural outcrossing was undertaken for two years at Morden and Portage la Prairie, Manitoba using the semi-dwarf character, which is due to a homozygous genotype with regard to a single recessive gene, as a marker. The semi-dwarf genotype was grown in 100 m rows running in four directions at 90 ° from a central 36 m² plot of a normal, tall variety. At maturity, seed samples were taken from the semi-dwarf population at designated intervals. At least 200 F₁ seeds from each sample were grown in the greenhouse to the 3-4 leaf stage and the proportion of tall plants was used to determine the percentage of outcrossing. Approximately 50% outcrossing occurred where the semi-dwarf plants were immediately next to the normal plants and then the proportion decreased with increasing distance. Although outcrossing occurred throughout the distance tested, there were no significant differences from 12 m onward from the pollen source. Lack of directional influence in outcrossing and a sharp reduction in the proportion of tall plants within a 3 m distance from the pollen source suggested that wind was not a major factor in dispersing the buckwheat pollen over long distances. Although a substantial cross pollination was limited to a few metres from the pollen source, nearly 1% outcrossing occurred even at 100 m. Since the distance in this experiment was limited to 100 m from the pollen source, the present results do not allow us to recommend the minimum isolation distance required to meet certain standards for buckwheat seed production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Rate and inheritance of cross-fertilization in faba bean (Vicia faba L.)

Summary To assess the feasibility for synthetic varieties of Vicia faba L., 108 faba bean genotypes were screened for cross-fertilization rate in five field experiments carried out in 1989 and 1990 at Wageningen.Significant genotypic differences were observed for outcrossing rate, which ranged from 1 to 55%. These percentages, however, are thought to be not high enough for a commercially feasible production of synthetic varieties.The inheritance of cross-fertilization rate, determined by testing the F₄ generation of a cross between two inbreds, both with high outcrossing rates, appeared to be of a polygenic nature, as was indicated by the wide variation in, and the continous distribution of, the cross-fertilization rates. Among the F₄ lines tested some had a significantly higher cross-fertilization rate than the parents. However, this difference was not found when these lines were retested, indicating that possibilities for successful selection for higher outcrossing rates might be limited.     

More precise map position and origin of a durable non-specific adult plant disease resistance against stripe rust (Puccinia striiformis) in wheat

Recently a major gene determining non-specific adult plant disease resistance against stripe rust (Puccinia striiformis) designated Yrns-B1 was mapped in wheat Triticum aestivum L. by using a cross between ‘Lgst. 79-74’ (resistant) and ‘Winzi’ (susceptible). Linkage to five Gatersleben wheat microsatellite (GWM) markers was discovered, previously mapped on chromosome arm 3BS. In the present study this map was improved by the incorporation of four additional GWM markers. QTL-analysis revealed high LOD values for the resistance at all nine loci, whereas the largest LOD (20.76) was found for the newly mapped marker Xgwm1329. Microsatellite analysis and resistance tests of a collection of old German/UK wheat varieties, including probable ancestors of ‘Lgst.79-74’ were carried out. A high coincidence of non-specific adult plant disease resistance against stripe rust and the presence of ‘Lgst. 79-74’ allele (117 bp) of the marker Xgwm533 was observed among the varieties tested. Linkage during the inheritance of both the resistance and the 117 bp allele of Xgwm533 was demonstrated. The probable origin of Yrns-B1 is discussed. Carriers of this resistance gene were grown on large areas since more than 100 years. To estimate the capability of Xgwm533 as a diagnostic marker for non-specific adult plant disease resistance against stripe rust, microsatellite analysis and resistance tests of a collection of Russian spring wheat varieties were performed. The 117 bp allele of Xgwm533 was found in about 35% of the Russian cultivars analysed, however, none of them possessed the expected disease resistance. Thus, the utilisation of Xgwm533 as diagnostic marker seems to be restricted to certain genepools.     

Mass selection for improved cold and density tolerance of two maize populations

Summary Data of planting and plant density are two cultural practices influencing grain yield of maize (Zea mays L.). Our study was designed to evaluate the usefulness of a mass selection scheme to improve cold and density tolerance of the BS2 and BS3 maize populations. Populations were planted at an early planting date and a high plant density, and three cycles of mass selection for well-filled ears on erect plants were conducted at earch of three Corn Belt locations (i.e., Waseca, MN; Ames, IA; Portageville, MO).Results showed that selection improved cold tolerance traits of BS3, but not of BS2. Mass selection did not increase density tolerance of either population at any location. Selection did not improve response to planting dates, although the early planting date did improve agronomic performance and grain yield of all entries. We concluded that mass selection at high plant densities and early planting dates at diverse geographical locations did not produce cycles adapted to specific environmental conditions.     

Tagging of four fertility restorer loci for wild abortive—cytoplasmic male sterility system in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) using microsatellite markers

Tagging of restorer genes for wild abortive (WA) CMS source by studying a 222 individual plants from a F₂ population of a cross between IR58025A × IR42686R. The restorer line IR42686R that was used in this study had been previously derived through random mating composite population (RMCP) involving 12 parents facilitated by IR36 genetic male sterility. Four Rf genes were tagged to simple sequence repeats (SSR) markers on chromosomes 1, 7, 10, 12 by recessive class analysis. The recombination frequency between a positive marker and Rf locus was calculated using maximum likelihood estimator assuming that all the 46 extremely sterile individual plants were homozygous at the targeted Rf locus. The recombination frequency between the marker and the restorer trait were converted to genetic distances using Kosambi function. A new Rf locus designated as Rf7 on chromosome 12 was found to be linked to RM7003 at a genetic distance of 13.3 cM (LOD 6.12). We report here first, a new molecular marker (RM 6344) linked to Rf4 locus on chromosome 7 that was previously mapped by trisomic analysis. RM443 and RM315 were flanking the Rf3 gene at a genetic distance of 4.4 (LOD 10.29) and 20.7 cM (LOD 3.98) on chromosome 1, respectively. The Rf6 was flanked on both side with SSR markers RM258 and RM591 at a genetic distance of 4.4 (LOD 10.29) and 23.3 cM (LOD 3.39) located on chromosome 10. The random mating composite population is an excellent breeding approach to develop superior restorer lines and for pyramiding different Rf genes of different CMS systems. Rf genes tagged with closely linked SSR markers would be facilitating marker assisted selection (MAS) in hybrid rice breeding program by reducing time and workload for identifying potential restorers. L. Bazrkar and A. J. Ali equally contributed to this work.     

Linkage relationships of genes for leaf morphology and double flowering in Matthiola incana

Summary The inheritance and linkage relationships of a leaf morphology gene of Matthiola incana were investigated. The allele for sinuate leaf shape, c, was found to be recessive to the allele for normal entire leaf, C. The c allele was tightly linked to the recessive allele for double flowering, s. The recombination frequency between the two loci was close to zero. The mode of inheritance of the C gene was in accordance with the hypothesis that a pollen lethal gene is responsible for the constant 1:1 segregation ratio of double-flowered (= male sterile) to single-flowered (= fertile) plants in most M. incana breeding lines. The sinuate leaf allele seemed to reduce the frequency and delay the flowering of double-flowered plants. The importance of the C gene as a double flowering marker in the cultivation and the breeding of M. incana is discussed.Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel. No. 1128-E, 1993 series.     

Allelic variation, sequence determination and microsatellite screening at the XGWM261 locus in Chinese hexaploid wheat (Triticum aestivum) varieties

Wheat microsatellite XGWM261, due to its closely linked to the dwarfing gene Rht8, has been adopted as the diagnostic molecular marker of Rht8. Screening 408 Chinese and 98 exotic varieties showed 13 allele variants in locus of XGWM261, with 6 alleles only to be found in Chinese varieties and 2 only in exotic varieties, respectively. Sequencing results of the 13 alleles revealed their absolute fragment sizes with 216, 212, 210, 206, 204, 202, 200, 196, 194, 192, 190, 174, and 164 bp, respectively. Allelic distribution analysis showed that the 204, 192, 174, and 164 bp alleles were prevailing in Chinese varieties, and the diagnostic 192 bp allele to Rht8 had a very high percentage in the Yellow and Huai River Valleys Facultative Wheat Zone than in the Northern Winter Wheat Zone in China. The GT → AC substitution at position 35 was found in 216, 200, and 174 bp alleles. Moreover, the AG insertion immediately at the end of CT-repeat region was also found in 216, 200, 174, and 164 bp alleles.     

不同花粉密度条件下光温敏雄性不育小麦BS366异交结实分析

研究了人工授粉和田间不同授粉方式条件下,光温敏雄性不育小麦BS366的异交结实情况。结果表明,在不育系花后不同时间人工饱和授粉条件下,其异交结实率呈先增后减的变化,花后第4天至第8天,异交结实率达80%以上,其中第5天为88%;穗中部异交结实能力最强,但异交结实粒数变异也最为明显;多次人工饱和授粉的异交结实率达94.7%,说明该不育系具有较高的异交结实潜力;花后天数与异交结实率的关系可以用方程y=-1.457 7x2 20.489x 12.472(R2=0.914 6**)描述,观察值与模拟值相差0～9%。田间不同授粉方式下,距恢复系1.5 m内,不育系异交结实变异较大,2.0 m以外无显著差异;距恢复系0.5 m内,不育系异交结实率较高,行间差异不显著;辅助授粉时,距恢复系1.0 m以内的不育系异交结实率和杂交种子重量的增幅最大,异交结实率最高可增加45%。光温敏雄性不育小麦BS366制种时播种宽度应控制2.0 m,并在花后8 d内,尤其是花后4～8 d进行辅助授粉,可实现较高异交结实率。     

Development of a SNLP marker from the Pi-ta blast resistance gene by tri-primer PCR

The Pi-ta gene from indica introgressed into japonica rice has been used to control the blast disease caused by the fungal pathogen Magnaporthe grisea (Herbert) Barr. (anamorph Pyricularia oryzae Cav.) worldwide. A single nucleotide length polymorphism (SNLP) was identified at the intron region of the Pi-ta gene to develop a codominant Pi-ta gene marker suitable for genotyping with an automated machine. The DNA primer specific to the resistant Pi-taallele was labeled with blue dye (FAM, 6-carboxyfluorescein) as a forward primer, the DNA primer specific to the susceptible pi-ta allele was labeled with green dye (HEX, 4,7,2′,4′,5′,7′-hexachloro-6-carboxyfluorescein) as another forward primer and the DNA primer identical to both Pi-ta/pi-ta alleles was unlabeled as the reverse primer for polymerase chain reaction (PCR). Using these three primers, a 181-bp blue peak in homozygous resistant and a green peak of 182–183 bp in homozygous susceptible, and both peaks in heterozygous plants were produced by PCR. The utility of marker was verified using a segregating F₂ population, inbred cultivated lines, dominant markers and pathogenicity testing. A codominant Pi-ta marker was thus developed for effective Pi-ta assisted selection for crop improvement. Using highly homologous competitive primers for allele detection by PCR can benefit the study of genome organization of the complex locus. This revised version was published online in August 2006 with corrections to the Cover Date.     

Outcrossing in lentil (Lens culinaris) depends on cultivar, location and year, and varies within cultivars

Knowledge about the degree of outcrossing is essential for onfarm crop management and breeding schemes and for lentil (Lens esculenta Medik.) only restricted evidence was available. Three varieties were investigated in two central European locations to foster research into on farm maintenance and development of cultivars. To determine outcrossing rates, the complete dominance of ‘orange cotyledons’ over ‘yellow cotyledons’ was used as a marker. The degree of outcrossing ranged from 0.06% to 5.12%. Results were strongly influenced by cultivar, year and location. The outcrossing rate of individuals also varied within cultivars, the extremes being 0% and 22.2%. There is evidence that the degree of outcrossing can be heritable. The potential for evolutionary changes in lentil cultivars can be considerable. Growing cultivars close together or as a mixture can lead to natural or even conscious selection and local adaptation. On the other hand, spatial separation may be necessary to avoid undesirable cross‐pollination.     

Gene flow in common bean (Phaseolus vulgaris L.)

This study was conducted in Brazil in order to assess the potential risk posed by gene escape from transgenic into non-transgenic plants and wild populations. A new methodology was applied to evaluate the gene flow between common bean cultivars, by means of a specially delineated experiment in two stages. The first stage consisted of the planting of one cultivar with violet flowers (BB) as pollen source (‘Diamante Negro’), and a receiver (‘Talismã’) with white flowers (bb), at different distances. The source was sown in the center of the area. The pollen receiver cultivar was sown, in concentrical squares around it. At maturity, the rows were sampled at varied distances from the source in the four cardinal directions. In the second stage, the sampled seeds of the previous stage were sown, and the percentage of outcrossing was evaluated during flowering through the presence of violet flowers (Bb). The highest frequency of natural hybrids, 0.136%, occurred at a distance of 0.5 m between the cultivars. The natural outcrossing rate was practically zero beyond a distance of 3.25 m.     

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.     

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.