Symbiotic variability in Vicia faba. 1. Genetic variation in the Rhizobium leguminosarum population

Summary Twenty isolates taken at random from indigenous populations of Rhizobium leguminosarum sampled near Aberystwyth were each inoculated into plants of a standard Vicia faba variety grown aseptically under conditions free of combined nitrogen. Plants in association with the individual rhizobium isolates exhibited large differences in dry matter yield, nitrogen content, efficiency of nitrogen utilization and date of first flower. The implications of these differences to the productivity and reliability of varieties of field beans are discussed.     

Symbiotic variability in Vicia faba. 3. Genetic effects of host plant,rhizobium strain and of host × strain interaction

Summary Six Vicia faba populations were grown in all possible combinations with six Rhizobium leguminosarum strains in order to estimate the relative importance of the three genetic components of symbiotic variability. Additive genetic effects of host genotype and rhizobium genotype accounted for only 8.9% and 11.8% of the total phenotypic variation. Non-additive variation attriabutable to specific host genotype × rhizobium genotype interactions was by far the largest component of variation and accounted for 73.8% of phenotypic differences. Therefore the greatest improvement in symbiotic nitrogen fixation is likely to arise from simultaneous selection of both symbionts.     

Cross-fertilization behaviour of some white flowering varieties of Vicia faba

Summary Applying a special design for maximalization of cross-fertilization and using flower colour as a marker, considerable differences in the percentage of cross-fertilization between white flowering Vicia faba varieties were observed. Part of these differences were due to differences in beginning of flowering in white and coloured varieties, part of them proves to be independent of this and, therefore appear to be characteristic for the variety. Interaction between mother and father varieties was absent.     

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.     

Genetics of resistance to 3 isolates of Ascochyta fabae on Faba bean (Vicia faba L.) in controlled conditions

Summary Genetics of resistance to Ascochyta fabae Speg. in Vicia faba L. was studied with a final objective to develop resistant faba bean varieties to Ascochyta blight. The study was conducted separately on 3 single spore isolates (AF10-2 and AF13-2 from Tunisia and AF4-3 from France) and belonging to different groups of virulence (GV1 and GV2). Important general combining ability (GCA) effects were found especially with isolates AF10-2 and AF4-3. Specific combining ability (SCA), although significant for the 3 isolates, was important only with AF13 -2, but less important than GCA. Additive gene effects were predominant to non-additive effects. Lines 29H and A8817 transmitted to their progenies resistance to the 3 isolates, whereas 14–12 and 19TB conferred resistance to their progenies only with isolates AF13-2 and AF4-3, respectively. In the material studied, resistance was generally controlled by dominant genes but also could be attributed to recessive genes although less frequent. Analysis of segregation in the F2 of 2 crosses between the resistant lines (A8817 and 29H) and the susceptible line (14–12) with isolate AF4-3 revealed dominant monogenic control at the level of leaves in the 2 resistant lines and, in addition, a recessive gene controlling resistance of stems. Non-allelic interactions were occasionally manifested and their origin appeared to be due to line 19TB. A recurrent selection scheme was proposed with the objective to develop improved open-pollination populations and synthetic varieties responding to the objective of the national Tunisian research programme on faba bean.     

Variation and covariation in Vicia faba L. populations of Mediterranean origins

Summary Variation and covariation in 15 Vicia faba L. ssp. major populations from Italy. Greece and Egypt were analyzed. Ten quantitative morphobiological characters of agronomical and nutritional interest were observed. Differences among countries of origin were highly significant for all characters except for lowest podded node. Phenotypic distances among populations, both within and among origins, estimated on canonical variates evidenced that the Egyptian populations from a distinct group and that a relevant degree of similarity exists among the Greek and the Italian meterial. The covariation structure within populations, which was studied by means of factor analysis, was comparatively simpler in the Egyptian material than in that form the other two Mediterranean countries. The differences evidenced can be attributed to different gene associations between characters and suggest the possibility of obtaining, through recombination, genotypes agronomically more favorable.Research work supported by C.N.R., Italy, special grant. Sub-Project 1, Paper No. 299.     

Controls over pollination and ovule fertilization in Vicia faba

Summary Variation in the frequencies of pollination and ovule fertilization was generated among faba bean (Vicia faba L.) genotypes in experiments which compared self-and open-pollination, natural and extended photoperiods, presence and absence of honey bees and a range of plant populations.There was a near-1:1 relationship between the frequencies of pollination of stigmas and the fertilization of at least one ovule within the pistil. Pollination was the major barrier to fertilization of the pistil, and ovules, since no mechanism was evident in the style to prevent pistil fertilization. The frequency with which single ovules were fertilized in pollinated pistils fell as their distance from the stigma increased, and averaged 83.5% overall.The frequency of pollination was most influenced by the level of autofertility possessed by the genotype, and the activity of pollinators. Provision of honey bees to plants in caged plots showed that high levels of pollination could be achieved under open-pollinated conditions, but this was also influenced by autofertility. Increasing plant population in the field had only a small effect on pollination, but extending photoperiod caused the levels and timing of self-pollination in three genotypes to vary.Large differences were found among genotypes in their early stigma receptivity, that is, the frequency of stigmas found to be pollinated at the erect standard stage, prior to the flower opening and being visited by bees. The implications of early stigma receptivity for producing new cultivars were discussed.     

Self-fertility and the distribution of seed yield in Vicia faba L.

Summary Five lines of field beans (Vicia faba L.) bred and selected for a high level of self-fertility have demonstrated heritable differences in distribution of seed yield on the reproductive nodes when grown under conditions of selfpollination. Fully self-fertile genotypes produce a large proportion of the seed yield on the lower nodes. The breeding and selection for both self-fertility and the distribution of seed yield is discussed.     

Interspecific hybridization between Vicia faba (L.) and Vicia narbonensis (L.): Early pod growth and embryo-sac development

Summary Fertilized embryo-sac development and pod growth was studied in one Vicia faba cultivar, one Vicia narbonensis population and their reciprocal crosses. The initial development of endosperm and embryo was at least four days faster in V. narbonensis than in V. faba. Pods and ovules developed also faster in V. narbonensis than in V. faba. The growth rate of the hybrid pods followed the growth rate of the mother species, but was slower than that of the pods from selfed flowers. In the cross V. narbonensis × V. faba the ovules stopped growing 9 days after pollination, while in the reciprocal cross they stopped growing 15 days after pollination. Hybrid embryo-sacs from V. faba × V. narbonensis were aborted before they reached the stage of 256 endosperm nuclei or 200 embryo cells. Selfed V. faba embryo-sacs reached this stage in less than 9 days after pollination. In the reciprocal cross the embryo-sacs were aborted before they reached the stage of 128 endosperm nuclei or 80 embryo cells. Selfed V. narbonensis embryo-sacs reached this stage at the 4th day after pollination. Given that at these stages the embryo has less than 200 cells it was concluded that an in-ovule embryo culture technique should be developed to obtain hybrid plants.     

Factors determining the performance of synthetics in Vicia faba L. 1. Heterogeneity,heterozygosity, and degree of cross-fertilization

Summary The relative performance of synthetic varieties in faba beans (Vicia faba L.) as compared with line varieties is the result of the joint effect of several factors. Factors investigated in the present study were (1) effects of heterogeneity and (2) heterozygosity on agronomic performance, (3) genetic variance for the degree of cross-fertilization among inbred lines and (4) genetic variance among homozygous and heterozygous genotypes for yield. The evaluation was based on multilocational trials in Germany with 36 inbred lines and derived entries with contrasting population structure (levels of heterozygosity and heterogeneity). When inbred lines and F₁-hybrids are included in the same mixture of genotypes, the resulting heterogeneity had a positive effect on yield. Relative mid-parent heterosis for yield was between 33% and 51%. The average degree of cross-fertilization was 53.5%, and its genetic standard deviation was 8.3%. The estimates for genetic standard deviation for yield among inbred lines were between 3.4 q/ha and 5.3 q/ha. The highest yielding synthetic variety is expected to outyield markedly the highest yielding line.     

Note on the presence of the sym-1 gene in Vicia faba hampering its symbiosis with Rhizobium leguminosarum

Summary A symbiotically defective mutant in Vicia faba is described. This character is controlled by a recessive gene named sym-1. Reciprocal graft experiments indicate that only the root system controls its expression. Depending on the strain of Rhizobium, the root phenotype can be either Fix⁺ Nod^- or Nod^-.     

Pattern analysis and genotypic x environment interactions in faba bean (Vicia faba L.) populations

Summary Twenty-nine populations of faba bean (Vicia faba L. subsp. major, equina and minor) of mediterranean origin were evaluated for the genetic variability of 12 biomorphological characters at four locations in Apulia and Basîli cata regions (Italy) in 1983–84.A pattern analysis appoach, based on classification and ordination procedures is presented for to characterize differences among population in mean performance and response across environments. Informations about G × E interaction were obtained.Research work supported by CNR, Italy. Special grant I.P.R.A. Sub-project 1. Paper N. 1099.     

Mutagenesis of faba bean (Vicia faba L.) and the identification of five different genes controlling no nodulation,ineffective nodulation or supernodulation

Summary Mutagenesis ofVicia faba L. using ethyl methanesulfonate has yielded three mutants (f13, f73, f245) with ineffective nodulation (Nod⁺Fix^–), one mutant (f48) unable to nodulate (Nod^–) and one supernodulating mutant (f32) having 3 to 5 times more nodules than the control in the absence of combined nitrogen, and expressing nitrate tolerant nodulation (Nod⁺⁺, Nts). All of the mutant phenotypes were controlled by single and distinct recessive genes designatedsyml tosym5, respectively. Grafting experiments showed that the (Nod⁺Fix^–) and (Nod^–) phenotypes were both under root genotype control and that the (Nod⁺⁺, Nts) phenotype was under shoot genotype control.     

Variation in traits affecting nodulation of common bean under intercropping with maize and sole cropping

Common bean (Phaseolus vulgaris L.), an important food crop in Europe, America, Africa and Asia, is thought to fix only small amounts of atmospheric nitrogen. It contributes significantly to the sustainability of traditional cropping systems because of the predominance of small-scale farmers who cultivate beans in those areas. The objectives of this work were to evaluate bush bean varieties under common agronomic cropping systems and to evaluate breeding lines under low N-fertility sole cropping and intercropping systems. The purpose of the study was to characterize the genotype and cropping system's variability in symbiotic and plant characters and to identify the most suitable genotypes to establish an effective symbiosis with indigenous strains of Rhizobium. No significant differences among the bush bean varieties evaluated under typical fertilization practices were observed for N₂-fixation and plant traits except for seed nitrogen. Significant differences among the bean lines studied under low N-fertilization conditions were detected for plant growth,plant component and N₂-fixation traits. A significant interaction of bean genotype x cropping system was found for number of nodules per plant and nodule moisture on the bush bean varieties studied, and for days to emergence, days to flowering, end of flowering, shoot length, root dry weight and shoot nitrogen on the bean lines evaluated. Nodulation parameters were correlated positively with the yield components, shoot and root parts and duration of flowering, and correlated negatively with seed crude protein, pod and seed dimensions and seed dry weight. These observations indicate that it may be possible to increase both the symbiotic N₂-fixation and seed yield through plant breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Present status and future strategy in breeding faba beans (Vicia Faba L.) for resistance to biotic and abiotic stresses

Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses.     

The identification of faba bean (Vicia faba L.) trisomics by translocation tester set

Summary Five subtelocentric primary trisomics of Vicia faba L. were isolated from crosses between asynaptic mutants and normal plants or translocation tester stocks. The extra chromosome of each trisomic was identified cytologically by critical multivalent association at metaphase I of meiosis.     

Genotype x Bradyrhizobium japonicum strain interactions in dinitrogen fixation and agronomic traits of soybean (Glycine max L. Merr.)

Summary Bradyrhizobium japonicum strain G49 has been the only inoculum used in French soils. Soybean (Glycine max L. Merr.) cultivars were selected and tested according to their performances with this rhizobial strain. The aim of the present study was to determine the consequences of strain substitution on N₂ fixation abilities of various genotypes. Three genotypes and cultivar Weber, in combination with B. japonicum strain G49 or SMGS1, were cultivated in pots and tested for nitrogenase activity under differing nitrogen nutrition conditions. The reliability of ARA (acetylene reduction activity) measurement for assessing symbiotic nitrogen fixation under the experimental conditions used was checked. Genotypic variability for symbiotic fixation activity was observed with each strain under soil culture conditions; important genotype x strain interactions were also involved. These results were corroborated for the protein yield and other yield component performances of the various genotype-strain associations. Thus, in France, the replacement of strain G49 with another one might result in the alteration of the relative agronomic performances of the soybean cultivars, since N₂ fixation is considered as a major factor of soybean productivity.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of faba bean <Emphasis Type="Italic">(Vicia faba</Emphasis> L.) using embryo axes

A system for the production of transgenic faba bean by Agrobacterium-mediated transformation was developed. This system is based upon direct shoot organogenesis after transformation of meristematic cells derived from embryo axes. Explants were co-cultivated with A. tumefaciens strain EHA105/pGlsfa, which harbored a binary vector containing a gene encoding a sulphur rich sunflower albumin (SFA8) linked to the bar gene. Strain EHA 101/pAN109 carrying the binary plasmid containing the coding sequence of a mutant aspartate kinase gene (lysC) from E. coli in combination with neomycinphosphotransferase II gene (nptII) was used as well. The coding sequences of SFA8 and LysC genes were fused to seed specific promoters, either Vicia faba legumin B4 promoter (LeB4) or phaseolin promoter, respectively. Seven phosphinothricin (PPT) resistant clones from Mythos and Albatross cultivars were recovered. Integration, inheritance and expression of the transgenes were confirmed by Southern blot, PCR, enzyme activity assay and Western blot.     

Prospects for commercialisation of F1 hybrid field beans Vicia faba L.

Summary Yields of F₁ hybrids averaged 22% more than the mean of their heavier-yielding inbred parents, and the advantage of hybrids over composite populations varied from 3 to 18%, more in winter than spring beans.From estimates of cost/benefit ratios that included the possibility of poor seedset on male steriles, on average 1 year in 10, it was concluded that hybrid seed might cost more than twice that of conventional varieties, but a 10% increase in yield on a 4t/ha crop would cover this extra cost of seed.Thus, hybrids could still be the more profitable type of variety provided seed rates are low and yield levels high. It is therefore still worthwhile to pursue the goal of stable cytoplasmic male sterility (CMS) and/or a control of the rate of increase in revertants. Some methods are suggested whereby it might be possible to reduce revertants, improve pollination or exploit heterosis other than with CMS.     

Differences in nitrogen fixation among field-grown red clover strains at different levels of 15N fertilization

Summary Genetic variation in fixed nitrogen (N) yield of red clover (Trifolium pratense L.) strains and cultivars was investigated using the ¹⁵N isotope dilution method under three regimes of N fertilization: 0.5, 30, and 60 N (kg N ha^–1 per cut). The yield of fixed N per cut (the mean of eight cuts over 2 production years) varied among the strains (progenies of crosses between inbred parents) from 148 to 443 mg per plant at 0.5 N, from 76 to 324 mg at 30 N, and from 69 to 300 mg at 60 N. There were significant and consistent strain differences in the percentage of clover N derived from the atmosphere (% N_dfa). However, %N_dfa was positively correlated with dry mass yield. Consequently, ranking of the strains according to fixed N yield reflected that of dry mass yield. There were only minor strain × N fertilizer interactions, suggesting that selection for enhanced N fixation can be carried out at a single rate of fertilizer N. For a selected pair of strains, the difference in yield of fixed N was confirmed in an Italian ryegrass-red clover mixture, both without and with the addition of N fertilizer (50 kg N ha^–1 per cut). Results with 7-week-old seedling plants in a growth chamber, although obtained in the presence of mineral N and with the isotope dilution method, did not adequately predict field performance. It is concluded that selection for dry matter or total N yield is likely to result in an enhanced yield of fixed N at any level of mineral N availability.Abbreviations % N_dfa percentage of clover nitrogen derived from the atmosphere by symbiotic nitrogen fixation - S₂-F₁-n progeny of pair cross between inbred parents obtained after two generations of selfing