The potential for breeding an improved lucerne-rhizobium symbiosis. 1. Assessment of genetic variation

Summary Six cultivars of lucerne (Medicago sativa L.) were grown in all possible combinations with eight strains of Rhizobium meliloti in order to assess genetic variation in symbiotic nitrogen fixation. Host genotype and Rhizobium genotype effects on nitrogen fixation were respectively 4.8% and 21.0% of total phenotypic variance. Genetic variation due to host cultivar × Rhizobium strain interactions accounted for a further 6.0% of phenotypic variance. The results indicated low heritability of general symbiotic effectiveness in the host with interaction effects being large enough to suggest that plant performance may be unpredictable with populations of R. meliloti in field soils. Joint regression analysis showed that about 50% of the interactive variation could be explained by generalized differences in the sensitivity of cultivars to alterations in the genotype of Rhizobium strains. One cultivar, Siriver, was relatively insentitive to changes in the Rhizobium genotype whilst still maintaining high average yield. The implications of the results for lucerne breeding 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.     

Symbiotic variability in Vicia faba. 2. genetic variation in Vicia faba

Summary Eight varieties of Vicia faba were examined in association with a standard strain of Rhizobium leguminosarum and also with the application of mineral nitrogen. Large differences in dry matter yield, nitrogen percentage and total nitrogen uptake were apparent between varieties and between rhizobium and mineral nitrogen treatments; the interaction was also significant. Genotypic differences in both the effectiveness of the symbiosis with this strain of rhizobium and the efficiency of utilization of nitrogen were thus demonstrated. The implications of this variability on the assessment of varieties and the possiblity of exploiting it by plant breeding techniques are discussed.     

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.     

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^-.     

Intraspecific Variation in Mean Endosperm Cell Cycle Time in Vicia faba (L.) and Interspecific Hybridization with Vicia narbonensis (L.)

The rate of early embryo-sac development was studied in seven faba bean (Vicia faba L.) cultivars grown in a controlled environment. The embryo-sac stage was determined from the number of endosperm nuclei per embryo-sac and the number of embryo cells during the first 12 days after pollination. Differences in early embryo-sac development were observed between the genotypes studied. In addition, five different V. faba×V. narbonensis crosses and reciprocals were made using genotypes with small differences in mean endosperm cell cycle time and genotypes with large differences. The percentage of success (pod set) in V. faba×V. narbonensis crosses ranged between 9 % and 59 % while in the reciprocals it ranged from 12 % up to 30 %. However, cytological studies showed that the high percentage of success (pod set) observed in the cross A-107 × A-202 was not associated with a higher percentage of interspecific hybrid embryos. The results indicate that genotypes of the two species with smaller differences in mean endosperm cell cycle time result neither in a higher percentage pod set nor in bigger hybrid embryos.     

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.     

Combining ability for nodulation in common bean (Phaseolus vulgaris L.) genotypes from Andean and Middle American gene pools

Ten F₁'s obtained from crosses among five common bean genotypes of Andean (WAF 15, Mineiro Precoce and Batatinha) and Middle American (BAT 304 and Ouro) gene pools were assessed for their combining abilities for root nodulation with Rhizobium tropici strain CIAT 899. The plants were grown under controlled conditions and evaluated for number of nodules per plant (NN), nodule dry weight (NDW), mean nodule weight (MNW) and plant fresh weight (PFW). The subdivision of the treatment effects on the general (GCA) and specific combining effects (SCA) were performed according to Griffing's diallel analysis method 2, model 1. The analyses of variance and estimates of quadratic components showed that non-additive gene effects were more important in the expression of NN and PFW, whereas additive gene effect was predominant for NDW and MNW. A close association was observed between high number of nodules and GCA. Generally, crosses involving parents of different gene pools yielded hybrids with high positive estimates of SCA for all assessed traits. The genotypes of Andean origin WAF 15 and Mineiro Precoce are the most promising parents for breeding programs to increase NN and NDW in common beans. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic specificity of nodulation

Summary A genetic system is proposed for establishing desirable strains of Rhizobium japonicum in soybean fields against the competition of aggressive but less productive indigenous strains. This system requires soybean cultivars which substantially exclude the indigenous strains while selectively admitting specific introduced strains. Rhizobium strains were identified which were infective, both in sand and in soil culture, with a host genotype (rj₁rj₁) which excludes most indigenous strains. Rhizobium infectivity was associated, for the first time, with the propensity to induce symptoms caused by rhizobitoxine production by the bacteria. The basis of this association is unknown. A possible enabling role for rhizobitoxine, or a precursor or derivative of rhizobitoxine, in the infection process is discussed.     

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.     

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 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.     

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.     

Contributions to the Analysis of Genotype × Environment Interactions: Comparison of Different Parametric and Non-parametric Tests for Interactions with Emphasis on Crossover Interactions

The objective of this study was to compare five statistical procedures (analysis of variance, the Azzalini/Cox test, the Hildebrand procedure, the Kubinger approach, and the de Kroon/van der Laan technique) for the analysis of genotype × environment interactions in cross‐classified data sets from cultivar performance yield trials with rows = cultivars and columns = environments (locations and/or years). The procedures Hildebrand, Kubinger and de Kroon/van der Laan are non‐parametric methods based on ranks, while analysis of variance and the Azzalini/Cox test proceed from the original absolute yield data. These very different statistical techniques were applied to extensive data sets from German official registration trials (1985–1989) with winter oilseed rape (Brassica napus L.), faba bean (Vicia faba L.), oat (Avena sativa L.), fodder beet (Beta vulgaris L.) and sugar beet (Beta vulgaris L.). The Azzalini/Cox and de Kroon/van der Laan methods are based on the crossover concept of interaction (different rank orders) while the other methods are based on the usual concept of interaction (deviations from additivity of main effects). For an analysis of usual interactions the procedures Hildebrand, Kubinger and analysis of variance are approximately equivalent. For the crossover concept of interaction, the Azzalini/Cox approach is recommended, especially if one is particularly interested in rank changes between environments within genotypes.     

Assessment of genetic divergence in salt tolerance of soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.) genotypes

A large number of soybean (Glycine max L.) genotypes of diverse growth habit and adaptive characters were used in the experiment. Soil salinity-induced changes in nine morpho-physiological characters of 30-day-old seedlings of 170 soybean genotypes were compared in the study. The first and second principal components (PC) of principal component analysis (PCA) results accounted for 97 and 2.5%, respectively, of the total variations of soybean genotypes. The variation for the first PC was composed mainly of relative total dry weight (DW), relative shoot dry weight, as well as petiole dry weight. There were four clusters distinguished in the cluster analysis. The genotypes in cluster IV performed better in respect to relative total dry weight and relative shoot dry weight and hence having salt tolerance. The genotypes clusters III performed very poorly and those of clusters II and I were moderate to poor. D2 analysis indicated that the clusters differed significantly from each other. Discriminant function analysis (DFA) again asserts strongly that more than 92% of the genotypes were correctly assigned to clusters. Both PCA and DFA confirmed that the relative total DW followed by shoot and petiole DW were the major discriminatory variables, and the root DW were the secondary important variables to distinguish genotypes into groups. In this study, multivariate analyses were used in identifying the soybean genotypes of desirable traits for salt tolerance.     

Variation in Nitrate Tolerance of Nitrogen Fixation in the Pea/Rhizobium Symbiosis

The short-term effects of nitrate on symbiotic nitrogen fixation were studied in six cultivars of pea (Pisum sativum L.) using two strains of Rhizobium leguminosarum. Plants grown symbiotically for three weeks, in order to eliminate the effect of NO₃ on nodule formation, were exposed to nil, 5 or 15 mM NO₃ for a period of seven days. Nodule mass, nitrogenase activity by acetylene reduction and the NO₂ and NO₃ concentrations in nodules were determined. Exposure to nitrate reduced the total acetylene reduction activity (ARA) in all cultivars, and a significant cultivar × nitrate treatment interaction was found. Nitrate influenced the components of N₂ fixation, nodule mass and specific ARA (ARA/mg nodule), differently within and among cultivars. Nodule mass was reduced in all cultivars, but a variation among cultivars in the degree of reduction was found. Nitrate also reduced the specific ARA to different extents in the cultivars- The specific ARA of nodules from NO₃-treated plants, expressed as per cent of the specific ARA of nodules from plants grown without NOT₃ was negatively correlated with the concentration of NO₃ in nodules. No relationship was found between the reduction of N₂ fixation and the nitrite concentration in nodules. The results indicate that it is possible to improve the nitrate tolerance of symbiotic N₂ fixation of the established pea/Rhizobium symbiosis by selection of suitable host plants.     

Differential response of Vicia faba L. genotypes to 2, 3, 5-triiodobenzoic acid (TIBA)

Summary Response to application of the growth regulator 2, 3, 5-triiodobenzoic acid (TIBA) was examined in the parents and F₁ progenies of a diallel cross of six inbred lines of Vicia faba. TIBA affected the genotypes differentially but in general gave shorter internodes, reduced apical dominance, increased pod set on the lower nodes and earlier seed maturity.     

Single gene mutation for green cotyledons as a marker for the embryonic genotype in faba bean, Vicia faba

A new spontaneous mutation is described in faba bean, Vicia faba L., characterized by a marked green colour in the entire cotyledon tissues of the mature seeds, while the wild phenotype is yellow. This seed character reflects the embryonic genotype and is determined by a single recessive allele named i1-1. It is distinct from the y gene, which codes for green testa. As is the case for gene y, the gene i1-1 displays no epistatic effects with the zero-tannin genes zt1 and zt2 which influence seed and flower colour in faba bean. In this allogamous species, such a mutation is a useful tool in cross-fertilization studies.     

Variation in Reaction of Faba Bean Lines to Different Accessions of Orobanche crenata Forsk.

Three pot experiments were earned our to investigate the variation among geographic accessions of Orobanche crenata Forsk. in their ability to attack different stocks of Vicia faba L. Orobanche accessions were quite different in their influence on the performance of genetic lines of the host. However, host parasite relationships appeared to be dependent upon environmental conditions. The usefulness or host genotypes for differentiating between parasite accessions was rather limited. The number of Orobanche shoots per host plant does not appear 10 be a precise indicator of host tolerance although it has been frequently used for this purpose.     

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.