Seed protein electrophoresis of the wild and cultivated species of selection faba of Vicia

Summary The seed protein profile obtained by disc electrophoresis was found to be a diagnostic characteristic of the broad bean, V. faba, and of its taxonomically wild relatives V. narbonensis, V. galilaea and V. hyaeniscyamus The marked difference in the seed protein profile of the cultivated species and the three wild species indicated that none of the latter can be considered as the wild progenitor of V. faba. The similarity in the profiles of V. galilaea and V. hyaeniscyamus was consistent with the morphological resemblance of these species and their similar ecological requirements.     

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.     

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.     

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.     

The sulphur-rich Brazil nut 2S albumin is specifically formed in transgenic seeds of the grain legume Vicia narbonensis

Summary Epicotyl explants were co-cultivated with Agrobacterium tumefaciens EHA101 to transfer a chimeric 2S albumin gene construct carried in the binary Ti plasmid vectors pGSGLUC1 or pGA472 into the grain legume Vicia narbonensis. This gene encoding the sulphur-rich Brazil nut albumin was under the control of either the CaMV 35S promoter which permits gene expression in all organs, or the Vicia faba legumin B4 promoter which elicits seed-specific gene expression. After callus formation and selection for kanamycin resistance, somatic embryos were induced which, in the case of transformation with the vector pGSGLUC1, were screened for GUS activity. Embryos that produced GUS were in addition analysed for 2S albumin formation. Selected transgenic embryos were cloned by multiple shoot regeneration. Rooted and fertile plants were obtained by grafting transgenic shoots on the appropriate seedlings. R₁ and R₂ generations were raised and analysed for GUS as well as 2S albumin gene expression.Expression of the 35S promoter/2S albumin gene fusion took place in all organs of the transgenic plants including the cotyledons of seeds, whereas seed-specific gene expression was found in transformants with the legumin promoter/2S albumin gene fusion. The 2S albumin accumulated in the 2S protein fraction of transgenic seeds and its primary translation product was processed into the 9 and 3 kDa polypeptide chains. The foreign protein was localised in the protein bodies of the grain legume. Analysis of the R₂ plants indicated Mendelian inheritance of the 2S albumin gene. In homozygous V. narbonensis plants the amounts of 2S albumin were twice that present in the corresponding heterozygous plants. Whereas only low level formation of the foreign protein was achieved if the gene was under the control of the 35S promoter, approximately 3.0% of the soluble seed protein was 2S albumin if seed-specific gene expression was directed by the legumin B4 promoter. Some of these transformants exhibited a three-fold increase in the methionine content of the salt-soluble protein fraction extracted from seeds.Abbreviations 35S cauliflower mosaic virus 35S protein gene - GUS -glucuronidase - NPTII neomycin phosphotransferase II - LeB4 Vicia faba legumin B4 gene - 2S albumin Brazil nut (Bertholletia excelsa) 2S albumin - ER endoplasmic reticulum - rER rough endoplasmic reticulum - HPLC high pressure liquid chromatography     

Inheritance and linkage of a gene for testa-imposed seed dormancy in faba bean (Vicia faba L.)

Seed dormancy in the grain legumes may interfere with germination in some situations, yet is not well described. Dormancy in faba bean (Vicia faba L.) was investigated to determine the mode of inheritance and whether the gene or genes controlling this trait are linked to other genes. Seeds produced by 235 recombinant inbred lines at the F₆ generation were used for this study. The parents were the cultivar ‘Optica’ and 172, an Afghanistan landrace which possesses extended seed dormancy. Germination was scored on a seed sample from each recombinant inbred line. Segregation patterns indicated that the trait is monogenic. This gene, named doz, is linked to a gene controlling anthocyanin and proanthocyanidin synthesis, sp-b, with the genetic distance estimated to be about 25 cM. No associations were detected between seed dormancy and pod dehiscence, seed lustre or mean seed weight. The implications of the persistence of a gene for seed dormancy in the gene pool of V. faba for the introgression of traits from landraces and cultivars possessing dormancy and for the origins of domesticated V. faba are discussed.     

Factors Affecting the Differentiation of Resistant Faba Bean Plants after Inoculation with Sclerotinia trifoliorum

Faba bean (Vicia faba L.) plants 3, 4, 5 and 6 weeks old were inoculated at different internodes by carrot root pieces colonized by the fungus Sclerotinia trifoliorum Eriks. The plants were left in the growth chamber for 72 h. In addition, eight faba bean cultivars, 4 weeks old, were inoculated by the same inoculum which was left on the stems for various lengths of time. When the inoculum was removed, the plants were scored for disease severity. The highest disease seventy was on the youngest tissues inoculated. Susceptible and partially-resistant cul-tivars could be differentiated effectively when the inoculum was removed 48 to 72 hours after the inoculation.     

Physiological and Molecular Characterization of Faba bean (Vicia faba L.) Genotypes for Adaptation to Drought Stress

Faba bean (Vicia faba L.) is one of the most important and drought sensitive grain legumes. Drought stress is thus one of major constraints in global faba bean production. In this study, twenty local and exotic faba bean genotypes were characterized on physiological and molecular basis. Seeds of faba bean genotypes (six per pot) were sown in poly venyl chloride pots. After seedling emergence, soil moisture was maintained at 100%, 50% and 25% of field capacity designated as well watered, moderate drought and severe drought, respectively. Drought stress significantly influenced the leaf area, leaf temperature, stomatal conductance, relative leaf water contents, grain yield and water‐use efficiency. Faba bean genotypes also differed for the leaf area, leaf temperature, relative leaf water contents, grain yield and water‐use efficiency. Faba bean genotypes Kamline and L.4 were better equipped to curtail water loss, maintain tissue water status, produce stable grain yield and had better water‐use efficiency under mild and severe drought stress, and may be used in breeding programmes. Amplified fragment length polymorphism markers showed high potential in detecting polymorphism and estimating genetic diversity among faba bean genotypes. Unweighted pair group method with arithmetic mean cluster analysis of the genotypes illustrated considerable association between molecular diversity, genetic background and geographic origin. In crux, high polymorphic rate and polymorphism information content values, together with the low genetic similarity observed among tested genotypes suggests a high level of heterogeneity, which may be used in breeding programmes to assemble different drought tolerance mechanisms in one genotype.     

Selection in Faba Bean for Tolerance to Broomrape, Orobanche crenata Forsk.

Six populations of faba bean (Vicia faba L.) were developed through various selection schemes to examine the influence of selection In improving tolerance to Orobanche crenata (Forsk.) in local faba bean stocks. All populations trace their origin to 209 farmers’ seed lots that were collected from Egypt. The entries of each population were grown during the 1982/1983 season in the Orobanche nursery located at Giza in Egypt. Comparison among means of the selected, populations showed that more progress in improving Orobanche tolerance could be achieved from combined selection between and within lots. Between-lots selection was less efficient than single-plant selection. Results, of this study show that considerable progress could be achieved from selection for Orobanche tolerance in local faba bean germplasm. There was sufficient residual genetic variation in the selected populations to expect substantial progress from further selection for most tolerance traits.     

Genetic variability of faba bean genotypes for chocolate spot (<Emphasis Type="Italic">Botrytis fabae</Emphasis>) resistance and yield

Faba bean (Vicia faba L.) has high utility as a food and soil fertility improving crop. One of the major fungal pathogens of faba bean is Botrytis fabae, the causative agent of chocolate spot. The disease affects significantly the leaf, stem, pod and seed of faba bean compromise its productivity in the smallholder farming sector. Nonetheless, there are limited resistant/tolerant faba bean varieties available and disease control technology options. Therefore, it was prudent to evaluate faba bean landraces for chocolate spot resistance. Fifty landraces together with ten improved varieties were evaluated both in the field and in the greenhouse under natural and artificial inoculation with previously selected aggressive Botrytis fabae isolate (Iso-016) from West Gojjam, in Ethiopia. There were highly significant differences (p?<?0.001) among the landraces for reaction to the disease and agronomic traits. Significant positive correlation was recorded between reaction of genotypes in the field and greenhouse disease data. The overall mean disease epidemics varied from 92.5 to 697.5 for the area under disease progress curve (AUDPC). The highest level of resistance was found in the ICARDA lines, ILB-4726, ILB-938 and BPL-710. Of all 18 landrace collections displayed significantly lower disease reaction than the susceptible check. However the resistance was moderate. The selected eighteen landraces will be recommended for use in breeding for chocolate resistance. Overall, resistance was highly heritable, suggesting that phenotypic selection can be exploited to improve chocolate spot resistance in faba bean varieties.     

Susceptibility of Faba Bean (Vicia faba L.) to Heat Stress During Floral Development and Anthesis

Experiments were conducted over 2 years to quantify the response of faba bean (Vicia faba L.) to heat stress. Potted winter faba bean plants (cv. Wizard) were exposed to temperature treatments (18/10; 22/14; 26/18; 30/22; 34/26 °C day/night) for 5 days during floral development and anthesis. Developmental stages of all flowers were scored prior to stress, plants were grown in exclusion from insect pollinators to prevent pollen movement between flowers, and yield was harvested at an individual pod scale, enabling effects of heat stress to be investigated at a high resolution. Susceptibility to stress differed between floral stages; flowers were most affected during initial green‐bud stages. Yield and pollen germination of flowers present before stress showed threshold relationships to stress, with lethal temperatures (t₅₀) ?28 °C and ~32 °C, while whole plant yield showed a linear negative relationship to stress with high plasticity in yield allocation, such that yield lost at lower nodes was partially compensated at higher nodal positions. Faba bean has many beneficial attributes for sustainable modern cropping systems but these results suggest that yield will be limited by projected climate change, necessitating the development of heat tolerant cultivars, or improved resilience by other mechanisms such as earlier flowering times.     

Early Embryo-Sac Development and Intraspecific Variation in the Mean Endosperm Cell Cycle Time in Vicia narbonensis (L.)

The early endosperm development in seven Vicia narbonensis populations was studied under a 23/ 16 ° C day/night temperature regime. Double fertilization occurred in less than 24 h after pollination. Endosperm divisions were synchronous until the 8th division of the endosperm nuclei. Aberrant endosperm nuclei were first observed alter the 8th division and their number increased after the 10th division. Endosperm nuclei continued to divide until the 12th day after pollination. During the following days a decrease in the number of endosperm nuclei was observed together with a substantial increase in the number of aberrants. Endosperm nuclei finally disintegrated. Neither cellularization nor starch deposition was observed in the endosperm. A positive correlation (r 0.69) was recorded between early pod length and the number of endosperm nuclei found in the ovules within each pod. In the populations studied the number of endosperm nuclei increased at a steady but different rate. This was followed by embryo development as well. It was concluded that intraspecific variation in the mean endosperm cell cycle time exists in Vicia narbonensis and that populations A 174 and A 202 should be more compatible to Vicia faba ev. ‘Polycarpe’ than population A 201.     

QTL detection for flowering time in faba bean and the responses to ambient temperature and photoperiod

Faba bean (Vicia faba L.) is a grain legume primarily used for animal feed and human food grown in a range of environments, globally. Time of flowering in faba bean is critical for adaptation to specific environments and is controlled largely by factors such as ambient temperature and photoperiod. The aim of this study was to investigate the genetic control of flowering time and the responses of flowering time to ambient temperature and photoperiod in faba bean. A bi-parental recombinant inbred line population (Icarus × Ascot) was evaluated over three years in field trials and three different controlled environments with varying temperatures and photoperiods. QTL analysis identified eight regions of co-localised QTLs associated with days to flowering, thermal time to flowering and node of first flower; on Chr-I.A/III/V, Chr-I.B.3, Chr-III.1, Chr-III.2, Chr-V.1 and Chr-V.2. Two of the detected regions are common with previously detected QTLs, up to two more are possibly common and the remaining four appear to be novel. For the first time, the associations of these QTLs with ambient temperature and photoperiod response were described. Candidate genes for some of the QTLs were identified using the associations with ambient temperature and photoperiod response together with knowledge extended from other legumes that have a syntenic relationship with faba bean.     

The Nutritive Value of Ten Inbred Lines of Faba Beans (Vicia faba L.) in Relation to their Content of Antinutritional Constituents and Protein Quality

Ten inbred lines of fabe beans (Vicia faba L.) selected according to their quality characters have been investigated for carbohydrates, proteins, and antinutritional compounds. Digestible energy, N-balance trials with growing rats comprising determination of the protein digestibility and biological value were used as criteria in connection with comprehensive chemical-biochemical analysis. The chemical composition of the ten lines showed a considerable diversity as cad the results from the rat crisis. However, the content of vicine and convicine in all or the investigated lines were below the level previously found to have effects on the nutritive value, Starch, protein and fibre were the quantitatively dominating seed constituents, and all showed great variation among the lines. The starch content was not correlated to the quality or nutritive value of the seed, whereas the protein content was negatively correlated to the biological value and net protein utilization. These correlations followed the content of essential amino acids, lysine, threonine and methionine in the faba bean proteins. The content of the sulfur-containing ammo acids cysteine and methionine are especially dominant factors for protein quality. A simple method for total sulphur determination was found nut 10 be a sufficiently reliable technique for evaluating the content of methionine and cysteine in the faba beans. Tannin, insoluble- and total dietary fibre are phenolic aromatic compounds which were negatively correlated with the faba bean quality as expressed by digestible energy, the protein digestibility and the biological value of the faba beans. The results obtained have also revealed, that it is not sufficient to consider tannin as a group in relation to the faba bean quality. We need to separate and evaluate the different types, of phenolics in relation to the variations in quality of faba beans. Some of the low molecular weight (LMW) phenolics in the faba beans seem 10 be involved in inhibitory effects on hydrolase enzymes, chymotrypsin and trypsin. Trypsin and chymotrypsin from different animals were different in their sensitivity to faba bean inhibitors and additional experiments are required to reveal details about these effects. It has also been revealed that fructosans Lind LMW carbohydrates (oligosaccharides) are important in relation to the quality of faba beans.     

Genetic Effects on Host × Strain Interaction in the Symbiosis of Vicia faba and Rhizobium leguminosarum

50 Vicia faba lines were inoculated with 4 strains of Rhizobium leguminosarum in all possible combinations under greenhouse conditions in 5 replications. A root-sterile method was used for the cultures. Characters such as plant height, leaf colour, number and colour of nodules, the dry weights of shoot, root and nodules, and the nitrogen content were recorded, after 10 weeks. It was demonstrated by analysis of variance that, for example, for the dry matter of the shoot only 1.9 % of the total phenotypic variance was caused by interaction between Vicia and Rhizobium genotypes, but 24.7 % Was conditioned by the faba bean genotypes and 73.4 % by the Rhizobium strains. Differences of Vicia faba genotypes in their reaction to different Rhizobium strains were detected by joint regression analysis.     

Introgression of resistance to late blight (Phytophthora infestans) from Solanum pinnatisectum into S. tuberosum using embryo rescue and double pollination

In order to investigate the agricultural potential of the genus Vicia, and identify traits associated with productivity and responsiveness to environment, 34 undomesticated Mediterranean accessions representing Section Narbonensis (V. johannis, V. narbonensis) and V. sativa were grown in five contrasting environments in northern Syria (growing season rainfall: 76–290 mm).Highly significant genotype × environment interactions were observed for all traits. For most of the components of yield, accession mean performance (productivity)was highly correlated with responsiveness across environments (r = 0.59–0.96), as defined by joint linear regressions. Thus high yielding genotypes tended to be relatively more productive than low yielding genotypes under conditions that favoured high yields. Regression analysis revealed that mean site yields were positively correlated to rainfall (r = 0.85) and its attendant effect on growing season length as measured by cumulative season temperature and phenology (r = 0.59–0.81).In order to examine yield related traits independently of taxonomy, genotypes were grouped into three categories using K-means clustering based on productivity and responsiveness of seed, hay and biological yield. Highly productive/responsive genotypes were tall with high harvest index, large seeds and low fecundity (seeds and pods per plant), whereas unproductive/unresponsive plants tended to be short, highly fecund, with small seeds and low harvest index. Principal components analysis showed that responsiveness, in terms of seed, hay and biological yields, was closely related to phenological plasticity. Thus highly productive/responsive genotypes were able to start flowering earlier than unproductive/unresponsive genotypes in early environments, but significantly later in late, higher rainfall environments. Plant growth habit was also related to yield responsiveness. In environments with little biomass production the proportion of erect plants was high in all three categories. In more favourable, high biomass environments, the proportion of erect plants in unproductive/unresponsive genotypes fell dramatically, but was unchanged among productive/responsive genotypes. We suggest that for unproductive/unresponsive genotypes competition for light is increased under optimal growth conditions. We argue that the optimal combination of fixed and responsive traits in high yielding genotypes results in a `compound interest-type' response to more favourable environments. Highly productive and responsive genotypes can capture resources more effectively than their low yielding counterparts, leading to a positive relationship between performance and responsiveness for most components of yield. Differences in productivity and responsiveness for seed, hay and biological yield reflected Vicia taxonomy, increasing in the following order from low to high: V. johannis, V. sativa, the small seeded V. narbonensis (salmonea, jordanica, affinis) V. n. var. narbonensis, and finally V. n. var.aegyptiaca. V. n. var. aegyptiaca showed the most agricultural potential, since the taxon contained all the properties of productive/responsive genotypes listed above, yielding >1 t/ha under extremely arid conditions (104 mm),and >2.5 t/ha on 290 mm rainfall, confirming its potential for dry environments. This revised version was published online in August 2006 with corrections to the Cover Date.     

Physiological and biochemical responses of two faba bean (Vicia faba L.) varieties grown in vitro to salt stress

Journal of Crop Science and Biotechnology - Faba bean (Vicia faba L.) is one of the most important legume crops worldwide. High salinity is a major constraint for faba bean productivity in many...     

Development and validation of a robust KASP marker for zt2 locus in faba bean (Vicia faba)

Faba bean has the potential to become a key food and feed protein crop in many areas of the world. The presence of tannins in its seed coat has limited the deployment of this crop as feed and food. The expression of either of the two recessive genes, zt1 and zt2, causes a great reduction of tannins from the seed coat and results in a white flower phenotype. Molecular markers linked to these loci are fundamental tools for speeding up the breeding of low-tannin varieties. The main aim of this study was to develop and validate a robust molecular marker linked to the zt2 locus. We used 176 recombinant inbred lines of the Disco/2 × ILB 938/2 cross at F₆ and genotyped those using 257 SNP (single nucleotide polymorphism) markers. An SNP marker associated with zt2 locus was found on faba bean chromosome 3 and was used to develop a high-throughput low-cost KASP (kompetitive allele-specific PCR) marker. The KASP marker can successfully discriminate low-tannin faba beans carrying zt2 from those carrying zt1 and wild-type alleles.     

Yield Stability in Faba Bean,Vicia faba L. 1. Variation Among Inbred Lines*

Cultivation of faba beans (Vicia faba L.) is hampered by poor yield stability. The genetic variation at the homozygous level for yield stability in the gene pool of the small-seeded and indeterminate European faba beans and the usefulness of auxiliary traits for the improvement of yield stability were investigated. The concept of stability, based on the regression technique, was applied. A sample of 36 faba bean lines was tested in 16 environments and a subsample of eight lines was tested in 11 additional environments. Significant differences were found between lines for yield stability parameters, but the repeatability of the results was limited. Early maturity correlated markedly (r = 0.51^**) with one of the yield stability parameters (deviation from regression). Although lodging resistance was not correlated with the stability parameters, it proved to be a safety factor for performance.     

Effects of Row Spacing, Support Plant Species and Support Plant Mixture Ratio on Seed Yield and Yield Characteristics of Hungarian Vetch (Vicia pannonica Crantz)

The effects of row spacing (17.5 or 35.0 cm), support plant species (barley or triticale) and the proportion of crops in mixtures (no support plant or support plant 20, 40 or 60 %, respectively) on the seed yield and yield characteristics of Hungarian vetch (Vicia pannonica Crantz) were investigated. Increasing the row spacing increased the seed yield of V. pannonica from 881.0 to 1248.0 kg ha^?1. On average, in a pure stand the seed yield of V. pannonica was 1141.0 kg ha^?1. In mixtures with barley and triticale, the seed yield of V. pannonica averaged 986.0 and 1143.0 kg ha^?1, respectively. In single mixed stands the seed yield of V. pannonica varied between 551.0 kg ha^?1 (60 % support plant barley) and 1603.0 kg ha^?1 (20 % support plant triticale). The yield advantage of V. pannonica in this triticale mixture was 40 % compared to the V. pannonica pure stand. With respect to the total yield in the mixture with 20 % triticale (1902.0 kg ha^?1) the yield advantage over the V. pannonica pure stand was as high as 65.1 %. In the mixed stands the number of seeds per pod and the thousand‐seed weight of V. pannonica were higher than in V. pannonica pure stands.