Developmental changes in plant resistance to water flow in Pisum sativum (L.)

Soil and plant resistance to water flow under field conditions in pea (Pisum sativum L.) plants were measured at six ages. Transpiration flux, leaf and soil water potentials were used to calculate the total resistance to water flow using the Ohm's law analogy. Plant resistance was estimated from the slope of the water potential difference () vs. transpiration (Q) relationship. Plant growth, root density and soil water content distribution were measured. Leaf area and root length both increased until the end of seed filling and decreased during seed maturation. Total resistance decreased with the transpiration flux in a non-linear relationship. Plant resistance estimated as the slope of the vs. Q regression line increased until pod filling and then decreased. The increased resistance to water flow during pod filling was associated with a 10% increase in cell wall thickness.     

Nitrogen contributions from faba bean (Vicia faba L.) reliant on soil rhizobia or inoculation

Background and Aims

Understanding the impact of soil rhizobial populations and inoculant rhizobia in supplying sufficient nodulation is crucial to optimising N₂ fixation by legume crops. This study explored the impact of different rates of inoculant rhizobia and contrasting soil rhizobia on nodulation and N₂ fixation in faba bean (Vicia faba L.).

Methods

Faba beans were inoculated with one of seven rates of rhizobial inoculation, from no inoculant to 100 times the normal rate of inoculation, sown at two field sites, with or without soil rhizobia present, and their nodulation and N₂ fixation assessed.

Results

At the site without soil rhizobia, inoculation increased nodule number and increased N₂ fixation from 21 to 129 kg shoot N ha^?1, while N₂ fixation increased from 132 to 218 kg shoot N ha^?1 at the site with high background soil rhizobia. At the site without soil rhizobia, inoculation increased concentrations of shoot N from 14 to 24 mg g^?1, grain N from 32 to 45 mg g^?1, and grain yields by 1.0 Mg (metric tonne) ha^?1. Differences in nodulation influenced the contributions of fixed N to the system, which varied from the net removal of 20 kg N ha^?1 from the system in the absence of rhizobia, to a net maximum input of 199 kg N ha^?1 from legume shoot and root residues, after accounting for removal of N in grain harvest.

Conclusions

The impact of inoculation and soil rhizobia strongly influenced grain yield, grain N concentration and the potential contributions of legume cropping to soil N fertility. In soil with resident rhizobia, N₂ fixation was improved only with the highest inoculation rate.     

Genomic regions associated with chocolate spot (Botrytis fabae Sard.) resistance in faba bean (Vicia faba L.)

Molecular Breeding - Maize (Zea mays L.) is the most important food crop in the world, with significant acreage and production across the globe. However, it is affected by low temperatures...     

Root aquaporins contribute to whole plant water fluxes under drought stress in rice (Oryza sativa L.)

Aquaporin activity and root anatomy may affect root hydraulic properties under drought stress. To better understand the function of aquaporins in rice root water fluxes under drought, we studied the root hydraulic conductivity (Lpr) and root sap exudation rate (Sr) in the presence or absence of an aquaporin inhibitor (azide) under well‐watered conditions and following drought stress in six diverse rice varieties. Varieties varied in Lpr and Sr under both conditions. The contribution of aquaporins to Lpr was generally high (up to 79% under well‐watered conditions and 85% under drought stress) and differentially regulated under drought. Aquaporin contribution to Sr increased in most varieties after drought, suggesting a crucial role for aquaporins in osmotic water fluxes during drought and recovery. Furthermore, root plasma membrane aquaporin (PIP) expression and root anatomical properties were correlated with hydraulic traits. Three chromosome regions highly correlated with hydraulic traits of the OryzaSNP panel were identified, but did not co‐locate with known aquaporins. These results therefore highlight the importance of aquaporins in the rice root radial water pathway, but emphasize the complex range of additional mechanisms related to root water fluxes and drought response.     

Quantitative genetic analysis of embryo heterosis in faba bean (Vicia faba L.)

Seeds, i.e. embryos, may be genetically different from either of their parents and moreover may express their own heterosis. The objective was to genetically analyse embryo heterosis for their own weight (i.e. seed weight) in comparison with their seedlings’ heterosis, taking the large-seeded crop (Vicia faba L.) as model. A specific diallel mating scheme was used, based on four parental lines, creating 76 seed genotypes in generations P, F₁, F₂ and BC. Mature seed weight was assessed for these embryo genotypes in 3 years at one German location, and young plant biomass yield of seedlings emerging from these seeds in two greenhouse experiments. The quantitative genetic analysis showed an average of 10.6% mid-parent heterosis for mature seed weight and 14.5% mid-parent heterosis for juvenile biomass. In both traits, the embryos contributed markedly and significantly via their own genes to the genetic variation. For mature embryo weight heterosis, apparently the parental difference in seed weight was decisive, whereas for juvenile biomass heterosis, genetic unrelatedness of parents had priority.     

Relationships between plant and soil water status in vine (Vitis vinifera L.)

Soil water status and its effect on plant water status are commonly evaluated for water stress diagnosis in annual crops. We investigated the application of this method to vineyards, using the fraction of transpirable soil water (FTSW) to characterise the soil water deficit experienced by the plant. The stability of the relationship between FTSW and predawn leaf water potential (Ψp) was analysed over two years (2000–2001), in two contrasted soils in vineyards in south eastern France, both planted with the cultivar Syrah, but grafted on different rootstocks (SO4 and 140Ru). FTSW was determined from soil moisture measurements performed with a neutron probe down to 2.5 m, under the rows and between the rows (3 replicates in each case). Vertical and horizontal variations in soil water content were analysed and the upper and lower limits of total vine’s transpirable soil water (TTSW) were calculated for each soil. The lower limit was also compared with the value of soil moisture content determined at −1.5 MPa in the laboratory. FTSW could be calculated for the soil depth analysed, without distinguishing horizontal position (row or inter-row). The lower limit of TTSW for vine was higher than the soil water content at −1.5 MPa, except in the upper horizons (0–0.2 m) which are prone to soil evaporation. A single relationship between Ψp and FTSW was obtained for the two vineyards and for the two years of measurement. This relationship was similar to that established by Lebon et al. (2003) on Gewürztraminer/SO4 in a vineyard in northern France. FTSW can therefore be used as an indicator of the water deficit experienced in vineyards, provided that TTSW is correctly estimated.     

Validation of QTLs for Orobanche crenata resistance in faba bean (Vicia faba L.) across environments and generations

Broomrape (Orobanche crenata Forsk.) is a major root–parasite of faba bean (Vicia faba L.), that seriously limits crop cultivation in the whole Mediterranean area. This parasitic weed is difficult to control, difficult to evaluate and the resistance identified so far is of polygenic nature. This study was conducted to identify genetic regions associated with broomrape resistance in recombinant inbred lines (RILs) and to validate their previous location in the original F₂ population derived from the cross between lines Vf6 and Vf136. A progeny consisting of 165 F₆ RILs was evaluated in three environments across two locations in 2003 and 2004. Two hundred seventy seven molecular markers were assigned to 21 linkage groups (9 of them assigned to specific chromosomes) that covered 2,856.7 cM of the V. faba genome. The composite interval mapping on the F₆ map detected more quantitative trait loci (QTL) than in the F₂ analysis. In this sense, four QTLs controlling O. crenata resistance (Oc2–Oc5) were identified in the RI segregant population in three different environments. Only Oc1, previously reported in the F₂ population, was not significant in the advanced lines. Oc2 and Oc3 were found to be associated with O. crenata resistance in at least two of the three environments, while the remaining two, Oc4 and Oc5, were only detected in Córdoba-04 and Mengíbar-04 and seemed to be environment dependent.     

Identification and characterization of NBS-LRR class resistance gene analogs in faba bean (Vicia faba L.) and chickpea (Cicer arietinum L.).

A PCR approach with degenerate primers designed from conserved NBS-LRR (nucleotide binding site-leucine-rich repeat) regions of known disease-resistance (R) genes was used to amplify and clone homologous sequences from 5 faba bean (Vicia faba) lines and 2 chickpea (Cicer arietinum) accessions. Sixty-nine sequenced clones showed homologies to various R genes deposited in the GenBank database. The presence of internal kinase-2 and kinase-3a motifs in all the sequences isolated confirm that these clones correspond to NBS-containing genes. Using an amino-acid sequence identity of 70% as a threshold value, the clones were grouped into 10 classes of resistance-gene analogs (RGA01 to RGA10). The number of clones per class varied from 1 to 30. RGA classes 1, 6, 8, and 9 were comprised solely of clones isolated from faba bean, whereas classes 2, 3, 4, 5, and 7 included only chickpea clones. RGA10, showing a within-class identity of 99%, was the only class consisting of both faba bean and chickpea clones. A phylogenetic tree, based on the deduced amino-acid sequences of 12 representative clones from the 10 RGA classes and the NBS domains of 6 known R genes (I2 and Prf from tomato, RPP13 from Arabidopsis, Gro1-4 from potato, N from tobacco, L6 from flax), clearly indicated the separation between TIR (Toll/interleukin-1 receptor homology: Gro1-4, L6, N, RGA05 to RGA10)- and non-TIR (I2, Prf, RPP13, RGA01 to RGA04)-type NBS-LRR sequences. The development of suitable polymorphic markers based on cloned RGA sequences to be used in genetic mapping will facilitate the assessment of their potential linkage relationships with disease-resistance genes in faba bean and chickpea. This work is the first to report on faba bean RGAs.     

Mapping of quantitative trait loci controlling broomrape (Orobanche crenata Forsk.) resistance in faba bean (Vicia faba L.).

Orobanche crenata Forsk. is a root parasite that produces devastating effects on many crop legumes and has become a limiting factor for faba bean production in the Mediterranean region. The efficacy of available control methods is minimal and breeding for broomrape resistance remains the most promising method of control. Resistance seems to be scarce and complex in nature, being a quantitative characteristic difficult to manage in breeding programmes. To identify and map the QTLs (quantitative trait loci) controlling the trait, 196 F2 plants derived from the cross between a susceptible and a resistant parent were analysed using isozymes, RAPD, seed protein genes, and microsatellites. F2-derived F3 lines were studied for broomrape resistance under field conditions. Of the 130 marker loci segregating in the F2 population, 121 could be mapped into 16 linkage groups. Simple interval mapping (SIM) and composite interval mapping (CIM) were performed using QTL Cartographer. Composite interval mapping using the maximum number of markers as cofactors was clearly the most efficient way to locate putative QTLs. Three QTLs for broomrape resistance were detected. One of the three QTLs explained more than 35% of the phenotypic variance, whereas the others accounted for 11.2 and 25.5%, respectively. This result suggests that broomrape resistance in faba bean can be considered a polygenic trait with major effects of a few single genes.     

CAPs markers to assist selection for low vicine and convicine contents in faba bean (Vicia faba L.)

The antinutritional factors (ANFs) present in Vicia spp. seeds are a major constraint to the wider utilization of these crops as grain legumes. In the case of faba bean (Vicia faba L.), a breeding priority is the absence vicine and convicine (v-c); responsible for favism in humans and for the reduced animal performance or low egg production in laying hens. The discovery of a spontaneous mutant allele named vc-, which induces a 10–20 fold reduction of v-c contents, may facilitate the process. However, the high cost and difficulty of the chemical detection of v-c seriously restricts the advances in breeding-selection. To identify random amplified polymorphic DNA (RAPD) markers linked to this gene, we have analysed an F₂ population derived from a cross between a line with high v-c content (Vf6) and the vc- genotype (line 1268). Quantification of v-c was done by spectrophotometry on the parents and the F₂ population (n = 136). By using bulked segregant analysis (BSA), two RAPD markers linked in coupling and repulsion phase to the allele vc- were identified and further converted into sequence characterized amplified regions (SCARs). Amplification of SCARS was more consistent, although the initial polymorphism between pools was lost. To recover the polymorphisms several approaches were explored. Restriction digestion with HhaI (for SCAR SCH01₆₂₀) and RsaI (for SCAR SCAB12₈₅₀) revealed clear differences between the parental lines. The simultaneous use of the two cleavage amplified polymorphism (CAP) markers will allow the correct fingerprinting of faba bean plants and can be efficiently used in breeding selection to track the introgression of the vc- allele to develop cultivars with low v-c content and improved nutritional value.Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

A comparison of minirhizotron,core and monolith methods for quantifying barley (Hordeum vulgare L.) and fababean (Vicia faba L.) root distribution

Root research has been hampered by a lack of good methods and by the amount of time involved in making measurements. The use of the minirhizotron as a quantitative tool requires comparison with conventional destructive methods. This study was conducted in the greenhouse to compare the minirhizotron technique with core and monolith methods in quantifying barley (Hordeum vulgare L.) and fababean (Vicia faba L.) root distribution. Plants were grown in boxes (80 cm long × 80 cm wide × 75 cm deep) in a hexagonal arrangement to minimize the effects of rooting anistrophy. Minirhizotron observations and destructive sampling to a depth of 70 cm using core and monolith methods were performed at the ripening growth stage. Total root length for the entire depth interval was generally higher in barley (159–309 m) than fababean (110–226 m). Significant correlation coefficients between monolith and core methods for root length density (RLD, cm cm^–3) was observed in both crops (p 0.01). A method and depth interaction showed no significant differences in fababean RLD distribution measured by core and monolith methods. However, the RLD was different for the uppermost 40 cm depth in barley. The relationship for RLD between minirhizotron and core methods was significant only in barley (r=0.77^*). For both crops, estimates of RLD in the top 10-cm layer by the minirhizotron technique were lower than those by core and monolith techniques. In contrast, estimates of RLD were higher in fababean at a depth >30 cm. Destructive sampling still remains the method to quantify root growth in the 0–10 cm soil layer. ei]B E Clothier     

Purification and characterization of trypsin inhibitor from seeds of faba bean (Vicia faba L.)

A trypsin inhibitor from seeds of faba bean (Vicia faba L.) was purified to near homogeneity as judged by native-PAGE with about 11 % recovery using ammonium sulphate fractionation, ion-exchange chromatography on DEAE-cellulose and gel filtration through Sephadex G-100. The inhibitor had a molecular weight of 18 kD as determined by SDS-PAGE and Sephadex G-100. The inhibitor inhibited trypsin and chymotrypsin to the extent of 48 and 12 %, respectively. The inhibtion was of non-competitive type with dissociation constant for the enzyme inhibitor complex in the region of 0.07 mg·ml⁻¹. The inhibtor was stable between pH 4 and 5. It completely lost its activity when heated at 125 °C for 1 h or at 100 °C for 2 h. The inhibitor also lost its activity on exposure to 2-mercaptoethanol. Based on these properties, it could be concluded that Vicia faba trypsin inhibitor belongs to Bowman-Birk type of inhibitors, as it has molecular weight lower than generally observed for Kunitz type inhibitors.     

Studies on pollination and fertilization in the field bean (Vicia faba L.)

The percentage of cross-fertilization in the field bean was found to be around 45%, and to be higher on the lower nodes of the plants than on the upper. No differences in the relative pollen tube growth rates of self and foreign pollen were observed. The different lines and genotypes investigated responded differently to open pollination, selfing with manual tripping, and selfing without tripping. Fertilizer application and date of sowing affected the number of flowers produced and the number of seeds set, but did not affect the ratio of seeds to flowers, i.e. the index of self fertility.     

Hydraulic architecture and water flow in growing grass tillers (Festuca arundinacea Schreb.)

The water relations and hydraulic architecture of growing grass tillers (Festuca arundinacea Schreb.) are reported. Evaporative flux density, E (mmol s^?1 m^?2), of individual leaf blades was measured gravimetrically by covering or excision of entire leaf blades. Values of E were similar for mature and elongating leaf blades, averaging 2·4 mmol s^?1 m^?2. Measured axial hydraulic conductivity, K_h (mmol s^?1 mm MPa^?1), of excised leaf segments was three times lower than theoretical hydraulic conductivity (K_t) calculated using the Poiseuille equation and measurements of vessel number and diameter. K_t was corrected (K_t*) to account for the discrepancy between K_h and K_t and for immature xylem in the basal expanding region of elongating leaves. From base to tip of mature leaves the pattern of K_t* was bell‐shaped with a maximum near the sheath–blade joint (≈ 19 mmol s^?1 mm MPa^?1). In elongating leaves, immature xylem in the basal growing region led to a much lower K_t*. As the first metaxylem matured, K_t* increased by 10‐fold. The hydraulic conductances of the whole root system, (mmol s^?1 MPa^?1) and leaf blades, (mmol s^?1 MPa^?1) were measured by a vacuum induced water flow technique. and were linearly related to the leaf area downstream. Approximately 65% of the resistance to water flow within the plant resided in the leaf blade. An electric‐analogue computer model was used to calculate the leaf blade area‐specific radial hydraulic conductivity, (mmol s^?1 m^?2 MPa^?1), using , K_t* and water flux values. values decreased with leaf age, from 21·2 mmol s^?1 m^?2 MPa^?1 in rapidly elongating leaf to 7·2 mmol s^?1 m^?2 MPa^?1 in mature leaf. Comparison of and values showed that ≈ 90% of the resistance to water flow within the blades resided in the liquid extra‐vascular path. The same algorithm was then used to compute the xylem and extravascular water potential drop along the liquid water path in the plant under steady state conditions. Predicted and measured water potentials matched well. The hydraulic design of the mature leaf resulted in low and quite constant xylem water potential gradient (≈ 0·3 MPa m^?1) throughout the plant. Much of the water potential drop within mature leaves occurred within a tenth of millimetre in the blade, between the xylem vessels and the site of water evaporation within the mesophyll. In elongating leaves, the low K_t* in the basal growth zone dramatically increased the local xylem water potential gradient (≈ 2·0 MPa m^?1) there. In the leaf elongation zone the growth‐induced water potential difference was ≈ 0·2 MPa.     

Phylogenetic multilocus sequence analysis of native rhizobia nodulating faba bean (Vicia faba L.) in Egypt

The taxonomic diversity of forty-two Rhizobium strains, isolated from nodules of faba bean grown in Egypt, was studied using 16S rRNA sequencing, multilocus sequence analyses (MLSA) of three chromosomal housekeeping loci and one nodulation gene (nodA). Based on the 16S rRNA gene sequences, most of the strains were related to Rhizobium leguminosarum, Rhizobium etli, and Rhizobium radiobacter (syn. Agrobacterium tumefaciens). A maximum likelihood (ML) tree built from the concatenated sequences of housekeeping proteins encoded by glnA, gyrB and recA, revealed the existence of three distinct genospecies (I, II and III) affiliated to the defined species within the genus Rhizobium/Agrobacterium. Seventeen strains in genospecies I could be classified as R. leguminosarum sv. viciae. Whereas, a single strain of genospecies II was linked to R. etli. Interestingly, twenty-four strains of genospecies III were identified as A. tumefaciens. Strains of R. etli and A. tumefaciens have been shown to harbor the nodA gene and formed effective symbioses with faba bean plants in Leonard jar assemblies. In the nodA tree, strains belonging to the putative genospecies were closely related to each other and were clustered tightly to R. leguminosarum sv. viciae, supporting the hypothesis that symbiotic and core genome of the species have different evolutionary histories and indicative of horizontal gene transfer among these rhizobia.     

Estimates of broad-sense heritability for seed yield and yield criteria in faba bean (Vicia faba L.)

Eight faba bean (Vicia faba L.) genotypes were grown at lowlands of the west-Mediterranean region of Turkey in order to estimate the broad-sense heritability for plant height, number of stems and pods per plant, seed yield, biological yield, 100-seed weight, days to flowering and maturity. The heritability for plant height, number of stems and pods per plant, seed yield, biological yield, 100-seed weight, days to flowering and maturity were estimated as 83%, 63%, 43%, 62%, 52%, 99%, 97% and 97%, respectively. It was found that seed weight was the least affected trait across changing environmental conditions and followed by days to flowering and maturity. On the other hand, number of pods per plant, biological and seed yields and number of stems per plant were the most affected traits versus environmental conditions.     

Nutritional and antinutritional attributes of faba bean (Vicia faba L.) germplasms growing in Bihar,India

Eleven germplasms of faba bean seeds from four agroclimatic regions of Bihar, India, have been investigated to estimate their nutritional (soluble protein, free amino acids, starch, reducing and non reducing sugar, total soluble sugar) and antinutritional (total extractable phenol and condensed tannin/proanthocyanidin) parameters. These parameters were found in varying concentration in all genotypes studied. The highest concentration of total extractable phenol and proanthocyanidin (condensed tannin) (2.56 and 1.59 % leucocyanidin equivalents respectively on dry matter basis) were found in Samastipur while the lowest from Patna (0.95 and 0.426 % leucocyanidin equivalent on dry matter basis). The different nutritional parameters were also found to be in variable concentration among different germplasms viz. total soluble protein ≈ 20–32 %, free amino acids ≈ 188–348 mg/100 g, starch ≈ 27–33 %, reducing sugars ≈ 85–188 mg/100 g, non reducing sugars ≈ 0.7–1.7 % and total soluble sugars ≈ 0.8–1.9 %.