Development of pea bacterial blight caused by Pseudomonas syringae pv. pisi in winter and spring cultivars of combining peas (Pisum sativum) with different sowing dates

Pea bacterial blight occurred by natural infection in a field trial on peas in 1995. Disease development in the winter cultivars Rafale, Frilene and Froidure was compared with that in the spring cultivars Baccara, Conquest and Bohatyr, each sown on six dates in October, November, December, mid-March, late March and April. Disease incidence had reached 100% plants affected in all treatments by mid-July. Disease severity was greater in winter-sown (October, November or December) than in spring-sown peas of each cultivar at each assessment. Significant (P < 0.05) differences in disease severity occurred between cultivars in the winter-sown plots in May and June and the spring cultivars were affected more severely than the winter cultivars. Comparison of areas under the disease progress curves for both disease incidence and severity also showed that the winter-sown peas were more affected by disease than spring-sown peas and that spring cultivars were more severely affected than winter cultivars. Yield was strongly correlated with disease severity. A linear regression model suggested that, for peas sown in October, November or December, a yield loss of 0.5 tha^-1 occurred for each 10% increase in canopy area affected by pea bacterial blight.     

Optimum plant densities for three semi-leafless combining pea (Pisum sativum) cultivars under contrasting field conditions

Yield and yield components of three semi-leafless pea (Pisum sativum) cultivars, of contrasting seed type/growth habit, were assessed at target planting densities of 40–140 plants/m² on nine sites over three years. Flat-topped parabolic/asymptotic yield/density relationships were obtained. The plant density required to maximise (p max) and optimise (p opt) yield differed between cultivars: Helka, small blue, p max 126 plants/m², p opt 101 plants/m²; Solara, large blue, p max 124 plants/m², p opt 94 plants/m²; and Countess, white-seeded, p max 104 plants/m², p opt 71 plants/m². Near-maximum yields were maintained between 70 and 140 plants/m² due to the ability of the pea crop to make compensatory increases in the number of pods per plant as density declined. Yield/density responses were influenced by site (e.g. soil type) more than by seasonal factors. The risk of yield reductions occurring at densities below 70 plants/m² was greater on a mineral soil than on a fertile organic soil. On the basis of agronomic and economic considerations, there was no evidence that target plant densities required to optimise yield should necessarily be higher for semi-leafless cultivars studied than for conventional leafed peas.     

Transmission of pea bacterial blight (Pseudomonas syringae pv. pisi) from seed to seedling: effects of inoculum dose, inoculation method, temperature and soil moisture

Quantitative analyses of the utilization of amino acids by Lactococcus lactis subsp. cremoris FD1 in yeast extract medium (YE) and in casein peptone medium (CP) have been performed. Both free and peptide-bound amino acids were measured. In the CP most amino acids are peptide-bound and some amino acids are virtually only present in peptides. Thirty-six per cent of all peptide bonds in CP are hydrolysed during fermentation (6·3 mmol peptide bonds per gram biomass formed) and there is a transition of the growth rate related ATP consumption Y _xATP (mmol ATP g biomass^-1) from 25 mmol g^-1 to 71 mmol g^-1 coincident with a decrease of the peptide consumption. In YE most of the amino acids are on the free form and only 26% of the peptide bonds are hydrolysed during fermentation (1·5 mmol peptide bonds per gram biomass formed). A constant Y _xATP= 38 mmol g^-1 prevails throughout the fermentation in YE.     

Association of dominant loci for resistance to Pseudomonas syringae pv. pisi with linkage groups II, VI and VII of Pisum sativum

Morphological characters, isoenzymes and recombinant inbred lines were employed to assign four loci for resistance to Pseudomonas syringae pv pisi to genetic linkage groups in Pisum sativum. A total of five morphological markers and 11 isoenzyme loci were screened in two independent F₂ P. sativum populations: Vinco × Hurst’s Greenshaft (V×HGS) and Partridge × Early Onward (P×EO). Mapping was also carried out in two recombinant inbred populations, unrelated to the F₂ populations. Previously reported linkage between resistance genes Ppi3 and Ppi4 was confirmed. Linkage was also detected between resistance gene Ppi2 and the isoenzyme locus Aldo (linkage group VII). The linked loci Ppi3 and Ppi4 were associated with a (linkage group II). A further resistance gene Ppi1 was associated with linkage group VI close to the hilum colour gene P1. RAPD markers tested in the cross P×EO were not well targeted; however, one marker, OPA-20_0.71, showed linkage to Ppi3. Received: 3 July 2000 / Accepted: 27 October 2000     

Effect of Heat-killed Bacteria on the Interaction of Pea with Pseudomonas syringae pv. pisi

The effect of simultaneous treatment with heat-killed and live bacteria on the responses of two pea cultivars, Early Onward and Hurst Green Shaft, to inoculation with two races, 1 and 2, of Pseudomonas syringae pv. pisi was investigated. Simultaneous application induced resistance in pea to the bacterial pathogen. The level of resistance response elicited in the host increased with increasing number of heat-killed cells in the inoculum. Heat-killed cells of neither race elicited the hypersensitive reaction. No symptoms were induced in plants of either cultivar from treatment of control plants with sterile distilled water or from treatment with heat-killed bacterial cells only. Simultaneous treatment with heat-killed and inoculation with live bacteria did not have any apparent effect on the trend of bacterial multiplication in vivo.     

Effect of ascochyta blight (Mycosphaerella pinodes) on yield components of single pea (Pisum sativum) plants under field conditions

Field studies were made in 1992 and 1993 to examine the yield components of pea inoculated with Mycosphaerella pinodes and those of healthy pea (sprayed with a mixture of flutriafol + chlorothalonil), in a split-plot design with the cv. Solara sown at different plant densities. Ascochyta blight was severe on leaves and on internodes of the basal part of the plants; pods had few lesions. The number and length of stems per plant were the same for diseased and healthy plants. The number of reproductive nodes and pods per stem were affected by disease only in 1993. In 1992 and 1993 respectively, disease caused reductions in the number of seeds per stem of 18% and 25%, and in seed size of 13.5% and 16.7%, compared with healthy plants. The harvest index and total biomass were lower in diseased than in healthy plants and seed yield was reduced by 40% in diseased plots. These results show a high relationship between the disease parameters (disease mean on stipules/nodes 8–18/ and on internodes/nodes 5–15/, percentages of stipules or internodes with a disease score 4, and percentage of stems encircled by lesions), plant density and yield reduction.     

Mating disruption of pea moth (Cydia nigricana) in organic peas (Pisum sativum)

We appraised mating disruption (MD) to control pea moth, Cydia nigricana (Fabricius) (Lepidoptera: Tortricidae), by assessing male attraction to monitor traps, larval pod infestation, and larval age structure in pheromone‐treated and untreated grain pea fields [Pisum sativum L. (Fabaceae)], over a 5‐year period. Cellulose pheromone dispensers were manually attached to the top shoots of pea plants and released 540 mg ha^?1 day^?1 synthetic pheromone E8,E10‐dodecadien‐1‐yl acetate in a first test series (2000–2001) and ca. 4 200 mg pheromone ha^?1 day^?1 in a second series (2004–2006). The dispensers had a half‐life of about 30 days. Although male attraction to pheromone monitoring traps was largely suppressed at the edges and within MD fields in both test series, MD treatments did not reduce pod infestation in the open field in 2000 and 2001. In the 2004–2006 series, larval damage reduction was achieved in the majority of the trials but overall MD efficacy in the open field was only 61% and not significant. In contrast, in field cages placed within the experimental sites and supplied with unmated pea moths, MD control was consistently high and significant. There were no obvious differences in the larval age distribution in all MD and control treatments, suggesting that infestations started and developed further similarly. As a univoltine species, C. nigricana larvae stay in the soil of pea fields for hibernation and pupate. The following year, emerging adults disperse and fly to the closest pea crop. Combined emergence site and pea crop treatments were conducted over 2 years to include this early migration phase of C. nigricana adults. However, the emergence site treatments did not enhance MD‐control efficacy. We conclude that mating activity was only prevented in cage tests, whereas substantial mating occurred during the transit phase outside the pheromone‐treated fields either within non‐crop vegetation and/or at the edges of pheromone‐treated pea fields orientated upwind. Thus, resulting gravid female entry can be regarded as the major constraint to reliable MD control.     

Effects of short-term waterlogging on the growth and yield of peas (Pisum sativum)

The effects of waterlogging the soil for up to 5 days on the growth of peas at different stages of development were investigated in glasshouse and outdoor experiments. During waterlogging the oxygen content of the soil declined to less than 2% within 2–3 days. Subsequently, leaf senescence was hastened, stem growth rate was slowed and yield was decreased. The effects on yield were mainly attributable to the development of fewer nodes and fewer pods. Waterlogging for 24 h or more just before flowering restricted growth and yield. The effects of waterlogging at earlier and later stages were less marked. Waterlogging at any period after flower buds were visible prevented further increase in root dry weight.     

Inheritance of seed yield and quality traits in peas (Pisum sativum L.)

Summary A half diallel analysis involving nine cultivars showed that additive as well as non-additive gene effects were important for the inheritance of seed yield per plant, 100-seed weights, protein content and potassium per cent. For remaining traits non-additive genetic components were important. Overdominance was observed for all traits except for 100-seed weight, which expressed partial dominance. Parents PMR-T10, EC21857, EC109182, T163 and EC109189 were good general combiners for seed yield, seed weight and quality traits. In general there was a good relationship between per se performance and the gca effects of the parents for all traits. Cross combinations such as LMR8 x EC109182,LMR8 x PMR-T10,LMR8 x EC21857,PMRT10 x EC21857 and P23 x EC21857 were found promising. The seed yield was positively correlated with other quality traits. Protein had a positive correlation with methionine and phosphorus. All the values of correlation co-efficients were non-significant except for yield with potassium, 100-seed weight and protein with methionine, indicating that yield and quality attributes can be improved simultaneously by simple selection procedures.     

Stimulation of nodulation in field peas (Pisum sativum) by low concentrations of ammonium in hydroponic culture

Although the inhibitory effects of high concentrations of mineral N (> 1.0 mM) on nodule development and function have often been studied, the effects of low, static concentrations of NH₄⁺ (< 1.0 mM) on nodulation are unknown. In the present experiments we examine the effects of static concentrations of NH₄⁺ at 0, 0.1 and 0.5 mM in flowing, hydroponic culture on nodule establishment and nitrogenase activity in field peas [Pisum sativum L. cv. Express (Svalöf AB)] for the initial 28 days after planting (DAP). Peas grown in the presence of low concentrations of NH₄⁺ had significantly greater nodule numbers (up to 4-fold) than plants grown without NH₄⁺. Nodule dry weight per plant was significantly higher at 14, 21 and 28 DAP in plants grown in the presence of NH₄⁺, but individual nodule mass was lower than in plants grown without NH₄⁺. The nodulation pattern of the plants supplied with NH₄⁺ was similar to that often reported for supernodulating mutants, however the plants did not express other growth habits associated with supernodulation. Estimates of N₂ fixation indicate that the plus-NH₄⁺ peas fixed as much or more N₂ than the plants supplied with minus-NH₄⁺ nutrient solution. There were no significant differences in nodule numbers, nodule mass or NH₄⁺ uptake between the plants grown at the two concentrations of NH₄⁺. Nodulation appeared to autoregulate by 14 DAP in the minus-NH₄⁺ treatment. Plant growth and N accumulation in the minus-NH₄⁺ plants lagged behind those of the plus-NH₄⁺ treatments prior to N₂ fixation becoming well established in the final week of the experiment. The plus-NH₄⁺ treatments appeared not to elicit autoregulation and plants continued to initiate nodules throughout the experiment.     

Comparative proteomic analysis of BTH and BABA-induced resistance in pea (Pisum sativum) toward infection with pea rust (Uromyces pisi)

Systemic acquired resistance (SAR) to Uromyces pisi in pea was studied by using a proteomic approach. Two-dimensional electrophoresis (2-DE) was used in order to compare the leaf proteome of two pea genotypes displaying different phenotypes (susceptible and partial resistance to the fungus), and in response to parasite infection under the effect of two inducers of SAR, BTH and BABA. Multivariate statistical analysis identified 126 differential protein spots under the experimental conditions (genotypes/treatments). All of these 126 protein spots were subjected to MALDI-TOF/TOF mass spectrometry to deduce their possible functions. A total of 50 proteins were identified using a combination of peptide mass fingerprinting (PMF) and MSMS fragmentation. Most of the identified proteins corresponded to enzymes belonging to photosynthesis, metabolism, biosynthesis, binding and defense response, whose behavior pattern was different in relation to susceptibility/resistance of the genotypes studied and to the BTH/BABA induction to pathogen response. Results obtained in this work suggested that plants could reduce their photosynthesis and other energy metabolism and enhance the production of defense-related proteins to cope the stress. On the other side, we postulated that resistance induced by the chemicals operates via different mechanisms: BABA inducer could act via phenolic biosynthesis pathway, whereas resistance provided by BTH inducer seems to be mediated by defense and stress-related proteins. The results are discussed in terms of response to rust under the effect of inducers.     

Response of tomato genotypes to bacterial speck (Pseudomonas syringae pv. tomato)

Two genotypes of tomato A 100 and Ontario 7710 which were inoculated separately with four strains of Pseudomonas syringae pv. tomato differed significantly in disease severity (susceptibility) to bacterial speck. At both concentrations of inoculum of each strain used (10⁷ and 10⁸ cfu/ml) A 100 appeared to be highly susceptible whereas Ontario 7710 showed very low or no susceptibility. The significant differences in virulence between strains and in response of tomato plants in three replicate experiments were found. Generally, concentration of inoculum 10⁷ cfu/ml was too low to induce consistent level of disease severity. The obtained results indicate the importance of consistent and favorable conditions for disease development in screening of tomato resistance to bacterial speck.     

Effects of pod infection by Mycosphaerella pinodes on yield components of pea (Pisum sativum)

Yield reduction of pea (Pisum sativum) due to various types of infections by Mycosphaerella pinodes on pods was assessed. A range of disease severities was created on pods of pea plants grown in the glasshouse, by painting the pods with different concentrations of spore suspensions, at three different pod development stages: lag phase, the beginning of seed filling (BSF) and mid-filling of the seeds. Seed number at harvest was reduced only if the pods were infected before BSF, as shown previously for whole plant infections. Pod infections led to individual seed weight (ISW) losses from zero (for late infections, at mid-filling) to 20% (for earlier infections and severe disease). Infection during the lag phase affected ISW by reducing seed growth rate, whereas infection at BSF tended to reduce the duration of seed filling. There was a linear relationship between the area under the disease progress curve and the percentage decrease in ISW. This model should be complemented by the effect of leaves and stem infections, in order to predict ISW losses in diseased crop conditions, in which epidemics occur on all aerial parts of the pea plant.     

The protective effect of desferrioxamine on paraquat-treated pea (Pisum sativum)

Paraquat, a widely used herbicide, is photoreduced by photosystem I to the monovalent cation radical, which in turn, can react quickly and efficiently with molecular oxygen to produce superoxide anion radicals. In the presence of redox-active iron (or copper) superoxide radicals can serve as a source for the more active species such as hydroxyl radicals. The present sludv investigated the possible mediatory role of iron in paraquat to xicity. The results demonstrate that desferrioxamme (0–150μM) a highiy specific iron chelator, reduces the loss of proteins (by 34–69%) and lipid peroxidation (by 31–96%) in paraquat treated leaf cuts. Dcsferrioxamine also protects malate dehydrogenase (61–70%) hydroxvpyruvate reductase (54–100%), and Ca²⁺-dependent ATPase (25–34%) against the paraquat-induced loss of their activity. It also induces an increase in glutathione reductase activity (by 188%). These results, together with those from other experiments concerning the effect of desferrioxamine on paraquat uptake by the leaf cuts, suggest that the protection by desferrioxamine arises from its specific iron chelanon properties, and lead to the conclusion that nan-protein-bound and redoxactive forms of iron pluy a role in the manifestation of paraquat toxicity in plants.     

Antigenic bacterial polysaccharides. 30. The structure of the polysaccharide chain of lipopolysaccharide of Pseudomonas syringae pv. syringae 281 (serogroup I)

Anomeric methyl 3-O-(D-mannopyranosyl- and L-rhamnopyranosyl)-beta-D-talopyranosides were synthesised by the stereoselective 1,2-cis- and 1,2-trans manno- and rhamnosylation of methyl 2,4,6-tri-O-acetyl-beta-D-talopyranoside, which has been prepared from methyl beta-D-galactopyranoside by a synthetic scheme including conversion of the C2 configuration. From 13C-NMR spectra of the disaccharides obtained the spectral alpha- and beta-effects of O3-glycosylation of talopyranose were determined.     

Tribology of the root cap in maize (Zea mays) and peas (Pisum sativum)

Frictional resistance to a penetrating body can account for more than 80% of the total resistance to penetration of soil. We measured the frictional resistance between growing root caps of maize and pea and ground and smooth glass surfaces, which was linearly correlated to load, allowing calculation of the coefficient of kinetic friction and adhesion. Coefficients of kinetic friction between the root caps and the ground and smooth glass surfaces were approximately 0.04 and 0.02, respectively, the first measurements of the frictional properties of root tips at rates approaching those of root elongation, and an order of magnitude smaller than those previously reported. Results suggest that roots are well designed for penetrating soil, and encounter only small frictional resistance on the root cap. These data provide important parameters for modelling soil stresses and deformation around growing root tips.     

Characterization of the metabolite produced by Mycosphaerella pinodes, the causal agent of mycosphaerella blight on field peas (Pisum sativum L.)

The metabolite produced by Mycosphaerella pinodes, the causal agent of mycosphaerella blight on field peas, was detected by thin layer chromatography (TLC) and was analyzed for its chemical and pathogenic characteristics. One blue dot was detected using 254nm UV light on TLC plate, and a spray of rho-anisaldehyde (110 degrees C, 30 min) also produced a blue dot. The solvent systems used for TLC analysis were ethyl acetate/water/acetone (5/2/5), chloroform/methanol/glacial acetic acid (19/10/2), toluene/ethyl acetate/90% formic acid (6/3/1), diethylether/methanol/water/90% formic acid (95/4/1/1), and bezene/methanol/acetic acid (24/2/1), with R(f) values (min-max) of 0.09-0.18, 0.88-0.95, 0.06-0.15, 0.39-0.47 and 0.05-0.12, respectively. The recovered metabolite from the TLC plate displayed UV absorption peaks at 212, 244, 250, 256 and 261 nm. The proposed formula of the main component of the metabolite was C16H12N3O6. The TLC-purified metabolite induced symptom of discoloration on detached pea leaves.     

Cultivar and Rhizobium strain effects on the symbiotic performance of pea (Pisum sativum)

The symbiotic performance of four pea ( Pisum sativum L.) cultivars in combination with each of four strains of Rhizobium leguminosarum was studied in growth chamber experiments in order to estimate the effects of cultivars, strains and cultivar × strain interaction on the variation in dry weight, N content and dry weight/N ratio. At harvest 63 days after planting, cultivars accounted for 75% of the variation in dry weight, while the Rhizobium strains accounted for 63% of the variation in N-content and 70% of the variation in dry weight/N ratio. Cultivar × strain interactions were statistically significant, but of minor quantitative importance, accounting for 5–15% of the total variation. Rhizobium strains also influenced the partitioning of N between reproductive and vegetative plant parts and between root and shoot biomass.     

Embryogenesis of the pea (Pisum sativum) I. The cytological environment of the developing embryo

Summary The structural relationships of the pea embryo to its immediate organic environment have been studied under the light and electron microscopes during a phase of development just preceding the period of rapid embryo growth. The following observations are reported: a) Following fertilization the suspensor elongates and displaces the embryo from the micropylar to the opposite end of the embryo sac that has, by this time, developed a large chamber that is eventually occupied by the cotyledons and a narrow tubular arm that contains the elongated suspensor and later the radicle of the enlarging embryo. b) The embryo and the suspensor are ensheathed by an extra-embryonic wall that subsequently becomes attached to the boundary wall of the embryo sac by means of crosslinking walls. These structures are essential in the precise positioning of the embryo within the embryo sac. c) The thin layer of endospermic cytoplasm that lines all extra-embryonic walls and the boundary of the embryo sac is highly motile and has certain characteristic ultrastructural features,e.g., large and intricate mitochondria, a dense population of ribosomes, a specialized form of smooth ER and an organelle that may be a type of plastid. d) The ovular tissue and the boundary wall of the embryo sac, particularly in the vicinity of the embryo, are structurally specialized. Relatively large intercellular spaces in the former are associated with a greatly increased surface of the boundary wall by means of extensive protrusions into the endospermic cytoplasm, many large and complex mitochondria are associated with these protrusions. It is suggested that this organization may indicate sites of nutrient entry into the embryo sac. Some ideas regarding the possible role of the described structures are discussed but it is emphasized that no experimental evidence is available at this stage to provide an unequivocal basis of interpretation.Supported by a grant from the Australian Research Grants Committee.     

Study of the factors influencing Agrobacterium-mediated transformation of pea (Pisum sativum L.)

Factors influencing the efficiency of Agrobacterium-mediated transformation of pea were tested using highly efficient, direct regeneration system. The virulence of three Agrobacterium strains (octopine LBA 4404, nopaline C58C1 and succinamopine, hypervirulent EHA 105) clearly varied giving 1 transgenic plant per 100 explants for LBA 4404, 2.2 for C58C1 and 8.2 for EHA 105. To test the efficacy of selection agents we used the hypervirulent EHA 105 strain carrying pGPTV binary vector with one of four different selection genes: nptII, hpt, dhfr or bar. The mean number of transgenic, kanamycin-resistant plants for two cultivars tested was 4.2 per 100 explants and was slightly higher than the number of phosphinothricin-resistant plants (3.6 plants per 100 explants). The proportion of transgenics among kanamycin-selected plants was also higher than among phosphinothricin-resistant plants (35% and 28% respectively). There was no regeneration on hygromycin or methotrexate media (transformation with hpt and dhfr genes). Acetosyringone had no apparent influence on efficiency of transformation with hypervirulent EHA 105 strain, however it did affect the rate of transformation when moderately virulent C58C1 was used. Recovery of transgenic plants was enhanced after application of 5-azacytidine. The presence of integrated T-DNA was checked by PCR and confirmed by Southern hybridization. T-DNA was stably transmitted to the next generation.