Foliar injury responses of 24 bean cultivars (Phaseolus vulgaris) to various concentrations of ozone

In the summer of 1981, symptoms on leaves of field grown beans (Phaseolus vulgaris) were noticed suggesting O₃ injury. In order to determine their relative sensitivity primary leaves of 24 bean cultivars were exposed to various concentrations of O₃. Differences in foliar injury responses between cultivars were affected by O₃ concentration. The cultivars Pros/Gitana and Stratego were the most sensitive, whereas Berna and Narda were the most insensitive. The symptoms observed in practice may be attributed to O₃. Sensitivity of Stratego primary leaves clearly depended on plant age. This cultivar will be tested as an indicator plant for O₃.     

Changes in susceptibility of bean leaves (Phaseolus vulgaris) to Sclerotinia sclerotiorum and Botrytis cinerea by pre-inoculative ozone exposures

The effects of ozone on the susceptibility of leaves ofPhaseolus vulgaris toSclerotinia sclerotiorum andBotrytis cinerea have been investigated. Seedlings of one ozone-sensitive (Pros) and five relatively ozone-insensitive cultivars (Gamin, Precores, Groffy, Narda, Berna) were exposed to different ozone concentrations (0, 120, 180 and 270 g m^–3) for 8 h. One day after the exposures, primary leaves were detached and immediately inoculated with spores of either pathogen suspended in water or in a 62.5 mM KH₂PO₄ (Pi) solution. Visible ozone injury differed between the cultivars and increased with increasing ozone concentration. On the leaves of non-exposed plants, spores of the pathogens suspended in water caused very few lesions, whereas fungal pathogenicity was stimulated by addition of Pi to the inoculum. Ozone-injured leaves of all cultivars exhibited lesions after inoculation of the leaves with the pathogens suspended in water, and the number of lesions was positively correlated with the level of ozone injury for either pathogen and cultivar. The increase in susceptibility of bean leaves in response to increasing ozone concentrations was greater forB. cinerea than forS. sclerotiorum when spores were suspended in water, but was similar when the spores were suspended in Pi.In general, the number of lesions following inoculation with spores in Pi increased with increasing ozone concentration. However, the number of lesions in the ozone-insensitive Groffy was reduced by an exposure to 120 g m^–3 but increased with higher concentrations. This pattern of susceptibility response to the pathogens was not found in the other ozone-insensitive cultivars and, thus, did not appear to be related to the inherent ozone-insensitivity in bean.     

Lipid metabolism in tomato and bean as a sensitive monitor for biocontrol of wilt diseases

The lipids metabolism of tomato and bean plants during biological control of wilt pathogens (Fusarium oxysporum f.sp.lycopersici andF. oxysporum f.sp.phaseoli, respectively) byBacillus subtilis was investigated. The interaction of wilt pathogens with both tomato and bean caused an imbalance and drastic reduction in total lipids, triacylglycerol, sterol and all phospholipd fractions except phosphatidic acid. The application of a formulated biocontrol agent,B. subtilis, eliminated the detrimental effect of both wilt pathogens and consequently prevented catabolism of lipid fractions in both tomato and bean. Moreover, the changes in the lipid fractions as a sensitive monitor for biocontrol of wilt diseases suggest a positive correlation between the application ofB. subtilis and improvement in the host metabolism towards anabolism. http://www.phytoparasitica.org posting Sept. 20, 2006.     

Phytoalexin production in French bean leaves infected by botrytis cinerea

French bean (Phaseolus vulgaris) leaves were inoculated with three pathogenic and two nonpathogenic isolates ofBotrytis cinerea and the infected tissues, containing either spreading lesions or lesions limited in size, were investigated for the presence of phytoalexins. In most cases phaseollin, phaseollidin, phaseollinisoflavan, the phaseollin metabolite, 6a-hydroxyphaseollin, and a few unidentified antifungal compounds were found; phaseollin was predominant. The concentration of phaseollin accumulating in leaves infected by the nonpathogenic isolate BC-5 was about twice as high as that in infections produced by pathogenic isolates. In contrast, leaves infected by the nonpathogenic isolate BC-6 only contained low concentrations of phaseollin.Our data do not provide strong evidence that phaseollin is the principal factor that limits lesion development.Samenvatting Bladeren van boon (Phaseolus vulgaris) werden geïnoculeerd met drie pathogene en twee niet-pathogene isolaten vanBotrytis cinerea, als gevolg waarvan zich uitbreidende lesies, resp lesies van beperkte omvang ontstonden. Deze lesies werden samen met een omringend groen gedeelte onderzocht op de aanwezigheid van fytoalexinen. Hiertoe werden deze stoffen geëxtraheerd uit drooggevroren bladmateriaal, en gezuiverd m.b.v. kolomchromatografie over Sephadex LH-20 (Tabel 1) en dunnelaag-chromatografie (Tabel 2). Kwantitatieve analyse vond plaats d.m.v. in situdensitometrie.In de meeste gevallen werden phaseolline, phaseollidine, phaseollineïsoflavaan, een omzettingsprodukt van phaseolline, nl. 6a-hydroxyphaseolline, en enkele onbekende fungitoxische stoffen aangetoond (Fig. 1). Phaseolline was het meest voorkomende fytoalexine (Fig. 2 en 3). De concentratie van phaseolline in bladeren geinfecteerd met het niet-pathogene isolaat BC-5 was ongeveer tweemaal zo hoog als in bladeren geïnfecteerd met een pathogeen isolaat (isolaat BC-1, BC-3 en BC-4). Daarentegen bevatten bladeren die met het niet-pathogene isolaat BC-6 waren geinfecteerd slechts lage concentraties phaseolline.De hoeveelheden phaseolline en 6a-hydroxyphaseolline, omgerekend per lesie, namen snel toe in zich uitbreidende lesies van isolaat BC-1, terwijl de toenamesnelheid van deze stoffen afnam in lesies van isolaat BC-5, die beperkt van omvang bleven (Fig. 4).De resultaten geven geen duidelijke aanwijzingen dat phaseolline de belangrijkste remmende factor zou zijn bij het beperkt blijven van lesies.     

The haustorial host-parasite interface in rust-infected bean leaves after high-pressure freezing

The haustorial fine structure of the bean rust fungus, Uromyces appendiculatus var. appendiculatus, was studied within the cells of its host, Phaseolus vulgaris. Results were obtained after high-pressure freezing and subsequent freeze-substitution or freeze-fracturing. Good preservation of leaf tissue after freeze-substitution needed cryoprotection with 8% methanol. For freeze-fracturing, no chemical treatment was applied. In addition to the organelles which are generally found in fungi after cryo-fixation, tubular-vesicular complexes were found in the cytoplasm. Both techniques revealed an extrahaustorial matrix of even width, surrounding the haustorial body. The extrahaustorial membrane was not undulated, and the side facing the plant cytoplasm was lined with a delicate fringe of well-stained material. The extrahaustorial membrane was nearly devoid of intramembrane particles. The host plasma membrane in infected tissue, especially the protoplasmic face, had fewer intramembrane particles than those in uninfected tissue. The haustorial plasma membrane contained many intramembrane particles.     

Broad bean stain virus: Identification,detectability with ELISA in faba bean leaves and seeds,occurrence in West Asia and North Africa,and possible wild hosts

During a survey of faba bean viruses in West Asia and North Africa a virus was identified as broad bean stain virus (BBSV) based on host reactions, electron microscopy, physical properties and serology. An antiserum to a Syrian isolate was prepared. With this antiserum the direct double antibody sandwich ELISA (DAS-ELISA) and dot-ELISA were very sensitive in detecting BBSV in leaf extracts, ground whole seeds and germinated embryos. Sensitivity was not reduced when the two-day procedure was replaced by a one-day procedure. Using ELISA the virus was detected in 73 out of 589 faba bean samples with virus-like symptoms collected from Egypt (4 out of 70 samples tested), Lebanon (6/44), Morocco (0/7), Sudan (19/254), Syria (36/145) and Tunisia (8/69). This is the first report of BBSV infection of faba bean in Lebanon, Sudan, Syria and Tunisia. Fourteen wild legume species indigenous to Syria were susceptible to BBSV infection, with only two producing obvious symptoms. The virus was found to be seed transmitted inVicia palaestina.Samenvatting Tijdens een inventarisatie van virussen in veld- of tuinbonen (Vicia faba) in het Midden-Oosten en Noord-Afrika werden virusisolaten verkregen, die op grond van toetsplantreacties en fysische eigenschappen en van elektronenmicroscopische en serologische waarnemingen werden herkend als het met zaad overgaande en door snuitkevers verspreide tuinbonezaadvlekkenvirus (broad bean stain virus).Met een nieuw antiserum, gemaakt tegen een Syrisch isolaat, bleek de aantoonbaarheid van het virus in bladextracten, vermalen zaden en gekiemde embryo's erg hoog te zijn, zowel bij toepassing van DAS-ELISA als dot-ELISA. De gevoeligheid van de methode werd niet geringer wanneer de procedure werd bekort van twee dagen tot één dag.Met ELISA werd het virus aangetoond in 73 van de 589 bladmonsters van veldboon, met symptomen die deden denken aan virusinfectie, verzameld in Egypte, Libanon, Marokko, Soedan, Syrië en Tunesië. Het virus werd aangetroffen in monsters uit alle genoemde landen behalve Marokko, maar het aantal uit dat land afkomstige monsters was slechts gering. Dit is de eerste maal dat het virus wordt gerapporteerd in Libanon, Soedan, Syrië en Tunesië.Veertien wilde soorten vlinderbloemigen uit Syrië bleken bij toetsing door inoculatie vatbaar voor infectie. Slechts twee ervan vertoonden symptomen. Het virus kan dus onzichtbaar voorkomen in de wilde vegetatie. InVicia palaestina ging het zelfs over met zaad.The project was financially supported by the Directorate General for International Cooperation (DGIS, Research and Technology Programme) of the Netherlands Minister for Development Cooperation.     

Increase in phenylalanine ammonia-lyase activity in bean leaves infected with tobacco necrosis virus

In bean leaves infected with tobacco necrosis virus the phenylalanine ammonia-lyase activity dramatically increases parallel with the necrotization process. At the same time the amount of proteins decreases, and the total amount of phenolics increases in the diseased tissues.Samenvatting In bonebladeren geïnfecteerd met tabaksnecrosevirus neemt tegelijk met het afstervingsproces de activiteit van fenylalanine ammonia-lyase drastisch toe. Gelijktijdig neemt in de zieke weefsels de hoeveelheid eiwitten af en de totale hoeveelheid fenolen toe.     

Formation of phytoalexins within and outside lesions of Botrytis cinerea in French bean leaves

Rapidly spreading lesions and lesions restricted in size developed in primary leaves of French bean (Phaseolus vulgaris) in response to infection byBotrytis cinera isolates BC-1 and BC-5, respectively. These isolates caused similar differential lesions in leaves of cucumber, flax, lettuce and tomato. To determine whether phytoalexin accumulation was correlated with the resistant reaction in bean leaves, accumulation of phytoalexins was examined in necrotic areas of both types of lesions and in their surrounding green tissues. Phaseollin was the predominant phytoalexin, both inside and outside lesions, whereas phaseollidin and sometimes also phaseollinisoflavan were always present in lower concentrations. Phaseollin accumulated earlier and to higher levels within and around lesions of isolate BC-5 than of isolate BC-1. Relatively low concentrations of phaseollin were detected in the more remote green areas, including the petiole, of leaves bearing a spreading lesion. The phaseollin metabolite, 6a-hydroxyphaseollin, was found only inside lesions and in a narrow zone around lesions of both types. The authors consider the possibility that the differing concentrations of phytoalexins in the infected tissues are not a determining factor for the differential interactions betweenB. cinerea and bean leaves, but are rather the result of it.     

Involvement of lipoxygenase,lipoxygenase pathway volatiles,and lipid peroxidation during the hypersensitive reaction of pepper leaves to Xanthomonas campestris pv. vesicatoria

Lipoxygenase activity and protein, production of lipid-derived volatiles, and lipid peroxidation levels were determined in pepper (Capsicum annuum L., cv. Early Calwonder-10R) leaves during the hypersensitive reaction induced by avirulent race 2 of Xanthomonas campestris pv. vesicatoria. Lipoxygenase activity increased during the collapse phase of the hypersensitive reaction (8 to 12 h after inoculation), and an increase in electrolyte leakage occurred. However, Western blot analysis revealed that lipoxygenase proteins decreased during the same period. When only one longitudinal half of a pepper leaf was inoculated with the avirulent bacterium race, a significant increase in lipoxygenase activity was observed in both inoculated and noninoculated leaf halves, 10 h after inoculation. In addition, lipoxygenase protein decreased in inoculated leaf halves, but remained unchanged in noninoculated ones. The evolution of some volatile compounds derived from the lipoxygenase pathway [(E,E)-2,4-hexadienal, 1-hexanol, 3-hexen-1-ol, 2,4-hexadienal and 2,4-eptadienal] and carotenoid degradation (α- and β-ionone) increased in the incompatible interaction during the collapse phase of the hypersensitive reaction. The level of the oxidative index (A₂₃₅/A₂₀₅) of leaf lipid extracts, determined to estimate lipid peroxidation, significantly increased in the advanced stage of the hypersensitive reaction. Furthermore, determination of the oxidative index in neutral lipid, glycolipid and phospholipid fractions showed that the oxidative index was significantly increased only in the glycolipid fraction. Lipoxygenase activity and protein, electrolyte leakage, volatiles and lipid peroxidation were not changed in pepper leaves inoculated with the virulent race 1 of X. campestris pv.vesicatoria during the time interval considered (2–12 h after inoculations). The hypothesis that a lipoxygenase with chloroplastic location is induced in the incompatible interaction, and which is responsible for the increase in lipid peroxidation is advanced.     

高浓度臭氧对春小麦膜脂过氧化和抗氧化系统的影响

利用开顶式气室(OTC)研究了高浓度O3对春小麦叶片膜脂过氧化程度、活性氧含量、抗氧化系统的影响。结果表明,随着臭氧浓度的升高,不同生育时期小麦叶片的相对电导率、MDA含量显著上升(P<0.01);穗粒重明显下降(P<0.05);O2-.产生速率和H2O2含量显著增加(P<0.05)。SOD活性、CAT活性、POD活性随生育进程和O3浓度升高而显著下降。而对抗氧化物如类Car含量、ASA含量的影响不显著。臭氧浓度增加,促使小麦叶片膜的伤害,引发了膜脂过氧化作用,产生了过多的活性氧自由基,破坏了抗氧化系统功能,影响了叶片的正常生理代谢。     

Penetration of bean leaves by aminotriazole as influenced by adjuvants and humidity

Penetration of bean leaves (Phaseolus vulgaris var. Canadian Wonder) by aminotriazole (3-amino-1,2,4-triazole) was greatly enhanced under high humidity conditions (ca 10% in 17 h at the low humidity level (LHL) compared with ca 80% in 2 h at the high humidity level (HHL)). The addition of polyoxyethylene 20 sorbitan monolaurate (polysorbate 20) to the spray fluid increased penetration on all occasions at the LHL. The increase obtained was found to be dependent on the concentration of polysorbate 20. The inclusion of polysorbate 20 at the HHL resulted in an increase in aminotriazole penetration at low polysorbate 20 concentrations 0.2–12.8 g/litre and a non-significant decrease over the aqueous control at a concentration of 40 g/litre. With additions of glycerol to the spray fluid, aminotriazole penetration was increased on all occasions at the LHL while at the HHL no concentration of glycerol was found to enhance penetration. The addition of polysorbate 20 (< 0.1 to 40 g/litre) to a spray solution containing glycerol (0.3 ml/litre) enhanced penetration at the HHL compared with the aqueous and glycerol controls. A polysorbate 20 plus glycerol combination (6.4 g+0.6 ml/litre) gave the same order of penetration (98.4 and 94.0%) at the HHL and LHL respectively. In both cases penetration exceeded that obtained with the corresponding polysorbate 20 and glycerol controls. Some of the practical implications of these findings are discussed.     

Penetration of bean leaves by asulam as influenced by adjuvants and humidity

Penetration of bean (Vicia faba var. Maris Bead) foliage by asulam was found to be affected differently by different surfactants. Some anionic and non-ionic surfactants brought about a significant increase compared with the aqueous control, others were similar to the aqueous control and one caused a significant reduction. High humidity increased asulam uptake and the effect was still further enhanced in the presence of Tween 20. High humidity was also found to influence asulam penetration in the presence of glycerol. With urea, uptake was greater under high humidity conditions and in the presence of Tween 20. A similar enhancement of penetration was found with potassium ethyl xanthate at lower concentrations (0–0.20%) in the presence of Tween 20. On the other hand, reduced uptake resulted at higher concentrations (0.40–1.00%). Tributyl phosphate brought about an increase in penetration at concentrations greater than 0.75%. However, both potassium ethyl xanthate and tributyl phosphate were shown to damage the leaf surface.     

Pectic enzymes associated with phosphate-stimulated infection of French bean leaves by Botrytis cinerea

Botrytis cinerea readily produced polygalacturonases and pectin esterases in shake cultures of Richards' medium containing orthophosphate and no pectinaceous material or galacturonic acid.In inoculum droplets containing glucose and KH₂PO₄ or glucose and Na-ATP, which were used to evoke a susceptible reaction in French bean leaves,B. cinerea produced pectic enzymes. However, in inoculum droplets containing glucose but lacking phosphate, used to evoke a resistant reaction, activities of pectic enzymes remained low. As the enzyme activities already increased during the penetration stage of the infection process, it is assumed that these phosphatestimulated activities of pectic enzymes are, at least partially, responsible for the phosphatestimulated susceptible interaction between French bean leaves andB. cinerea.Electrophoresis in pectin-polyacrylamide gels showed that two polygalacturonases with a high isoelectric point value were associated with the penetration stage of the infection process.Samenvatting Botrytis cinerea vormde vlot polygalacturonasen en pectineësterasen in schudculturen in Richards' medium dat wel orthofosfaat maar geen pectineachtig materiaal of galacturonzuur bevatte.In inoculumdruppels met glucose +KH₂PO₄ of met glucose + Na-ATP, waarmee een vatbare reactie in bonebladeren kon worden opgewekt, vormdeB. cinerea verschillende pectolytische enzymen. In inoculumdruppels met glucose maar zonder fosfaat, waarmee een resistente reactie werd teweeggedbracht, bleeff de activiteit van pectolytische enzymen gering.Aangezien de enzymactiviteiten al begonnen toe te nemen tijdens de binnendringingsfase van het infectieproces, wordt verondersteld dat deze door fosfaat gestimuleerde activiteiten van pectolytische enzymen althans ten dele verantwoordelijk ziijn voor de door fosfaat gestimuleerde vatbare interactie tussen bonebladeren enB. cinerea.Elektroforese in pectine-polyacrylamidegels liet de toename zien van twee polygalacturonasen met een hoog isoëlektrisch punt tijdens de binnendringingsfase van het infectieproces.