Effects of Ca2+ and Mg2+ on Botrytis cinerea and Penicillium expansum in vitro and on the biocontrol activity of Candida oleophila

Calcium salts have been reported to play an important role in the inhibition of postharvest decay of apples and in enhancing the efficacy of postharvest biocontrol agents. Therefore, the present study was conducted in order to examine and compare the effects of calcium and magnesium salts on the germination and metabolism of the postharvest pathogens Botrytis cinerea and Penicillium expansum , and to determine the effects of these salts on the biocontrol activity of two isolates (182 and 247) of the yeast Candida oleophila. Increasing concentrations of CaCl₂ (25–175 mM) resulted in decreased spore germination and germ-tube growth of both pathogens. The greatest effect was observed in the case of B. cinerea. The inhibitory effect could be overcome by the addition of glucose to the germination medium. MgCl₂ (25–175 mM) had no effect on germination or germ-tube growth of either pathogen, indicating that the calcium cation rather than the chloride anion was responsible for the inhibition. The pectinolytic activity of crude enzyme obtained from the culture medium of both pathogens was also inhibited by 25–175 mM CaCl₂, with the greatest effect on the crude enzyme from P. expansum. Biocontrol activity of isolate 182 was enhanced by the addition of 90 or 180 MM CaCl₂, whereas there was no effect on the biocontrol activity of isolate 247. This was apparently due to the inability of isolate 247 to proliferate in apple wounds. It is postulated that enhanced biocontrol activity of isolate 182 of the yeast C. oleophila in the presence of Ca²⁺ ions is directly due to the inhibitory effects of calcium ions on pathogen spore germination and metabolism, and indirectly due to the ability of isolate 182 to maintain normal metabolism in the presence of"toxic" levels of calcium.     

Biological evidence for a 1:1 Ca2+:glyphosate association in deposit residuals on the leaf surface of barley

It has long been known that calcium ion antagonizes glyphosate, but it was not clear whether the stoichiometry of their interaction is 1:1 or 1:2. Two independent methods were used to determine which stoichiometry was the most probable. First, dose–response curves of barley ( Hordeum vulgare L.) plants treated with glyphosate were determined in the presence of 0, 1.25, 2.5, 5 and 10 mM CaCl₂. The doses of 'free' glyphosate (=not inactivated by calcium ion) were computed using the assumptions of 1:1 and 1:2 stoechiometries. The response curves were redrawn as a function of 'free' glyphosate. Analysis showed that the 1:2 hypothesis could be rejected, whereas the 1:1 hypothesis was highly probable. Second, kinetics of glyphosate penetration into barley leaves were determined in the presence of 0, 2.5 and 10 mM CaCl₂. The concentrations of 'free' glyphosate were computed as above. The kinetics of glyphosate penetration at these concentrations were established and were compared to the kinetics of glyphosate penetration in the presence of CaCl₂. Again, the 1:2 hypothesis was rejected, whereas the 1:1 hypothesis was more probable. These results strongly suggest that the stoichiometry of the Ca²⁺:glyphosate association is 1:1 in deposit residuals on the leaf surface.     

Control of vascular-streak dieback of cocoa with triazole fungicides and the problem of phytotoxicity

The effectiveness of a range of systemic fungicides against vascular-streak dieback of cocoa, caused by Oncobasidium theobromae , was evaluated over 2 years in Papua New Guinea. Monthly foliar sprays of the triazole fungicides hexaconazole, tebuconazole and triadimenol all gave good protection against the disease. The relative efficacy of these chemicals was, in order, tebuconazole (EC₅₀ 0–90 mg a.i. per plant), hexaconazole (EC₅₀ 2.35 mg a.i. per plant), triadimenol (EC₅₀ 4.17 mg a.i. per plant). However, the relative severity of plant growth inhibition induced by these compounds had the same ranking. Growth inhibition was reduced by extending the interval between treatments, but this also reduced the efficacy of disease control. Propiconazole stem paints (EC₅₀ 2502 mg a.i. per plant) were markedly less effective than these sprays. Neutralized phosphorous acid sprays and paints showed no activity against the disease.     

The global regulator GacA of Pseudomonas fluorescens CHA0 is required for suppression of root diseases in dicotyledons but not in Gramineae

Pseudomonas fluorescens strain CHA0 suppresses various plant diseases caused by soil-borne fungi. The pseudomonad produces the antimicrobial metabolites 2,4-diacetylphloroglucinol (Phl), pyoluteorin (Plt) and hydrogen cyanide, which are important for disease suppression, as well as the siderophores pyoverdine (Pvd), salicylic acid (Sal) and pyochelin (Pch). In the current work, a derivative of CHA0 with a mutation in the global regulator gene gacA (GacA⁻), which is unable to produce Phl, Plt and HCN, failed to protect the dicotyledonous plants cress and cucumber against damping-off caused by Pythium ultimum . In contrast, the GacA⁻ mutant could still protect the Gramineae wheat and maize against damping-off mediated by the same strain of P. ultimum , and wheat against take-all caused by Gaeumannomyces graminis . However, the GacA⁻ mutant overproduced Pch and Pvd. To gain more insight into disease protection afforded by the GacA⁻ mutant, a GacA⁻ Pvd⁻ double mutant (strain CHA496) was constructed by gene replacement. Strain CHA496 overproduced Pch and Sal compared with CHA0 and protected wheat against P. ultimum and G. graminis , whereas cress and cucumber were not protected. Addition of FeCl₃ repressed Pch and Sal production by strain CHA496 in vitro and impaired the protection of wheat in soil microcosms. In conclusion, a functional gacA gene was necessary for the protection of dicotyledons against root diseases, but not for that of Gramineae. Results indicated also that Pch and/or Sal were involved in the ability of the GacA⁻ Pvd⁻ mutant of CHA0 to suppress root diseases in Gramineae.     

K84的生物防治效果与土壤杆菌Ti质粒类型的关系

用K84对我国不同寄主来源的68个Agrobacterium tumefaciens菌株进行生物防治试验,结果表明:K84对我国桃树菌株的防治效果显著,对新疆的啤酒花菌株也有效;但对浙江萧山的部分啤酒花菌株无效;对甜菜菌株的防治效果不佳;对葡萄菌株基本上无效。对致病菌株的质粒类型的研究结果表明:桃树、啤酒花和甜菜菌株基本上都是胭脂碱(Nopaline)型质粒、葡萄菌株基本上是章鱼碱(Octopine)型质拉。试验中还发现不少胭脂碱型菌株不能被K84所防治。此外,在甜菜和毛白杨菌株中也发现了对细菌素A84(agrocin84)敏感,但不能被K84防治的菌株。     

Effects of nitrogen, phosphorus, potassium and calcium nutrition on strawberry anthracnose

The effects of a range of concentrations of four nutrients – nitrogen, phosphorus, potassium and calcium – in fertilizer solutions on the severity of anthracnose on strawberry cv. Nyoho cultivated under a noncirculation hydroponics system were determined after inoculation with Colletotrichum gloeosporioides . Crop growth and tissue nitrogen, phosphorus, potassium and calcium contents of the entire above-ground parts of the plant were also investigated. Elevated nitrogen and potassium concentrations in the fertilizer solution increased disease severity in contrast to phosphorus and calcium. Treatment with either NH₄ or NO₃ nitrogen was not significantly different. The dry weight of the strawberry plants increased significantly with elevated concentrations of nitrogen ( R ² = 0·9078) and phosphorus ( R ² = 0·8842), but was not influenced by the elevated amounts of potassium ( R ² = 0·8587) and calcium ( R ² = 0·6526) concentrations.     

Genetic analyses of resistance of the wheat differential cultivars Carstens V and Spaldings Prolific to two races of Puccinia striiformis

Genetic analysis of resistance of wheat seedlings to two races of Puccinia striiformis was conducted on F₁, F₂ and F₃ generations from crosses Carstens V (CV) × Lee, Spaldings Prolific (SPA) × Lee and CV × SPA. F₂ generations from crosses of CV and SPA with Strubes Dickkopf (SD) were also studied. The plants were classified into six resistance classes and analysed by factorial correspondence analysis and nonhierarchical classification. The two P. striiformis isolates tested were a French isolate of race 43E138 and a Lebanese isolate of race 2E16, selected for the differences in their virulence spectra for the common differential cultivars Strubes Dickkopf and Nord Desprez. Resistance of CV and SPA was recessive and dominant to races 43E138 and 2E16, respectively. CV possessed three or four resistance genes, one of them being expressed with both races. Two genes of CV had a cumulative effect for resistance to 43E138 and two or three gave dominant resistance to 2E16. SPA had three resistance genes, all of which gave resistance to 2E16 and two of which also gave resistance to 43E138. SPA had one gene in common with CV for resistance to both races. Furthermore, the gene for resistance to race 2E16 in CV and SPA was allelic with a gene in SD, and was probably Yr25 .     

Smoke pollution and the effects of root-knot nematodes on the growth of eggplants

Single and combined effects of smoke pollution and the root-knot nematode, Meloidogyne incognita race 1, were studied on eggplant in 1988 and 1989 at two sites (K1 and K2) 1 and 2 km respectively away from a coal-fired thermal power plant, and a control site at the Department of Botany, Aligarh Muslim University (AMU). Mean concentrations of SO₂, NO₂ and suspended particulate matter (SPM) were respectively 172, 95 and 626 μg/m³ in 1988 and 144, 97 and 556 μg/m³ in 1989 at K1; and 258, 150 and 344 μg/m³ in 1988 and 226, 113 and 293 μg/m³ in 1989 at K2. Peak levels of the gases and SPM were recorded in the early (12.00–15.00) and late (15.00–18.00) afternoon respectively. Concentrations of the pollutants at a control site were very low. Marginal browning appeared on the leaves of eggplants grown at site K2 whether or not they were infected by the nematodes. The galling caused by nematodes was severe at both polluted sites but egg mass production was inhibited at K2. Egg laying was significantly reduced at both polluted sites. M. incognita significantly reduced plant growth, yield and leaf pigment production of eggplant at the control and polluted sites. Most non-infected plants also showed significant reductions in these plant varieties i.e. from the effects of pollution at K2. Foliar concentrations of sulphur were significantly increased at both polluted sites, being greater in infected plants. The interaction of smoke pollution and M. incognita infection led to greater suppressions at the polluted sites, which were mostly significant at K1. At the polluted sites, leaves had more and longer trichomes; also the number and size of stomata were decreased but their openings were wider especially in infected plants at the two polluted sites.     

油菜花叶病毒15号和菸花叶病毒间干扰作用的研究

 TMV与YMV₁₅在系统感染寄主——菸(Nicotiana tabacum)上的相互干扰作用是很微弱的。同时接种二病毒时,在感染的早期(接种后2-3天) YMV₁₅占优势,在此时间以后TMV的浓度就超过YMV₁₅。二病毒以相同的侵染力同时接种心叶菸(Nicotiana glutinosa)时,则大部分的局部斑点是由YMV₁₅形成的。上述结果表明,YMV₁₅竞争侵染点的能力较TMV为大。试验并证明完整的YMV₁₅及其蛋白都能干扰TMV侵染菜豆(Phaseoluss vulgaris),因而这种干扰似决定于病毒蛋白质部分对于侵染点附着的特异性。     

Phage sensitivity in relation to pathogenicity and virulence of the cotton bacterial blight pathogen of Sudan

At least two pathotypes of Xanthomonas campestris pv. malvacearum are now known to exist in Sudan. The pre-Barakat (race 1) and post-Barakat (race 2) pathogens have been shown to exhibit different host specificity. The former is pathogenic and highly aggressive on only cultivars with no resistance genes or with the B₂ and/or B₃ resistance factors, while the latter can infect the B₆ cultivars also. Race 2 in Sudan, which was previously reported to infect all the standard differentials, produced milder angular leaf spot symptoms and occasionally restricted vein infection. Moreover, it exhibited reduced growth in planta compared with race 1.
Bacteriophage studies revealed that the two races are quite distinct in their phage sensitivity. Race 1 can be lysed by only three, or rarely four, of the six phages used for typing, while race 2 is sensitive to all of them. The present study suggests that phage 7 may be the type-determining phage for race 2. Race 2 strain mutants resistant to phage 3 or 4 were found to be sensitive to phage 7 and pathogenic to both Acala and Barakat, although showing marked attenuation of virulence. However, mutants resistant to phage 2 or 7 were insensitive to all the phages and although they retained their pathogenicity to Acala, they either lost the ability to infect Barakat or produced a hypersensitive reaction. The resistance of all mutants was found to be due to failure to adsorb the homologous phage, indicating a change in the cell wall. The association of this with the attenuation of virulence suggests that bacterial wall components may function as virulence determinants in Xanthomonas campestris pv. malvacearum.     

Premiers résultats de lutte biologique par les souches K84 et K1026 contre Agrobacterium radiobacter var. tumefaciens, agent du crown-gall des rosacées fruitières et du rosier au Maroc

Une collection de 206 souches marocaines d' Agrobacterium radiobacter var. tumefaciens a été testée vis-à-vis des souches antagonistes Agrobacterium radiobacter K₈₄ et K₁₀₂₆ in vitro . Ce test d'antagonisme a montré que la sensibilité des souches varie en fonction de la souche antagoniste, du biovar et de la plante-hôte d'origine. La K₁₀₂₆ s'est avérée meilleure que la K₈₄ aussi bien par le nombre et par le diamètre plus élevé. Des essais au champ ont montré une bonne efficacité de la souche antagoniste K₈₄ en seaux ou en sol naturellement infesté. La réussite de la lutte biologique avec la K₈₄ varie entre 70% et 95% sur GF677, rosier, amandier amer et pêcher par rapport aux témoins sans K₈₄, alors que l'abricotier s'est montré résistant au crown-gall aussi bien en sol inoculé qu'en sol naturellement infesté. Nous pensons généraliser cette lutte dans tous les foyers marocains à crown-gall avec une souche mutante de la K₈₄, la K₁₀₂₆, qui a donné aussi une satisfaction meilleure in vitro dans d'autres pays méditerranéens comme l'Espagne, la France et l'Italie.     

Possible involvement of gibberellins and ethylene in Orobanche ramosa germination

Summary Six inhibitors acting at different stages of gibberellin biosynthesis, applied during conditioning of Orobanche ramosa seeds, reduced subsequent germination in the presence of GR₂₄ (a strigol analogue). Ethylene seems to be involved in the induction of germination of conditioned seeds by GR₂₄, as inhibitors of its synthesis or action, applied to conditioned seeds, also strongly reduced induction of germination by GR₂₄. Exogenous ethylene did not induce germination of conditioned seeds, but 2-chloroethylphosphonic acid was able to do so. When inhibitors of gibberellin biosynthesis were applied to conditioned seeds in the presence of GR₂₄, they inhibited germination. These same inhibitors also strongly inhibited germination of conditioned Striga hermonthica seeds in response to GR₂₄; this inhibitory effect was reversed by the addition of 2-chloroethylphosphonic acid. The effect of these inhibitors on S. hermonthica , in which ethylene is a necessary mediator of germination induction by GR₂₄, strongly suggests that ethylene synthesis is also required for the induction of O. ramosa seed germination by GR₂₄. These growth regulators, which inhibit the two steps of germination in O. ramosa , could be useful for the development of methods for early season control of this parasite.     

Involvement of soilborne Phytophthora species in Central European oak decline and the effect of site factors on the disease

A survey was made on the occurrence of soilborne Phytophthora species in 35 oak stands on a range of geologically different sites in Bavaria. The most widespread species were P. quercina , P. cambivora and P. citricola . Seven other Phytophthora species were isolated infrequently. The fine root systems of 106 healthy and 111 declining mature trees of Quercus robur and Q. petraea were intensively investigated. The results indicate that, depending on the site conditions, at least two different complex diseases are referred to under the name 'oak decline'. On sites with a mean soil pH (CaCl₂)  3·5 and sandy-loamy to clayey soil texture Phytophthora spp. were commonly isolated from rhizosphere soil, and highly significant correlations existed between crown transparency and various root parameters. Oaks with P. quercina or other Phytophthora spp. in their rhizosphere had markedly higher levels of fine root damage than oaks without Phytophthora spp., and were subject to a relative risk of severe crown symptoms of 2·1 and 2·8, respectively. In contrast, in stands with sandy to sandy-loamy soils and a mean soil pH  3·9, Phytophthora spp. were not found. In these stands, correlations between crown transparency and various root parameters were either less significant or not significant. It is concluded that Phytophthora species are strongly involved in oak decline on sandy-loamy to clayey sites with a mean soil-pH (CaCl₂)  3·5.     

解淀粉芽胞杆菌XJ5挥发性物质抑菌活性及对苹果褐腐病防效测定

解淀粉芽胞杆菌Bacillus amyloliquefaciens XJ5挥发性物质对苹果褐腐病菌Monilinia fructigena等多种病原真菌具有良好的拮抗活性。通过固相微萃取-气相色谱质谱联用仪（Solid-phase microextraction-Gas chromatography mass spectrometry,SPME-GC-MS）对菌株XJ5产生的挥发性物质进行检测,利用平板对扣法测定挥发性化合物的抑菌活性,并离体接种评价菌株XJ5挥发性物质对苹果褐腐病的防效。结果表明,XJ5菌株挥发性物质对9种植物病原真菌的抑菌率在47.9%~84.8%,其中对苹果褐腐病菌的抑菌率最高为84.8%。SPME-GC-MS检测及抑菌活性测定表明,菌株XJ5分泌的主要挥发性物质为十二醛（Dodecanal）,具有抑菌活性的挥发性物质主要为2-壬醇（2-Nonanol）、2-乙基己醇（2-Ethyl-1-hexanol）和2-十一醇（2-Undecanol）。其中2-壬醇抑制苹果褐腐病菌的EC₅₀为3.43 μg/mL,且XJ5菌液离体熏蒸与2-壬醇离体熏蒸均可有效抑制苹果褐腐病菌丝生长及病斑扩展,其对苹果褐腐病的离体防效分别在14.4%~71.2%和66.4%~97.6%,与对照差异显著。     

Factors affecting the control of Pythium ultimum damping-off of sugar beet by Pythium oligandrum

Application of Pythium oligandrum to a soil-based compost as a mycelial suspension (5 × 10² CFU g⁻¹ of dry compost) and oospore alginate pellets (10⁵ oospores/g of dry compost) controlled pre- and postemergence damping-off of sugar beet caused by Pythium ultimum to a level similar to metalaxyl seed treatment. Oospore seed treatments and aqueous suspensions of oospores applied to compost failed to control disease. Problems in the use of P. oligandrum oospore inocula for the control of damping-off were highlighted. It was shown that treatment of oospores with cellulase (20 g L⁻¹) increased germination approximately three-fold in comparison to untreated spores. Untreated and cellulase pretreated oospores were subsequently evaluated as seed treatments for their ability to control damping-off of sugar beet. The highest rate of pretreated oospores (10⁴ oospores/seed) gave levels of emergence and establishment in infested compost that were not significantly different from the uninfested controls, whereas seed treatment with untreated oospores gave no significant reduction in disease. In a trial carried out in a controlled environment to assess the effect of pH (4.5–8.0), P. oligandrum (10⁴ cellulase pretreated oospores/seed) was shown to control pre- and postemergence damping-off of sugar beet at pH 7.0 and 7.5 only.     

Virulence for faba bean and production of ascochitine by Ascochyta fabae

The production of ascochitine by seven isolates of Ascochyta fabae accounted for the toxicity of culture filtrates of the fungus to cells isolated from leaves of Vicia faba. The LD₅₀ value for cells from cultivars that were susceptible, tolerant or resistant to the fungus was similar i.e. 3·0 × 10⁻⁵ m , 3·8 × 10⁻⁵ m and 2·9 × 10⁻⁵ M, respectively. Ascochitine affected neither the germination of seeds nor the growth of mature plants at 5·17 × 10⁻⁴ m but caused necrosis and wilting of plant cuttings at 2·5 × 10⁻⁴ m and 5·10⁻⁴ m . There was no association between virulence of 16 isolates of A. fabae for three cultivars of V.faba and the production of ascochitine in vitro. One isolate produced no ascochitine in vitro and yet was the most virulent for two of three cultivars. The toxin could not be extracted from infected plants.     

Studies on the inheritance of resistance to metalaxyl in Phytophthora infestans

Single-oospore progeny from matings between field isolates of Phytophthora infestans either highly resistant or sensitive to metalaxyl were analysed for fungicide sensitivity in vitro, mating type and, in some cases, allozyme variation at the locus for glucosephosphate isomerase ( GPI-1. ) In each cross the majority of first-generation (F₁) progeny showed intermediate sensitivity to metalaxyl. Frequency distributions were skewed towards sensitivity and a few progeny were either wholly sensitive or resistant phenotypes. Allozyme analysis of F₁ progeny from a cross between parents of Dutch and Mexican origin showed that c. 10% were selfs of both parents and of parental phenotype for metalaxyl sensitivity. The selfs of the A2 but not the Al parent segregated for mating type. Two backcross generations to the metalaxyl-resistant Dutch parent gave unexpected phenotypic frequency distributions and aberrant ratios for genotypes at the GPI-I locus.
Progeny of another backcross between an F₁ isolate of intermediate sensitivity and its sensitive Egyptian parent gave a 1:1 ratio for sensitive: intermediate phenotypes. F₂ progeny from a related sibmating between intermediate phenotypes segregated in a ratio close to 1:2:1 for sensitive: intermediate: resistant phenotypes. Segregation was also observed among sexual progeny of an intermediate self-fertile isolate from the backcross generation but not among progeny of a resistant self-fertile phenotype. Mating type segregated in both cases.
These data are consistent with resistance to metalaxyl in P. infestans being governed by a single nuclear locus exhibiting incomplete dominance. This hypothesis is discussed in relation to the incidence of resistance to metalaxyl in natural populations of P. infestans.     

Inheritance of partial resistance to race 2 of Albugo candida in canola-quality mustard (Brassica juncea) and its role in resistance breeding

The inheritance of partial resistance to race 2 of Albugo candida was studied in a canola-quality line of Brassica juncea . This partially resistant line was crossed with the susceptible B. juncea cultivar Commercial Brown. F₁, F₁(reciprocal), F₂, BC and doubled haploid generations from the cross were inoculated with a zoospore suspension of race 2 to study segregation of partial resistance. The partially resistant phenotype appeared to be controlled by a single dominant gene that has variable expression. This partial resistance can have implications in breeding for disease resistance against white rust, as adult plants did not develop hypertrophic growth or stagheads under greenhouse and field conditions.     

AN AUTORADIOGRAPHIC STUDY OF THE DISTRIBUTION OF 14C LABELLED ASSIMILATES AT DIFFERENT STAGES OF DEVELOPMENT OF OXALIS LATIFOLIA H.B.K. AND O. PES-CAPRAE L.

Summary. An autoradiographic study of the distribution of ¹⁴C-labelled assimilates, derived from ¹⁴CO₂ fed to the aerial parts, was made at ten different developmental stages of Oxalis latifolia H.B.K. and Oxalis pes-caprae L. Changing patterns of distribution of labelled assimilates to the underground parts of both species were revealed. These patterns are discussed in the context of the timing of control by defoliation and by foliageapplied herbicides.
Etude autoradiographique de la distribution des métabolites marqués an ¹⁴C à différents stades de développement de l'Oxalis latifolia H.B.K. et de l' O. pes-caprae L.