Systemic Migration and Establishment of Xanthomonas campestris pv. pruni in Plum Leaves and Twigs

The systemic migration of Xanthomonas campestris pv. pruni (Xcp) through vascular bundles of leaves and twigs of plum was investigated. A rifampicin-resistant strain of Xcp was inoculated into leaves located midway from the tip of new green twigs of Golden King plum trees in a glasshouse. High numbers of the pathogen were recovered 4 and 8 weeks after inoculation from sections of uninoculated and symptomless veins, petioles, and twig tissue. Symptoms of bacterial spot developedwithin 8 weeks on main and secondary veins of uninoculated leaves located as far as 13 cm from the inoculated leaf on the same twig. Weekly isolation indicated the constant presence of Xcp in apparently unaffected shields of twig tissue obtained from a naturally infected Golden King orchard. Xcp apparently enters plum twigs through veins of infected leaves and migrates systemically through twigs to leaves.     

Severe Outbreak of Pseudomonas syringae pv. avellanae on Hazelnut in Italy

Pseudomonas syringae pv. avellanae , the causal agent of bacterial canker of hazelnut, has been isolated for the first time in Italy. Biochemical tests were consistant with the type-strain of the microorganism and whole-cell protein profiles of the studied isolates were similar to those of the type-strain. When artifically inoculated, either in summer or in autumn, the isolates incited the same symptoms as those observed in the field.
The main symptoms are the failure of the buds to swell or their withering after breaking in spring; the chlorosis of the foliage and rapid withering of the leaves that remain attached on the twigs during summer; dark brown discolouration of the bark and cambium, necrosis in the primary roots and the final dieback of the plant. The pathogen killed more than 1,800 adult hazelnut plants in one orchard within a period of five years. The main differences of the symptomatology between bacterial canker and bacterial blight of hazelnut, caused by Xanthomonas campestris pv. corylina , and the "seccume" of the hazelnut are briefly discussed.     

Reduction of olive knot disease by a bacteriocin from Pseudomonas syringae pv. ciccaronei

A bacteriocin produced by Pseudomonas syringae pv. ciccaronei, used at different purification levels and concentrations in culture and in planta, inhibited the multiplication of P. syringae subsp. savastanoi, the causal agent of olive knot disease, and affected the epiphytic survival of the pathogen on the leaves and twigs of treated olive plants. Treatments with bacteriocin from P. syringae pv. ciccaronei inhibited the formation of overgrowths on olive plants caused by P. syringae subsp. savastanoi strains PVBa229 and PVBa304 inoculated on V-shaped slits and on leaf scars at concentrations of 10(5) and 10(8) CFU ml(-1), respectively. In particular, the application of 6,000 arbitrary units (AU) of crude bacteriocin (dialyzed ammonium sulfate precipitate of culture supernatant) ml(-1) at the inoculated V-shaped slits and leaf scars resulted in the formation of knots with weight values reduced by 81 and 51%, respectively, compared to the control, depending on the strains and inoculation method used. Crude bacteriocin (6,000 AU ml(-1)) was also effective in controlling the multiplication of epiphytic populations of the pathogen. In particular, the bacterial populations recovered after 30 days were at least 350 and 20 times lower than the control populations on twigs and on leaves, respectively. These results suggest that bacteriocin from P. syringae pv. ciccaronei can be used effectively to control the survival of the causal agent of olive knot disease and to prevent its multiplication at inoculation sites.     

Erwinia salicis in Cricket Bat Willows: Rate of Movement of the Bacterium and the Production of Symptoms in Young Trees and Shoots

The lack of movement of Erwinia salicis and the erratic development of symptoms of watermark disease in young rooted leafy cuttings of the cricket bat willow, Salix alba var. caerulea , grown in a controlled environment is described. Erwinia salicis remained in the zone of inoculation for 3 h after inoculation. Fifty-five % of inoculations resulted in the bacterium being isolated on glycerol nutrient agar after 6 months but in the development of none of the classical symptoms of the disease. The maximum penetrations of wood by E. salicis were 24 cm in 24 h in 4·5-year-old wood, 20 cm in 24 h in 2·5-year-old wood and 6 cm in 6 d in 1·5-year-old wood. The pathogen was not isolated from anywhere other than the wood of the original cutting, and did not penetrate into shoots existing before or developing after inoculation, in 6 months. Greatest numbers of E. salicis were present in the 1-cm length of wood including the inoculation site at all time intervals studied. Results using a saprophyte, Serratia marcescens , were similar to those obtained using E. salicis insofar as distribution of the bacteria in the wood with time was concerned. Although E. salicis could still be isolated from test inoculations after 6 months, only 7% of inoculated small trees developed typical watermark symptoms in the wood, though a further 45% showed red leaf symptoms. Movement of the pathogen over longer periods needs study, and older trees need to be artificially inoculated. Young, artificially inoculated cricket bat willow trees, up to 5 years of age are resistant to the systemic spread of E. salicis , and of other bacteria. Symptom expression is very erratic, despite the genetic homogeneity of the host material. The use of very young shoots as propagating material should be tested as a means of controlling the disease.     

Reduction of Olive Knot Disease by a Bacteriocin from Pseudomonas syringae pv. ciccaronei

A bacteriocin produced by Pseudomonas syringae pv. ciccaronei, used at different purification levels and concentrations in culture and in planta, inhibited the multiplication of P. syringae subsp. savastanoi, the causal agent of olive knot disease, and affected the epiphytic survival of the pathogen on the leaves and twigs of treated olive plants. Treatments with bacteriocin from P. syringae pv. ciccaronei inhibited the formation of overgrowths on olive plants caused by P. syringae subsp. savastanoi strains PVBa229 and PVBa304 inoculated on V-shaped slits and on leaf scars at concentrations of 10⁵ and 10⁸ CFU ml⁻¹, respectively. In particular, the application of 6,000 arbitrary units (AU) of crude bacteriocin (dialyzed ammonium sulfate precipitate of culture supernatant) ml⁻¹ at the inoculated V-shaped slits and leaf scars resulted in the formation of knots with weight values reduced by 81 and 51%, respectively, compared to the control, depending on the strains and inoculation method used. Crude bacteriocin (6,000 AU ml⁻¹) was also effective in controlling the multiplication of epiphytic populations of the pathogen. In particular, the bacterial populations recovered after 30 days were at least 350 and 20 times lower than the control populations on twigs and on leaves, respectively. These results suggest that bacteriocin from P. syringae pv. ciccaronei can be used effectively to control the survival of the causal agent of olive knot disease and to prevent its multiplication at inoculation sites.     

BACTERIAL CANKER OF STONE-FRUITS III. INOCULUM CONCENTRATION AND TIME OF INOCULATION IN RELATION TO LEAF-SCAR INFECTION OF CHERRY

Leaf scars on the fruiting spurs of the cherry varieties Roundel (high resistance) and Napoleon (low resistance) were inoculated with Pseudomonas mors-prunorum on four separate occasions in the autumn, using, on each occasion, the same range of five different inoculum concentrations. The results, recorded the following year, showed that the percentage diseased spurs (disease incidence) and the severity of the disease symptoms (disease severity) both increased with inoculum concentration.
The median threshold concentration of inoculum (T. C. 50), defined as the concentration necessary to give 50% diseased spurs, varied with time of inoculation, but on all occasions was considerably higher for Roundel than for Napoleon.
In another experiment the leaf scars at the nodes of the current year's growth, inoculated at weekly intervals throughout the autumn, were found to be susceptible from the beginning of September to the latter part of October. During this period disease incidence varied considerably with time of inoculation. There was evidence that this variation was related to two factors which influenced the numbers of bacteria penetrating into leaf scars, namely, (1) the rate of evaporation of the infection drop, and (2) the rate of suction of inoculum into the vessels of the leaf traces.
The experiments provided evidence of a long infection period beginning early in the autumn. It is suggested, therefore, that the timing of bactericidal sprays in the autumn be advanced and that the present concept of 'protective' sprays in disease control be replaced by one based on the eradication of external sources of inoculum.     

Occurrence of Pseudomonas avellanae (Psallidas) Janse et al. and related pseudomonads on wild Corylus avellana trees and genetic relationships with strains isolated from cultivated hazelnuts

Surveys in submediterranean forests of central Italy were carried out during 1996–98 to verify the possible presence of bacterial canker caused by Pseudomonas avellanae in wild hazelnut trees ( Corylus avellana L.). Wilted twigs were noticed several times especially in summer. In other cases, wild C. avellana trees growing near to hazelnut orchards appeared completely wilted. Isolates that were pathogenic to C. avellana , showing a different degree of virulence, were obtained in both situations. Biochemical, physiological and nutritional tests as well as the comparison of whole-cell protein profiles, revealed the presence of 16 isolates identical to P. avellanae reference strains that had previously been isolated in the same area and five deviating isolates. Repetitive-PCR genomic fingerprinting performed by using BOX (Box elements), ERIC (Enterobacterial Repetitive Interkingdom Consensus) and REP (Repetitive Extragenic Palindromic) primer sets and analysed by means of upgma , revealed the existence of two main groups of pseudomonads pathogenic to C. avellana . Group A includes P. avellanae strains isolated in northern Greece and central Italy as well as the isolates obtained from the wild C. avellana trees grown near the cultivated hazelnut orchards. Group B includes strains previously isolated in northern, southern and other areas of central Italy as well as the isolates obtained from C. avellana wild trees showing twig dieback. Control measures should be taken to avoid the spread of bacterial canker of hazelnut in the forests of central Italy.     

Preliminary Investigations about the Mode of Transmission and Spread of Xanthomonas campestris pv. graminis on Forage Grasses

Extensive studies showed that no disease was caused when seeds of different forage grasses were inoculated with Xanthomonas campestris pv. graminis. The disease could easily be induced by infecting the plants in the root system, leaves or flower. The inoculation site in the leaf proved to be of vital importance for the development of the disease. Wilting symptoms were quickly induced when the pathogen was inoculated near the leaf base. Plants in root-contact with diseased plants showed disease symptoms. It is not known whether these symptoms were caused by the bacteria or by toxins released by nearby diseased plants. Cross inoculation trials on different grass varieties revealed that different pathovars exist in the group of xanthomonads, pathogenic to forage grasses. Some have a broad host range whereas others are more limited to a single plant genus. Field trials suggest that in Belgian climatic conditions, the losses caused by bacterial wilt are rather limited.     

Assimilatverteilung in Sommergerstensorten mit unterschiedlicher Resistenz gegenüber dem Echten Mehltau (Erysiphe graminis f. sp. hordei)

Distribution of assimilates in cultivars of spring barley with different resistance against powdery mildew (Erysiphe graminis f. sp. hordei) Transport and distribution of radioactive labelled assimilates in spring barley cultivars with different degrees of resistance to powdery mildew were studied after ¹⁴CO₂-treatment of single leaves. Plants of the cultivars ‘Amsel’ (susceptible), ‘Asse’ (adult plant resistant), and ‘Rupee’ (resistant) were analyzed at the vegetative growth stage (5. leaf unfolded) and the generative growth stage (anthesis). At the vegetative growth stage the assimilate export from the mildew inoculated 5. leaf of ‘Amsel’ and ‘Rupee’ is decreased; in ‘Asse’, there is no considerable change of assimilate distribution due to infection. At the generative growth stage the assimilate export from the infected flag leaf of ‘Amsel’ is reduced when the fungus, is sporulating. In the cultivar ‘Asse’ the assimilates are bound at the infection site until the seventh day after inoculation, then the transport of assimilates to the ear is increased. In ‘Rupee’ mildew inoculation causes an enhanced assimilate transport to the ear. The changes in assimilate distribution due to mildew inoculation are discussed with respect to the different types of host-parasite-interactions and the source-sink-activities in the different cultivars.     

亚热带常绿阔叶林木本植物幼树阶段抽枝展叶的权衡关系

亚热带常绿阔叶林植物幼树阶段适应林内生境并开枝散叶是其长成大树的一个重要过程, 植物一年内多次抽枝的现象及其在抽枝展叶过程中小枝伸长、枝茎增粗与叶面积的增加优先顺序及其内在驱动机制还有待进一步研究。该研究对青城山常绿阔叶林木本植物多次抽枝发生比例进行了调查, 并以茶(Camellia sinensis)、细枝柃(Eurya loquaiana)、短刺米槠(Castanopsis carlesii var. spinulosa)、润楠(Machilus nanmu)和大叶山矾(Symplocos grandis) 5种植物的幼树为研究对象, 比较分析了植物在多次抽枝中小枝和叶片生长动态及适应策略的差异。结果显示: 1)一、二次抽枝分别开始于春季(4月)和夏末(8月下旬), 小枝水平上二次抽枝率乔木小于灌木, 常绿植物小于落叶植物。2)一次抽枝小枝枝长、单叶面积, 小枝直径和叶片数量(除大叶山矾外)均高于二次抽枝, 但二次抽枝单叶面积相对生长速率均高于一次抽枝, 二次抽枝叶片比叶质量(LMA)的增长速率高于一次抽枝。3)一次抽枝小枝枝长、叶片数量、小枝直径(除细枝柃和短刺米槠外)和总叶面积(除短刺米槠外)最大相对生长速率均高于二次抽枝, 且大部分物种最大相对生长速率出现在抽枝开始的第一、二周。4)两次抽枝中, 物种先侧重于叶片的生长, 其次是小枝枝长的生长, 最后是小枝直径的增粗。单叶面积和总叶面积皆随着小枝枝长和小枝直径的增加呈显著的异速生长关系, 表明叶片的增长速度大于小枝。单叶面积与叶片数呈显著大于1的异速生长关系, 暗示单叶面积的增长速度大于叶片数的增加速度。小枝枝长与小枝直径也呈显著大于1的异速生长关系, 揭示小枝枝长的增长速度大于小枝直径。综上所述, 两次抽枝过程中, 植物枝叶的优先生长顺序反映了植物为获取更多的资源(尤其是光源)而形成特定的抽枝展叶策略; 二次抽枝单叶面积相对生长速率和LMA增长速率高于一次抽枝, 这可能与植物即将面临的昆虫取食和气温降低压力有关。因此, 了解植物抽枝策略对于理解物种生态适应机制, 揭示物种生活史过程中存在的权衡关系具有重要的理论意义。     

Apoplexy of Peach Trees Caused by Pseudomonas viridiflava

A sudden apoplexy of young nectarine trees was observed during early autumn 1995 in Piedmont (northern Italy). At collar level, extensive wet necrosis of the tissues beneath the bark was observed. When the infection girdled the trunk, the tree died. LOPAT tests, fluorescence on single-carbon sources and comparison of whole-cell protein profiles with type-strains indicated that the bacterial isolates belong to Pseudomonas viridiflava and P. syringae pv. syringae. In the pathogenicity tests, the first bacterium incited symptoms similar to those observed in the field. This is the first record of P. viridiflava as a pathogen of peach.     

Some Etiological Characteristics of Xanthomonas campestris pv. sesami and Disease Reaction of Different Sesame Cultivars

Abstract Isolates of X. campestris pv. sesami used in the present work behaved almost similarly on the basis of their morphological, biochemical and physiological characterization. The studies failed to differentiate any strains within the isolates.
Pathogenicity of the isolates to sesame plants was proved by inoculation experiments. When other plant species were inoculated infection did not occur.
The levels of resistance of some naturally selected sesame cultivars were investigated following artificial infection with the pathogen. Seven cultivars proved resistant, three ( cvs Tozi 3, S-76F₂-22 and K 112) were highly resistant.     

Scanning Electron Microscopy of Pseudomonas syringae pv, morsprunorum on Sweet Cherry Leaves

Scanning electron microscopy indicated that sub-stomatal cavities on sweet cherry leaves are “protected sites” which shelter resident populations of Pseudomonas syringae pv. morsprunorum. Bacteria entered the stomata, multiplied in the cavities and emerged in a mass 6 days after inoculation. There were no visible symptoms, suggesting that the pathogen colonized the host in “sub-clinical” numbers to generate populations which were then released onto the leaf surface under suitable conditions.     

Pre symptomatic detection of wheat leaf rust in the susceptible cv Skalmeje and the resistant cv Esket by means of UV laser-induced fluorescence

In modern agriculture there is a great demand for a rapid and objective screening method for stress resistance, because so far, the resistance of new cultivars is tested in time- and money consuming field experiments. Based on fluorescence ratios, and lifetime of fluorophores measured by fluorescence spectroscopy, we have postulated that an early discrimination of susceptible and resistant wheat cultivars to the leaf rust pathogen Puccinia triticina can be accomplished. As representative for leaf rust resistant and leaf rust susceptible wheat genotypes the cultivars Esket and Skalmeje, respectively, were chosen. Plants were grown under controlled environment conditions and inoculated with the leaf rust pathogen at the second-leaf-stage by single-droplet application. Fluorescence measurements were carried out from two to four days after inoculation (dai) by using a compact fibre-optic fluorescence spectrometer with nanosecond time-resolution. Experimental results indicated that UV laser-induced spectral characteristics as well as determination of fluorescence lifetime are suited to detect leaf rust two dai. For this purpose several ratios and wavelength can be considered. In general, the tested cultivars showed distinct responses to the pathogen development. In this context the ratio F451/F687 measured three dai and mean lifetimes at 500 nm and 530 nm are suited to differentiate the resistant Esket from the susceptible Skalmeje genotypes.     

Nutritional similarity between leaf-associated nonpathogenic bacteria and the pathogen is not predictive of efficacy in biological control of bacterial spot of tomato

It has been demonstrated that for a nonpathogenic, leaf-associated bacterium, effectiveness in the control of bacterial speck of tomato is correlated with the similarity in the nutritional needs of the nonpathogenic bacterium and the pathogen Pseudomonas syringae pv. tomato. This relationship was investigated further in this study by using the pathogen Xanthomonas campestris pv. vesicatoria, the causal agent of bacterial spot of tomato, and a collection of nonpathogenic bacteria isolated from tomato foliage. The effects of inoculation of tomato plants with one of 34 nonpathogenic bacteria prior to inoculation with the pathogen X. campestris pv. vesicatoria were quantified by determining (i) the reduction in disease severity (number of lesions per square centimeter) in greenhouse assays and (ii) the reduction in leaf surface pathogen population size (log(10) of the number of CFU per leaflet) in growth chamber assays. Nutritional similarity between the nonpathogenic bacteria and X. campestris pv. vesicatoria was quantified by using either niche overlap indices (NOI) or relatedness in cluster analyses based upon in vitro utilization of carbon or nitrogen sources reported to be present in tomato tissues or in Biolog GN plates. In contrast to studies with P. syringae pv. tomato, nutritional similarity between the nonpathogenic bacteria and the pathogen X. campestris pv. vesicatoria was not correlated with reductions in disease severity. Nutritional similarity was also not correlated with reductions in pathogen population size. Further, the percentage of reduction in leaf surface pathogen population size was not correlated with the percentage of reduction in disease severity, suggesting that the epiphytic population size of X. campestris pv. vesicatoria is not related to disease severity and that X. campestris pv. vesicatoria exhibits behavior in the phyllosphere prior to lesion formation that is different from that of P. syringae pv. tomato.     

Detection of maize streak virus antigens over time in different parts of maize plants of a sensitive and a so-called tolerant cultivar by ELISA

Maize streak virus (MSV) capsid antigens were detected over time in different parts of maize plants of a sensitive (INRA508) and a so-called tolerant (“tolerant”) cultivar (IRAT297) using a direct or an indirect double antibody sandwich ELISA. Based on three types of experiments, it was shown that the antigens were distributed in the plant according to the age of the tissues. When the virus was inoculated on a particular leaf of 18-day old plants with infective Cicadulina mbila, only the young leaves above the inoculated one were positive by ELISA but not the older ones below. The antigens could not be detected in the inoculated leaf. At day 3 after inoculation, the antigens were detected in the sheath and/or in the whorl of the third leaf above the inoculated one but not in the oldest part of the leaf, the unfolded lamina. Plants of the sensitive cultivar were inoculated at 9 days with C. mbila deposited in the whorl. At 23 h after inoculation, the antigens were detected in the sheath but not in the whorl which was found to be positive only at 32 h. On the basis of these results, a hypothesis of the mode of virus infection is proposed. Our results contribute to a better understanding of the relationship between the age of the plant at inoculation and yield loss as well as secondary infection. By transmission tests with C. mbila, it was shown that virus could only be acquired from leaves exhibiting symptoms. Virus concentrations were measured in plant samples by ELISA using a range of dilutions of purified virus. The virus concentrations were higher in the sensitive than in the “tolerant” cultivar, but no difference in antigen distribution was observed between the two cultivars. The “tolerant” cultivar appeared to be resistant to virus multiplication.     

Common resistance in red raspberry to Botrytis cinerea and Didymella applanata, two pathogens occupying the same ecological niche

Comparisons were made between Botrytis cinerea and Didymella applanata, which occupy the same ecological niche on canes of red raspberry. Isolates of B. cinerea from diverse localities within the British Isles were pathogenic to the SCRI selection M30 when internodal wounds on young canes were inoculated. A single inoculation frequently caused the buds at several nodes to fail the following spring. Differences in the lengths of the stem lesions that formed indicated differences in isolate pathogenicity, but these were not related to isolate origin. Bud suppression and lateral shoot failure also occurred when petioles of cvs Mailing Orion, Mailing Jewel and Glen Clova were wound inoculated with B. cinerea up to late August. The relative resistance of seven cultivars to three isolates of each pathogen was determined. Principal components analysis of data from five estimates of resistance to each pathogen showed that 40% of the variation described a common resistance to the two diseases. Further analysis showed that cvs Chilcotin and Meeker had the strongest common resistance and that cvs Glen Prosen and Mailing Jewel had the weakest. The remaining variation described cultivar differences in relative bud length after petiole inoculation with either pathogen, and differences in the relative importance of spring and autumn symptoms. Only 7% of the variation indicated independent resistance to the two pathogens and this was not influenced by cultivar differences.     

A Laboratory Leaf Assay of Carrot Susceptibility to Botrytis cinerea

Laboratory experiments were conducted to develop an assay for evaluation of carrot (Daucus carota L.) leaf reaction to Botrytis cinerea Pers. ex Pers. The detached carrot leaflets were inoculated with the colonized agar plugs attached to the cut surface of the midrib or by spraying conidial suspension. The estimation of the infected leaflet area expressed as the area under the disease progress curve enabled the discrimination between carrot genotypes differing in their susceptibility to the pathogen. Evaluation based on the measurement of the length of the midrib lesion did not allow such differentiation. The inoculation of leaf surface with suspension of conidia was less reliable and the development of the disease symptoms was not reproducible. The established assay using colonized plugs enables a fast assessment of carrot leaf susceptibility to grey mould and can be particularly useful for a non‐destructive, preliminary evaluation of precious and limited source materials.     

Relationship between Symptom Development and Actual Sites of Infection in Leaves of Anthurium Inoculated with a Bioluminescent Strain of Xanthomonas campestris pv. dieffenbachiae

The infection process of bacterial blight of anthurium was monitored with a bioluminescent strain of Xanthomonas campestris pv. dieffenbachiae. The relationship between symptom expression on infected leaves (assessed visually) and the extent of bacterial movement within tissues (evaluated by bioluminescence emission) varied among anthurium cultivars. In several cultivars previously considered susceptible on the basis of symptom development alone, bacterial invasion of leaves extended far beyond the visually affected areas. In other cultivars previously considered resistant, bacterial invasion was restricted to areas with visible symptoms. In three cultivars previously considered resistant, leaves were extensively invaded by the bacterium, and yet few or no symptoms were seen on infected leaves. The pathogen was consistently recovered from leaf sections emitting bioluminescence but not from sections emitting no light. At an early stage of infection, no significant differences in the percentages of infected areas as determined by visual assessment were observed in any of the cultivars. However, differences among cultivars were detected by bioluminescence as the disease progressed, because bacterial invasion was not always accompanied by symptom expression. In susceptible cultivars, the advancing border of infection was 5 to 10 cm inward from the margins of the visible symptoms and often reached to the leaf petiole even when symptoms were visible in <10% of the total leaf area. Comparisons of anthurium cultivars in which a nondestructive method was used to quantify the severity of leaf infection by a bioluminescent pathogen have enabled us to evaluate susceptibility and resistance to bacterial blight accurately. Such evaluations will be of importance in breeding resistant cultivars for disease control.