Detection of latent infection of Erwinia chrysanthemi and Pseudomonas caryophylli in carnation1

Three methods for detecting latent infections of Erwinia chrysanthemi and Pseudomonas caryophylli in carnation stems were studied. The same procedure was used, in all three methods, to prepare the samples and to extract and concentrate bacteria. The samples consisted of 100–500, 1-cm-long, pieces of carnation stems. Under the standard procedure, the samples were homogenized in maceration fluid, then centrifuged at low speed to eliminate the larger tissue fragments and centrifuged again at high speed to obtain a final pellet with a maximum concentration of the bacteria to be detected. Immunofluorescence staining (IFAS), direct isolation (DI) and immunoenzymatic test (ELISA) were all applied to the final pellets. IFAS had a sensitivity threshold equal to 8.65 × 10⁴ and 3.33 × 10⁴ bacteria per 100 stem pieces for E. chrysanthemi and P. caryophylli. In simulated latently infected lots IFAS could detect down to 0.2% infected pieces for both pathogens, both at high and at low infection level. Although DI had a similar theoretical sensitivity threshold for both bacteria, it was less successful when applied to simulated latently infected lots. ELISA had the highest theoretical sensitivity threshold for both bacteria and as a consequence could only detect them in simulated latently infected lots with a high infection level. The comparative results are discussed in relation to the applicability of the methods to commercial lots of carnation.     

Characteristics of immunofluorescence microscopy and of dilution-plating to detect Pseudomonas syringae pv. phaseolicola in bean seed lots and for risk assessment of field incidence of halo blight

Routine laboratory testing of 710 bean seed lots from various origins forPseudomonas syringae pv. phaseolicola (Psp) with immunofluorescence microscopy (IF) showed that 27.5% of the seed lots (five subsamples of 1000 seeds tested per sample) contained two or more IF-positive cells in a total of 500 microscope fields (magnification 500×). Simultaneously performed dilution-platings of IF-positive subsamples on King's medium B confirmed presence of Psp for one-third of these IF-positive seed lots. The grey area of disagreement between both laboratory tests was studied by comparison of test data and by field trials.The number of IF-positive cells per subsample was positively correlated with isolation and identification of Psp (R=0.85). The detection level of IF was ca. 10² Psp cells per ml of undiluted subsample extract. The detection level of Psp by isolation on King's medium B was variable, being inversely related with the saprophyte to Psp ratio. The high sensitivity of IF was in part due to high percentages of dormant or dead IF-positive cells in the sample extract. Field trials over two years with 10 000 seeds per seed lot, showed disease incidence for 9 of the 22 seed lots. Of ten IF-positive lots with five positive subsamples per sample, nine were positive in the field test plot (the negative lot gave primary infection spots of Psp when used for commercial growing). By isolation, seven of these ten IF-positive lots were positive. Of the five IF-positive lots with two or less positive subsamples, isolation and field trial were both negative. Based on data on seed transmission from literature, field incidence was unlikely for these five samples in a 10 000 seeds field trial. All seven IF-negative lots were negative in the field trial. The value of IF and isolation for indexing bean seed lots for Psp is discussed.This study was carried out at the Centre for Plant Breeding and Reproduction Research (CPRO-DLO), Binnenhaven 1, P.O. Box 16, 6700 AA Wageningen the Netherlands, to which address correspondence should be addressed.     

Detection of Corynebacterium michiganense in tomato seed lots1

Corynebacterium wilt of tomato causes severe losses in France and Belgium. The disease can be seed-transmitted and sanitary selection of seedlots is a direct prophylactic control method. It must be selective, objective and sensitive and must provide information on the pathogenicity of the bacteria detected. A method is proposed that associates immunofluorescence (IF) staining with bioassay and isolation. In a preliminary comparison of inoculation on different host plants, tomato seedlings with two or four leaves were the most sensitive. Artificially and naturally contaminated seed extracts were IF-screened and confirmed by isolation of the pathogen from inoculated seedlings. The detection method consists of maceration of tomato seeds in PBS-cycloheximide buffer, filtration and centrifugation of the seed extract, and IF screening of the pellets with two antisera. Pellets with positive or suspect IF reading (threshold 5 × 10³ cells in the pellet) are inoculated into tomato seedlings incubated at 25°C. Within 30 days the seedlings are inspected for typical wilt symptoms and isolation experiments are carried out from the vessels. The method has been tested on commercial seedlots.     

Epiphytic Curtobacterium flaccumfaciens strains isolated from symptomless solanaceous vegetables are pathogenic on leguminous but not on solanaceous plants

In Iran, during 2013–16, 16 Gram‐positive corynebacteria‐like strains were recovered from the epiphytic parts of solanaceous vegetables including eggplant, pepper and tomato. The strains were recovered accidentally as a result of monitoring for other bacterial pathogens in solanaceous fields. The strains were phenotypically different from Clavibacter michiganensis strains. Although none of the strains were pathogenic on their host of isolation or on any other solanaceous plants, 12 out of 16 strains were pathogenic on common bean, cowpea, mung bean and soybean. Colonization by strains was observed on maize, zucchini, faba bean, honeydew melon, rapeseed, sugar beet and sunflower plants under greenhouse conditions. In PCR tests, the primer pair CffFOR2/CffREV4, specific for Curtobacterium flaccumfaciens pv. flaccumfaciens, enabled the amplification of the appropriately sized fragment in 12 out of 16 strains, and all 12 strains were pathogenic on dry beans. Phylogenetic analysis, using the gyrB and recA genes, showed all 16 bacterial strains clustered within several pathovars of C. flaccumfaciens. A nonpathogenic yellow‐pigmented strain (Xeu15) was clustered with the type strains of C. flaccumfaciens pv. betae and C. flaccumfaciens pv. oortii. Bacteriocin profiling assays revealed no significant differences among the pathogenic and nonpathogenic strains. Host range and population dynamics of four representative strains on 17 plant species showed population build‐up of the strains only on common bean, cowpea, wheat and red nightshade plants. The results provide important insights into the possible role of nonhost plants as reservoirs of plant pathogenic bacteria, which has important implications in plant disease epidemiology and management.     

Contamination of bean seeds by <Emphasis Type="Italic">Xanthomonas axonopodis</Emphasis> pv. <Emphasis Type="Italic">phaseoli</Emphasis> associated with low bacterial densities in the phyllosphere under field and greenhouse conditions

Xanthomonas axonopodis pv. phaseoli and its variant fuscans are the causal agents of common bacterial blight of bean. Production of seeds is recommended in arid climates with the use of pathogen-free seeds. However, contamination of seeds still occurs in these seed production areas. To verify if low contamination levels of sown seeds could explain these field contaminations, we used seeds that were naturally contaminated with CFBP4834-R, a rifamycin-resistant X. axonopodis pv. phaseoli fuscous strain, to contaminate field plots at different rates. We also inoculated seeds to verify some parameters of plant colonization and seed transmission. In growth chambers, seedling contamination was always successful from seeds contaminated with CFBP4834-R having population sizes greater than 1 × 10³ CFU seed⁻¹ and were not successful below 1 × 10² CFU seed⁻¹. In the greenhouse, the efficiency of contamination of seeds was not significantly different between contaminated plants that had a low or a high CFBP4834-R population size and reached between 40% and 52% whatever the origin of the inoculum (aerial or seed-borne). In field experiments, under low relative humidity, plots with 0.1–0.003% contamination rates or plots sown with seeds that were inoculated with low CFBP4834-R population sizes (1 × 10² and 1 × 10⁴ CFU seed⁻¹) led to an asymptomatic colonization of bean during the entire growing season with low CFBP4834-R population sizes. Seeds were contaminated both in primary and secondary foci. The contamination of seeds without symptom expression during the growing season represents a risk for eventual disease outbreaks.     

Detection of Clavibacter michiganensis ssp. michiganensis in tomato seeds by immunofluorescence microscopy and dilution plating

A method for detectingClavibacter michiganensis ssp.michiganensis in tomato seeds was evaluated. The method is based on rapid screening of tomato seed lots using indirect immunofluorescence staining (IF), followed by dilution plating of IF positive seed lots. Different polyclonal antisera, prepared againstC. michiganensis ssp.michiganensis were tested for their specificity using IF. All strains ofC. michiganensis ssp.michiganensis tested reacted with the polyclonal antisera. Two of nine saprophytic isolates from tomato seeds were positive with the antisera as well as with the control normal serum, but cells of these isolates were distinct in shape from cells ofC. michiganensis ssp.michiganensis.For extraction of the pathogen from the seed, seeds were either blended with a stomacher or soaked at 4–6 °C. The stomacher method yielded more fluorescent cells in IF than 24 h soaking of seed samples. However, soaking of seeds for 48 h generally yielded less saprophytes and overall higher numbers ofC. michiganensis ssp.michiganensis colonies in dilution plating when compared to blending by a stomacher. SCM medium was generally more selective than KBT and modified CNS medium. However, the efficacy of the medium was dependent on the seed lot and/or extraction method used. Confirmation of suspected colonies with YDC (yeast-dextrose-carbonate medium), IF and a pathogenicity test on tomato seedlings proved to be highly reliable (P>0.95). For routine testing of seed lots it is recommended to screen tomato seed lost after soaking seeds for 24 h at 4–6 °C with IF, followed by plating of IF-positive seed lots on modified CNS and SCM after soaking seeds for an additional 24 h.     

绿磺隆、醚苯磺隆及其体外代谢物对乙酰乳酸合成酶的离体抑制作用

利用异源表达于酵母细胞中的小麦细胞色素P450cDNA(CYP71C6v1)研究了磺酰脲类除草剂绿磺隆、醚苯磺隆的代谢作用。结果表明,代谢产物5-羟基绿磺隆和5-羟基醚苯磺隆能够抑制乙酰乳酸合成酶(ALS酶)活性,且代谢产物与母体化合物绿磺隆、醚苯磺隆抑制ALS酶活性的IC50值差异小,但是代谢产物在茎叶喷雾小麦和菜豆时,均未表现出活性。绿磺隆及其代谢产物抑制小麦ALS酶活性的IC50值分别为7.1×10-9和7.9×10-9mol/L,抑制菜豆ALS酶活性的IC50分别为3.6×10-9和4.1×10-9mol/L;醚苯磺隆及其代谢产物抑制小麦ALS酶活性的IC50分别为4.6×10-9和5.3×10-9mol/L,抑制菜豆ALS酶活性的IC50分别为4.7×10-9和4.9×10-9mol/L。结果表明,在磺酰脲类分子苯环5位上进行结构改造,有可能得到高活性的化合物。     

Direct evidence of surface infestation of seeds and tubers by Plasmodiophora brassicae and quantification of spore loads

Using quantitative PCR, DNA of Plasmodiophora brassicae, the causal agent of clubroot, was detected and quantified on canola, pea and wheat seeds, as well as on potato tubers, all harvested from clubroot‐infested fields in Alberta, Canada. Quantifiable levels of infestation were found on seven of the 46 samples analysed, and ranged from <1·0 × 10³ to 3·4 × 10⁴ resting spores per 10 g seeds; the vast majority (80–100%) of resting spores on these samples were viable, as determined by Evan’s blue vital staining. However, the levels of infestation found were generally lower than that required to cause consistent clubroot symptoms in greenhouse plant bioassays. While the occurrence of P. brassicae resting spores on seeds and tubers harvested from clubroot‐infested fields suggests that seedborne dissemination of this pathogen is possible, practices such as commercial seed cleaning may be sufficient to effectively mitigate this risk.     

A quantitative real‐time PCR assay for detection of Colletotrichum lindemuthianum in navy bean seeds

Bean anthracnose is a seedborne disease of common bean (Phaseolus vulgaris) caused by the fungal pathogen Colletotrichum lindemuthianum. Using seed that did not test positive for the pathogen has been proven to be an effective strategy for bean anthracnose control. To quantify the extent of anthracnose seed infection, a real‐time PCR‐based diagnostic assay was developed for detecting C. lindemuthianum in seeds of the commercial bean class navy bean. The ribosomal DNA (rDNA) region consisting of part of the18S rDNA, 5^.8S rDNA, internal transcribed spacers (ITS) 1, 2 and part of the 28S rDNA of seven races of C. lindemuthianum, 21 isolates of Colletotrichum species and nine other bean pathogens were sequenced with the universal primer set ITS5/ITS4. Based on the aligned sequence matrix, one primer set and a probe were designed for a SYBR Green dye assay and a TaqMan MGB (minor groove binder) assay. The primer set was demonstrated to be specific for C. lindemuthianum and showed a high sensitivity for the target pathogen. The detection limit of both assays was 5 fg of C. lindemuthianum genomic DNA. To explore the correlation between the lesion area and the DNA amount of C. lindemuthianum in bean seed, seeds of the navy bean cultivar Navigator with lesions of different sizes, as well as symptomless seeds, were used in both real‐time PCR assays.     

A competition index for Orobanche crenata Forsk effects on broad bean (Vicia faba L.)

The influence of broomrape (Orobanche crenata Forsk) infection levels on commercial broad bean (Vicia faba L.) crops was investigated and an equation relating yield losses to O. crenata density was derived. O. crenata growth was weakly and negatively related to the final height or shoot number of the broad bean plant. O. crenata decreased seed yield mainly by reducing the number of pods. The number of seeds per pod was also decreased by O. crenata infection when competition occurred at late-crop growth stages when most of the pods had already developed. Bean unit seed weight was only weakly affected by infection. Correlation coefficients between O. crenata number and broad-bean seed yield per plant were significant at P=0·05 and greater than 0·70 in most locations. On average, about four O. crenata per broad-bean plant decreased seed yield by half. An estimation of the percentage of losses in any field can be calculated from the equation: %loss = 100 × 0·124 × OcN. where OcN is the average final number of emerged O. crenata per plant.     

Interpretation of the EC method for the detection of latent Corynebacterium sepedonicum infections in potato1

The EC method for the detection of latent ring-rot infections (Corynebacterium sepedonicum) consists essentially of an indirect immunofluorescence test and a pathogenicity test on eggplant (EP). When interpreting the results of this method, care should be taken that: (a) eggplants are grown at 21°C. At 28–29°C the detection level of the EP was increased 10² to 10³-fold to 10⁵ to 10⁶ cells ml^-1 and latent infections (which may go unnoticed) occurred with 10⁴ to 10⁵ cells ml^-1; (b) reisolations are made from symptomless eggplants grown at the optimum temperature (21°C) as latent infections can also occur in these plants at concentrations of 10² to 10³ cells ml^-1; (c) potato tuber extracts are tested without freezing or stored freeze-dried or in a proper protective agent. Freezing at -20°C in maceration buffer of cells of C. sepedonicum and Pseudomonas solanacearum (the latter was included for comparison) reduced their numbers by 90–98% after one day. This could easily cause viable cell numbers to drop below the detection level of the pathogenicity test.     

Effectiveness of Coniothyrium minitans and Trichoderma atroviride in suppression of sclerotinia blossom blight of alfalfa

The effects of the mycoparasites Coniothyrium minitans and Trichoderma atroviride on the suppression of alfalfa blossom blight caused by Sclerotinia sclerotiorum were evaluated under indoor and field conditions. When T. atroviride (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (9·0 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) were applied to detached young alfalfa florets, T. atroviride effectively inhibited saprophytic growth of S. sclerotiorum, whereas C. minitans showed no inhibition under the same conditions. When T. atroviride (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) or C. minitans (6·9 × 10⁴ conidia/floret) + S. sclerotiorum (6·0 × 10³ ascospores/floret) was applied to young alfalfa petals in vivo just after pollination, the percentage of pod formation was higher for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum, and the percentage of pod rot was lower for T. atroviride+S. sclerotiorum than that for C. minitans+S. sclerotiorum. However, when they were applied to senescent petals attached to developing pods of alfalfa at 9·2 × 10⁴ conidia/floret together with S. sclerotiorum at 4·5 × 10³ ascospores/floret at 14 days after pollination, C. minitans was more effective than T. atroviride in suppressing sclerotinia pod rot and seed rot of alfalfa. Field experiments showed that three applications of C. minitans (5·4 × 10⁶ conidia mL⁻¹) or T. atroviride (5·4 × 10⁶ conidia mL⁻¹) at a 7-day interval to blossoms of alfalfa effectively suppressed sclerotinia pod rot in two out of three annual trials. Coniothyrium minitans effectively suppressed sclerotinia seed rot in all three years, whereas T. atroviride was not effective against seed rot in any of the trial years. The efficacy of C. minitans was not significantly different (P > 0·05) from benomyl (250 µg ai mL⁻¹). This study suggests that C. minitans has potential as a biocontrol agent to control blossom blight of alfalfa caused by S. sclerotiorum.     

Real‐time quantitative PCR assays for the rapid detection and quantification of Fusarium oxysporum f. sp. phaseoli in Phaseolus vulgaris (common bean) seeds

Fusarium oxysporum f. sp. phaseoli (Fop) is a devastating pathogen that can cause significant economic losses and can be introduced into fields through infested Phaseolus vulgaris (common bean) seeds. Efficient seed health testing methods can aid in preventing long‐distance dissemination of this pathogen by contaminated seeds. In order to improve detection of Fop in seed, a rapid, accurate and sensitive real‐time PCR assay (qPCR) protocol was developed for detection of Fop in common bean seeds. Seed lots of seven cultivars with infection incidence ranging from 0·25 to 20% were prepared by mixing known amounts of Fop‐infected seeds with Fop‐free seeds. Direct comparisons between SYBR Green and TaqMan qPCR methods were performed using primers based on the Fop virulence factor ftf1. The primers developed in this study produced a 63 bp product for highly virulent strains of Fop but did not produce an amplicon for nonpathogenic or weakly pathogenic isolates of F. oxysporum from P. vulgaris or other hosts. Under optimized conditions, both qPCR assays detected Fop infection at low levels (0·25%); however, the results suggest the TaqMan assay was more reliable at quantification than the SYBR Green assay. Linear regression models were fitted to the relationships between results of qPCR assays and infection incidence, but the models differed among cultivars. Fungal biomass per seed differed among cultivars and was related to seed size. The results indicate that the TaqMan assay developed in this study is a useful tool for the detection and quantification of Fop in bean seeds.     

Comparison of extraction procedures for detection of Xanthomonas axonopodis pv. phaseoli in common bean seed

Xanthomonas axonopodis pv. phaseoli (Xap) is an important seedborne pathogen of Phaseolus vulgaris. Accurate seed health testing methods are critical to protect seed quality and meet phytosanitary requirements. Currently employed selective media‐based methods include several variations in extraction procedures. In order to optimize pathogen extraction from seeds, the influence of different extraction steps on the sensitivity of Xap detection was assessed. Seeds were inoculated by vacuum infiltration with Xap to achieve inoculum levels from 10¹ to 10⁵ CFU per seed; one contaminated seed was mixed into 1000‐seed subsamples of uncontaminated P. vulgaris seeds. Thirty subsamples of 1000 seeds were tested using each different extraction procedure. These included soaking whole seeds in sterilized saline phosphate buffer, either overnight at 4°C or for 3 h at room temperature, with or without vacuum extraction, and either with or without concentrating the seed extract by centrifuging. Seed extract dilutions were cultured on semiselective agar media MT and XCP1. The percentages of positive subsamples were compared to measure the effects of each extraction step on detection sensitivity. Vacuum extraction and centrifugation of seed extracts increased sensitivity; the highest sensitivity was obtained with the 3 h vacuum extraction followed by centrifugation. These results were confirmed with naturally infested seeds; Xap was detected in 48 of 70 samples using the 3 h vacuum extraction with centrifugation, whereas only 35 of 70 field samples tested positive using overnight soaking, a significant difference. The results suggest that these steps would be valuable modifications to the current method approved by the International Seed Testing Association (ISTA).     

A high temperature requirement for after ripening of imbibed dormant Poa annua L. seeds

Freshly harvested seeds of Poa annua L. collected in south Louisiana were stored in moist soil at seven temperatures between 5°C and 35°C. At monthly intervals, seed lots were removed and germinated at each of the seven temperatures. Seed were dormant for at least 1 month at all test temperatures. Seeds stored for 2 months at 30 and 35°C showed conditional dormancy; there was 100% germination at 10 or 15°C, and poorer germination at 5 or 20°C. Seeds started to lose viability after 2 months at 35°C and were dead after 7 months. In seeds stored at 10–30°C, there were increased percentages and a wider range of germination temperatures as storage time or storage temperatures increased. Seeds stored at 10°C remained dormant for 9 months, but by 12 months of storage the seeds germinated only at 5 or 10°C. Nearly all seeds stored at the same temperatures in air dry soil remained dormant for 6 months, regardless of storage temperature. These results differ from other reports of low temperatures breaking seed dormancy in Poa annua L. and suggest an adaptation to subtropical climates.     

Evaluation of seedborne bacterial pathogens on common bean cultivars grown in central Anatolia region,Turkey

Bacterial diseases of bean cause economically important losses worldwide. The most important method for managing bacterial diseases on bean is the use of pathogen-free seed. In this study, 198 different dry bean seed samples of six different cultivars including Dermason, Cali, Sira, Battal, Bombay and Seker, were collected from 12 provinces of Central Anatolia Region of Turkey. All were tested to investigate the seedlots as primary inoculum sources of the major bacterial diseases. The data revealed that 22,72 %, 13,63 %, 11,11 %, 1,51 % and 0.5 % of seed samples tested were contaminated with five seedborne bacterial pathogens, Pseudomonas savastanoi pv. phaseolicola (Psp), Pseudomonas syringae pv. syringae (Pss), Xanthomonas axonopodis pv. phaseoli (Xap), X. axonopodis pv. phaseoli var. fuscans (Xapf) and Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), respectively. All bacterial strains isolated were identified based on morphological, physiological, biochemical, molecular and pathogenicity tests. The results showed that Psp and Pss were found together on cv. Cali; Psp and Xap on cv. Dermason and cv. Sira; and Pss and Xap on cv. Seker, cv. Dermason, and cv. Cali. Therefore, the results in the present study suggested that evaluation and selection of pathogen-free seeds are very important for preventing the spread of pathogens and effective management of seed borne bacterial diseases prevalent in bean growing regions; in addition to implementation of integrated crop production strategies such as crop rotation, sanitation, seed treatment, tolerant/resistant cultivar selection and proper bactericide application.     

Seed bank and other demographic parameters of broomrape (Orobanche crenata Forsk.) populations in faba bean (Vicia faba L.)

A study on the population dynamics of broom-rape (Orobanche crenata Forskal) in faba bean (Viciafaba L.) was conducted in two locations for 8 years. The O. crenata seed bank increased every year in the 20-cm depth arable layer until it reached approximately 4 million (M) seeds m^?2. Seed viability ranged between 53% and 68%, approximately half of the seeds remaining dormant. About 3 × 10^?3% of the seed bank became attached to the root system of the faba beans. Only 9% of the attached broomrapes developed and emerged from the soil, possibly reflecting high levels of intraspecific competition. Maximum broomrape seed production for a population of 53 emerged broomrapes per m² was approximately 4 M seeds m^?2. About 43% of the seeds produced were not incorporated into the soil, most probably as a result of their degradation, decomposition or dispersion. Stock semencier et autres paramètres démographiques de I'orobanche crénelée (Orobanche crenata Forsk.) dans des cultures de féverolle (Vicia faba L.) Une étude de la dynamique de populations d'orobanche crénelée (Orobanche crenata Forsk.) dans des cultures de féverolle (Viciafaba L.) a été menée pendant 8 années sur deux sites. Le stock semencier de O. crenata a augmenteéchaque année dans la couche (20 cm) arable du sol jusqu'à atteindre 4 millions (M) de graines m^?2. La viabilité des graines était comprise entre 53 et 68%, approximativement la moitié d'entre elles présentaient une dormance. Environ 3 × 10^?3% du stock semencier adhérait au système racinaire de la féverolle. Seulement 9% des orobanches adhérantes se développaient et levaient, ce qui pourrait refléter une compétition intraspécifique importante. La production maxi-male de graines par une population de 53 orobanches levées au m²était d'environ 4 M graines m^?2. Environ 43% des graines produites n'étaient pas incorporées au sol, très prob-ablement en raison de leur dégradation, de leur décomposition ou de leur dispersion. Bodensamenbank und andere populationsde-mographische Parameter der Sommerwurz Orobanche crenata Forsk. in Bestanden der Acker-Bohne (Vicia faba L.) Bei einer Untersuchung der Populationsdy-namik der Sommerwurz Orobanche crenata Forsk. in Beständen der Acker-Bohne (Vicia faba L.) an 2 Orten über 8 Jahre hin nahm die Bodensamenbank in der Pflugzone von 20 cm Tiefe jedes Jahr zu, bis sie rund 4 Millionen Samen m^?2 erreichte. Die Lebensfähigkeit der Samen lag bei 53 bis 68%, etwa die Hälfte war dormant. Ungefähr 3 × 10^?3% kamen mit den Bohnenwurzeln in Berührung. Nur 9% davon entwickelten sich und wuchsen heran, worin vielleicht der hohe Grad intraspezifischer Konkurrenz zum Ausdruck kommt. Die höchste Zahl gebildeter Samen eines Sommer-wurzbestands von 53 Pflanzen m^?2 war rund 4 Millionen. Etwa 43% der gebildeten Samen gelangten nicht in den Boden, wahrscheinlich weil sie zerstört oder verweht wurden.     

The influence of soil and other physical factors on the antifungal activity of carbendazim against Rhizoctonia solani

The inhibition of the growth of Rhizoctohia solani in vitro by carbendazim was maximal at 20°C and at pH 8. In pot tests using mung bean, maximum protection against ‘damping-off’, caused by R. solani, was obtained when seeds were treated with carbendazim. at 1 g a. i. kg^?1 (as a wettable powder) and sown in river sands of pH 7 and 8, kept at a 20°C. Better disease control was obtained in soils kept moist by frequent watering than in soils under water stress. Disease control was best in sandy soil and least in clay loam. The implications of these results, for the antifungal efficacy of carbendazim under diverse soil conditions, are discussed.     

Survival of Curtobacterium flaccumfaciens pv. flaccumfaciens in weeds

Weeds are important alternative hosts of pathogens, responsible for the survival and spread of phytopathogenic bacteria. Our study evaluated the potential of weeds as hosts of Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), causal agent of bacterial wilt, one of the main diseases of common beans. Cff survival was evaluated in the phyllosphere and in the rhizosphere of 21 weeds, in four experiments under field conditions, during the years 2018 and 2019. The aerial part of the plant was inoculated by spraying bacterial suspension (10⁷ cfu/ml) of Cff, while the soil of the growing pots was infested with the same suspension. Cff survival was evaluated every 7 days, for 70 days. The identity of the bacterium was confirmed by PCR with the specific primers CffFOR2 and CffREV4, from strains recovered from all samples. Principal component analysis (PCA) showed that high temperatures and rainfall reduced Cff survival in the phyllosphere, while high temperatures reduced the survival of the bacterium in the rhizosphere. Our results demonstrated that Amaranthus viridis (family Amaranthaceae), Conyza bonariensis, Emilia fosbergii, Galinsoga parviflora, Gnaphalium purpureum (Asteraceae), Raphanus sativus, Lepidium virginicum (Brassicaceae), Commelina benghalensis (Commelinaceae), Ipomoea triloba (Convolvulaceae), Cyperus rotundus (Cyperaceae), Senna obtusifolia (Fabaceae), Digitaria insularis (Poaceae), Nicandra physalodes, and Solanum americanum (Solanaceae) are potential hosts for Cff. Their eradication in common bean fields is recommended, especially in fields with a history of bacterial wilt occurrence.