Note: Survey of seedborne fungi of sudanese cultivars of onion,with new records

Through incubation of onion (Allium cepa L.) seed samples on potato dextrose agar and moist filter papers at 28±2°C, 32 species and four varieties belonging to 19 genera of fungi were determined as seedborne in this crop. Among them, 23 species are new reports to the mycoflora of onion seeds.Aspergillus (11 species and three varieties, ∼42.1% of the total colony count of fungi) was the most prevalent genus:A. niger (∼19.7%) was found at the highest rate in the seeds, followed byA. parasiticus (∼17.3%,A. oryzae (∼17.3%) andA. flavus (∼14.5%). The genusAspergillus was followed byPenicillium (∼12.4%),Sclerotium (∼7.1%),Fennellia, Rhizopus, Chaetomium, Drechslera andAlternaria (∼12.4%),Fusarium, Emericella andByssochlamys (∼7.1%), whereas the remaining eight genera displayed a low level of infection (∼10%). Twenty-seven species, three varieties and 12 genera represent Hyphomycetes; four genera, three species and one variety — Ascomycotina; one genus and one species — Zygomycotina; two genera and two species — other Deuteromycotina; and one genus — Oomycetes. Some species, which are known to cause devastating pre- and postharvest diseases to onion crops, were recovered from the seeds of this crop, suggesting the high possibility of their transmission by seed. http://www.phytoparasitica.org posting July 14, 2004.     

Implications of bacterial contaminated seed lots and endophytic colonization by Pseudomonas fuscovaginae on rice establishment

The economic impact of seedborne bacterial diseases on rice production provides a major motivation for research on seed health. This paper reports on the endophytic growth of a rifampicin‐marked strain of the seedborne rice pathogen Pseudomonas fuscovaginae. The bacterium was found in most tested seeds indicating that, even without visible discolouration, seed transmission is possible. Crushed discoloured seeds contained more bacterial cells than did non‐crushed discoloured seeds. These bacteria were released during seed soaking, contaminating clean seed and lowering seed germination. Cells of a rifampicin‐resistant strain of P. fuscovaginae, which had been inoculated onto rice seeds, were subsequently recovered from different growth stages and from different rice tissues, thereby indicating endophytic colonization. These results have implications for seedling establishment, as symptomless seeds do not assure disease‐free seeds, and the presence of seedborne bacteria results in poor germination and poor seedling establishment. Elimination of seedborne bacteria by soaking in sodium hypochlorite can increase seed germination. This could be used in developing control strategies, and, if practised regularly, reduce entry of seedborne disease‐causing organisms into crops, resulting in lower disease pressure.     

Determination of seedborne fungi and detection of aflatoxins in sudanese fenugreek seeds

Following incubation of fenugreek(Trigonella foenum- graecum L.) seeds on potato dextrose agar and moist filter papers at 28±2°C, 59 species and 11 varieties belonging to 21 genera of fungi were determined as seedborne in fenugreek crops. Among these isolates, 45 species and 9 varieties are new records for this crop; and two species are new to the mycoflora of Sudan:Aspergillus stellifer andEmericella variecolor. The genusAspergillus (15 species and 8 varieties) is the most prevalent, followed byDrechslera (3 species),Rhizopus (3 species),Alternaria andFusarium (6 species each),Emericella (4 species and 2 varieties)Cladosporium andPenicillium (4 species each),Chaetomium (3 species) andCurvularia (3 species and one variety). The remaining 11 genera displayed low level of infection. Of the common pathogens of fenugreek plants,Fusarium oxysporum (2.13%) was recovered from the seeds of this crop. Thin layer Chromatographic analysis of chloroform extracts of 13 seed samples showed that two samples were naturally contaminated with aflatoxins B₁, B₂, G₁ and G₂ (7.5–35.2 μg/kg). http://www.phytoparasitica.org posting Nov. 4, 2001.     

Yucca schidigera extract: a potential biofungicide against seedborne pathogens of sorghum

Treatment of seeds with an aqueous extract of yucca (YE), Yucca schidigera, was evaluated for antifungal activity against seedborne pathogens as well as its effect on seed germination and seedling growth of sorghum (Sorghum bicolor). The antifungal effect of YE was observed against Leptosphaeria sacchari (syn. Phoma sorghina) when the extract was applied at 2·5 and 10% concentrations. At 10% concentration, YE significantly reduced not only the incidence of L. sacchari, but also that of Fusarium spp., Cochliobolus lunatus (syn. Curvularia lunata) and Cladosporium spp. The effect of 10% YE on seedborne fungi was broader than the fungicide fludioxonil, particularly with regard to Fusarium. Furthermore, the number of normal, healthy‐looking seedlings increased in a dose‐responsive manner with YE treatment. Seedling vigour was also stimulated by YE but no correlation was observed with the concentrations tested. Under glasshouse conditions, the treatment of seeds with 10% YE increased the emergence of seedlings and plant height and reduced the number of seedlings with crown rot compared to negative controls and saponin. The positive effect was similar to the effect obtained with fungicide‐treated seeds. Treatment of seeds with synthetic saponin inhibited seedborne fungi less effectively and also negatively affected germination and vigour of the seedlings, compared to the treatment with YE. The results demonstrate an agronomic potential for the use of YE as a biofungicide for seed treatment of sorghum. The difference between the antifungal and the vigour‐stimulating effects of YE warrants further investigation.     

Pea seed-borne mosaic virus: occurrence in faba bean (Vicia faba) and lentil (Lens culinaris) in West Asia and North Africa,and further information on host range,transmission characteristics,and purification

In a survey for viruses of cultivated legumes in West Asia and North Africa, pea seed-borne mosaic virus (PSbMV) was found in faba bean, lentil and pea. Using ELISA, it was detected in 107 out of 1554 faba bean samples and 40 out of 496 lentil samples with virus-like symptoms collected in Algeria, Egypt, Ethiopia, Jordan, Lebanon., Libya, Morocco, Sudan, Tunisia and Turkey.A pea isolate (SP9-88) from Syria was further characterized. Out of 57 plant species tested, 35 were found susceptible, 19 of which are newly reported hosts of the virus. The virus was transmitted efficiently in the non-persistent manner by five aphid species, especiallyMyzus persicae. Purification from systemically infected faba bean plants yielded 10–15 mg of purified virus per kg of infected tissue. Sap-inoculation of the food and forage legume species chickpea, faba bean, lentil, pea,Vicia narbonensis, V. sativa, Lathyrus ochrus andL. sativus at flowering stage led to 66.0, 40.5, 44.6, 49.2, 31.7, 7.5, 35.7 and 12.0% yield loss, respectively, and to seed-transmission, rates of 0.7, 6.0, 10.8, 1.1, 0.3, 0.2 and 0.4%, respectively. No transmission was detected in chickpea seed embryo axes. However, the virus was detected in the seed coat of SPbMV-infected chickpea at an estimated rate of 1.81%.     

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).     

Effects of neemgard on phytophagous and predacious mites and on Spiders

The effects of Neemgard, an acaricidal and fungicidal formulation obtained from neem (Azadirachta indica) seed kernels, on the phytophagous miteTetranychus cinnabarinus, the predacious mitePhytoseiulus persimilis, and the predatory spiderChiracanthium mildei, were investigated in laboratory experiments. Neemgard was highly toxic toT. cinnabarinus but had no toxic effect onC. mildei orP. persimilis. Another neem formulation—the insecticidal Neemix 45—caused conspicuous repellency, but no mortality, inT. cinnabarinus.     

Seedborne infection ofAscochyta rabiei in chickpea and its transmission to aerial plant parts

Ascochyta blight of chickpea (Cicer arietinum L.), caused byAscochyta rabiei (Pass.) Labrousse, was found to be both externally as well as internally seedborne in nature, with the external infection being predominant. The pathogen was located primarily on or in the seed coat and in very few cases penetrated into the cotyledons and rarely, to the embryo. Seedling infection resulted from both externally as well as internally seedborne inoculum. The mode of fungal transmission from seed to the foliar plant parts was strictly non-systemic in nature.     

Evidence for seed transmission and symptomless growth of Ramularia collo‐cygni in barley (Hordeum vulgare)

Ramularia collo‐cygni (Rcc) is becoming an increasing problem for barley growers across Europe. However, the life cycle of the pathogen is only slowly being elucidated. In this study, Rcc DNA was detected in a number of harvested seed samples from 1999 to 2010, with mean levels peaking in winter barley samples in 2009. A number of experiments were carried out to determine whether the pathogen could move from barley seed to seedlings, and also from seed through the developing plant and into the subsequent generation of seed, both in controlled experiments and in field trials. Results from testing of seed indicated that the fungus is widespread at the end of the growing season in harvested grain samples and can be transmitted to developing plants from infected seed stock. Examination of infected seedlings did not reveal the presence of spores but fungal structures were found within the leaf. The location of the fungus within seed was examined, with Rcc DNA found in both embryo and non‐embryo tissue. The implications for barley production of the pathogen being seedborne are discussed.     

Effect of biological seed treatment withCylindrocarpon olidum var.olidum on control of common bunt (Tilletia laevis) of wheat

The goals of the present study were to determine ifCylindrocarpon olidum var.olidum exerts antibiotic activity on teliospore germination ofTilletia laevis in vitro and to evaluate the efficacy of a seed treatment with this antagonist against artificial seedborne infection in the field under natural cropping conditions. Germination of bunt teliospores was completely inhibitedin vitro on water agar medium supplemented with the antagonist’s culture filtrate. In field experiments seed application reduced the incidence of common bunt from 82.9% to 40.4%, and from 81.4% to 42.0% in 1995/1996 and 1997/1998, respectively.C. olidum var.olidum may have a potential againstT. laevis and is worthy of further research on its use as a biocontrol agent for common bunt. http://www.phytoparasitica.org posting July 14, 2005.     

Management of bakanae and bacterial seedling blight diseases in nurseries by irradiating rice seeds with atmospheric plasma

The control of seedborne rice seedling diseases in the seed beds is important to avoid epidemics in rice nurseries and paddies, which may result in severe yield loss. Recently, irradiation with plasma containing electrons, creating positive or negative ions and neutral species, has been shown to have an antimicrobial effect, probably via generation of reactive oxygen species. This study examines whether two seedborne rice seedling diseases, bakanae disease caused by the fungal pathogen Fusarium fujikuroi, and bacterial seedling blight caused by Burkholderia plantarii, are suppressed by irradiation of infected rice seeds with atmospheric plasma. Seed germination and seedling growth were not inhibited in plasma‐treated healthy seeds. When F. fujikuroi‐infected rice seeds were irradiated with plasma after being immersed in sterile distilled water, bakanae disease severity index and the percentage of plants with symptoms were reduced to 18.1% and 7.8% of non‐irradiated control, respectively, depending on the duration of plasma irradiation. The bacterial seedling blight disease index was also reduced by plasma irradiation in vacuum‐inoculated seeds to 38.6% of the non‐irradiated control, and in infected seeds harvested from spray‐inoculated heads of rice plants to 40.1% of the control. Therefore, plasma irradiation seems to be effective in controlling two independent seedborne rice seedling diseases.     

A method for detection of seedborne bacterial diseases in tomato seeds

A method for detection and quantitative estimation of tomato seedborne pathogenic bacteria has been developed. It enables detection in a 7 g tomato seed sample of as few as ten colony-forming units per gram tomato seeds of the following seedborne pathogens of tomato:Pseudomonas syringae pv. tomato,Pseudomonas corrugata, Xanthomonas campestris pv.vesicatoria, andClavibacter michiganense subsp.michiganense. With representative seed samples, the method employs dry grinding, weighing, bacterial extraction and quantitative calculation on selective or semi-selective medium. The efficiency of this method was tested by diluting pathogen-free seed lots with naturally or artificially infested tomato seeds. This procedure enables one to determine the minimal threshold of pathogen which can be detected by this method on media, in comparison with the percentage of diseased seedlings developed from the same seed lots in the growth chamber or in the greenhouse.     

De Invloed van de Uitwendige Omstandigheden Bij Het Optreden van Boonenziekten

Summary The influence of environmental conditions on diseases of the bean (Phaseolus) were studied last summer on the field and at the laboratory. The diseases due to deficiencies of anorganic matter were accompanied by distinct symptoms during June and July, but later on could scarcely be recognized. The dry and warm conditions during both these months were the cause of great damage due to red spider and plant lice. The virus diseases which are transmitted by lastnamed insects, were very much spread, in particular„curlmosaic” (Phaseolus virus 1), „yellow mosaic” or „steengrauw” (Phaseolus virus 2?). „Stipple-streak” (Nicotiana virus 11?) was partly masked during the heat season, but showed in August characteristic redbrown „ringspot” on the pods. This virus disease has wrongly been taken for a bacterial blight. The haloblight, however, is seedborne and gave rise to a heavy attack e.g. on „Ceka” beans in August. In June and July the disease could hardly be discovered in the fields. Further some „rust”- and „foot” diseases have been studied, which are often troubled withUromyces appendiculatus andColletotrichum Lindemuthianum. It was shown thatAscochyta Boltshauseri andAscochyta phaseolorum, both seedborne diseases, caused at first a foot- and stem-anthracnose, which was of little importance in midsummer; but in August the disease spread over the leaves and pods, causing brownish spots that flow together and give a reduction of quality and in yield. The footrots due to Fusarium sp., Sclerotinia sclerotiorum and Botrytis cinerea eem also to increase during cool moist weather.

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Laboratorium voor Mycologie en Aardappelonderzoek, Dec. ’41.     

Multiple virus infections of lablab [<Emphasis Type="Italic">Lablab purpureus</Emphasis> (L.) Sweet] in Nigeria

Leaf samples of Lablab purpureus collected from two agroecological zones of Nigeria—the northern guinea savanna zone (NGSZ) and the derived savanna zone (DSZ)—were infected with viruses when serologically indexed against available antisera. Approximately 31.1 and 81.1% of the leaf samples collected from the NGSZ and DSZ, respectively, were infected. Seven viruses were found: Bean common mosaic virus (BCMV), Cowpea aphid-borne mosaic virus (CABMV), Cucumber mosaic virus (CMV), Cowpea mottle virus (CPMoV), Cowpea severe mosaic virus (CPSMV), Southern bean mosaic virus (SBMV) and Tobacco mosaic virus (TMV) were detected from samples collected from NGSZ, while CMV, CPMoV, Cowpea mosaic virus (CPMV) and CPSMV were detected from samples from DSZ.     

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.     

First report of blueberry red ringspot disease caused by <Emphasis Type="Italic">Blueberry</Emphasis><Emphasis Type="Italic">red</Emphasis><Emphasis Type="Italic">ringspot</Emphasis><Emphasis Type="Italic">virus</Emphasis> in Japan

Virus-like symptoms—red ringspots on stems and leaves, circular blotches or pale spots on fruit—were found on commercial highbush blueberry (Vaccinium corymbosum) cultivars Blueray, Weymouth, Duke and Sierra in Japan. In PCR testing, single DNA fragments were amplified from total nucleic acid samples of the diseased blueberry bushes using primers specific to Blueberry red ringspot virus (BRRV). Sequencing analysis of the amplified products revealed 95.7–97.7% nucleotide sequence identity with the BRRV genome. This paper is the first report of blueberry red ringspot disease caused by BRRV in Japan. The nucleotide sequence data reported in this paper are available in the GenBank/EMBL/DDBJ database as accessions AB469884 to AB469893 for BRRV isolates from Japan.     

Phytotoxic effects of fumonisin B1 on cowpea seed

The cultivation of cowpea (Vigna unguiculata) plays a vital role in the livelihood of many subsistence farmers and rural communities in tropical and subtropical countries. The seeds are prone to fungal infestation and mycotoxin contamination during sub-optimal storage conditions. Fumonisin B₁ (FB₁), produced byFusarium proliferatum, has been detected in cowpea seeds. Surface-disinfected seeds were imbibed in sterile distilled water amended with FB₁ at various concentrations. Percentage germination was determined according to the International Seed Testing Association rules. All the toxin concentrations significantly decreased seed germination and the two highest concentrations — 50 and 100μg ml⁻¹ FB₁ — inhibited root and shoot elongation. FB₁-treated embryonic tissues evinced compaction of the protoplasm and separation of the plasmalemma from the cell wall. Lipid bodies accumulated, which seemed to be lining the cell wall. This is the first study to demonstrate the phytotoxic effects of FB₁ on cowpea seeds.     

Effect of seed treatment of<Emphasis Type="Italic">Arachis hypogaea</Emphasis> with<Emphasis Type="Italic">Bacillus subtilis</Emphasis> on nodulation in biocontrol of southern blight (<Emphasis Type="Italic">Sclerotium rolfsii</Emphasis>) disease

The employment of formulatedBacillus subtilis for peanut seeds (Arachis hypogaea cv. ‘Shulamit’) counteracted the destructive effects of the seedborne pathogenSclerotium (Athelia)rolfsii on the nodulation, leghemoglobin and nitrogenase activity of peanuts. Moreover, the changes in crop vigor index, total nitrogen content and survivability of bothRhizobium spp. andB. subtilis have been related to compatibility and even an occasional synergism between them. http://www.phytoparasitica.org posting Nov. 12, 2006.     

应用蛋白宏阵列快速检测西瓜细菌性果斑病菌

蛋白阵列(Protein array)也叫蛋白芯片,是生物芯片(Biochip)的一种,主要利用抗原/抗体可特异性结合原理制备而成。按阵列样点大小,蛋白阵列又可分为微阵列(Microarray)和宏阵列(Mac-roarray)两类。微阵列集成度极高,可进行高通量