Fate of Clavibacter michiganensis ssp. sepedonicus, the Causal Organism of Bacterial Ring Rot of Potato, in Weeds and Field Crops

Crops and weeds were tested for their ability to host Clavibacter michiganensis ssp. sepedonicus (Cms), the causal agent of bacterial ring rot in potato. Ten crops grown in rotation with potato in Europe, namely maize, wheat, barley, oat, bush bean, broad bean, rape, pea and onion and five cultivars of sugar beet were tested by stem and root inoculation. About 6–8 weeks after inoculation, Cms could be detected in most crops except onion and sugar beet, in larger numbers in stems (10⁵–10⁶ cells/g of tissue) than in roots (≤10³ cells/g of tissue) in immunofluorescence cell‐staining (IF). Cms was successfully re‐isolated only from IF‐positive stem samples of maize, bush bean, broad bean, rape and pea, but not from roots. Twelve solanaceous weeds and 13 other weeds, most commonly found in potato fields in Europe, were tested in IF as hosts of Cms by stem and root inoculations. Only in Solanum rostratum, a weed present in northern America, Cms persisted in high numbers (10⁸ cells/g tissue) in stems and leaves, where it caused symptoms. In the other solanaceous weeds, Cms persisted at low numbers (approximately 10⁵ cells/g of tissue) in stems but less so in roots. The bacteria could be frequently re‐isolated from stem but not from root tissues. In 2 consecutive years, plants from 14 different weed species were collected from Cms‐contaminated potato field plots and tested for the presence of Cms by dilution plating or immunofluorescence colony‐staining (IFC), and by AmpliDet RNA, a nucleic acid‐based amplification method. Cms was detected in roots but not in stems of Elymus repens plants growing through rotten potato tubers, and in some Viola arvensis and Stellaria media plants, where they were detected both in stems and roots, but more frequently by AmpliDet RNA than by IFC.     

Widespread distribution of Beet necrotic yellow vein virus in Iranian sugar beet industry

Beet necrotic yellow vein virus (BNYVV) is the most devastating pathogen of sugar beet worldwide. This virus has been reported in the majority of sugar beet growing regions of Iran as well. For the present study, we collected samples from different sugar beet varieties with suspected symptoms of BNYVV from the main important sugar beet growing regions in eight provinces of Iran. Infection of collected samples to BNYVV was tested by ELISA and RT-PCR. Upon testing of 167 collected samples of BNYVV suspected through ELISA and RT-PCR, 115 (68.9%) were infected. Different incidences of BNYVV through surveyed provinces may represent the presence of diverse infective viral sources or resistance genes in tested sugar beet varieties which need further attempts to develop control strategies. Results also showed that BNYVV has been recently distributed throughout some surveyed regions. Otherwise, trace infection or resistance to BNYVV infection in some varieties of distinct regions may represent proper sources of resistance to BNYVV.     

The unconventional geminivirus Beet curly top Iran virus: satisfying Koch's postulates and determining vector and host range

Beet curly top Iran virus (BCTIRV) is a geminivirus with unusual genomic organisation, recently reported in Iran, infecting sugarbeet and a few other plant species. Although three BCTIRV sequences have been reported, demonstration that BCTIRV DNA is the causal agent of the disease was missing. A full‐length genomic DNA was obtained from symptomatic leaves of sugarbeet collected in the Sivand area of Iran, and its nucleotide sequence was determined (BCTIRV‐Siv, 2845 nt). To satisfy Koch's postulates, an infectivity assay was developed by inserting a 1.4‐mer of BCTIRV‐Siv DNA in Agrobacterium tumefaciens and using it in agroinoculation experiments. The cloned viral DNA was capable of infecting sugarbeets, reproducing the leaf curling and vein enations observed in the field. These results demonstrate that the single DNA component of BCTIRV is sufficient for infectivity. Host range studies indicated that some economically important crops can be affected, such as spinach, tomato and sweet pepper, as well as important laboratory plants including Nicotiana benthamiana, Arabidopsis thaliana and Jimson weed. Circulifer haematoceps, the dominant leafhopper species present in sugarbeet fields in Iran, was successfully used to transmit the disease. The availability of an infectious clone will facilitate extended host range studies, to determine the potential risks to other crops, as well as genetic studies on this unusual member of the family Geminiviridae.     

Growth of crop roots in relation to soil moisture extraction

Growth of the roots of sugar beet, potato and barley in the field was observed through glass panels and related to changes in soil moisture measured by a neutron probe during 1969–71. The depth of observed root growth was generally related to, but 10–15 cm deeper than, the maximum depth of soil-moisture extraction. On average of three years, sugar beet, potato and barley used water from the top 23, 33 and 45 cm soil respectively by the beginning of June, and from the top 70, 68 and > 100 cm soil by the end of June. Maximum soil drying in each horizon gave an in situ measure of available water capacity, and showed that sugar beet and barley eventually extracted similar amounts of water from each horizon, but potatoes extracted less, especially from below 60 cm. Between 30 and 100 cm deep, the in situ available water capacity (per 10 cm soil) progressively decreased from 16 to 10, 15 to 5 and 16 to 8 mm under sugar beet, potato and barley respectively. The calculated soil-moisture deficit (potential evapotranspiration minus rainfall) and measured soil moisture deficit were not related early in the growing period before the crops established much leaf cover.     

Trade-off between foliage and tuber resistance to Phthorimaea operculella in wild potatoes

Beet leafhopper, Circulifer tenellus (Baker) (Homoptera: Cicadellidae), is the only known North American vector of beet curly top virus (Geminiviridae), which causes major economic losses in a number of crops including sugar beet, tomato, beans, and peppers. Beet curly top virus is a phloem-limited, persistently transmitted, circulative geminivirus. The strain/species of curly top virus used in this study is the CFH strain, also referred to as beet severe curly top virus (BSCTV). The direct current (DC) electrical penetration graph technique was used to determine the specific stylet penetration behavior associated with inoculation of BSCTV. Viruliferous leafhoppers were allowed to feed on healthy 3–4-week-old sugar beet plants until specific electrical penetration graph waveforms were produced, at which point feeding was artificially terminated. A series of comparisons between leafhoppers that produced different combinations of waveforms clearly implicated waveform D1 as the only waveform correlated with inoculation of BSCTV. All successful inoculations contained waveform D1, and 56 out of 64 leafhoppers that produced waveform D1 successfully inoculated test plants. Eighty-five leafhoppers did not produce waveform D1 and none of these inoculated BSCTV. While the occurrence of waveform D1 appears to be necessary for BSCTV inoculation, there was no correlation between duration of waveform D1 and inoculation success rate. The correlation of waveform D1 and BSCTV inoculation found in this study implies that waveform D1 is associated with phloem salivation.     

Investigation of the Effects of Phosphate Fertilizer Application on the Heavy Metal Content in Agricultural Soils with Different Cultivation Patterns

The use of phosphate fertilizers is essential in agriculture, because they supply farmland with nutrients for growing plants. However, heavy metals might be included as impurities in natural materials and minerals, so heavy metals can also be present in phosphate fertilizers or other chemical fertilizers. The aim of this work was to assess the heavy metal content and contamination status of agricultural soils in the Hamadan province of Iran used for the cultivation of different crops, including cucumber, potatoes, and sugar beet. Surface soil samples were collected and analyzed to determine the total concentration of specific elements (As, Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn), before the pollution index was calculated for each element. Soils used for the cultivation of the three types of crop were not contaminated with As, Cr, Cu, Pb, or Zn. However, the pollution indices for Cd were 1.1, 4.4, and 3.8 in cucumber, potato, and sugar beet fields, respectively, which indicated moderate, high, and high levels of contamination, respectively. Soils from potato and sugar beet fields were heavily contaminated with Cd, which may have resulted from long-term overuse of phosphate fertilizers.     

Arable weeds identified as new sources of beet mosaic potyvirus in Greece

Field and laboratory studies were carried out between 1992 and 1994 to assess the potential for arable weeds to act as reservoirs of beet mosaic potyvirus (BtMV) in fields where sugar beet is cultivated for seed. 933 weed samples were collected and tested by biological methods, while a representative number of weeds were tested by serological methods. The results showed that 97 samples, representing 14 weed species from eight families, were infected with BtMV. The following weeds were found to be naturally infected with BtMV for the first time: Bilderdykia convolvulus, Nonea pulla, Cerastium glomeratum, Galium aparine, Conyza (Erigeron) spp., Fumaria officinalis, Heliotropium europaeum and Rumex spp. The most common weeds found to be infected with BtMV in high percentage were members of the Papaveraceae.     

Beet curly top virus from phloem exudate of Amsinckia douglasiana

Amsinckia douglasiana when infected with beet curly top virus (BCTV) produces more exudate, which is highly infective, than any other known host. Attempts were made to purify BCTV from the phloem exudate of infected Amsinckia douglasiana by differential centrifugation and sucrose density gradient fractionation. A_260/280 ratio of virus preparations was 1.58; S values were 74 and 147. Infectivity was distributed among several fractions during density gradient purification. Electron microscopy revealed the presence of pentagonal and hexagonal particles of 14–23 nm, some of which were paired. The virus was insensitive to DNAse, RNAse, and trypsin when these enzymes were tested individually but infectivity was substantially decreased when the virus was subjected to trypsin treatment followed by DNAse, suggesting that DNA is associated with the beet curly top virus.     

Diet selection by hares (Lepus europaeus) in arable land and its implications for habitat management

Populations of European hares (Lepus europaeus) have experienced a dramatic decline throughout Europe in recent decades. European hares are assumed to prefer weeds over arable crops, and weed abundance was reduced by the intensification of agriculture. Therefore, modern agriculture has been blamed as a major factor affecting European hare populations. However, it is questionable whether European hares select weeds at all, as previous studies had major methodological limitations. By comparing availability and use of plants with Chesson’s Electivity Index, we investigated whether the European hare actually feeds selectively on different plants in arable land. Food availability and use were dominated by cultivated crops (e.g. winter wheat, spring barley and sugar beet). Diet selection analysis revealed that in autumn and winter, European hares predominantly preferred cultivated crops (winter wheat) and food items provided by hunters (tubers of sugar beet and carrot). In spring and summer, apart from soy, only weeds (e.g. clover and corn poppy) were positively selected, especially after cereal crops were harvested. We suggest that the decline in European hare populations throughout Europe was facilitated by the decrease in weed abundance. Wildlife-friendly set-asides in arable land have the potential to reconcile the European Union’s Common Agricultural Policy with wildlife conservation.     

Molecular Characterization and Potential Insect Vector of a Phytoplasma Associated with Garden Beet Witches' Broom in Yazd, Iran

In 2002, garden beet witches’ broom (GBWB) phytoplasma was detected for the first time in garden beet plants (Beta vulgaris L. ssp. esculenta) in Yazd, Iran. Nested polymerase chain reaction (PCR) and restriction fragment length polymorphic (RFLP) analysis of PCR‐amplified phytoplasma 16S rDNA were employed for the detection and identification of the phytoplasma associated with garden beet. A phytoplasma belonging to subgroup 16SrII‐E, in the peanut witches’ broom group (16SrII), was detected in infected plants. Asymptomatic plant samples and the negative control yielded no amplification. The result of analysis of the nucleotide sequence of a 1428 bp fragment of 16S rDNA gene from GBWB phytoplasma (GenBank accession number DQ302722 ) was basically consistent with the classification based on RFLP analysis, in which GBWB phytoplasma clustered with phytoplasmas of the 16SrII‐E subgroup. A search for a natural phytoplasma vector was conducted in Yazd in 2004, in an area where garden beet crops had been affected since 2002. The associated phytoplasma was detected in one leafhopper species, Orosius albicinctus, commonly present in this region. The leafhopper O. albicinctus was used in transmission tests to determine its vector status for the phytoplasma associated with GBWB. Two of eight plants that had been fed on by O. albicinctus, showed mild symptoms of GBWB including stunting and reddening of midveins. A phytoplasma was detected in the two symptomatic test plants by PCR using universal primers and it was identified by RFLP as the GBWB phytoplasma. This finding suggests O. albicinctus is a vector of the GBWB phytoplasma.     

Economic consequences for UK farmers of growing GM herbicide tolerant sugar beet

Weed control is important and one of the more expensive inputs to sugar beet production. The introduction of genetically modified herbicide tolerant (GMHT) sugar beet would result in a major saving in weed control costs in the crop for growers, including control of problem weeds such as perennial weeds and weed beet. However, there would be other economic consequences of growing GMHT beet, some of which would manifest themselves in other parts of the rotation, such as the previous crop, the cereal stubbles that proceed most beet crops, soil tillage and spray application. The average national saving for UK sugar beet growers if they could use the technology would be in excess of £150 ha^?1 yr^?1 or £23 million yr^?1, which includes reductions in agrochemical use of c. £80 ha,^?1 yr^?1 or £12 million yr^?1. However, for some growers, the gains would be much larger and for a few, less than these figures. The possible cost savings are sufficiently large that they could ensure that sugar beet production, with its regionally important environmental benefits as a spring crop, remains economically viable in the UK post reform of the EU sugar regime.     

Effects of increasing weed-beet density on sugar-beet yield and quality

Weed beets are an increasing problem in many sugar-beet crops in many countries. At present about one sugar-beet field in four in England is infested with weed-beet seed. Control in other crops can be achieved using selective herbicides but in sugar beet the weed beets, many of which are of annual habit, are not easily controlled and often compete with the crop. Experiments were done to quantify the yield loss caused by weed beet in sugar-beet crops. Transects were laid out across three fields in 1985 and 1986 and plots located thereon to include the range of weed-beet densities found in the field. Weed beet did not affect the concentration of sugar (sucrose), potassium, sodium, α amino nitrogen or invert sugar in the crop beets. Root and sugar yields were progressively reduced by increasing densities of weed beet. A rectangular hyperbola described the data slightly better than an asymptotic model. There was no indication of a threshold density of weed beet below which there was no yield loss, which averaged 11.7% for each weed beet plant/m². This corresponds to an average 0.6% sugar yield loss for each 1% of bolted weed beet in the root crop up to 100%, which is similar to the reported losses resulting from bolters in the root crop.     

The use of monoclonal antibodies to detect beet mild yellowing virus and beet western yellows virus in aphids

Information on infectivity of the aphids which invade sugar beet root crops each Spring is required for forecasting incidence and providing advice on control of virus yellows. Monoclonal antibodies, produced in the USA to barley yellow dwarf virus (BYDV) and in Canada to beet western yellows virus (BWYV), were used to distinguish between sugar-beet-infecting strains of the luteovirus beet mild yellowing virus (BMYV), and the non-beet-infecting strains of the closely-related BWYV in plant and aphid tissue. Totals of 773 immigrant winged Myzuspersicae and 124 Macrosiphum euphorbiae were caught in water traps in a crop of sugar beet between 25 April and 5 August 1990. Using the monoclonal antibodies and an amplified ELISA, 67%M. persicae and 19%M. euphorbiae were shown to contain BWYV; 8%M. persicae and 7%M. euphorbiae contained BMYV. In studies with live winged aphids collected from the same sugar beet field during May, 25 of 60 M. persicae and two of 13 M. euphorbiae transmitted BWYV to the indicator host plant Montia perfoliata; two M. persicae and two M. euphorbiae transmitted BMYV. In another study three of 65 M. persicae and one of three M. euphorbiae in which only BWYV was detected, transmitted this virus to sugar beet.     

Populations of Rhizoctonia solani in soil under crops in rotation with sugar beet

Using a soil debris isolation method, populations of Rhizoctonia solani were monitored over a 4 -yr period in four fields which were initially cropped to sugar beet and in which four areas of Rhizoctonia crown rot diseased beets (DA) and four areas of apparently healthy beets (AH) had been selected and precisely located. Soil from these areas was assayed during the subsequent crops, which included sugar beet, tomato, cucumber, maize and soybean. No significant differences in colony counts were found between the soils in DA and AH on any of 30 sampling dates. R. solani population counts were, in general, quite low, except under sugar beet and following tomato harvest. Areas of diseased beet and high R. solani soil populations that developed in subsequent sugar beet crops did not necessarily coincide with the previously selected diseased areas. High R. solani populations developed from parasitic activity on sugar beet or saprophytically on tomato crop residues. Of the other crops, both maize and soybean may have slightly increased the low R. solani residual populations in soil. The monitoring of R. solani populations in the season prior to, and during the early season of sugar beet cropping did not provide a basis for forecasting disease in fields or sites within fields. The initiation of disease patches in these sugar beet fields was therefore governed by factors other than inoculum density.     

Outcrossing potential between 11 important genetically modified crops and the Chilean vascular flora

The potential impact of genetically modified (GM) crops on biodiversity is one of the main concerns in an environmental risk assessment (ERA). The likelihood of outcrossing and pollen‐mediated gene flow from GM crops and non‐GM crops are explained by the same principles and depend primarily on the biology of the species. We conducted a national‐scale study of the likelihood of outcrossing between 11 GM crops and vascular plants in Chile by use of a systematized database that included cultivated, introduced and native plant species in Chile. The database included geographical distributions and key biological and agronomical characteristics for 3505 introduced, 4993 native and 257 cultivated (of which 11 were native and 246 were introduced) plant species. Out of the considered GM crops (cotton, soya bean, maize, grape, wheat, rice, sugar beet, alfalfa, canola, tomato and potato), only potato and tomato presented native relatives (66 species total). Introduced relative species showed that three GM groups were formed having: a) up to one introduced relative (cotton and soya bean), b) up to two (rice, grape, maize and wheat) and c) from two to seven (sugar beet, alfalfa, canola, tomato and potato). In particular, GM crops presenting introduced noncultivated relative species were canola (1 relative species), alfalfa (up to 4), rice (1), tomato (up to 2) and potato (up to 2). The outcrossing potential between species [OP; scaled from ‘very low’ (1) to ‘very high’ (5)] was developed, showing medium OPs (3) for GM–native relative interactions when they occurred, low (2) for GMs and introduced noncultivated and high (4) for the grape‐Vitis vinifera GM–introduced cultivated interaction. This analytical tool might be useful for future ERA for unconfined GM crop release in Chile.     

Characterization of Iranian Pectobacterium carotovorum Strains from Sugar Beet by Phenotypic Tests and Whole-cell Proteins Profile

Thirty isolates of Pectobacterium carotovorum from soft rot‐affected sugar beet plants in the Fars province of Iran were characterized phenotypically and by analysis of whole‐cell protein electrophoresis patterns. The isolates were found to be heterogeneous based on the results of physiological and biochemical tests and protein profiles. The results of numerical analysis of phenotypic characteristics and protein patterns showed that only 27% of the collected isolates (phenon 4) could be identified as P. betavasculorum when compared with reference strains. Strains of the first, second, third and fifth phenon shared similar characters with those of P. carotovorum subsp. carotovorum, P. betavasculorum and P. carotovorum subsp. odoriferum, but were distinct from these subspecies. Inoculation of phenon 4 isolates into wounded sugar beet petioles led to black streaking, root rot and vascular necrosis. Other isolates were incapable of causing systemic symptoms in inoculated plants.     

An aggregation attractant for the sugar-beet weevil, Bothynoderes punctiventris

During our screening studies, attractiveness of a ternary mixture of synthetic Grandlure I [racemic cis‐1‐methyl‐2‐(1‐methylenethenyl)‐cyclobutane ethanol], Grandlure II [(Z)‐2‐(3,3‐dimethyl)cyclohexylidene ethanol], and Grandlure III–IV [(Z)‐ and (E)‐2‐ochtodenal; (Z)‐ and (E)‐(3,3‐dimethyl)cyclohexylidene acetaldehyde] for the sugar‐beet weevil, Bothynoderes punctiventris Germar (Coleoptera: Curculionidae), was observed in field‐trapping tests at several sites in Hungary and Serbia. The mixture attracted both males and females. Later tests revealed that of the components in the ternary mixture, only Grandlure III–IV were responsible for attraction, and the addition of Grandlures I or II in varying percentages had no influence on trap captures. Traps baited with 50–50 000 µg of Grandlure III–IV on rubber or polyethylene dispensers yielded high catches of weevils. When testing synthetic samples enriched in the respective geometrical isomer, Grandlure IV had a tendency of catching more weevils, but differences were not significant from lower catches by a 1:1 Z:E blend or Grandlure III. In gas chromatography–flame ionization detection/electroantennographic detection studies, antennae of both female and male weevils were more responsive to the (E)‐ than to the (Z)‐isomer suggesting a more important role for Grandlure IV. Efforts to verify the presence of Grandlure III or IV in volatiles collected from either sex of live sugar‐beet weevils or body washings with pentane remained inconclusive. Traps baited with Grandlure III–IV can now be used as sensitive and powerful trapping tools in the control of the sugar‐beet weevil.     

Needle nematodes (Longidorus spp.) and stubby-root nematodes (Trichodorus spp.) harmful to sugar beet and other field crops in England

Longidorus attenuatus produces galls at the tips of roots of field crops, including sugar beet, growing in alkaline, sandy soils in eastern England. L. elongatus produces similar, but often larger, galls on the roots of sugar beet and other crops in sandy soils in the W. Midlands and in Fen peats. Trichodorus spp. cause ‘stubby root’ of sugar beet and can feed on many field crops. Seven species of Trichodorus were found in sandy soils in eastern England. L. attenuatus, L. elongatus and Trichodorus spp. aggregate around roots and stunt sugar beet and other crop plants. L. attenuatus is commoner below plough depth than in the topsoil, whereas T. cylindricus, T. pachy-dermus and T. anemones are more abundant in the topsoil. These nematodes cause some forms of ‘Docking disorder’.     

Detection of Ralstonia solanacearum in Soil and Weeds from Commercial Tomato Fields Using Immunocapture and the Polymerase Chain Reaction

A detection assay for Ralstonia solanacearum in soil and weeds was developed by combining immunocapture and the polymerase chain reaction (IC‐PCR). Anti‐R. solanacearum polyclonal antibodies were produced in a white female rabbit and Dynal^® super‐paramagnetic beads were coated with purified immunoglobulinG (IgG). Using IC‐PCR, the 718 bp target DNA was amplified at a detection threshold of approximately 10⁴ colony‐forming units (CFU) bacteria per millilitre of suspension. DNA was not amplified in soil suspensions derived from autoclaved and non‐autoclaved soils, which contained R. solanacearum at 1–10⁵ CFU/g soil. However, a positive PCR result was obtained when bacteria in the soil suspensions were first enriched in nutrient broth. IC‐PCR detected R. solanacearum in tomato stems 24 h after inoculation by stem puncture with a suspension containing approximately 10⁵ CFU/ml. IC‐PCR detected the bacterium in 28 of 55 (51%) weeds and 10 of 32 (31%) soil samples. Of the weeds, Physalis minima, Amaranthus spinosus and Euphorbia hirta had the highest incidence of infection. R. solanacearum was not detected in soil taken from fallow fields, but it was discovered in some weed species. Symptomless tomato and pepper plants collected from the fields in which tomato bacterial wilt had previously occurred were found to contain R. solanacearum. These discoveries suggest that weeds and latent hosts may play a role in the survival of R. solanacearum between cropping cycles.     

Rhizobium radiobacter conjugation and callus-independent shoot regeneration used to introduce the cercosporin export gene cfp from Cercospora into sugar beet (Beta vulgaris L.)

Leaf spot disease caused by Cercospora is responsible for crop and profitability losses in sugar beet crops in the US and worldwide. The cfp gene that encodes a protein that exports phytotoxic cercosporins from Cercospora was conjugally transferred to sugar beet using Rhizobium radiobacter (Agrobacterium tumefaciens), to improve Cercospora-induced leafspot resistance. Conditions for shoot regeneration were optimized to increase regeneration/transformation efficiencies. Low-light and room-temperature conditions were favorable to sugar beet regeneration without callus when cytokinin had been added to the tissue culture medium. Using this procedure adventitious shoots from leaf pieces were obtained in a simple, one-step regeneration procedure. T7, a cfp-transgenic clone verified by PCR with gene-specific primers, is being propagated for leaf spot disease resistance evaluation.