Measurement of the adenosine triphosphate content of cysts of Globodera rostochiensis as a method for assessing the efficacy of fumigant nematicides

The measurement of the adenosine triphosphate (ATP) content of samples of 50 cysts of Globodera rostochiensis collected from 40 fields in the Netherlands has been tested as a method for determining the mortality of the nematodes achieved by fumigation of these field soils. The technique used is based on bioluminescent photometry and it has been fully described previously. The ATP assessment has been compared with results obtained by the advisory service in the Netherlands using hatching tests based on samples of 100 cysts from the same field soils. The ratio of the values before and after soil treatment showed that the methods usually provided similar estimates of mortality. The relationship between ATP and hatched juveniles was also similar from pre-treatment and post-treatment samples. The amount of ATP, calculated from these results, which is equivalent to the maximum post-treatment hatch allowed for a successful treatment in the Netherlands was used in conjunction with the mortality estimates to assess the success of the fumigation. On this basis the two methods disagreed in the overall assessment of fumigation in only five of the 40 fields. This frequency of disagreement is not significantly greater than expected from the variability of either method. The ATP technique clearly offers a reliable and relatively inexpensive method for estimating the efficacy of fumigant nematicides and is much more rapid than the alternative hatching tests or egg counts. Therefore, it could replace these in situations where the grower wanted a rapid assessment on the success, or otherwise, of his fumigation.     

The adenosine triphosphate content of cysts of Globodera pallida and G. rostochiensis as a possible quantitative estimate of field populations

A rapid method is described for estimating the number of juveniles of Globodera pallida and G. rostochiensis in cysts collected from field soils. The technique uses a photometer to measure the adenosine triphosphate (ATP) content of the cysts by bioluminescence. Results for ten to twelve soil samples of 100 g taken from each of four fields showed that the number of juveniles that emerged when the samples were placed in potato root diffusate was significantly related to ATP content of a second series of cyst samples from these fields. Cysts from one field gave a higher ATP content per hatched juvenile than was recorded for the remaining three fields. There are several, untested, explanations of this discrepancy, but some evidence suggests that the hatching test and not the ATP method may have given an unreliable estimate of the viable egg content of cysts from this field. Further work may result in an enhanced precision and increased rapidity in estimating field populations of cyst nematodes by this technique.     

Screening for resistance to potato cyst nematodes using bioluminescent photometry

The measurement of the adenosine triphosphate (ATP) content of cysts by bioluminescent photometry has been used as a screen for assessing the resistance of five potato clones to Globodera rostochiensis Rol and G. pallida Pa3. The multiplication rates (P_f/P_i) of the nematodes are based on the ratio of ATP levels for the cyst inoculum and their progeny. These have been compared to the corresponding ratios based on conventional visual counts of eggs and juveniles. The use of recoverable standard volumes of cysts to inoculate pots has allowed an assessment of the unhatched proportion (C_p) of juveniles from the parental cysts by both methods and an estimate of the corrected multiplication rate (P/P^l-Cp)). This is an accurate assessment of resistance for a clone and the use of standard inocula and the ATP method will enable the criterion adopted for establishing resistance to be measured rapidly and precisely.     

Survival of soft rot coliforms, Erwinia carotovora subsp. carotovora and E. carotovora subsp. atroseptica in soil in Scotland

An enrichment method was used to monitor Erwinia carotovora in soil or the rhizosphere of different crops and weeds in 17 fields with different cropping histories on three farms. The bacteria were detected in all fields not cropped with potatoes, although not consistently, and the mean annual frequency of detection was generally low (< 10%). Fields in which potatoes were grown were extensively contaminated after harvest in September but contamination declined over the winter to very low levels by early summer in the following year. Contamination level tended to rise in some fields without potatoes regardless of their cropping history but for only a short time during autumn and winter. The bacteria were no more frequent in rhizosphere soil of any of the weeds or crops examined, with the exception of brassicas, than in bare soil. In fields where more than 16 months had elapsed since cropping with potatoes, 91% of erwinia isolates obtained were E. carotovora subsp. carotovora , the remainder being E. carotovora subsp. atroseptica. The bacteria were shortlived in soil and in the rhizospheres of inoculated field and pot grown crop and weed plants. Longevity was greater in dry (10% moisture) than in wet (21% moisture) soil and decreased as temperatures rose, particularly above 25°C. Survival was best in association with brassica plants, moderate on grasses and cereals, and least on potatoes and weeds. E.c. carotovora survived better than E.c. atroseptica. Because survival of the bacteria in soil is apparently restricted, their presence in fields could be attributed to recurrent introductions from different sources.     

Effect of crop rotation on populations of Epitrix tuberis (Coleoptera: Chrysomelidae) in potato

The effect of crop rotation on populations of tuber flea beetle, Epitrix tuberis Gentner, in potatoes was investigated using data supplied by an integrated pest management (IPM) company and Geographic Information System software and conventional statistical methods. Using combined 1995 and 1996 data, beetles of the overwintered and F1 generations in both the interior and edges of potato fields showed a significant linear increase with an increase in the preceding consecutive years (0, 1, and 2 years) that the current years' crop was planted to potatoes. Populations were significantly higher in nonrotated fields compared with rotated fields. Both the percentage of the cropping region requiring insecticidal control of tuber flea beetles and the cost of insecticides per hectare of potatoes grown increased linearly with an increase in the number of previous years planted to potatoes. Not practicing crop rotation resulted in a 4.2-7.3% increase in the cropping region requiring insecticidal control of tuber flea beetles. The cost of controlling beetles in potato fields planted to potatoes for 3 consecutive years was up to $20/ha greater than potatoes rotated from the preceding year. Beetle counts from the interior of rotated potato fields never exceeded threshold levels when field edges also were below threshold. It is concluded that sampling of overwintered beetles in interior sites of rotated fields could be abandoned, and only 1 monitoring scout rather than 2 would be necessary to monitor a field during this time. From these results, we concluded that rotating potato crops would reduce spray costs to the farmer and monitoring costs to IPM companies.     

Changes in the adenine nucleotide content of cysts of Globodera rostochiensis associated with the hatching of juveniles

The content of AMP, ADP and ATP within single cysts of Globodera rostochiensis (60–80 fig dry weight) was determined as ATP to an accuracy of ± 10^-11 mol by a bioluminescent technique, after microenzymic methods had been used to phosphorylate AMP and ADP to ATP. Results for a total of 120 cysts showed that a change occurs in the adenylate energy charge of their contents after they have been exposed to potato root diffusate. Cysts in water had a mean adenylate energy charge of 0–63 (s.E. ± 0–04), but a randomly selected group of cysts after 24 h treatment with potato root diffusate had a significantly lower mean of 0–49±0–04. In a second, similar experiment, cysts in diffusate for only 8 h had an energy charge of 0*55 ± 0–03, but this value was not significantly less than the corresponding mean of o-6i ±0–03 of cysts that remained in water. The results indicate an effect on the metabolism of the unhatched juveniles that occurs too soon after the addition of diffusate to be directly due to any increase in locomotor activity. Apparently, the primary action of the hatching factor had affected many juveniles within 24 h of the addition of potato root diffusate to the cyst.     

The behaviour of populations of potato cyst eelworm Heterodera rostochiensis Woll. Towards resistant potato varieties derived from Solarium tuberosum ssp. andigena Juz & Buk

Many populations of Heterodera rostochiensis mostly from the main potato growing areas of England and Wales, were tested with eelworm-resistant potato varieties (andigena hybrids). Except in the Eastern Region, where resistant varieties were planted alongside commercial varieties in infested fields, tests were done in pots inoculated with a standard number of cysts extracted from a single soil sample from each field. The tests showed that England and Wales could be divided into areas according to the percentage of cysts(< 10%, 11–70%,> 70%) produced on the resistant compared with a commercial non-resistant variety. On the peat soils of the East Anglian fens and in Bedfordshire, Essex and Kent (area A) most populations produce few cysts on resistant varieties. In much of the East Midlands and Yorkshire (area D), populations produce many cysts on resistant varieties which behave much as ordinary susceptible potatoes. Populations in the rest of England and Wales (areas B, C) vary and in many districts every field must be tested separately. If the pathotypes occurring in Britain had separate origins in South America, the present distribution in England and Wales is probably due to chance introductions dispersed with seed and following the patterns of trade in seed potatoes. As all commercial varieties, before the release of eelworm-resistant ones, were about equally susceptible to the different pathotypes, the varieties themselves probably played little part in determining the present distribution of pathotypes.     

Selection of a rapidly maturing population of Globodera rostochiensis by continuous cultivation of early potatoes in Ayrshire, Scotland

Development of three G. rostochiensis populations was compared after 2, 3, 4, 5 and 13 wk at 12 and 15 °C on cv. Pentland Crown. The Jameston Farm (JF) and Skelmorlie populations originated from Ayrshire, Scotland, the former where early potatoes are grown annually, the latter from a farm where potatoes are grown in a conventional manner. The Feltwell population was originally from Norfolk, England. At 12 °C, nematodes from the JF population penetrated roots and developed more quickly than nematodes from the other populations. The JF population also produced the most cysts, which were the largest and contained most embryonated eggs. At 15 °C, differences between the populations were less marked, but JF nematodes invaded roots quickest and produced cysts with most eggs. These results are consistent with the hypothesis that the annual practice of early planting and harvesting to control losses caused by G. rostochiensis in Ayrshire is selecting a nematode population adapted genetically to the procedure.     

Testing potatoes bred from Solarium vernei for resistance to the white potato cyst-nematode, Heterodera pallida

Three clones of potatoes bred from Solanum vernei were tested for their resistance to the white potato cyst-nematode, Heterodera pallida, both by root-ball counts and by estimates of total cysts produced. Two susceptible clones were used as controls. Root-ball counts suggested a higher degree of resistance than that based on total cysts produced. There were differences in pathogenicity between the two populations. The degree of resistance found was such that in practice the resistant clones would be best used in conjunction with nematicide treatment in a system of integrated control.     

Effect of cropping systems on cereal stemborers in the cool‐wet and semi‐arid ecozones of the Amhara region of Ethiopia

1 Field experiments were conducted on maize and sorghum at three locations in the Amhara state of Ethiopia to determine the effects of mixed cropping on stemborer infestation, borer natural enemies and grain yields. In the cool‐wet ecozone of western Amhara, sole maize was compared with maize intercropped with faba bean, mustard, potatoes and cowpea. In the semi‐arid ecozone of eastern Amhara, the trial was conducted on both maize and sorghum with the companion crops haricot bean, sesame, cowpea and sweet potatoes. 2 The results showed that the predominant borer species in western and eastern Amhara were, respectively, Busseola fusca and Chilo partellus. In Addis Zemen, western Amhara, maize intercropped with mustard and potatoes had significantly lower pest numbers and percent tunnelling than other intercrops and the maize monocrop during the vegetative stage. In eastern Amhara, the cropping system did not significantly affect pest densities but damage to stem, ear or heads tended to be greatest when cereals were intercropped with sweet potatoes. 3 Parasitism of C. partellus by the braconid Cotesia flavipes was greater on maize than sorghum, and on maize it was greater with sweet potatoes than in other intercrops or sole maize. Cocoon mass number per plant did not vary significantly between treatments. 4 There were significant differences between treatments in yields of both sorghum and maize (per plant and per unit area) with the lowest yields observed when they were intercropped with a tuber crop. 5 The results suggest that simultaneous planting of the crop species selected has little advantage over monocropped maize.     

A method for estimating the contribution of seed potatoes, machinery and soil tare in field infestations with potato cyst nematodes on a national scale

In order to make a cost benefit analysis for the management of the potato cyst nematodes Globodera rostochiensis and G. pallida, we developed a method to estimate the relative importance of three basic distribution channels of potato cyst nematodes: seed potatoes, machinery and soil tare. The baseline is determined by the area planted with potatoes, the area infested with potato cysts, the proportion of resistant potato cultivars and the distribution of cysts trough different channels. This quantification forms a basis for the evaluation of the effects of different control measures for potato cyst nematode on a national scale. The method can be useful as an example for application in other countries.     

On cyst colour changes, bionomics and distribution of potato cyst-eelworm (Heterodera rostochiensis Woll.) pathotypes in the East Midlands

Pot tests and field trials in 1966 confirmed earlier observations that pathotype A potato cyst-eelworm (Heterodera rostochiensis Woll.) showed marked colour differences from pathotypes B and C during development on the roots of susceptible potatoes. In hatching tests started in July, newly formed brown cysts of pathotype A released four times as many larvae as those of pathotypes B and C, without entering an immediate diapause; the latter produced larvae after a rest period of 18–21 days. Subsequent second-generation cyst production in glass-tube culture was greater from pathotype A. Field observations suggested that all pathotypes mature at about the same time and give rise to only one flush of cysts on both early and maincrop potatoes in the growing season. Cyst colour surveys in commercial crops, and pot and field trials using resistant potato selections, confirmed that pathotypes B and C are both dominant and widespread in the East Midlands.     

Influence of nematicide, resistant and susceptible potato cultivars and bare fallow on the population dynamics of Globodera rostochiensis Woll. Rol under field conditions in Spain

The susceptible potato cultivar Désirée and the resistant cv. Diamant were cultivated on untreated and nematicide-treated plots on two fields in northern Spain infested with Globodera rostochiensis Rol. Initial and final infestations of cysts and viable eggs were assessed. Multiplication rates differentiated more accurately the effects of the different treatments than final number of cysts and eggs, due to the elimination of some of the errors associated with variability of the initial infestation. Many new cysts were observed on all plots where susceptible potatoes had been grown, whether they had been treated with nematicide or not, although significantly fewer cysts were actually found on the nematicide-treated plots. With the resistant cultivar no significant effects of the nematicide were detected even though there was a noticeable reduction of viable cyst contents over and above the effect of the resistant cultivar.     

Endophytic bacterial systems governing red clover growth and development

Endophytic competent bacteria capable of promoting both beneficial and detrimental growth responses in red clover (Trifolium pratense L.) were recovered from three adjacent areas of farmland each having a different cropping history — continuous red clover, continuous potatoes (Solanum tuberosum L.) or a 2 yr rotation of red clover and potatoes. The population composition of these rhizobacteria was altered by the various crop sequences. The greatest instance of significant growth responses (beneficial or detrimental) occurred with those bacteria derived from the clover-potato soil, suggesting increased interactive ‘competition’ among bacterial populations at the ‘interface’ between different crop rotations. Whether bacterial strains promoted or inhibited growth appeared to depend on the cropping history and prior exposure of pre-bacterised clover plants to the natural microflora in the peat-based growing media. The interaction between bacterial colonists influenced plant trait expression to the degree that some characteristics were completely masked. Improvements in plant growth were interpreted as an allelopathic side-effect of the competition between endophytes for the same ecological niche, from which the plant inadvertently benefits.     

Soil Fungal Diversity and Community Composition in Response to Continuous Sweet Potato Cropping Practices

Soil fungi are extremely important for maintaining soil health and plant production in agricultural systems. Currently, the effect of continuous cropping of sweet potato on soil fungal communities and physiochemical parameters has not been well documented. In the present study, four sweet potato fields consecutively monocultured for 1, 2, 3, and 4 years were selected to investigate the effect of monoculture on soil fungal communities through Illumina MiSeq sequencing. Continuous cropping of sweet potatoes dramatically altered the fungal community composition, whereas fungal diversity was almost unchanged. Ascomycota and Basidiomycota were the most abundant phyla in all soil samples, accounting for 32.59% and 21.14% of the average relative abundance, respectively. The abundance of some potential pathogens, such as Ascobolus spp, specifically Ascobolus stercorarius, and some unknown fungi increased significantly as the sweet potato monoculture period increased, and their presence were highly positively correlated with disease incidence. In contrast, Basidiomycota, Bullera, Fusarium and Trichocladium most likely play roles as antagonists of sweet potato disease development, as their relative abundance decreased significantly over time and were negatively correlated with disease incidence. Redundancy and correlation analyses revealed that soil pH and organic carbon content were the most important factors driving these changes. Our findings provided a dynamic overview of the fungal community and presented a clear scope for screening beneficial fungi and pathogens of sweet potato.     

不同大豆连作年限对黑土细菌群落结构的影响

大豆连作导致作物产量下降、病原微生物富集和土壤退化等问题日趋严重。然而,目前关于大豆连作对土壤细菌群落结构组成及多样性分布的影响及发生机制尚不清楚。采用高通量测序技术,对大豆连作(不同年限)和大豆-玉米轮作下的黑土细菌16S rRNA基因进行测序分析。结果表明:轮作5年(CR5)和13年长期连作(CC13)处理显著增加了土壤pH、全氮(TN)、全磷(TP)和速效养分(AN、AP和AK)含量。与短期连作相比,CR5和CC13处理均提高了细菌群落的OTUs数量、PD值、Chao1指数和Shannon指数。聚类分析图谱结果显示细菌群落结构组成受到轮作和连作年限的双重影响,而土壤pH、TN、TP、AN、AP和AK是细菌群落结构发生变化的主要驱动因子(P0.05)。此外,VPA分析发现上述土壤因子中,土壤pH对细菌群落结构变化的贡献度最大。本研究证明大豆长期连作提高了土壤养分含量和细菌群落的丰富度和多样性指数,从分子生物学的角度证实大豆长期连作在一定程度上改善了土壤环境,为大豆连作障碍的研究提供了理论依据。     

Effect of the trap crop Solanum sisymbriifolium and two biocontrol fungi on reproduction of the potato cyst nematode,Globodera pallida

The potato cyst nematode, Globodera pallida, is one of the most important pests of potato worldwide. Owing to regulatory considerations and potential environmental impact, control options for this nematode are becoming increasingly limited. Solanum sisymbriifolium and biological control agents offer viable alternative options for controlling G. pallida. Therefore, experiments were conducted to determine the effect of the nematode trap crop S. sisymbriifolium, alone or in combination with the biocontrol agents Trichoderma harzianum or Plectosphaerella cucumerina, on population decline of G. pallida. Experiments were conducted for three different ‘cropping systems’: potato (Solanum tuberosum), S. sisymbriifolium, or soil only (fallow), each followed by a potato crop. Soil was amended with P. cucumerina, T. harzianum or left unamended, and then infested with nematodes at a rate of five eggs g^?1 of soil. After 16 weeks in the greenhouse, plants were removed and the soil containing cysts was refrigerated at 4°C for 8 weeks, and then planted to potato. Cysts of G. pallida were counted after an additional 16‐week period. The P_f/P_i of G. pallida was significantly reduced by 99% in potato following S. sisymbriifolium compared to both the potato‐following‐fallow and the potato‐following‐potato treatments. Amendment of soil with T. harzianum significantly reduced P_f/P_i of G. pallida by 42–47% in the potato‐following‐potato but not in either the potato‐after‐fallow nor in the potato‐after‐S. sisymbriifolium cycles which supports evidence that the plant species may play a role in the biocontrol activity of this fungus. Addition of the fungus P. cucumerina resulted in a 64% decrease in P_f/P_i in the potato‐following‐fallow in one experiment, and an 88% decrease in P_f/P_i in potato‐following‐potato but the decrease in P_f/P_i was not consistent over all experiments. However, both biocontrol fungi resulted in lower numbers of progeny cysts after an initial 16‐week incubation with potato. To look at the effect of varied population density of the nematode on efficacy of S. sisymbriifolium to reduce G. pallida populations, potato, S. sisymbriifolium, or barley were planted into soil infested with G. pallida at rates of 5, 20 or 40 eggs g^?1 soil applied as cysts (20, 80 or 160 cysts pot^?1). After 16 weeks, numbers of cysts produced in each treatment were determined for each infestation rate. No new cysts were recovered from either S. sisymbriifolium or barley treatments, confirming that neither plant is a host for G. pallida. High numbers of cysts were recovered with potato. Soil from each treatment (containing original cysts and newly‐formed cysts when present) were then planted with potato. After an additional 16 weeks, few cysts were found in the potato‐after‐ S. sisymbriifolium treatments regardless of initial infestation rate. When potato followed barley, numbers of cysts were similar to those found after a single cycle of potato, indicating that the barley crop had no effect on the survival of initial inoculum. Overall, these results suggest that S. sisymbriifolium has potential to significantly reduce G. pallida populations, and also that the cropping system (i.e. the sequence of non‐host and host plants) may play a significant role in the efficacy of fungal biological control agents.     

Plant–parasite coevolution: A weak signature of local adaptation between Peruvian Globodera pallida populations and wild potatoes

Plant–parasite coevolution has generated much interest and studies to understand and manage diseases in agriculture. Such a reciprocal evolutionary process could lead to a pattern of local adaptation between plants and parasites. Based on the phylogeography of each partner, the present study tested the hypothesis of local adaptation between the potato cyst nematode Globodera pallida and wild potatoes in Peru. The measured fitness trait was the hatching of cysts which is induced by host root exudates. Using a cross‐hatching assay between 13 populations of G. pallida and root exudates from 12 wild potatoes, our results did not show a strong pattern of local adaptation of the parasite but the sympatric combinations induced better hatching of cysts than allopatric combinations, and there was a negative relationship between the hatching percentage and the geographical distance between nematode populations and wild potatoes. Moreover, a strong effect of the geographic origin of root exudates was found, with root exudates from south of Peru inducing better hatching than root exudates from north of Peru. These results could be useful to develop new biocontrol products or potato cultivars to limit damages caused by G. pallida.     

Weed species diversity and community composition in cropping practices at two intensity levels – a six-year experiment

Alternative cropping practices can be expected to increase the diversity and change the species composition of weed communities. In a six-year field experiment we compared species diversity and species composition in the weed community of cereal fields cultivated by conventional and by low-input cropping methods. In the conventional cropping both mineral fertilizers and herbicides were applied but in the low-input cropping the fertlizer was manure and herbicides were avoided altogether. The level of nitrogen fertilization was higher in conventional cropping. A five-year crop rotation comprising rye, oat-pea, barley and two-year grasslands was used in both cropping methods. We found that the species pool was larger (42 vs 35 species) and the average number of species higher (20.2 vs 13.1 species) in low-input than in conventional cropping. This was the case both in the data pooled across crops and in each crop separately. Herbicides affected species diversity and species composition more than did nitrogen fertilization. Species susceptible to the herbicides applied declined in abundance, which resulted in more skewed dominance patterns in the communities of conventional fields than in those of low-input fields. The crop affected the species composition of weed communities more than did the cropping method. Our results suggest that weed species diversity could be promoted by using low-intensity cropping practices.