Mustard biofumigation disrupts biological control by Steinernema spp. nematodes in the soil

Mustard green manures or seed meal high in glucosinolates, which produce a natural biofumigant upon incorporation into the soil, form an alternative to synthetic fumigants. However, the non-target impacts of these biofumigants in the field are unclear. We examined the effectiveness of soil incorporation of Brassica carinata seed meal both in controlling the plant-parasitic Columbia root-knot nematode (Meloidogyne chitwoodi), and on the biological control exerted by the entomopathogenic nematodes Steinernema feltiae and Steinernema riobrave on root-knot nematodes and the Colorado potato beetle (Leptinotarsa decemlineata). Singly, both the seed meal and Steinernema spp. reduced root-knot nematode damage to potato tubers and increased marketable tuber yields. However, there was a negative interaction between the two bioagents such that their combination did not further improve suppression of plant-parasitic nematodes. Thus, mustard seed meal applications harmful to the target root-knot nematode also disrupted the ability of Steinernema spp. to act as biocontrol agents. Further, we observed modest disruption of the biological control of potato beetles following biofumigation. But, the potato beetles were less likely to lay eggs on potato plants grown in mustard-amended soil, suggesting a counteracting benefit of mustard application. Multiple, complementary controls must be integrated to replace the very effective pest suppression typical of synthetic soil fumigants. Our study suggests significant interference between biofumigation and biocontrol agents in the soil, presenting challenges in combining these two environmentally friendly approaches to managing plant-parasitic nematodes and other pests.     

土壤灭菌-生物有机肥联用对连作马铃薯及土壤真菌群落结构的影响

甘肃省中部沿黄灌区是全国重要的加工型马铃薯生产基地,然而因集约化生产带来的连作障碍问题已经严重影响到当地马铃薯种植业的健康发展。结合田间试验和相关的室内分析,从马铃薯块茎产量和品质、植株生理特征和土壤真菌群落结构等角度,初步评估土壤灭菌和生物有机肥联用(Ammonia Disinfection plus Bio-organic Fertilizer Regulation,ABR)对马铃薯连作障碍的防控效果。同对照相比,ABR处理的块茎产量和商品薯率分别显著增加约71.1%—152.1%和39.2%—53.3%,但块茎化学品质变化不大。ABR处理叶绿素含量和根系活力较CK均显著增加,而叶片和根系丙二醛含量显著下降。PCR-DGGE分析发现,ABR处理显著影响了马铃薯连作土壤的真菌群落结构,表现为真菌群落的多样性指数较CK相比显著下降。ABR处理还有效抑制了土传病害的滋生,植株发病率和收获后的病薯率较CK分别显著下降约67.2%—82.2%和69.1%—70.5%。采用Real-time PCR评估连作土壤中3种优势致病真菌的数量变化,显示ABR处理下立枯丝核菌、茄病镰刀菌和接骨木镰刀菌的数量在生育期内较CK均有不同程度的下降。综合来看,土壤灭菌和生物有机肥联用技术在防控甘肃省中部沿黄灌区马铃薯连作障碍上具有较大的应用潜力,而对土传病害的抑制和微生物群落结构的改善是其主要的作用机理。     

Inoculum Density and Population Dynamics of Suppressive and PathogenicStreptomycesStrains and Their Relationship to Biological Control of Potato Scab

SeveralStreptomycesstrains are capable of suppressing potato scab caused byStreptomyces scabies.Although these strains have been successful in the biocontrol of potato scab in the field, little is known about how populations of pathogenicStreptomycesin the potato rhizosphere are influenced by inoculation of the suppressive strains. The effects of inoculum densities of pathogenic and suppressiveStreptomycesstrains on their respective populations on roots and in rhizosphere soil were examined during the growing season. The relationships between inoculum density or rhizosphere population densities and disease severity were also investigated. Populations of suppressiveStreptomycesstrain 93 increased significantly on roots with increasing inoculum dose. At its highest inoculum dose, the suppressive strain reached a population density greater than 10⁶CFU/g root 14 weeks after planting. The ability of the suppressive strain to increase its populations with increasing inoculum density was hindered at high inoculum doses of the pathogen, suggesting that density-dependent competitive interactions may be occurring between the two antagonists. Strain 93 was most effective at preventing scab early in the growing season (8 weeks after planting), when tubers were most susceptible to the scab disease. Population densities of the suppressive strain in soil were more highly negatively correlated with scab severity than were populations on roots, suggesting that rhizosphere soil rather than potato roots may be the primary source of inoculum of the suppressive strain for tubers.     

The Relative Importance of Seed-borne Inoculum to Common Scab Disease of Potato and the Efficacy of Seed Tuber and Soil Treatments for Disease Control

The importance of both seed and soil-borne inoculum in the epidemiology of common scab disease under Australian conditions was clearly demonstrated. In field trials the severity of disease in harvested potatoes was directly related to the severity of disease on the planted seed tubers. Chemical seed dressing treatments were assessed for common scab disease control under field conditions in four trials over 5 years. Where seed treatments were applied to both diseased and visibly clean seed pieces significantly more disease was found in tubers harvested from diseased seed than the corresponding clean seed treatment. In all but one trial, the treatments applied to diseased seed significantly reduced the incidence of common scab. Fluazinam, flusulfamide (at elevated rates), fenpiclonil, pentachloronitrobezene and mancozeb seed treatments were particularly effective as seed dressing treatments. Applications to visibly clean seed failed to significantly diminish disease levels below that found on untreated seed. Preliminary investigations of some chemical soil treatments gave disappointing levels of control.     

Effects of rotation length, fungicide treatment of seed tubers and nematicide on diseases and the quality of potato tubers

In 1982 – 88, potatoes were grown in 2-, 4- and 6-course rotations with spring barley on a field infested with Globodera rostochiensis. Severity of stem canker and black scurf increased with increasing frequency of previous potato crops, and seed tuber treatment with tolclofos-methyl became less effective in controlling diseases. This suggested that previous crops had increased the amounts of soil-borne inoculum of Rhizoctonia solani. Oxamyl soil treatment increased stem canker in one year and decreased black scurf in four years. Seed tuber treatment with imazalil or prochloraz decreased stem base infection by Polyscytalum pustulans and skin spot and silver scurf on tubers. Black dot was prevalent on tubers in all years and was not affected by seed tuber treatment or previous cropping. Oxamyl increased black dot and common scab in five years and decreased % tuber dry matter in six years. Cysts of G. rostochiensis were found attached to Désirée but not to Maris Piper tubers in August. At harvest tubers of both cultivars were affected by superficial pitting and its severity was related to soil populations of G. rostochiensis at planting. This damage was controlled by oxamyl. It is suggested that the pitting developed from holes made in the tuber skin at larval invasion. In 1989, Désirée seed tubers and healthy mini tubers were planted in all plots and severity of stem canker and black scurf increased with increasing proximity of previous potato crops and with the number of previous crops. Black dot on stems and tubers was not affected by previous cropping but was much less severe in a plot that had not grown potatoes during the seven years of the experiment. The severity of common scab generally decreased as the number of preceding potato crops increased.     

Root, shoot tissues of Brassica juncea and Cereal secale promote potato health

Brassica species are increasingly being used as cover crops to suppress soil-borne diseases in potato cropping systems. Experiments were conducted in controlled environments and in the field to evaluate the effects of cover crop root or shoot or a combination of root and shoot tissues on potato root and tuber health. In a lab assay we examined the extent to which volatile compounds released from tissues of two cover crop species, rye (Cereale secale L.) and oriental mustard (Brassica juncea L.), could inhibit mycelium growth of two important potato diseases, Rhizoctonia solani and Pythium ultimum. Twenty-four hours into the lab assay, volatile compounds from all residues suppressed fungal growth. After 48 h, marked suppression of hyphal growth continued in the presence of mustard residues but not in the presence of rye tissues or the control without tissues. A 75 L volume container experiment evaluated the effect of incorporating different quantities of mustard shoot and root tissues (none, comparable to field level and fourfold field level) into R. solani and P. ultimum infested soil on potato growth, root health and tuber disease. In the container study, incorporating mustard shoots at the highest dose increased potato yield by 54% and reduced disease rating to 2.3 compared to a severe rating of 4.4 in the control. In the field trial, potato growth, root health and tuber disease levels were evaluated in plots where disease management involved either incorporation of mustard or rye cover crop roots, shoots and whole plants (roots plus shoots) or standard farmer practice of a fumigated fallow as a control. White root tissue was used as a health indicator, and averaged 58 and 78% in the fumigated control and mustard cover crop treatments, respectively. The highest healthy root tissue status (91%) was recorded where whole plants of mustard were incorporated. In contrast to the visual assessment of root and tuber health, tuber yield in the field was not influenced by cover crop treatment. Across experiments, the incorporation of or exposure to whole mustard plants was consistently effective at suppressing soil-borne fungi and promoting healthy roots and tubers, especially at higher rates of biomass. Mustard should be managed so as to maximize incorporated biomass for effective biofumigation. Multipurpose management requiring removal of mustard shoots is incompatible with promoting potato rhizosphere health.     

Chemical techniques for control of stem canker and black scurf (Rhizctonia solani) disease of potatoes

Thiabendazole, iprodione and benodanil were tested for control of stem canker and black scurf disease of potatoes using seed tubers with or without black scurf planted in soil infested or not with Rhizoctonia solani . Dormant seed tubers were treated with fungicide dusts, dips or sprays and before planting dusts were applied to soil or to sprouts on seed tubers.
Plants grown from seed tubers with black scurf had more stem canker, stolon infection and Thanatephorus cucumeris and progeny tubers had more black scurf than from seed without black scurf. Infesting soil with R. solani increased black scurf but not stem canker or stolon infection.
Fungicide treatment of seed tubers controlled all phases of the disease and sprout and soil treatments also had significant effects. Numbers of tubers in July were usually increased by treatment although by harvest many small tubers (>3 cm) had been resorbed. Fungicide treatment substantially increased yield only when shoot tips were pruned by R. solani .
Regression analyses showed that stolon pruning, Thanatephorus and black scurf were significantly related to stem canker, and numbers of tubers in July but not in September were inversely related to stolon pruning.
The significance of sources of inoculum and the effects of controlling disease on tuber production are discussed in relation to methods of fungicide use.     

Decreased severity of potato common scab after foliar sprays of 3,5 -dichlorophenoxyacetic acid, a possible antipathogenic agent

Glasshouse tests on the potato cultivars Majestic and Maris Bard measured the effects of single early foliar sprays of ring-substituted phenoxyacetic acids (0.9 × 10–³ M) on the incidence of potato common scab, caused by soil-borne Streptomyces scabies. The most effective compound was 3,5-dichlorophenoxyacetic acid (3,5-D), which decreased scab by about 90%; its action was preventative, early sprays being more effective than late However, it slightly decreased yield, and increased the number of tubers per plant and the proportion of deformed tubers. The trichloro- and tetrachloro- acids containing the 3,5-dichloro- group also decreased scab, though to lesser extents; the most active of these was the 2,3,4,5-tetrachloro- acid, which had the same side effects as 3,5-D. The herbicide 2,4-D almost halved scab incidence, but seriously decreased yield. Other acids tested had little or no effect on scab. In tests against S. scabies in culture, 3,5-D was no more toxic than other disubstituted acids which were inactive, or weakly active, against the disease in vivo. 3,5-D may be an ‘antipathogenic agent’, preventing scab development by altering tuber metabolism.     

The Influence of the Inoculum Source of Rhizoctonia solani on Development of Black Scurf on Potato

Rhizoctonia solani, the causal agent of stem canker and black scurf on potato, survives as sclerotia on tubers, in soil and in plant residues. The objective of the present study was to evaluate the importance of inoculum source on disease development. Disease‐free minitubers and seed tubers contaminated with low levels of R. solani were planted in fumigated or artificially inoculated growth mixture in greenhouse experiments. Black scurf incidence and severity were significantly higher when the inoculum was present in both seed tubers and soil, compared with either of them separately. The severity of disease symptoms on the subterranean parts of the plant also were significantly higher in plots where both seed tubers and soil were contaminated, compared with plots where the inoculum source was either the seed tubers or the soil. Thus, both major sources of inoculum, seed tubers and soil, are important in disease development. However, when both sources are present, black scurf incidence and severity are increased, leading to economical damage to tuber yield and quality. Additional results from field trials support these findings. Disease incidence and severity on daughter tubers were correlated with levels of contamination in seed tubers and soil. When seed tubers and soil were heavily infested, the levels of black scurf incidence and severity on daughter tubers were very high; when seed tuber and soil infestation were very low, black scurf incidence and severity on progeny were also lower. Disease levels were reduced by in‐furrow fungicide treatment, but were less effective when the initial levels of the fungus on the seed tubers and in the soil were high.     

Effect of chitosan and a biocontrol streptomycete on field and potato tuber bacterial communities

Four treatments applied on potato seed tubers i.e. talc only, chitosan only, Streptomyces melanosporofaciens strain EF-76 in talc and S. melanosporofaciens strain EF-76 in chitosan were compared for their efficiency to reduce common scab incidence. As previously reported the joint application of the geldanamycin-producing S. melanosporofaciens strain EF-76 and chitosan was effective to control common scab of potato. Nevertheless, field application of S. melanosporofaciens EF-76 did not allow the selection of geldanamycin-resistant actinomycetes (GRA) in the bulk soil during the potato growing season. The number of GRA on harvested potato tubers was, however, significantly higher in treatments that contained chitosan than in other treatments suggesting that chitosan might promote the establishment of the antagonistic actinomycete on progeny tubers. Biolog EcoPlates were used to determine the metabolic profiles of the bacterial soil communities. A permutation MANOVA analysis detected significant differences within the metabolic profiles of the bacterial communities at the potato flowering period but not at the beginning of the season or a week before harvest. A combination of S. melanosporofaciens EF-76 and chitosan thus represents a promising tool against common scab, with low short-term impact on soil bacterial communities.     

Dry rot of potato [Fusarium caeruleum (Lib.) Sacc.]. Investigation on the sources and time of infection

Inoculation with soil samples proved that the fungus causing dry rot is frequently present in field soils in Cheshire and in soil adhering to imported seed tubers. The fungus was viable in soils having a wide range of p H values and in fields which had not grown potatoes for 5-6 years. Other sources of infection include lofts, used sacks, seed boxes, diseased tubers, and knives used for cutting seed potatoes.
Bruised tubers stored in heavily contaminated boxes developed much dry rot; far less disease occurred in unbruised tubers. In boxes containing own-saved seed, healthy tubers in contact with diseased ones remained sound. Bruised tubers in contact with, or contaminated by, diseased tubers contracted dry rot. Cutting seed with a contaminated knife increased the disease seven-fold.
Inoculation of tubers attached to the parent plant showed that little or no infection occurred before lifting. In field trials severe dry rot developed in several varieties 6-8 weeks after lifting.
The results are discussed in relation to seed treatment.     

Utilization of poultry manure when cultivating potatoes in the southern steppe of the Republic of Bashkortostan

The goal of the research is to determine the effect of pretreated poultry manure and irrigation on the yield and quality of potatoes (Solanum tuberosum) grown in fields located on the southern steppe of the Republic of Bashkortostan. Field experiments were repeated every three years. During vegetation, potato leaves and tubers were studied at the full blossom, leaves’ decay, and potato harvest. The results showed that applying pretreated poultry manure at the rate of 120 t/ha favored higher tuber weight both on the rainfed and irrigated plots (from 0.23 to 0.82 kg/plant and from 0.24 to 1.02 kg/plant, respectively). On the rainfed and irrigated plots where poultry mature was not applied (control fields) the tubers’ weight ranged from 0.08 to 0.31 kg/plant and from 0.16 to 0.50 kg/plant, respectively. Upon application of 40 t/ha of manure, under irrigation, the highest marketable value of tubers was 78%. On the rainfed plots the same value was 72% when applying poultry manure at a dose of 120 t/ha. Soil moisture monitoring showed that the potatoes did not get adequate water during the growing season. When the soil moisture on irrigated plots was 70% less than its minimum water capacity, potato plantings were watered.     

Organic manuring for potato scab control and its relation to soil manganese

The suggestion that organic amendment of soil controls scab by reducing insoluble soil manganese to soluble forms toxic to Streptomyces scabies was investigated in pot and field experiments by assessing numbers of manganese-reducing micro-organisms in soil. Dried grass meal (2 tons/acre) gave significant control of scab but increases in soluble manganese were insufficient to account for this control. Populations of micro-organisms in soil increased after adding grass meal but decreased again by the time tubers were initiated and scab infection began. The results suggest that manganese reduction could affect scab if soil contained easily reducible manganese, because dried grass meal increased the numbers of manganese-reducing micro-organisms.     

Determination of Factors Associated with Natural Soil Suppressivity to Potato Common Scab

Common scab of potatoes is a disease, which is difficult to manage due to complex interactions of the pathogenic bacteria (Streptomyces spp.) with soil, microbial community and potato plants. In Bohemian-Moravian Highlands in the Czech Republic two sites (Vyklantice and Zdirec) were selected for a study of common scab disease suppressivity. At both sites, a field with low disease severity occurs next to one with high severity and the situation was regularly observed over four decades although all four fields undergo a crop rotation. In the four fields, quantities of bacteria, actinobacteria and the gene txtB from the biosynthetic gene cluster of thaxtomin, the main pathogenicity factor of common scab, were analyzed by real-time PCR. Microbial community structure was compared by terminal fragment length polymorphism analysis. Soil and potato periderm were characterized by contents of carbon, nitrogen, phosporus, sulphur, calcium, magnesium, and iron. Quality of organic matter was assessed by high performance liquid chromatography of soil extracts. The study demonstrated that the suppressive character of the fields is locally specific. At Zdirec, the suppressivity was associated with low txtB gene copies in bulk soil, while at Vyklantice site it was associated with low txtB gene copies in the tuberosphere. The differences were discussed with respect to the effect of abiotic conditions at Zdirec and interaction between potato plant and soil microbial community at Vyklantice. Soil pH, Ca soil content or cation concentrations, although different were not in the range to predict the disease severity. Low severity of common scab was associated with low content of soil C, N, C/N, Ca and Fe suggesting that oligotrophic conditions may be favorable to common scab suppression.     

Effect of organic soil amendments on damping-off of lettuce caused by Corticium praticola

Ten organic amendments were added to unsterile soil which was contaminated 14 days later with Corticium praticola and sown with lettuce seeds. Substantial increases in final stands of seedlings were obtained with grass meal, bran and wood cellulose. Corn and barley meal, linseed cake and fish meal decreased final stands; molassine meal, potato starch and peptone had relatively little effect. Seedlings grown with wood cellulose were very chlorotic and stunted. Up to 30% of lettuce seeds sown in soil which, 180 days earlier, had been amended with corn meal and contaminated with C. praticola became colonized by the fungus. None was colonized in unamended soil or in soil amended with grass meal. Ninety days after amendment and contamination fewer seeds were colonized in soil amended with grass meal than in unamended soil. The amendment of soil with grass meal was as effective as thiram seed treatment in protecting lettuce seedlings against C. praticola and grass meal was particularly effective in reducing both the numbers of seedlings attacked and the survival of the fungus in the soil.     

Studies on the lenticel development, surface microflora and infection by common scab (Streptomyces scabies) of potato tubers growing in wet and dry soils

The mechanisms of control of potato common scab (Streptomyces scabies) by irrigation were investigated in 5 years by studying the growth and surface microflora of young potato tubers developing in wet or dry soils. Direct examination of the tuber surface by light or scanning electron microscopy showed that the newly formed internodes near the apex, which carry stomata, had a very sparse microflora. In dry soil the older internodes, which carry lenticels at a susceptible stage, were increasingly colonised by actinomycete hyphae and discrete, although sometimes large, bacterial colonies. In wet soil, actinomycetes were rarely seen on tuber surfaces but bacteria were generally scattered over them, differences not always shown by isolation from periderm pieces onto water agar. When dry scab-infested soil was wetted, scabs did not subsequently develop on the two youngest tuber internodes (A-1, A-2), which may be an example of disease escape rather than inherent resistance of the stomata. The scab control achieved in wet soil was probably caused by some form of microbial antagonism, but whether through competition or antibiosis was not established.     

Chemical control of potato cyst-nematode in sandy clay soil

In sandy clay soil in Bedfordshire, potato cyst-nematode (Heterodera rostochiensis Woll.) was controlled and yields of Majestic potatoes greatly increased when i-2g aldicarb, 4-0 g fensulphothion or 10-3 g diazinon, disulphoton, ‘Isolan’ (i-isopropyl-3-methyl-5-pyrazolyl dimethylcarba-mate) or thionazin per m² were incorporated in the top soil before potatoes were planted. Diethyl phosphorothioates controlled H. rostochiensis better than dimethyl phosphates. One hundred and eleven g methyl bromide, 47-6 g carbon disulphide or 45-2 g D-D per m² applied to the soil and covered with polyethylene sheeting greatly increased the yield of potatoes, but also increased the number of nematodes in the soil. Estimates of the number of larvae able to invade potato roots in treated soil after harvest and of cysts, total eggs or live eggs selected the same range of treatments as effective in controlling H. rostochiensis.     

Effects of foliar sprays of daminozide on the incidence of potato common scab

Single foliar sprays of the growth retardant daminozide (1.5–12 g/l) approximately halved the incidence of common scab, caused by soil-borne Streptomyces scabies, on potted potato plants in the glasshouse. Two analogues of daminozide (N-dimethylaminomaleamic and N-(dimethylamino)-methylsuccinamic acids) also decreased scab, but others were inactive. Of 22 other unrelated growth regulators and translocated chemicals tested as foliar sprays, only gibberellic acid (0.1 g/l) decreased scab incidence, but many of the tubers were distorted. Chlormequat chloride and chlorphonium chloride, as root treatments, were inactive. In other experiments with daminozide, scab incidence was decreased after application to soil. In tests with two plants per pot, spraying one of each pair decreased its rate of stem extension, but did not affect the other, indicating that little or no daminozide passed into the soil from the roots of the sprayed plant. The decrease in scab brought about by foliar sprays was not altered by varying their timing during the period before symptom development (1 to 5 wk after potting). In agar plate tests, daminozide was only weakly toxic to S. scabies. It is concluded that daminozide probably decreased scab by altering the physiology of the plants, so that scab symptoms did not develop.     

Development of a Genotyping Method for Potato Scab Pathogens Based on Multiplex PCR

Scab disease significantly damages potato and other root crops. Streptomyces scabiei, S. acidiscabiei, and S. turgidiscabiei are the best-known causal agents of this disease. We have developed a novel genotyping method for these potato scab pathogens using multiplex PCR, whose benefits include rapid and easy detection of multiple species. We designed a species-specific primer set (6 primers, 3 pairs) for the 16S rRNA genes and 16S–23S ITS regions of these potato scab pathogens. The specificity of the primer set was confirmed by testing 18 strains containing potato scab pathogens, other Streptomyces species, and strains of other genera. The application of the developed method to potato field soil and potato tissue samples resulted in the clear detection and identification of pathogens. Since this method is applicable to a large number of environmental samples, it is expected to be useful for a high-throughput analysis of soil and plant tissues of scab disease.     

Incorporating granular nematicides in soil to control potato cyst-nematode, Heterodera rostochiensis

Incorporating either Du Pont 1410 or Nemacur P at 11-2 kg a.i./ha in peaty loam topsoil in Spring, controlled potato cyst-nematodes (Heterodera rostochiensis) to 20 cm deep as well as did 5.6 kg incorporated in Winter before ploughing followed by another dose of 5.6 kg incorporated in the seedbed in Spring. In pots Du Pont 1410 remained effective after several months incubation in soil at 5 or 10 ^oC. Dazomet at 440 kg/ha incorporated in the topsoil in Winter (220 kg before ploughing and 220 kg after ploughing) did not control the nematodes as consistently as 5.6 or 11.2 kg a.i./ha of Du Pont 141 o or Nemacur P, even when the dazomet-treated plots were covered with Polythene sheeting to prolong fumigation. In large containers, aldicarb at 45 mg a.i./13 l soil increased the yield of Arran Banner potatoes as much when incorporated to 13 cm deep in moderately infested peaty loam as when incorporated to 25 or 38 cm deep, but not as much as when all the soil (to 51 cm deep) was treated. Treating the soil to 13 cm deep did not control the nematodes 13–25 cm deep even though some of the nematicide was probably leached into this layer. In field plots, the nematodes were better controlled when Du Pont 1410 or Dowco 275 was rotavated into the top 10 cm than into the top 20 cm of a peaty loam soil. Rotavating soil twice instead of once after applying aldicarb, Du Pont 1410 or Dowco 275 to the soil surface did not increase nematode control. Although small amounts of aldicarb incorporated into the topsoil in Spring controlled the nematodes, the same amounts concentrated in the seed furrows, just before susceptible potatoes were planted in them, did not.