A Plant Health Care Program for Brambles in the Pacific Northwest

Pratylenchus and Xiphinema species have been associated with decline and mortality of brambles (Rubus species) in the Pacific Northwest of the United States. These nematodes cause direct feeding damage and (or) transmit viruses that result in poor fruit quality and plant decline. A nematode management program has been developed by the author to minimize chemical use and nematode-induced damage while optimizing fruit production. Nematode management is an integral part of a total plant health care program in which foliar and soil pests, plant stresses, and fertility are managed.     

Effect of Introduced Pseudomonas fluorescens Strains on Soil Nematode and Protozoan Populations in the Rhizosphere of Wheat and Pea

Abstract Previous studies have shown that inoculation of pea seeds with Pseudomonas fluorescens strains F113lacZY or F113G22 increased mineralization of organic nitrogen in the rhizosphere. In contrast, inoculation of the same strains onto wheat seeds reduced mineralization of N from organic residues incorporated into soil. In the present study, we report on a likely explanation of this phenomenon, which appears to be governed by the effect of plant-microbe interactions on bacterial-feeding nematodes and protozoa. In soil microcosm tests, inoculation of pea seeds with Pseudomonas fluorescens strains F113lacZY or F113G22 resulted in an increase in the number of nematodes and protozoa in the rhizosphere as compared to noninoculated controls. This trend was repeated using a model sand system into which the bacteriophagous nematode Caenorhabditis elegans was introduced. It was subsequently found that non-inoculated germinating pea seeds exerted a nematicidal effect on C. elegans, which was remedied by inoculation with either strain F113lacZY or F113G22. This suggests that nematicidal compounds released by the germinating pea seeds were metabolized by the microbial inoculants before they affected nematode populations in the spermosphere or rhizosphere of pea. In contrast, inoculation of wheat plants resulted in significantly lower nematode populations in the rhizosphere, whereas protozoan numbers were unaffected. No nematicidal effects of inoculated or noninoculated wheat seeds could be found, suggesting that microfaunal populations were affected at a later stage during plant growth. Because of their key roles in accelerating the turnover of microbially immobilized N and organic matter, plants that support a larger microfaunal population are likely to benefit from a higher availability of inorganic nitrogen. Therefore, an understanding of plant-microbe interactions and their effects on soil microfaunal populations is essential in order to assess the effects of microbial inocula on plant mineral nutrition. Received: 27 May 1999; Accepted: 15 July 1999; Online Publication: 17 December 1999     

Effectiveness of Hirsutella minnesotensis and H. rhossiliensis in control of the soybean cyst nematode in four soils with various pH,texture, and organic matter

The parasitism of soybean cyst nematode, Heterodera glycines, by the fungi Hirsutella rhossiliensis and Hirsutella minnesotensis and their biocontrol effectiveness against the nematode were investigated in four soils with various pH, texture, and organic matter. Fungal parasitism was assayed in the soils in 25 mL vials. As expected, percentage of H. glycines second-stage juveniles (J2) parasitized by either fungus increased with increasing number of fungus-colonized J2 initially added into the soils. Parasitism of J2 by the fungi was negatively related with soil pH. Both positive and negative relationships with fungal parasitism were observed for soil sandiness and organic matter. In greenhouse study, both fungi at 0.2–0.8 g fresh mycelium of liquid culture per 0.3 L pot and 1% corn-grits culture effectively reduced nematode population density. The relationship between biocontrol effectiveness and the soil factors depended on fungal species and inoculation levels. In general, percentage reduction of egg population density in the soil was negatively correlated with soil pH and positively correlated with sandiness. There was no or weak correlation between egg reduction and organic matter. The percentage of J2 parasitized by the fungi 2 months after planting did not correlate with the soil factors. Plant growth was better in the two soils with intermediate pH and sand than the soil with high pH and low sand or with low pH and high sand. It appeared that soil pH and/or texture are important in influencing biocontrol effectiveness, but further studies are needed to determine the effect of individual factors because they are correlated.     

Bacillus-based bionematicides: development,modes of action and commercialisation

Agricultural crops are severely damaged by root-knot nematodes causing extensive financial losses globally. Historically, agrochemicals have been the preferred method to combat these pests; however, threats to humans and the environment posed by these agrochemicals led to the need for developing new biocontrol agents. Importantly, the latter should adhere to biosafety regulations while being highly effective. Root-knot nematodes live in soil and thus the use of rhizobacteria such as Bacillus for biocontrol development have shown potential. Although various Bacillus species have been tested in this capacity, little is known about their secondary metabolites and the mechanisms of action responsible for their nematicidal activity. If these secondary metabolites can be qualitatively and quantitatively characterised, metabolic features could be synthetically engineered and used to combat root-knot nematodes. Although there is great potential for bionematicides, the commercialisation and development of such products can be difficult. This review summarises the importance of Bacillus species as natural antagonists of root-knot nematodes through the production of secondary metabolites. It provides an overview of the significance of root-knot nematodes in agriculture and the advances of chemical nematicides in recent years. The potential of Bacillus species as biocontrol agents, the known mechanisms of action responsible for the nematicidal activity demonstrated by Bacillus species, non-target effects of biocontrol agents and the commercialisation of Bacillus-based bionematicides are discussed.     

Resporulation of Metarhizium anisopliae granules on soil and mortality of Tenebrio molitor: Implications for wireworm management in sweetpotato

In Australia, sweetpotato (Ipomoea batatas L.) is vulnerable to root feeding insect pests such as wireworms (e.g., Agrypnus spp.). The number of registered insecticides to control these insect pests is limited and often pest pressure, for example by wireworms, is severe close to harvest, further limiting what insecticides can be applied. Incorporating biological control agents such as entomopathogenic fungi (e.g., Metarhizium anisopliae) into integrated pest management programmes may be feasible in sweetpotato. M. anisopliae has been shown to be effective in controlling more than 200 insects and it is able to reside and grow in the rhizosphere and rhizoplane, suggesting that M. anisopliae could be a promising candidate against soil insect pests. In the study presented here, M. anisopliae was formulated into calcium alginate granules fortified with nutrients. The resporulation of the fungal granules was tested on four different soil types in the laboratory. The biocontrol efficacy of the resulting fungal growth was also examined using larval mealworms, Tenebrio molitor as a model insect in the laboratory and the glasshouse. Our results indicated that sterilised soil favoured optimal fungal resporulation, although different soil types did not have a significant effect on fungal resporulation. The resulting fungal resporulation and growth on sterilised soil caused high mortality (up to 76%) of larval mealworms in the glasshouse, whereas the fungal granules applied to non-sterile soil demonstrated poor resporulation that led to low mortality (13%) of larval mealworms. The result of this study indicates that the manipulation of microbial populations in field soil is required to enhance the fungal growth and potential insect control against wireworms in the field.     

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.     

Rate of consumption of Western flower thrips pupae by the soil-dwelling mite Hypoaspis sclerotarsa (Acari: Laelapidae)

Western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), are one of the most serious pests of horticultural crops worldwide. Despite one third of its lifecycle being in the soil, the potential for biological control of WFT in the soil is poorly understood and requires further elucidation. A number of studies report that polyphagous predatory mites prey on pupal stages of WFT in the soil, but little has been done on consumption rates. Therefore, we designed a laboratory study to examine the rate of consumption of WFT pupae in potting media, by the soil-dwelling mite Hypoaspis sclerotarsa (Costa). Five predator densities were evaluated (0, 2, 4, 6 and 8) against four densities of WFT prey (5, 10, 15 and 20 pupae). Pupal consumption was assessed at 2 hourly intervals over a six-hour period. The study confirmed that H. sclerotarsa did consume WFT pupae and that the rate of consumption increased with increasing density of H. sclerotarsa. The rate of consumption also increased with the density of WFT pupae. However, this was not consistent because, as the numbers of WFT pupae increased, so did the ratio of WFT pupae remaining to those consumed, increase. This paper is the first report of H. sclerotarsa in Kenya, and of its potential as a biocontrol agent of WFT. Further studies are now needed to understand interaction of foliar and soil dwelling predatory mites (H. sclerotarsa) for control of WFT under field conditions.     

Application of PSX biocontrol preparation confers root-knot nematode management and increased fruit quality in tomato under field conditions

PSX is a combination of biocontrol bacteria that can potentially prevent and control soil-borne diseases for a variety of crop cultivars. In this study, we investigated the utility of PSX in controlling root-knot nematodes in tomato under field conditions. The application of PSX reduced the severity of disease caused from Meloidogyne incognita by 63–69% and increased tomato yield by 31.5–39%. Furthermore, to investigate the effect of PSX treatment on tomato fruit quality, we quantified the soluble sugar, titratable acid, soluble solids, and vitamin C contents in fruit postharvest. We demonstrated that PSX treatment improved tomato fruit quality. Finally, we also showed that the total nitrogen (N), available N, potassium, and organic matter contents in the soil increased after PSX treatment. PSX is a promising biocontrol preparation that can provide beneficial effects to tomato growers, including biological control of root-knot disease, plant growth promotion, enhanced tomato fruit quality, and increased levels of organic fertilisers in the soil.     

Biological control of nematodes: Soil amendments and microbial antagonists

Summary Organic matter amendments to soil can be used to manage phytoparasitic nematodes. The most effective amendments are those with narrow C:N ratios and high protein or amine-type N content. For soil with 1.0% (w/w) organic matter amendment there is a direct relation between extent of nematode control and the N content of amendments. A special group of amendments are those containing chitinous materials. Chitin addition to soil results in stimulation of a select microflora capable of degrading the polymer. Several microbial species are known to destroy the eggs of phytonematodes (Meloidogyne spp). Organic matter can be modified by addition of specific compounds or by inoculation with particular microbial species to produce an amendment that will induce suppressiveness.     

Entomopathogenic nematodes control Plum Sawflies (Hoplocampa minuta and H. flava)

Plum sawflies are among the most damaging pests of European plum. Current control strategy implies insecticide application. Three species of entomopathogenic nematodes (EPN), Steinernema feltiae Filipjev, S. carpocapsae Weiser and Heterorhabditis bacteriophora Poinar were tested under laboratory and field conditions to assess effectiveness against larval and adult stages. Laboratory tests resulted in up to 100% mortality of last instar larvae before construction of a cocoon. However, the nematodes were not able to penetrate the cocoon. Foliar application did not result in plum sawflies larvae infestation by EPNs. Under field conditions, the nematodes reduced the number of emerging adults by application against sawfly larvae in the previous year before migration into the soil for overwintering by 62%–92%. Application of the nematodes against adults just before their anticipated emergence resulted in reduction of fruit infestation up to 100%. Mean results of 5 trials using caged trees were 47.8% with S. feltiae, 56.3% with S. carpocapsae and 62.9% with H. bacteriophora. In open field trails, control of adults obtained with S. feltiae at 0.5 million nematodes/m² was 98.2 and 67.8% and at 0.25 million m⁻² 41.7 and 41.2%. Forecasting adult emergence and optimal soil moisture conditions are essential for success of the nematode application.     

Development of biological soil disinfestations in Japan

Biological soil disinfestations (BSDs) were developed separately in Japan and in The Netherlands as an alternative to chemical fumigations. In Japan, it was developed based on the knowledge of irrigated paddy rice and upland crop rotation system that was rather tolerant of soil-borne disease development. The methods consist of application of easily decomposable organic matter, irrigation, and covering the soil surface with plastic film, thereby inducing anaerobic (reductive) soil conditions and suppressing many soil-borne pests including fungi, bacteria, nematodes, and weeds. The methods are widely used by organic farmers in the area where residences and agricultural fields are intermingled. To note one advantage of these methods, maintenance of soil suppressiveness to Fusarium wilt of tomato was suggested, while soil treated with chloropicrin became conducive to the disease. Suppression of soil-borne fungal pathogens by BSDs might be attributed to anaerobicity and high temperature, organic acids generated, and metal ions released into soil water. Contributions of respective factors to suppression of respective pathogens might be diverse. Presumably, these factors might vary on the fungal community structure in BSD-treated soil. These factors also work in paddy fields. Therefore, the BSDs developed in Japan are probably a method to raise the efficacy of paddy–upland rotation through intensive organic matter application and through maintenance of a strongly anaerobic (reductive) soil condition.     

Pathogenicity of 17 isolates of entomophagous nematodes (Steinernematidae and Heterorhabditidae) for the ticks Amblyomma variegatum (Fabricius), Boophilus microplus (Canestrini) and Boophilus annulatus (Say)

Entomopathogenous nematodes are well known biocontrol agents of insects. They live in the superficial layer of the soil, a location where ticks accomplish their molt and where they oviposit their eggs, making them, theoretically, the preys of infective larvae of nematodes. Seventeen strains of entomopathogenous nematodes: eight strains of Steinernema and nine strains of Heterorhabditis were placed in contact with each of the free living stages of three tick species: Amblyomma variegatum, Boophilus microplus and B. annulatus. The first two species were resistant to all the nematode strains that were tested, whereas B. annulatus was susceptible to all of them. Ovipositing females were more susceptible than females during the preoviposition period. There were no anatomical differences between the two species of Boophilus which can account for such differences in their susceptibility. The use of nematodes to control some species of ticks should be considered.     

Assessing nitrogen fluxes from roots to soil associated to rhizodeposition by apple (<Emphasis Type="Italic">Malus domestica</Emphasis>) trees

The mass transfer from root to soil by means of rhizodeposition has been studied in grasses and forest trees, but its role in fruit trees is still unknown. In this study, N fluxes from roots to soil were estimated by applying a ¹⁵N mass balance technique to the soil–tree system. Apple (Malus domestica) trees were pre-labelled with ¹⁵N and then grown outdoors in 40 L pots for one vegetative season in (1) a coarse-textured, low organic matter soil, (2) a coarse-textured, high organic matter soil, and (3) a fine-textured, high organic matter soil. At tree harvest the ¹⁵N abundance of the soils was higher than at transplanting, but the total amount of ¹⁵N present in the tree–soil system was similar at transplanting and tree harvest. The soils had a strong effect on N fluxes from and to the soil. In the fine-textured soil, 11% of the total plant-derived nitrogen was transferred to the soil, compared with 2–5% in the two coarse-textured soils. Rhizodeposition was higher in the fine soil (18% of the primary production) than in the coarse-textured soils, whereas higher soil organic matter depressed rhizodeposition. Nitrogen uptake was almost double in the coarse-textured, high organic matter soil versus the other soils. Our results indicate that belowground primary productivity is significantly underestimated if based on root production data only. Rhizodeposition represents a major process, whose role should not be underestimated in carbon and nitrogen cycles in orchard ecosystems.     

Dynamics of a Subterranean Trophic Cascade in Space and Time

Trophic cascades, whereby predators indirectly benefit plant biomass by reducing herbivore pressure, form the mechanistic basis for classical biological control of pest insects. Entomopathogenic nematodes (EPN) are lethal to a variety of insect hosts with soil-dwelling stages, making them promising biocontrol agents. EPN biological control programs, however, typically fail because nematodes do not establish, persist and/or recycle over multiple host generations in the field. A variety of factors such as local abiotic conditions, host quantity and quality, and rates of movement affect the probability of persistence. Here, we review results from 13 years of study on the biology and ecology of an endemic population of Heterorhabditis marelatus (Rhabditida: Heterorhabditidae) in a California coastal prairie. In a highly seasonal abiotic environment with intrinsic variation in soils, vegetation structure, and host availability, natural populations of H. marelatus persisted at high incidence at some but not all sites within our study area. Through a set of field and lab experiments, we describe mechanisms and hypotheses to understand the persistence of H. marelatus. We suggest that further ecological study of naturally occurring EPN populations can yield significant insight to improve the practice and management of biological control of soil-dwelling insect pests.     

Isolation,characterization and pathogenicity of bacteria from Rhynchites bacchus (Coleoptera: Rhynchitidae)

The leafroller weevil beetle (Rhynchites bacchus L., Coleoptera: Rhynchitidae) is one of the most serious pests of apple, plum, apricot, cherry and peach (nectarine) fruit worldwide. The aim of this study was to isolate and characterize pathogenic bacteria from this pest for possible use in biocontrol, and to determine their pathogenicity. R. bacchus were collected from the Eastern Black Sea Region of Turkey in 2007. Based on their morphological, physiological and biochemical features, five different bacterial isolates were obtained from adults and larvae. In addition, 16S rRNA gene was amplified and sequenced to confirm isolate identification. According to the numerical and molecular data, the culturable bacterial flora of R. bacchus was determined as Bacillus licheniformis (Rb1), Serratia marcescens (Rb2), Enterobacter hormaechei (Rb3), Paenibacillus sp. (Rb4), and Enterobacter sp. (Rb5). Isolate Rb2 produced the highest mortality (73%) against larvae within 10 days after inoculation (P<0.05). The others, Rb1, Rb3, Rb4 and Rb5, caused 20, 13, 26 and 13% mortality in the larvae within the same period. These results indicate that S. marcescens Rb2 seems to be the most promising biocontrol agent againts this pest.     

食细菌线虫对土壤微生物量和微生物群落结构的影响

线虫与微生物的相互作用研究往往是在悉生培养体系 (gnotobiotic microcosm)中进行 ,为了研究在自然或开放土壤条件下土壤线虫与微生物的相互作用 ,作者在开放盆栽体系中接种土壤食细菌线虫 (原小杆线虫 ,Protorhabditis sp) ,研究在小麦不同生育期、在有和无根系作用下食细菌线虫对土壤微生物量和微生物群落结构的影响。结果表明 :接种线虫分别使 SMBC、SMBN、SMBP提高了 2 6 .4 %、32 .9%、2 1.8% ,这种促进作用除个别无根系土和非根际土处理外 ,均达到显著性差异。根际土中的 SMBC、SMBN、SMBP>非根际土 >无根系土。从方差解释比例 v来看 ,SMBN受线虫的影响最大 (v=2 4 % )、其次是 SMBC(v=16 % )、然后是 SMBP(v=12 % ) ,线虫对 SMBC的促进作用在根际土中最突出。接种线虫对土壤细菌、真菌和放线菌的数量有明显的影响。在苗期的无根系土和根际土中 ,接种线虫显著降低了细菌的数量、特别在根际土中尤为突出 ,但在其它处理中却增加了细菌的数量。接种线虫对真菌和放线菌数量的促进作用比对细菌更为明显 ,接种线虫后真菌和放线菌数量的总平均值分别比未接种提高了 4 8.5 %和 6 8.2 % ,而细菌数量的总平均值没有变化。细菌数量与微生物量 C相关散点图表明二者相关性在根际土、非根际土和无根系土中均未     

Vesicular-arbuscular mycorrhizal effectiveness in an acid soil amended with fresh organic matter

A greenhouse investigation was undertaken to determine the influence of fresh organic matter on the formation and functioning of vesicular-arbuscular mycorrhizal symbiosis in Leucaena leucocephala grown in an acid aluminum-rich ultisol. In soil not amended with fresh organic matter or lime, plants failed to grow. Mycorrhizal infection level, mycorrhizal effectiveness measured in terms of pinnule P content of L. leucocephala leaves and dry matter yield of the legume increased with increase in fresh organic matter. Although VAM colonization level and dry matter yield of L. leucocephala were significantly higher if the test soil was limed (7.2 cmole OH^–) than if amended with fresh organic matter, the latter was as effective as lime in off-setting the detrimental effect of aluminum on mycorrhizal effectiveness. The lower mycorrhizal colonization level and the lower dry matter yield noted in the soil treated with fresh organic matter appears to be related to the inadequacy of Ca in the soil amended with fresh organic matter. These observations are supported by the low calcium status of soil and plant tissues in the absence of lime. It is concluded that while fresh organic matter, in appropriate amounts, could protect sensitive plants and VAM symbiosis against Al toxicity in acid soils, maximum mycorrhizal inoculation effects are not likely to be attained unless the soils are also amended with Ca.Contribution from Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No 3740.     

Dual inoculation with Aspergillus fumigatus and Glomus mosseae enhances biomass production and nutrient uptake in wheat (Triticum aestivum L.) supplied with organic phosphorus as Na-phytate

In a pot experiment, wheat was grown for 50 days in two heat-sterilized low-phosphorus (P) soils supplied with organic P as Na-phytate. Seed inoculation with the phosphatase-producing fungus (PPF) Aspergillus fumigatus or soil inoculation with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae increased shoot and root dry weight and root length, phosphatase activity in the rhizosphere and shoot concentrations of P and to a lesser extent of K and Mg. As a rule, the greatest effects on those parameters were most in the combined inoculation treatment (PPF + VAM). Shoot concentrations of Cu and Zn were only enhanced by VAM, not by PPF. At harvest, depletion of organic P in the rhizosphere soil increased in the order of: sterilized soil < PPF < VAM < PPF + VAM which corresponded with the enhanced P concentrations in the plants. The results demonstrate that organic P in form of Na-Phytate is efficiently used by VAM and that use of organic P can be increased by simultaneous inoculation with phosphatase-producing fungi.     

Long‐term influence of sod cutting depth on the restoration of degraded wet heaths

Temperate heaths have an unfavorable conservation status in most European biogeographical regions. Increasing nitrogen levels promote competitive grass species such as Molinia caerulea, which is a main threat to heathland conservation in Europe. This article investigates the long‐term influence of sod cutting and the resulting changes in soil properties on the heath composition, integrity, and structure. In 15 nature reserves across the northern half of Belgium, we used (1) a large number of plots (203); (2) a broad range of sod cut depths (2–40 cm), and (3) a temporal dimension that describes how long the effects of sod cutting persist (census up to 19 years after sod cutting). Multivariate analyses were used in order to explore the influence of sod cut depth and time after sod cutting on the soil and vegetation properties. There was a positive relationship between sod cut depth and soil pH and water level, and a negative relationship with Al³⁺, NH₄⁺, and total organic matter (TOM). However, only a limited number of typical (target) species appeared after sod cutting, and then only weakly. Most of the time they remained a minor component of the restored vegetation. Moreover, M. caerulea reappeared and its cover significantly increased during the years following sod cutting. Although we were able to show that sod cut depth has a differential effect on soil properties and vegetation recovery, it also appeared that sod cutting does not restore wet heaths in the long term when applied in regions with high nitrogen deposition.     

Biological control potential of native entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) against Spodoptera cilium (Lepidoptera: Noctuidae) in turfgrass

In laboratory studies, we demonstrated that five native entomopathogenic nematode species/isolates caused 100% mortality of Spodoptera cilium larvae, a soil surface-feeding pest of turfgrass. At 25 infective juveniles/cm² applied to sod, two selected Turkish species, Steinernema carpocapsae and Heterorhabditis bacteriophora (Sarigerme isolate), averaged 77% and 29% larval mortality, respectively.