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Biotic stresses (fungi, bacteria, insects, weeds, etc.) are some of the most important causes of the decrease in the quality and quantity of crops that could become an emergency due to a noteworthy increase in the world population. Thus, to overcome these problems, massive use of chemical pesticides has been carried out with heavy consequences for environmental pollution and food safety. An eco-friendly alternative can be using natural compound-based biopesticides with high efficacy and selectivity. Some bacterial lipodepsipeptides (tolaasins I, II, A, D, and E and WLIP together with hexacetyl- and tetrahydro-tolaasin I and WLIP methyl ester) and cyclic dipeptides (cyclo(l-Pro-l-Tyr), cyclo(d-Pro-l-Tyr), cyclo(l-Pro-l-Val), and cyclo(l-Pro-l-Leu)) were assayed against several pathogenic bacteria and fungi of important agrarian plants. Lipodepsipeptides showed strong growth inhibition of all microorganisms tested in the range of 0.1-0.8 μg/mL, while cyclodipeptides, despite preserving this ability, showed a noteworthily reduced antimicrobial activity being active only in the range of 15-900 μg/mL. Among the lipodepsipeptides and cyclic dipeptides assayed, tolaasin d and cyclo(l-Pro-l-Tyr) (also named maculosin-1) appeared to be the most toxic compounds. Some structure-activity relationships of lipodepsipeptides were also discussed along with their practical application as biopesticides in agriculture.

Brassicaceae family includes an important group of plants of great scientific interest, e.g., the model plant Arabidopsis thaliana, and of economic interest, such as crops of the genus Brassica (Brassica oleracea, Brassica napus, Brassica rapa, etc.). This group of plants is characterized by the synthesis and accumulation in their tissues of secondary metabolites called glucosinolates (GSLs), sulfur-containing compounds mainly involved in plant defense against pathogens and pests. Brassicaceae plants are among the 30% of plant species that cannot establish optimal associations with mycorrhizal hosts (together with other plant families such as Proteaceae, Chenopodiaceae, and Caryophyllaceae), and GSLs could be involved in this evolutionary process of non-interaction. However, this group of plants can establish beneficial interactions with endophytic fungi, which requires a reduction of defensive responses by the host plant and/or an evasion, tolerance, or suppression of plant defenses by the fungus. Although much remains to be known about the mechanisms involved in the Brassicaceae-endophyte fungal interaction, several cases have been described, in which the fungi need to interfere with the GSL synthesis and hydrolysis in the host plant, or even directly degrade GSLs before they are hydrolyzed to antifungal isothiocyanates. Once the Brassicaceae-endophyte fungus symbiosis is formed, the host plant can obtain important benefits from an agricultural point of view, such as plant growth promotion and increase in yield and quality, increased tolerance to abiotic stresses, and direct and indirect control of plant pests and diseases. This review compiles the studies on the interaction between endophytic fungi and Brassicaceae plants, discussing the mechanisms involved in the success of the symbiosis, together with the benefits obtained by these plants. Due to their unique characteristics, the family Brassicaceae can be seen as a fruitful source of novel beneficial endophytes with applications to crops, as well as to generate new models of study that allow us to better understand the interactions of these amazing fungi with plants.

The honeybee Apis mellifera is an important pollinator insect. Any pathogenic infection in this beneficial insect is undesirable. In this study, bacterial diversity in beehives was investigated to determine the potential of pathogenic bacteria in honeybees. To do this, bacterial isolations were carried out from dead and diseased adult bees collected from 9 districts in Ordu Province in Turkey.Twenty species of pathogenic bacteria, 18 of which were nonsporeforming Staphylococcus lentus, Klebsiella oxytoca, Citrobacter freundii, Leucanostoc mesenteroides ssp. cremoris, Kocuria rosea, Kocuria kristinae, Sphingomonas paucimobilis slashline, Burkholderia cepacia, Leucanostoc mesenteroides ssp. dextranicum,Hafnia alvei, Escherichia coli, Aeromonas salmonicida, Citrobacter braakii, Pantoea agglomerans, Streptococcus equi ssp. zooepidemicus, Staphylococcus pseudintermedius, Staphylococcus lugdunesis and Staphylococcus vitulinus and 2 sporeforming Bacillus licheniformis and Paenibacillus polymyxa, were isolated and identified from the honeybees. The infectivity of these bacteria were also documented with bioassay experiments on the healthy bees.The highest insecticidal effectwas determinedwith Bacillus licheniformis 84% , Escherichia coli 84% , and Streptococcus equi ssp. zooepidemicus 80% on the adult honeybees.This result confirms that the honeybee contains a very large number of bacterial species and that the majority of them are pathogenic for the species in Turkey. In addition, some of the entomopathogenic bacteria used for biological control can have negative impact on this economically important insect.

Red pepper, one of the major economic crops in Korea, is being affected by anthracnose disease caused by Colletotrichum acutatum. To control this disease, an antagonistic bacterial strain, Bacillus subtilis YGB36 identified by 16S rDNA sequencing, physiological and biochemical analyses is used as a biological control agent. In vitro screening revealed that the strain YGB36 possess strong antifungal activity against the pathogen Cylindrocarpon destructans. The strain exhibited cellulase, protease, amylase, siderophore production and phosphate solubility. In vitro conidial germination of C. acutatum was most drastically inhibited by YGB36 cell suspensions (106 cfu/ml) or culture filtrate. Development of anthracnose symptoms was reduced on detached immature green pepper fruits by treatment with cell suspensions, and its control value was recorded as 65.7%. The YGB36 bacterial suspension treatment enhanced the germination rate of red pepper seeds and promoted root development and growth under greenhouse conditions. The in vitro screening of fungicide and insecticide sensitivity test against YGB36 revealed that the bacterial growth was not affected by any of the insecticides, and 11 fungicides out of 21 used. Collectively, our results clearly suggest that the strain YGB36 is considered as one of the potential biocontrol agents against anthracnose disease in red pepper.

Khon Kaen Agriculture Journal, 50, 6, 1683-1700

To manage stem canker disease on royal poinciana, actinobacterial isolates were used as biological control agents (BCAs) based on their strong in vitro inhibitory effects against Neoscytalidium dimidiatum. Streptomyces griseorubens UAE2 and Streptomyces wuyuanensis UAE1 had the ability to produce antifungal compounds and cell-wall-degrading enzymes (CWDEs). Only S. griseorubens, however, restored the activity of 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCD). In vivo apple fruit bioassay showed that lesion development was successfully constrained by either isolates on fruits inoculated with N. dimidiatum. In our greenhouse and container nursery experiments, S. griseorubens showed almost complete suppression of disease symptoms. This was evident when the preventive treatment of S. griseorubens significantly (p < 0.05) reduced the numbers of conidia of N. dimidiatum and defoliated leaves of royal poinciana seedlings to lesser levels than when S. wuyuanensis was applied, but comparable to control treatments (no pathogen). The disease management of stem canker was also associated with significant (p < 0.05) decreases in ACC levels in royal poinciana stems when S. griseorubens was applied compared to the non-ACCD-producing S. wuyuanensis. This study is the first to report the superiority of antagonistic actinobacteria to enhance their effectiveness as BCAs not only for producing antifungal metabolites and CWDEs but also for secreting ACCD.

Simple Summary Two egg parasitoid species (Trichogramma mwanzai and Trichogrammatoidea lutea) from parasitized eggs of fall armyworm (FAW) Spodoptera frugiperda in Zambia were identified by using a combination of both molecular and morphological characters. We compared the parasitism capabilities of the two species with three native Chinese trichogrammatid species (T. ostriniae, T. leucaniae and T. japonicum) using 0- to 2-day-old FAW eggs. Both parasitoid species accepted eggs of all ages tested and completed their development successfully. Trichogrammatoidea lutea females showed the highest parasitism rate of host eggs among the five tested species. T. mwanzai had the shortest developmental time on all test age eggs. Of the five parasitoid species reared on FAW eggs, T. lutea performed the best overall, while T. japonicum was the worst performing of the parasitoids. Abstract The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a noctuid moth native to the tropical and subtropical Americas that has successfully invaded Africa and Asia, where it is has become a serious threat to food security as a pest of cereals and other crops. Biological control is an environmentally friendly means of combating the pest and contributes to an integrated pest management approach. In our study, two egg parasitoid species (Trichogramma mwanzai and Trichogrammatoidea lutea) found in parasitized fall armyworm eggs in Zambia were identified by using a combination of both molecular and morphological characteristics. To evaluate their potential and efficiency on 0- to 2-day-old fall armyworm eggs, we compared their parasitism capabilities with three Trichogramma species native to China (T. ostriniae, T. leucaniae and T. japonicum) under laboratory conditions. The results showed that both parasitoid species would accept 0-, 1- and 2-day-old fall armyworm eggs, and complete their development successfully. Trichogramma mwanzai and T. lutea preferred parasitizing 0- and 1-day-old eggs over 2-day-old eggs. Trichogrammatoidea lutea females supplied with fall armyworm eggs produced the highest parasitism rate of host eggs among the five tested species, while T. mwanzai had the shortest developmental time on all test age eggs. In general, T. lutea was the best performing of the five species when reared on fall armyworm eggs, while T. japonicum was the worst. There were no significant differences, however, in percent emergence in the five test species when reared on fall armyworm eggs.

Crown gall disease in grapevine is caused by pathogenic strains of Rhizobium vitis with a tumor-inducing (Ti) plasmids. A nonpathogenic strain, VAR03-1 of R. vitis, has been isolated from the grapevine root of nursery stock and it was shown to act as a biological control agent to crown gall disease. Its disease-suppressive effect was observed even when it was coinoculated with the pathogen in a 1:1 ratio. Here, we present the complete genome data of R. vitis VAR03-1, assembled by sequencing reads obtained by both PacBio and Illumina technologies with annotation. This genome sequence could contribute to investigations of the molecular basis underlying the biocontrol activity as well as the root-colonization ability of this bacterial strain. [Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Streptomyces padanus PMS-702 strain produces a polyene macrolide antibiotic fungichromin and displays antagonistic activities against many phytopathogenic fungi. In the present study, experimental formulations were assessed to improve the production of fungichromin, the efficacy of PMS-702 on the suppression of sporangial germination, and the reduction of cucumber downy mildew caused by Pseudoperonospora cubensis. PMS-702 strain cultured in a soybean meal-glucose (SMG) medium led to low levels of fungichromin accumulation and sporangial germination suppression. Increasing medium compositions and adding plant oils (noticeably coconut oil) in SMG significantly increased fungichromin production from 68 to 1,999.6 μg/ml. Microscopic examination reveals that the resultant suspensions significantly reduced sporangial germination and caused cytoplasmic aggregation. Greenhouse trials reveal that the application of PMS-702 cultural suspensions reduced downy mildew severity considerably. The addition of Tween 80 into the synthetic medium while culturing PMS-702 further increased the suppressive efficacy of downy mildew severity, particularly when applied at 24 h before inoculation or co-applied with inoculum. Fungichromin at 50 μg/ml induced phytotoxicity showing minor necrosis surrounded with light yellowish halos on cucumber leaves. The concentration that leads to 90% inhibition (IC90) of sporangial germination was estimated to be around 10 μg/ml. The results provide a strong possibility of using the S. padanus PMS-702 strain as a biocontrol agent to control other plant pathogens.

Microbiomes composition, diversity, and variability into a collection of suppressive composts were investigated for effective biological control of soil-borne phytopathogens. Pyrosequencing resulted be a reliable and faster method for characterizing fungal and bacterial microbiomes into composts derived from a varied feedstock of different composition, origin and provenience. Differences in taxonomic structure assessed by bioinformatics analyses were related to feedstock origin. Green composts derived from agro-waste and agroindustrial co/byproducts provided the most varied microbiomes either related to suppression of Rhizoctonia damping-off in bean and Verticillium wilt in eggplant, either to control of Phytium damping-off in cucumber and Phytophthora root rot in tomato. On the other hand, composted municipal solid wastes and co-composted cow manure with household waste prevalently given a most specific microbiota related to suppression of Fusarium wilt in melon.

Tuta absoluta (Meyrick), a key pest of tomato, is quickly spreading over the world and biological control is considered as one of the control options. Worldwide more than 160 species of natural enemies are associated with this pest, and an important challenge is to quickly find an effective biocontrol agent from this pool of candidate species. Evaluation criteria for control agents are presented, with the advantages they offer for separating potentially useful natural enemies from less promising ones. Next, an aggregate parameter for ranking agents is proposed: the pest kill rate km. We explain why the predator's intrinsic rate of increase cannot be used for comparing the control potential of predators or parasitoids, while km can be used to compare both types of natural enemies. As an example, kill rates for males, females and both sexes combined of three Neotropical mirid species (Campyloneuropsis infumatus (Carvalho), Engytatus varians (Distant) and Macrolophus basicornis (Stål)) were determined, taking all life-history data (developmental times, survival rates, total nymphal and adult predation, sex ratios and adult lifespan) into account. Based on the value for the intrinsic rate of increase (rm) for T. absoluta and for the kill rate km of the predators, we predict that all three predators are potentially able to control the pest, because their km values are all higher than the rm of the pest. Using only km values, we conclude that E. varians is the best candidate for control of T. absoluta on tomato, with C. infumatus ranking second and M. basicornis last.

Ecologists study how populations are regulated, while scientists studying biological pest control apply population regulation processes to reduce numbers of harmful organisms: an organism (a natural enemy) is used to reduce the population density of another organism (a pest). Finding an effective biological control agent among the tens to hundreds of natural enemies of a pest is a daunting task. Evaluation criteria help in a first selection to remove clearly ineffective or risky species from the list of candidates. Next, we propose to use an aggregate evaluation criterion, the pest kill rate, to compare the pest population reduction capacity of species not eliminated during the first selection. The pest kill rate is the average daily lifetime killing of the pest by the natural enemy under consideration. Pest kill rates of six species of predators and seven species of parasitoids of Tuta absoluta were calculated and compared. Several natural enemies had pest kill rates that were too low to be able to theoretically reduce the pest population below crop damaging densities. Other species showed a high pest reduction capacity and their potential for practical application can now be tested under commercial crop production conditions., Centro de Estudios Parasitológicos y de Vectores

Pesticides of chemical synthesis have mainly been used to control pests, diseases and adventitious plants up until now. However, it has been shown that some pesticides can remain in the soil for long periods of time, thus affecting the development of organisms in the rhizosphere as well as human health, which are two of the most noteworthy side effects. The aim of this research was to analyze the compatibility of autochthonous Trichoderma strains with different synthetic fungicides, acaricides, insecticides (including an entomopathogenic fungus) and herbicides. Sulfur encouraged the growth of all autochthonous strains assayed, and the combination Trichoderma-B. bassiana did not disturb their growth. So, the combination of the autochthonous Trichoderma strains with these organic pesticides will be a positive strategy to apply in the field to control pests and some diseases. Conventional pesticides modified the development of all autochthonous Trichoderma strains, demonstrating that not only do they affect weeds, fungus or pests but also rhizosphere microorganisms. In conclusion, conventional pesticides indiscriminately used to control pests, diseases and weeds could reduce the development of autochthonous Trichoderma strains, especially fungicides and herbicides.

Bacillus velezensis 83 was isolated from mango tree phyllosphere of orchards located in El Rosario, Sinaloa, México. The assessment of this strain as BCA (biological control agent), as well as PGPB (plant growth-promoting bacteria), were demonstrated through in vivo and in vitro assays. In vivo assays showed that B. velezensis 83 was able to control anthracnose (Kent mangoes) as efficiently as chemical treatment with Captan 50 PH™ or Cupravit hidro™. The inoculation of B. velezensis 83 to the roots of maize seedlings yielded an increase of 12% in height and 45% of root biomass, as compared with uninoculated seedlings. In vitro co-culture assays showed that B. velezensis 83 promoted Arabidopsis thaliana growth (root and shoot biomass) while, under the same experimental conditions, B. velezensis FZB42 (reference strain) had a suppressive effect on plant growth. In order to characterize the isolated strain, the complete genome sequence of B. velezensis 83 is reported. Its circular genome consists of 3,997,902 bp coding to 3949 predicted genes. The assembly and annotation of this genome revealed gene clusters related with plant-bacteria interaction and sporulation, as well as ten secondary metabolites biosynthetic gene clusters implicated in the biological control of phytopathogens. Despite the high genomic identity (> 98%) between B. velezensis 83 and B. velezensis FZB42, they are phenotypically different. Indeed, in vitro production of compounds such as surfactin and bacillomycin D (biocontrol activity) and γ-PGA (biofilm component) is significantly different between both strains.

To manage stem canker disease on royal poinciana, actinobacterial isolates were used as biological control agents (BCAs) based on their strong in vitro inhibitory effects against Neoscytalidium dimidiatum. Streptomyces griseorubens UAE2 and Streptomyces&nbsp, wuyuanensis UAE1 had the ability to produce antifungal compounds and cell-wall-degrading enzymes (CWDEs). Only S. griseorubens, however, restored the activity of 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCD). In vivo apple fruit bioassay showed that lesion development was successfully constrained by either isolates on fruits inoculated with N. dimidiatum. In our greenhouse and container nursery experiments, S. griseorubens showed almost complete suppression of disease symptoms. This was evident when the preventive treatment of S. griseorubens significantly (p &lt, 0.05) reduced the numbers of conidia of N. dimidiatum and defoliated leaves of royal poinciana seedlings to lesser levels than when S. wuyuanensis was applied, but comparable to control treatments (no pathogen). The disease management of stem canker was also associated with significant (p &lt, 0.05) decreases in ACC levels in royal poinciana stems when S. griseorubens was applied compared to the non-ACCD-producing S. wuyuanensis. This study is the first to report the superiority of antagonistic actinobacteria to enhance their effectiveness as BCAs not only for producing antifungal metabolites and CWDEs but also for secreting ACCD.

RESUMEN Botrytis cinerea es un patógeno capaz de atacar a su hospedador empleando sus estrategias de ataque de múltiples formas, y, cuando se dan las condiciones favorecedoras de la infección, se ponen de manifiesto síntomas que producen grandes pérdidas en los cultivos, como ocurre en la enfermedad de la podredumbre gris u oídio. En este proyecto se realizó un cribado de microorganismos capaces de inhibir el crecimiento del fitopatógeno y así suprimir la enfermedad. Para ello, se emplearon 34 cepas de una colección de Bacillus subtilis con el fin de evaluar su capacidad antagonista frente a B. cinerea, ya que esta bacteria produce moléculas bioactivas capaces de combatir el oídio. El potencial de cada una de las cepas fue evaluado mediante diversos ensayos. Se seleccionaron las cepas que mostraron mayor capacidad antifúngica mediante enfrentamientos duales en placa entre una alícuota de cultivo celular de la bacteria antagonista y el hongo, y entre una alícuota de extracto acelular y el hongo, y, además, se realizó un enfrentamiento en microplaca con pocillos entre la alícuota de cultivo del fitopatógeno y extracto acelular de B. subtilis. Tras esto, en condiciones controladas de laboratorio, se aplicaron cultivos celulares y extractos acelulares sobre hojas de pepino infectadas por el fitopatógeno como medida preventiva, evaluando los daños de manera cualitativa. Los resultados mostraron que la mayoría de las cepas seleccionadas presentaron una incidencia directa, es decir, cuando el enfrentamiento se producía entre el hongo y la alícuota de cultivo celular de la bacteria antagonista, siendo el porcentaje de inhibición del crecimiento del agente fitopatógeno mayor. Sin embargo, resultó interesante la capacidad de determinados extractos para frenar la enfermedad. En definitiva, Bacillus subtilis es un potente agente de control biológico (ACB) con capacidad biopesticida frente a la enfermedad de oídio. ABSTRACT Botrytis cinerea is a pathogen capable of attacking its host using its attack strategies in multiple ways, and, when favorable conditions for infections occur, symptoms that produce large losses in crops are revealed, as happens in the disease of gray rot or powdery mildew. The aim of this project was to screen from a collection of Bacillus subtilis the microorganisms that were capable to inhibit the growth of the plant pathogen and suppress the disease. For this, 34 strains were used to evaluate their antagonistic capacity against B. cinerea, since these microorganisms produce bioactive molecules capable of fighting powdery mildew. The potential of each of the strains was evaluated through various tests. The strains showing the highest antifungal capacity were selected by means of dual plaque assays between an aliquot of cell culture of the antagonistic bacteria and the fungus, and between an aliquot of acellular extract and the fungus, and, in addition, a microplate confrontation with wells between the culture aliquot of the phytopathogen and acellular extract of B. subtilis. After this, in controlled laboratory conditions, cell cultures and acellular extracts were applied to cucumber leaves that were later infected by the plant pathogen as a preventive measure, evaluating the damage in a qualitative way. The results showed that most of the selected strains had a direct incidence. When the confrontation occurred between the fungus and the aliquot of cell culture of the antagonistic bacteria, being the percentage of the phytopathogen growth inhibition greater. However, the efficacy to stop the disease by some extract was remarkable. Definitely, Bacillus subtilis is a powerful biological control agent (BCA) with biopesticidal capability against powdery mildew disease.

The genus Stenonabis Reuter (Hemiptera: Heteroptera: Nabidae: Nabinae) from Korea is reviewed. Stenonabis uhleri Miyamoto, 1964 is reported from the Korean Peninsula for the first time. Morphological information such as descriptions and diagnoses for the Korean species with male and female genital illustrations are provided, and some biological information is also given. Additionally, we provide a key to the Stenonabis species in the Far East.

The Aureobasidium pullulans L1 and L8 strains are known as efficient biocontrol agents against several postharvest fungal pathogens. In order to better understand the mechanism of action underneath the antifungal activity of L1 and L8 strains, yeast cell filtrates grown at different times were evaluated in vivo against Monilinia laxa on peach. Lesion diameters on peach fruit were reduced by L1 and L8 culture filtrates of 42.5% and 67% respectively. The ability of these filtrates to inhibit M. laxa conidia germination and germ tube elongation was studied by in vitro assays. The results showed a 70% reduction of conidia germination for both strains while for germ tube elongation, it was 52% and 41% for L1 and L8 culture filtrates respectively. Finally, the activity of cell wall hydrolytic enzymes such as chitinase and glucanase in cell filtrates was analysed and the expression of genes encoding these activities was quantified during yeast growth. From 24 h onward, both culture filtrates contained β,1-3,glucanase and. chitinase activities, the most pronounced of which was N-β-acetylglucosaminidase. Gene expression level encoding for these enzymes in L1 and L8 varied according to the strain. These results indicate that L1 and L8 strains culture filtrates retain the yeast antagonistic activity and suggest that the production of hydrolytic enzymes plays an important role in this activity.

BACKGROUND: Bacillus amyloliquefaciens strain CPA‐8 has been described as an effective biocontrol agent to control brown rot in stone fruit for both preharvest and postharvest applications. However, no information about the environmental fate and behaviour of this strain under field conditions is available. RESULTS: The dispersion of the CPA‐8 application was evaluated using water‐sensitive papers, and complete coverage was observed on the leaves of treated trees, while

The development of biological control methods for exotic invasive pest species has become more challenging during the last decade. Compared to indigenous natural enemies, species from the pest area of origin are often more efficient due to their long coevolutionary history with the pest. The import of these well‐adapted exotic species, however, has become restricted under the Nagoya Protocol on Access and Benefit Sharing, reducing the number of available biocontrol candidates. Finding new agents and ways to improve important traits for control agents (“biocontrol traits”) is therefore of crucial importance. Here, we demonstrate the potential of a surprisingly under‐rated method for improvement of biocontrol: the exploitation of intraspecific variation in biocontrol traits, for example, by selective breeding. We propose a four‐step approach to investigate the potential of this method: investigation of the amount of (a) inter‐ and (b) intraspecific variation for biocontrol traits, (c) determination of the environmental and genetic factors shaping this variation, and (d) exploitation of this variation in breeding programs. We illustrate this approach with a case study on parasitoids of Drosophila suzukii, a highly invasive pest species in Europe and North America. We review all known parasitoids of D. suzukii and find large variation among and within species in their ability to kill this fly. We then consider which genetic and environmental factors shape the interaction between D. suzukii and its parasitoids to explain this variation. Insight into the causes of variation informs us on how and to what extent candidate agents can be improved. Moreover, it aids in predicting the effectiveness of the agent upon release and provides insight into the selective forces that are limiting the adaptation of indigenous species to the new pest. We use this knowledge to give future research directions for the development of selective breeding methods for biocontrol agents.

The release of large quantities of microorganisms to soil for purposes such as pest control or plant growth promotion may affect the indigenous soil microbial communities. In our study, we investigated potential effects of Metarhizium brunneum ART2825 on soil fungi and prokaryota in bulk soil using high-throughput sequencing of ribosomal markers. Different formulations of this strain, and combinations of the fungus with garlic as efficacy-enhancing agent, were tested over 4 months in a pot and a field experiment carried out for biological control of Agriotes spp. in potatoes. A biocontrol effect was observed only in the pot experiment, i.e. the application of FCBK resulted in 77% efficacy. Colony counts combined with genotyping and marker sequence abundance confirmed the successful establishment of the applied strain. Only the formulated applied strain caused small shifts in fungal communities in the pot experiment. Treatment effects were in the same range as the effects caused by barley kernels, the carrier of the FCBK formulation and temporal effects. Garlic treatments and time affected prokaryotic communities. In the field experiment, only spatial differences affected fungal and prokaryotic communities. Our findings suggest that M. brunneum may not adversely affect soil microbial communities., Formulated Metarhizium brunneum has small and temporal effects on fungal communities in a pot experiment and no effects were detected in the field.

Adult spider mite females that are aware of predatory mites can reduce predation by dispersing from patches invaded by predators or by shifting their oviposition site onto webs where eggs are predated less than on leaf surfaces. Therefore, the stealthiness of predatory mites stalking spider mite females should largely determine the success of spider mite control. This study examined the stealthiness of three predatory mites: Phytoseiulus persimilis Athias-Henriot, Neoseiulus californicus McGregor, and Neoseiulus womersleyi Schicha. The first two species have been commercialized as spider mite biological control agents, whereas the last species is a native predator of spider mites in Japan. We measured two indices of predator stealthiness in bioassays using Tetranychus kanzawai Kishida as prey: the proportion of prey mites that dispersed from their patches in response to predator intrusion and the proportions of prey eggs laid on webs in response to predator eggs. Both indices resulted in the same ranking, with P. persimilis ranking highest and N. womersleyi lowest. The greater stealthiness of P. persimilis eggs compared with N. womersleyi eggs seemed to be due to the lower amount of detectable substances on the egg surface. We also measured this index of stealthiness for European and Japanese Tetranychus urticae Koch populations and found that they had the same stealthiness ranking as measured using T. kanzawai. Therefore, we predict that the stealthiness of predatory mites is consistent across prey species and could be an effective index, along with conventional indices, for evaluating predatory mites as spider mite biological control agents.

Plant protection via disease-suppressive bacteria in desert farming requires specific biological control agents (BCAs) adapted to the unique arid conditions. We performed an ecological study of below-ground communities in desert farm soil and untreated desert soil, and based on these findings, selected antagonists were hierarchically evaluated. In contrast to the highly specific 16S rRNA fingerprints of bacterial communities in soil and cultivated medicinal plants, internal transcribed spacer profiles of fungal communities were less discriminative and mainly characterised by potential pathogens. Therefore, we focused on in vitro bacterial antagonists against pathogenic fungi. Based on the antifungal potential and genomic diversity, 45 unique strains were selected and characterised in detail. Bacillus/Paenibacillus were most frequently identified from agricultural soil, but antagonists from the surrounding desert soil mainly belonged to Streptomyces. All strains produced antibiotics against the nematode Meloidogyne incognita, and one-third showed additional activity against the bacterial pathogen Ralstonia solanacearum. Altogether, 13 broad-spectrum antagonists with antibacterial, antifungal and nematicidal activity were found. They belong to seven different bacterial species of the genera Bacillus and Streptomyces. These Gram-positive, spore-forming bacteria are promising drought-resistant BCAs and a potential source for antibiotics. Their rhizosphere competence was shown by fluorescence in situ hybridisation combined with laser scanning microscopy.

Invasive weeds are threatening ecosystem function and productivity all over the world by outcompeting desirable vegetation and reducing species diversity. One option for long-term control of such weeds is biological control using natural insect enemies. Such a program has been developed for management of Dalmatian toadflax (Linaria dalmatica (L.) Miller (Plantaginaceae)) in North America using a stem-mining weevil, Mecinus janthiniformis To��evski and Caldara (Coleoptera: Curculionidae). Although widely effective in northern regions, such as in British Columbia and in the American northwest, this insect has been slow to suppress Dalmatian toadflax in southern most regions of their current range, including areas in Utah and Colorado, and little is known of the limiting factors leading to slow weed suppression in these areas. Using field assessments of insect and plant activity over two growing seasons at several sites, this study aimed to provide degree-day and calendar-date descriptions of insect phenology. In addition, dead, overwintered Dalmatian toadflax stems were dissected to determine overwintering mortality of weevil adults before spring emergence, and living stems were dissected to determine development stages and mortality of weevils during summer development. Degree-day and calendar-date based models independently resulted in consistent trends in weevil phenology between sites and between years in this study, although the degree-day model is likely to be most useful for purposes of predicting weevil life cycle timing. Interestingly, the sexes differed in their phenology in that males consistently emerged from overwintering sites and were found on Dalmatian toadflax stems considerably earlier than females in the spring. Females as well as males tended to peak in abundance on stems in late-May when Dalmatian toadflax stems reached full maturity. Overall mortality of M. janthiniformis during a lifecycle was low for all samples; approximately 83% of adults successfully emerged from overwintered stems in the following spring, and greater than 65% of larvae survived to adulthood before overwintering. This resulted in >50% of weevils surviving larval development, overwintering, and spring emergence as adults. The majority of M. janthiniformis deaths (51%) resulted from parasitism by chalcidoid wasps during summer development to adulthood. These parasitoid wasps, and also M. janthiniformis adults, likely created the peculiar exit holes that were observed in live Dalmatian toadflax stems during the summer. Although M. janthiniformis populations were slow to provide effective control of Dalmatian toadflax at sites in Utah, this study indicates that the phenology and survivorship of M. janthiniformis individuals in Utah are well suited for successful biocontrol. If given enough time to build populations, M. janthiniformis appears to be capable of providing effective Dalmatian toadflax control in southern regions, but other limiting factors, such as precipitation and host plant quality, should be considered in future studies to explain slow weed suppression. Phenology models and estimates of mortality of M. janthiniformis generated by this study at sites in Utah may be helpful in implementing future biocontrol programs of Dalmatian toadflax.

SUMMARY Pseudomonas corrugata is the causal agent of tomato pith necrosis yet it is also used in the biological control of plant pathogenic bacteria and fungi. Potentially it could be used in other fields, such as the production of commercial biomolecules with a wide range of application and including bioremediation. This review reports the multiple characteristics of the bacterium, highlights its known molecular features and speculates on the possible underlying mechanisms of action. Taxonomy: Bacteria; Proteobacteria; γ subdivision; order Pseudomonadales; family Pseudomonadaceae; genus Pseudomonas. Microbiological properties: Gram-negative, oxidase-positive, non-spore forming rods; non-fluorescent on King's B medium; produces wrinkled and rarely smooth colonies on yeast peptone glucose agar or nutrient dextrose agar; yellow to brown diffusible pigments are frequently produced. Disease symptoms: The typical symptom on tomato is necrosis and/or hollowing of the pith of the stem; the syndrome determines loss of turgidity of the plant, hydropic/necrotic areas and long conspicuous adventitious roots on the stem. Biological control agent:In vitro assessed against plant pathogenic fungi and bacteria, as well as the phytotoxin indicator microorganims Rhodotorula pilimanae and Bacillus megaterium; in vivo used against pre- and post-harvest plant pathogens.

The fungal species Trichoderma harzianum is widely used as a biological agent in crop protection. To verify the continued presence of this fungus on plant roots manually inoculated with T. harzianum strain T22, a Q-PCR was designed using specific probes for this particular strain. To develop these molecular diagnostic tools, genome mining was first carried out to retrieve putative new regions by which different strains of T. harzianum could be distinguished. Subsequently, Sanger sequencing of the L-aminoacid oxidase gene (aox1) in T. harzianum was applied to determine the mutations differing between various strains isolated from the Trichoderma collection of Koppert Biological Systems. Based on the sequence information obtained, a set of hydrolysis probes was subsequently developed which discriminated T. harzianum T22 strains varying in only a single nucleotide. Probes designed for two strains uniquely recognized the respective strains in Q-PCR with a detection limit of 12,5 ng DNA. Titration assays in which T. harzianum DNA from distinct strains was varied further underscored the specificity of the probes. Lastly, fungal DNA extracted from roots of greenhouse cultured tomato plants was analyzed using the probe-based assay. DNA from T. harzianum strain T22 could readily be identified on roots of greenhouse reared tomato plants inoculated with varying concentrations up to one week after treatment with a detection limit of 3e6 colony forming units of T. harzianum T22. We conclude that the Q-PCR method is a reliable and robust method for assessing the presence and quantity of T. harzianum strain T22 in manually inoculated plant material. Our method provides scope for the development of DNA based strain specific identification of additional strains of Trichoderma and other fungal biological control agents.

Previous studies have described the development of control methods against bacterial wilt diseases caused by Ralstonia solanacearum. This review focused on recent advances in control measures, such as biological, physical, chemical, cultural, and integral measures, as well as biocontrol efficacy and suppression mechanisms. Biological control agents (BCAs) have been dominated by bacteria (90%) and fungi (10%). Avirulent strains of R. solanacearum, Pseudomonas spp., Bacillus spp., and Streptomyces spp. are well-known BCAs. New or uncommon BCAs have also been identified such as Acinetobacter sp., Burkholderia sp., and Paenibacillus sp. Inoculation methods for BCAs affect biocontrol efficacy, such as pouring or drenching soil, dipping of roots, and seed coatings. The amendment of different organic matter, such as plant residue, animal waste, and simple organic compounds, have frequently been reported to suppress bacterial wilt diseases. The combined application of BCAs and their substrates was shown to more effectively suppress bacterial wilt in the tomato. Suppression mechanisms are typically attributed to the antibacterial metabolites produced by BCAs or those present in natural products; however, the number of studies related to host resistance to the pathogen is increasing. Enhanced/modified soil microbial communities are also indirectly involved in disease suppression. New promising types of control measures include biological soil disinfection using substrates that release volatile compounds. This review described recent advances in different control measures. We focused on the importance of integrated pest management (IPM) for bacterial wilt diseases.

Fusarium , a genus of filamentous fungi, has many species which serving as important pathogens to many diseases in crops. Till today, there have not been effective and efficient control methods for such fungi. Recently, scientists agree that application of biological agents is a tactful choice. Development of hypovirulent strains of fungus as biocontrol agents is very limited. This research was aimed to find hypovirulent isolates of Fusarium from field as biological agents. A hundred isolates of Fusarium from chili were collected in Boyolali, Central Java. Morphological characterization revealed that isolates performed varied colony phenotypes. Based on colony phenotype pattern, isolates were classified into five groups. From each group, one hypovirulent isolate was selected based on colony growth rate on potato dextrose agar media. The selected hypovirulent isolates were used for virulence assay in apple. The result showed that there were four hypovirulent isolates i.e.: B6, C15, D19, and E20 isolates. Total RNA extraction of the identified hypovirulent isolates revealed the existence of viral RNA in C15 isolate. Based on the existence of viral RNA in C15 isolate, the hypovirulent traits were due to mycoviral infection, whereas the hypovirulent traits performed by the other three were due to genetic factors.

To search for a useful biological control agent and to determine the optimal rearing conditions for the green lacewings Mallada desjardinsi and Chrysoperla nipponensis, thermal effects on development, survival, and adult body size were examined at seven constant temperatures (15.0, 17.5, 20.0, 22.5, 25.0, 27.5, and 30.0°C) with a photoperiod regime of 16L : 8D (h). The developmental zero of M. desjardinsi larvae was higher than that of C. nipponensis larvae. The developmental zeros of C. carnea and C. nipponensis in earlier reports were similar to that of C. nipponensis in the present study. On the other hand, that of M. desjardinsi in earlier studies was somewhat higher than that of M. desjardinsi in the present study. A lower survival rate of C. nipponensis was observed in the cocoon stage at 15.0°C. On the other hand, lower survival rates of M. desjardinsi were observed in the egg stage at 30.0°C and in the cocoon stage at 15.0, 27.5 and 30.0°C. The body size of both green lacewings was affected by temperature throughout the range tested. Smaller body sizes of M. desjardinsi and C. nipponensis adults were observed at 15.0, 27.5 and 30.0°C. These results indicate that C. nipponensis has a wider optimum range of temperature for immature development than M. desjardinsi.

The complete nucleotide sequence of mitochondrial genome of the important mycoparasitic fungus Clonostachys rosea was determined using the next-generation sequencing technology. The circular molecule is 40,921 bp long with a GC content of 27.90%. Gene prediction revealed 42 genes encoding 15 conserved proteins, 25 tRNAs, the large and small ribosomal RNAs. All genes are located on the same strand. It is found to be similar to the previously sequenced mitochondrial genomes of Acremonium chrysogenum and Nectria cinnabarina. The differences lie in the copy number of trnG-UCC and locations of trnN-GUU and cox2. The phylogenetic analysis confirmed C. rosea as a sister taxon of A. chrysogenumin (Bionectriaceae). The mitochondrial genome of C. rosea will contribute to the understanding of phylogeny and evolution of Hypocreales.

Background The fungus Pochonia chlamydosporia parasitizes nematode eggs and has become one of the most promising biological control agents (BCAs) for plant-parasitic nematodes, which are major agricultural pests that cause tremendous economic losses worldwide. The complete mitochondrial (mt) genome is expected to open new avenues for understanding the phylogenetic relationships and evolution of the invertebrate-pathogenic fungi in Hypocreales. Results The complete mitogenome sequence of P. chlamydosporia is 25,615 bp in size, containing the 14 typical protein-coding genes, two ribosomal RNA genes, an intronic ORF coding for a putative ribosomal protein (rps3) and a set of 23 transfer RNA genes (trn) which recognize codons for all amino acids. Sequence similarity studies and syntenic gene analyses show that 87.02% and 58.72% of P. chlamydosporia mitogenome sequences match 90.50% of Metarhizium anisopliae sequences and 61.33% of Lecanicillium muscarium sequences with 92.38% and 86.04% identities, respectively. A phylogenetic tree inferred from 14 mt proteins in Pezizomycotina fungi supports that P. chlamydosporia is most closely related to the entomopathogenic fungus M. anisopliae. The invertebrate-pathogenic fungi in Hypocreales cluster together and clearly separate from a cluster comprising plant-pathogenic fungi (Fusarium spp.) and Hypocrea jecorina. A comparison of mitogenome sizes shows that the length of the intergenic regions or the intronic regions is the major size contributor in most of mitogenomes in Sordariomycetes. Evolutionary analysis shows that rps3 is under positive selection, leading to the display of unique evolutionary characteristics in Hypocreales. Moreover, the variability of trn distribution has a clear impact on gene order in mitogenomes. Gene rearrangement analysis shows that operation of transposition drives the rearrangement events in Pezizomycotina, and most events involve in trn position changes, but no rearrangement was found in Clavicipitaceae. Conclusions We present the complete annotated mitogenome sequence of P. chlamydosporia. Based on evolutionary and phylogenetic analyses, we have determined the relationships between the invertebrate-pathogenic fungi in Hypocreales. The invertebrate-pathogenic fungi in Hypocreales referred to in this paper form a monophyletic group sharing a most recent common ancestor. Our rps3 and trn gene order results also establish a foundation for further exploration of the evolutionary trajectory of the fungi in Hypocreales. Electronic supplementary material The online version of this article (doi:10.1186/s12866-015-0341-8) contains supplementary material, which is available to authorized users.

O objetivo deste trabalho foi avaliar os parâmetros biológicos do ácaro predador Phytoseiulus macropilis na ausência de alimento por períodos de um a seis dias. Foram avaliados: o número de ovos por fêmea ao dia; o número total de ovos; a longevidade de fêmeas; e a razão sexual dos descendentes. Fêmeas que ficaram por mais de três dias sem alimento apresentaram queda em todas variáveis biológicas avaliadas. O predador P. macropilis somente se estabelecerá em campo se as manchas ocupadas pelo ácaro-rajado não estiverem muito distantes umas das outras. The objective of this work was to access the biological parameters of the predator mite Phytoseiulus macropilis in the food absence for periods of one or more days. Data evaluated were: egg number per female per day; total number of eggs; longevity of females; and the offspring sex rate. Female in starvation for three or more days showed decrease in all biological variables. P. macropilis only will settle in the field if patch populations occupied by spider mite are not very distant from each other.

The sweet potato weevil (Cyclas formicarius Fabr.) remains a serious threat to sweet potato (Ipomoea batatas Poir.) production and is considered the most destructive pest of sweet potatoes in the field and storagein the Philippines. Chemical control of the weevil is seldom practiced by farmers because they find it too costly, it may increase the chance for pesticide resistance, and because of public concern of its effectson non-target organisms. The use of biological controls such as entomopathogenic nematodes (EPN) could offer an effective, economical, and environmentally-friendly alternative management of the weevil. This study determined the occurrence and distribution of entomopathogenic nematodes in selected sweet potato growing areas in the Philippines. Using soil from 13 sweet potato growing areas, EPNs were recovered using the insect baiting method. Morbid insect larvae were suspended in sterile water for 48 h, and the suspension was examined under a stereomicroscope for the presence of EPN. Out of 47 samples collected from the 13 sweet potato production areas, 39 (82%) were positive for the presence of EPNs. Preliminary identification of the EPNs through morphological characters showed that they belonged to Rhaditida: Heterorhabditidae and Steinernematidae. This is the first report on the occurrence of EPNs in sweet potato fields in the Philippines, and their distribution strongly supports the possibility of utilizing them in an IPM management approach as biological agents against the sweet potato weevil. Morphometric and molecular-based identification and pathogenicity studies are underway.

Rehabilitate forest and farm critical must special attention and need of the new method that practical, quick and cheap. Method of seed ball is its alternative method with consideration referred, then this research bent on for (1) know seed amount every seed ball that give percentage grows and plants growth sengon the best (2) know size of seed ball diameter the best to percentage grow and plants growth sengon; (3) know appropriate type of farm closing to percentage grow and plants growt sengon; (4) know interaction influence between seed amount every ball, level of seed ball, type of the best farm closing to percentage grow and plants growth sengon at the site. This research is executed start month September 2012 up to final December 2012 in district Tinggimoncong Gowa Regency. This research uses random design complete pattern factorial that consisted of 3 factor that is seed amounts every seed ball as first factor with 4 levels that is 5 seeds, 7 seeds, 9 seeds, and 11 seeds every seed ball, second that is large size factor its diameter of seed ball that consisted of 4 levels that is 2.5 cm, 4.0 cm, 5.5 cm, and 7.0 cm, third factor that is farm closing that consisted of 4 levels that is opened (grass), coppice, beams, and cleaned (disc with diameter 20 cm). The three of this treatment factor are combined until got counted 4 x 4 x 4 = 64 treatment units. Every treatment unit consisted of 3 seed balls and repeated 3 times until required 64 x 3 x 3 = 576 seed balls. Variable that perceived is time starts germinate and percentage germinates every week during a month first, plants growth (high and leaf amount) every month after second month for 2 months. Research result indicates that treatment of land cover has an effect on reality to germination percentage, leaf amount, and high plants as for seedball diameter, seed amount and interaction between land cover and seedball diameter have an effect on reality to percentage of germination and leaf amount. Treatment of grass land cover gives influence is done well by germination percentage, leaf amount, and high plants. Seedball diameter 7,0 cm and seed amount 11 seeds give influence is done well by germination percentage. Combination of grass land cover treatment and diameter of seed ball 7,0 cm give best influence.

The majority of Coccinellidae are beneficial predators and they have received considerable research attention because of their potential as biological control agents. Indeed the role of coccinellids as predators of pest insects has been a major factor in the movement of coccinellids between countries. The commercial production of coccinellids by biological control companies and local producers led to a rapid increase in distribution thoughout the 1990's. To date, 13 alien coccinellid species have been documented in Europe; 11 of these are alien to Europe (two are alien to Great Britain and Sweden but native within Europe). The distribution of alien coccinellids in Europe mirrors the biogeographical distribution and patterns of introduction. Some species have dispersed widely; Harmonia axyridis has spread rapidly from countries where it was deliberately introduced to many others across Europe. The ecological and economic impacts of alien coccinellids are not well documented. In this chapter we provide an overview of the temporal and spatial patterns of alien coccinellids in Europe.

Field surveys were carried out in four agroecological zones to assess the geographic distribution, host range and perennation of Cotesia sesamiae and Cotesia flavipes in cultivated and natural habitats in Kenya. The distribution of the two Cotesia species in different ecological regions was most affected by the suitability of the local stem borer species for parasitoid development, and temperature, as both species were found in localities dominated by their suitable host(s) where temperature favoured their occurrence. Fourteen years after its release, C. flavipes has maintained a high level of specificity to its target host Chilo partellus on maize and sorghum in cultivated habitats and on Sorghum arundinaceum in natural habitats. Cotesia flavipes appeared to be an appropriate biological control agent against C. partellus in eastern Africa, with minimal or no effects on non-target hosts in different habitats. Conversely, C. sesamiae lacked host specificity in different habitats, as its stem borers or host plants varied with both locality and habitat type. Perennation by both Cotesia species occurred mainly in cultivated habitats. Furthermore, natural habitats played a role in sustaining some individuals of C. flavipes during both rainy and dry seasons. These areas acted as refuges for C. flavipes, but not for C. sesamiae, because its hosts were scarce on natural host plants. The availability of these Cotesia species across seasons was mainly influenced by the presence of actively feeding stem borers on cereal plants during different seasons, as well as the duration of the dry season in different localities.

WOS: 000233851700006 The effects of the endoparasitic fungus Hirsutella minnesotensis on populations of Meloidogyne hapla from Michigan (MI), Rhode Island (RI), Connecticut (CT), Lyndonville, New York (NYL), Geneva, New York (NYG), and Wisconsin (WI) were studied in the greenhouse. Twenty-day-old tomato (cv. Rutgers) seedlings were inoculated with either 0 or 600 eggs of each nematode population mixed with either 0, 0.02, or 0.1 g of fresh H. minnesotensis mycelium 0.1 L-1 of soil in pots containing 0.5 L of soil, and maintained at 25 +/- 2 degrees C for 2 months. No effect of the fungal treatments and nematode treatments on tomato plant heights and shoot dry weights was observed. While all M. hapla populations were suppressed by H. minnesotensis, the degree to which each population was affected varied slightly. Across fungal treatments and nematode populations, the fungus reduced total number of nematodes in roots by 61-98%, with the highest for NYG and RI, intermediate for NYL and CT, and lowest for MI and WI populations. The study demonstrated that H. minnesotensis may be used as a potential suppressor of M. hapla in vegetable production systems in the Great Lakes Region.

Oenopia conglobata is one of the most common ladybird species in urban green areas of the Mediterranean region. We have obtained data about its prey acceptability and prey preferences. In a laboratory experiment, we investigated the acceptability of seven aphid and one psyllid species as prey for this coccinellid: the aphids Chaitophorus populeti, Aphis gossypii, Aphis craccivora Monelliopsis caryae, Eucallipterus tiliae, Aphis nerii (on white poplar, pomegranate, false acacia, black walnut, lime, and oleander, respectively), and the psyllid Acizzia jamatonica on Persian silk tree. These species are abundant in urban green areas in the Mediterranean region. In addition, we tested the acceptability of Rhopalosiphum padi on barley, an aphid species easily reared in the laboratory. We also tested preferences of the predator in cafeteria experiments with three aphid species and one aphid and the psyllid. Adults and larvae of the coccinellid accepted all of the preys offered, except A. nerii, with a clear preference for M. caryae. The predator also showed preference for M. caryae when it was offered in a cafeteria experiment with other aphid species or with the psyllid. The aphid R. padi obtained a good acceptability and could be used for rearing O. conglobata in the laboratory. We thank Noemí Bel for help in the aphid collections and the development of the experiment, and Verena Rösch for English revision. This work was partially funded by the project UdL Impuls X10020 (Universitat de Lleida—Santander Bank).