Effects of Low Doses of a Novel dsRNA-based Biopesticide (Calantha) on the Colorado Potato Beetle.

The Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) is a destructive pest of the cultivated potato, Solanum tuberosum. Members of this species are well-suited to agricultural habitats because of a suite of physiological adaptations and their ability to evolve resistance to multiple insecticides. Recently, a novel double-stranded RNA (dsRNA) insecticide (Calantha, active ingredient ledprona) has been demonstrated as an effective tool to manage Colorado potato beetle populations through RNA interference (RNAi). Previous studies have demonstrated the lethality of the high doses of ledprona but had not assessed possible effects of low doses that may happen due to product degradation in the environment, incomplete spray coverage, and foliage growth. Exposure of fourth instar larvae to low concentrations of ledprona interfered with their pupation. Exposure of adults significantly reduced their mobility after seven days, as well as their fertility. Reproductive effects were stronger in females, especially when exposed before reaching sexual maturity. The observed effects of low doses of ledprona may aid in the overall management of Colorado potato beetles by reducing the size of resident populations, inhibiting beetle movement within and between fields, and reducing the population growth rate.

Discovery of novel natural products for mosquito control.

Vector control plays a key role in reducing the public health burden of mosquito-borne diseases. Today's vector control strategies largely rely on synthetic insecticides that can have a negative environmental impact when applied outdoors and often become inefficient because of the mosquitoes' ability to develop resistance. An alternative and promising approach to circumvent these challenges involves the implementation of insecticides derived from nature (biopesticides) for vector control. Biopesticides can constitute naturally occurring organisms or substances derived from them that have lifespan-shortening effects on disease vectors such as mosquitoes. Here we present the discovery and evaluation of natural product-based biological control agents that can potentially be developed into biopesticides for mosquito control. We screened a natural product collection comprising 390 compounds and initially identified 26 molecules with potential ability to kill the larval stages of the yellow fever mosquito Aedes aegypti, which is responsible for transmitting viruses such as dengue, Zika, chikungunya and yellow fever. Natural products identified as hits in the screen were further evaluated for their suitability for biopesticide development. We show that a selection of the natural product top hits, bactobolin, maytansine and ossamycin, also killed the larval stages of the malaria-transmitting mosquito Anopheles gambiae as well as the adult form of both species. We have further explored the usefulness of crude extracts and preparations from two of the best candidates' sources (organisms of origin) for mosquitocidal activity, that is extracts from the two bacteria Burkholderia thailandensis and Streptomyces hygroscopicus var. ossamyceticus.

Óleos essenciais e extratos vegetais como ferramentas alternativas ao controle químico de larvas de Aedes spp, Anopheles spp e Culex spp / Essential oils and plant extracts as alternative tools for chemical control of Aedes spp, Anopheles spp, and Culex spp

Objetivos: Realizar um levantamento das contribuições científicas produzidas entre 2017 e 2021 acerca do efeito larvicida de óleos essenciais e extratos vegetais no controle de Aedes spp, Anopheles spp e Culex spp. Métodos: de setembro a outubro de 2022, foi realizado um levantamento de artigos científicos publicados entre os anos de 2017 e 2021, nas bases de dados Portal Periódicos Capes, Scielo, Science Direct e Scopus. Foram utilizados os descritores "larvicide", "essential oil" e "plant extracts" com a interposição do operador boleano "AND". Resultados: inicialmente, foram obtidos 246 artigos, dos quais 110 foram excluídos (68 não estavam disponíveis na íntegra e 42 apareceram em mais de uma base de dados). Dos 136 artigos restantes, 36 foram excluídos por não terem realizado ensaio larvicida. Dos 100 artigos remanescentes, 63 foram excluídos por não mencionarem valores de CL50, enquanto 3 não especificaram a estrutura vegetal de obtenção dos produtos naturais, restando, portanto, 34 artigos para análise. Foram utilizadas 57 espécies vegetais para a obtenção dos produtos vegetais utilizados contra larvas de Aedes spp; 11 espécies nos ensaios contra Anopheles spp, e 36 espécies nos ensaios contra Culex spp. Os óleos essenciais predominaram nos ensaios contra Aedes spp, enquanto os extratos, contra Anopheles spp. A maior parte dos produtos testados exibiu CL50 < 100 ppm. Conclusão: a atividade larvicida demonstrada por uma grande variedade de extratos vegetais e óleos essenciais representa uma alternativa promissora ao tradicional controle químico feito à base de inseticidas sintéticos em programas de manejo integrado de vetores.

Objectives: Conduct a survey of the scientific contributions produced between 2017 and 2021 on the larvicidal effect of essential oils and plant extracts in the control of Aedes spp, Anopheles spp, and Culex spp. Methods: from September to October 2022, a survey was carried out of scientific articles published between 2017 and 2021 in the Portal Periódicos Capes, Scielo, Science Direct, and Scopus databases. The descriptors "larvicide", "essential oil" and "plant extracts" were used with the Boolean operator "AND". Results: initially, 246 articles were obtained, of which 110 were excluded (68 were not available, and 42 appeared in more than one database). Of the remaining 136 articles, 36 were excluded because they did not perform a larvicide assay. Of the 100 remaining articles, 63 were excluded for not mentioning LC50 values, while three did not specify the plant structure for obtaining natural products, thus leaving 34 articles for analysis. A total of 57 plant species were used to obtain plant products used against Aedes spp larvae; 11 species in the tests against Anopheles spp, and 36 species in the tests against Culex spp. Essential oils predominated in the tests against Aedes spp, while extracts against Anopheles spp. Most of the products tested exhibited an LC50 < 100 ppm. Conclusion: the larvicidal activity demonstrated by a wide variety of plant extracts and essential oils represents a promising alternative to traditional chemical control based on synthetic insecticides in integrated vector management programs.

Identification of antibacterial metabolites produced by a marine bacterium Halobacillus marinus HMALI004.

AIMS: This study examined and characterized the extract for metabolites of Halobacillus marinus HMALI004 to understand their antibacterial activities against opportunistic marine pathogens, that is, Vibrio parahaemolyticus and Vibrio cholerae. METHODS AND RESULTS: The bacterial strain HMALI004 was characterized as H. marinus, and an antibacterial spectral test revealed its inhibition against two opportunistic marine pathogens (V. parahaemolyticus and V. cholera). Fermentation broth of strain HMALI004 was subjected to column chromatography and high-performance liquid chromatography to separate antibacterial substances. Two compounds were successfully isolated and identified as 1H-pyrrole-2-carboxylic acid and 4-chloro-1H-pyrrole-2-carboxylic acid by mass spectrometry (MS) and nuclear magnetic resonance. The minimal inhibition concentration (MIC) values of 1H-pyrrole-2-carboxylic acid and 4-chloro-1H-pyrrole-2-carboxylic acid for V. parahaemolyticus were 25 µg/ml, while their MIC values for V. cholerae were 50 and 100 µg/ml, respectively. The reactive oxygen species (ROS) production of two pathogen strains treated with 1H-pyrrole-2-carboxylic acid and 4-chloro-1H-pyrrole-2-carboxylic acid were detected to investigate the antimicrobial mechanism. The results suggested that 4-chloro-1H-pyrrole-2-carboxylic acid exerted enhanced ROS production in V. parahaemolyticus, whereas 1H-pyrrole-2-carboxylic acid had a weaker effect. Both compounds caused a significant rise in ROS production in V. cholerae, causing severe damage to the cell wall and cytoplasm, leading to cell death. CONCLUSIONS: The bacterium H. marinus HMALI004 was isolated from a shrimp pond and was found to produce antimicrobial compounds, which could inhibit the growth of opportunistic marine pathogens V. parahaemolyticus and V. cholerae by increasing ROS. SIGNIFICANCE AND IMPACT OF THE STUDY: Successfully isolated antibacterial-producing strain, H. marinus HMALI004, and its antimicrobial compounds could be used as biological control agents for marine pathogens.

Specialized Metabolites from the Allelopathic Plant Retama raetam as Potential Biopesticides.

To cope with the rising food demand, modern agriculture practices are based on the indiscriminate use of agrochemicals. Although this strategy leads to a temporary solution, it also severely damages the environment, representing a risk to human health. A sustainable alternative to agrochemicals is the use of plant metabolites and plant-based pesticides, known to have minimal environmental impact compared to synthetic pesticides. Retama raetam is a shrub growing in Algeria's desert areas, where it is commonly used in traditional medicine because of its antiseptic and antipyretic properties. Furthermore, its allelopathic features can be exploited to effectively control phytopathogens in the agricultural field. In this study, six compounds belonging to isoflavones and flavones subgroups have been isolated from the R. raetam dichloromethane extract and identified using spectroscopic and optical methods as alpinumisoflavone, hydroxyalpinumisoflavone, laburnetin, licoflavone C, retamasin B, and ephedroidin. Their antifungal activity was evaluated against the fungal phytopathogen Stemphylium vesicarium using a growth inhibition bioassay on PDA plates. Interestingly, the flavonoid laburnetin, the most active metabolite, displayed an inhibitory activity comparable to that exerted by the synthetic fungicide pentachloronitrobenzene, in a ten-fold lower concentration. The allelopathic activity of R. raetam metabolites against parasitic weeds was also investigated using two independent parasitic weed bioassays to discover potential activities on either suicidal stimulation or radicle growth inhibition of broomrapes. In this latter bioassay, ephedroidin strongly inhibited the growth of Orobanche cumana radicles and, therefore, can be proposed as a natural herbicide.

New Sorbicillinoids with Tea Pathogenic Fungus Inhibitory Effect from Marine-Derived Fungus Hypocrea jecorina H8.

Four new dimeric sorbicillinoids (1-3 and 5) and a new monomeric sorbicillinoid (4) as well as six known analogs (6-11) were purified from the fungal strain Hypocrea jecorina H8, which was obtained from mangrove sediment, and showed potent inhibitory activity against the tea pathogenic fungus Pestalotiopsis theae (P. theae). The planar structures of 1-5 were assigned by analyses of their UV, IR, HR-ESI-MS, and NMR spectroscopic data. All the compounds were evaluated for growth inhibition of tea pathogenic fungus P. theae. Compounds 5, 6, 8, 9, and 10 exhibited more potent inhibitory activities compared with the positive control hexaconazole with an ED50 of 24.25 ± 1.57 µg/mL. The ED50 values of compounds 5, 6, 8, 9, and 10 were 9.13 ± 1.25, 2.04 ± 1.24, 18.22 ± 1.29, 1.83 ± 1.37, and 4.68 ± 1.44 µg/mL, respectively. Additionally, the effects of these compounds on zebrafish embryo development were also evaluated. Except for compounds 5 and 8, which imparted toxic effects on zebrafish even at 0.625 µM, the other isolated compounds did not exhibit significant toxicity to zebrafish eggs, embryos, or larvae. Taken together, sorbicillinoid derivatives (6, 9, and 10) from H. jecorina H8 displayed low toxicity and high anti-tea pathogenic fungus potential.

Comparison of Mineral Oil, Insecticidal, and Biopesticide Spraying Regimes for Reducing Spread of Three Potato virus Y Strains in Potato Crops.

In-field management of Potato virus Y (PVY) faces challenges caused by the changing availability and environmental acceptability of chemical agents to control aphid vectors of the virus and by proliferation of PVY strains with different symptoms and rates of spread. From 2018 to 2020, foliar spray treatments were compared in field experiments in New Brunswick, Canada, to measure effectiveness at reducing spread of PVYO, PVYN:O, and PVYNTN strains. Mineral oil, insecticide, combined oil and insecticide spray, and a biopesticide (i.e., LifeGard WG) were compared. Insecticide-only and mineral oil-only treatments were not effective, but several combined oil and insecticide treatments and biopesticide treatments significantly reduced PVY spread. The biopesticide was proportionately more effective with recombinant PVYN:O and PVYNTN strains, possibly by exciting the plant's hypersensitive resistance response, caused naturally only in cultivar 'Goldrush' by PVYO. Pesticide residue analysis showed that mineral oil increased the retention of pyrethroid insecticide in the potato foliage longer than with insecticide applied alone, which may explain the beneficial synergistic effect of combined sprays for reducing PVY spread. Tuber yields were generally unchanged in chemical insecticide treatments but were slightly lower in biopesticide treatment. The cost per PVY treatment was competitive across all effective treatments, including biopesticide; however, there was some revenue loss from lower yield with the biopesticide. This biopesticide is certified organic, however, and thus a small premium on the price for organic production could offset this yield deficit.

Bio-efficacy of Soil Actinomycetes and an Isolated Molecule 1,2-Benzenedicarboxylic Acid from Nonomuraea sp. Against Culex quinquefasciatus Say and Aedes aegypti L. Mosquitoes (Diptera: Culicidae).

Vector-borne diseases such as filariasis and dengue that contribute significantly to disease burden, death, poverty, and social frailty are still a major public healthcare problem worldwide. Currently, synthetic chemicals have been used in mosquito control programs. However, repeated use of chemical insecticides causes environmental pollution and harmful effects on non-target organisms. Therefore, alternative ecofriendly sources from biological source are urgently needed to manage mosquitoes. In this respect, the present study was aimed to evaluate mosquito larvicidal and pupicidal activities of 22 crude extracts of soil actinomycetes on Culex quinquefasciatus and Aedes aegypti and to identify the active molecule. Briefly, the crude ethyl acetate extract and fractions were tested at 62.5, 125, 250, and 500 ppm and 2.5, 5.0, 7.5, and 10.0 ppm concentrations on larval and pupal stages of Cx. quinquefasciatus and Ae. aegypti. The larval and pupal mortality was assessed after 24 h of treatment. Among the 22 isolates screened, Nonomuraea sp. VAS-16 exhibited significant larvicidal and pupicidal activities against the tested mosquito species. Among the 18 fractions screened, fraction-6 showed strong larvicidal and pupicidal activities with the LC50 and LC90 values of 9.1, 18.7, 9.82, and 22.85 ppm against the larvae and LC50 and LC90 values of 10.5, 23.1, 12.3, and 24.13 ppm against the pupae of Cx. quinquefasciatus and Ae. aegypti, respectively. Fascinatingly, the isolated compound 1,2-benzenedicarboxylic acid from fraction-6 at 0.5, 1.0, 1.5, and 2.0 ppm concentration recorded lower LC50 and LC90 values of 4.27, 14.90, 4.67, and 11.90 ppm against the larvae and LC50 and LC90 values of 4.58, 12.06, 5.36, and 13.07 ppm against the pupae of Cx. quinquefasciatus and Ae. aegypti, respectively. On the other hand, the compound recorded less ovicidal activity of 11.0% and 10.3% at 2 ppm against the eggs of Cx. quinquefasciatus and Ae. aegypti, respectively. The present study clearly shows that the crude extract and the compound from Nonomuraea sp. VAS-16 can be used as an effective biopesticide in integrated mosquito management program.

Biological Activity of Essential Oils of Four Juniper Species and Their Potential as Biopesticides.

The objective of this study was to assess the biological activity of essential oils (EOs) of four Juniperus species obtained via two different distillation methods and their potential as biopesticides. The studied factors were juniper species (Juniperus communis L., J. oxycedrus L., J. pygmaea C. Koch., and J. sibirica Burgsd), plant sex (male (M) and female (F)), and distillation method (hydrodistillation via a standard Clevenger apparatus (ClevA) and semi-commercial (SCom) steam distillation). The hypothesis was that the EO will have differential antioxidant, antimicrobial, and insecticidal activities as a function of plant species, plant sex, and distillation method. The two distillation methods resulted in similar EO composition within a given species. However, there were differences in the EO content (yield) due to the sex of the plant, and also differences in the proportions of some EO components. The concentration of α-pinene, ß-caryophyllene, δ-cadinene and δ-cadinol was dissimilar between the EO of M and F plants within all four species. Additionally, M and F plants of J. pygmaea, and J. sibirica had significantly different concentrations of sabinene within the respective species. The EOs obtained via ClevA extraction showed higher antioxidant capacity within a species compared with those from SCom extraction. All of the tested EOs had significant repellent and insecticidal activity against the two aphid species Rhopalosiphum padi (bird cherry-oat aphid) and Sitobion avenae (English grain aphid) at concentrations of the EO in the solution of 1%, 2.5%, and 5%. The tested EOs demonstrated moderate activity against selected pathogens Fusarium spp., Botrytis cinerea, Colletotrichum spp., Rhizoctonia solani and Cylindrocarpon pauciseptatum. The results demonstrate that the standard ClevA would provide comparable EO content and composition in comparison with SCom steam distillation; however, even slight differences in the EO composition may translate into differential bioactivity.

Essential oils of Bursera morelensis and Lippia graveolens for the development of a new biopesticides in postharvest control.

Fruit and vegetable crops that are not consumed immediately, unlike other agricultural products, require economic and time investments until they reach the final consumers. Synthetic agrochemicals are used to maintain and prolong the storage life of crops and avoid losses caused by phytopathogenic microorganisms. However, the excessive use of synthetic agrochemicals creates health problems and contributes to environmental pollution. To avoid these problems, less toxic and environment-friendly alternatives are sought. One of these alternatives is the application of biopesticides. However, few biopesticides are currently used. In this study, the biopesticide activity of Bursera morelensis and Lippia graveolens essential oils was evaluated. Their antifungal activity has been verified in an in vitro model, and chemical composition has been determined using gas chromatography-mass spectrometry. Their antifungal activity was corroborated in vitro, and their activity as biopesticides was subsequently evaluated in a plant model. In addition, the persistence of these essential oils on the surface of the plant model was determined. Results suggest that both essential oils are promising candidates for producing biopesticides. This is the first study showing that B. morelensis and L. graveolens essential oils work by inhibiting mycelial growth and spore germination and are environment-friendly biopesticides.

Mechanism of Action of Endophytic Fungi Hypocrea lixii and Beauveria bassiana in Phaseolus vulgaris as Biopesticides against Pea Leafminer and Fall Armyworm.

Endophytic fungal isolates Hypocrea lixii F3ST1 and Beauveria bassiana G1LU3 were evaluated for their potential to endophytically colonize and induce active compounds in Phaseolus vulgaris, as a defense mechanism against pea leafminer (Liriomyza huidobrensis) and fall armyworm (Spodoptera frugiperda). Endophytic colonization was achieved through seed inoculation with the volatile emissions from P. vulgaris plants being analyzed using GC-MS. The crude extracts of P. vulgaris obtained using methanol and dichloromethane were assayed against leafminer and fall armyworm larvae using leaf dipping and topical application, respectively. The two isolates successfully colonized the entire host plant (roots, stems, and leaves) with significant variation (p < 0.001) between fungal isolates and the controls. The results showed qualitative differences in the volatile profiles between the control plants, endophytically colonized and insect-damaged plants attributed to fungal inoculation and leafminer damage. The crude methanol extracts significantly reduced the percentage pupation of 2nd instar leafminer larvae (p < 0.001) and adult-flies emergence (p < 0.05). The survival of the 1st instar fall armyworm larvae was also significantly reduced (p < 0.001) compared to the controls. This study demonstrated the high potential of endophytic fungi H. lixii and B. bassiana in inducing mainly specific defense compounds in the common bean P. vulgaris that can be used against pea leafminer and fall armyworm.

Efficacy of Noni (Morinda citrifolia L.) Ethanolic Leaf Extract Against German Cockroach (Blattella germanica L.).

<b>Background and Objective:</b> The ethanol extract of noni leaves (<i>Morinda citrifolia</i> L.) can be used as insecticides to control populations of German cockroaches that have been resistant to synthetic insecticides. This study aimed to determine the potency of the ethanol extract of noni leaves to kill and repel German cockroaches and affect the amount of food consumed. <b>Materials and Methods:</b> The methods used in this study were the contact toxicity test, the repellency test and the food consumption test. The noni leaves extract and German cockroach populations were provided in the laboratory. <b>Results:</b> The noni leaves extract concentration of 20% (residue of 3.14 mg cm² ¹) was very effective in killing the standard population and effective in killing the field population of German cockroaches. The sub-lethal concentration noni leaves extract of 0.36% (residue 0.056 mg cm² ¹) and 1.08% (residue 0.169 mg cm² ¹) was very high grade as repellent of German cockroaches. The sub-lethal concentration of noni leaves extract did not inhibit the amount of food consumption in German cockroach populations. <b>Conclusion:</b> Leaves of noni plants can be used as bioinsecticides to control German cockroach populations that have been resistant to commercial insecticides.

Sensitivity of Botrytis cinerea Isolates Complex to Plant Extracts.

New agricultural strategies aim to reduce the use of pesticides due to their damage to the environment and humans, and the caused resistance to pathogens. Therefore, alternative sources of antifungal compounds from plants are under investigation lately. Extracts from plants have a wide composition of chemical compounds which may complicate the development of pathogen resistance. Botrytis cinerea, causing grey mould, is an important horticultural and ornamental pathogen, responsible for the relevant yield and quality losses. B. cinerea isolated from a different plant host may differ in the sensitivity to antifungal substances from plants. Assessing the importance of research covering a wide range of pathogens for the rapid development of biopesticides, this study aims to determine the sensitivity of the B. cinerea isolate complex (10 strains) to plant extracts, describe morphological changes caused by the extract treatment, and detect differences between the sensitivity of different plant host isolates. The results showed the highest sensitivity of the B. cinerea isolates complex to cinnamon extract, and the lowest to laurel extract. In contrast, laurel extract caused the most changes of morphological attributes in the isolates. Five B. cinerea isolates from plant hosts of raspberry, cabbage, apple, bell pepper, and rose were grouped statistically according to their sensitivity to laurel extract. Meanwhile, the bell pepper isolate separated from the isolate complex based on its sensitivity to clove extract, and the strawberry and apple isolates based on their sensitivity to cinnamon extract.

Field Experiment Effect on Citrus Spider Mite Panonychus citri of Venom from Jellyfish Nemopilema nomurai: The Potential Use of Jellyfish in Agriculture.

Jellyfish are rich in resources and widely distributed along coastal areas. As a potential approach to respond to jellyfish blooms, the use of jellyfish-derived products is increasing. The citrus spider mite (Panonychus citri) is one of the key citrus pests, negatively impacting the quality and quantity of oranges. Due to the resistance and residue of chemical acaricides, it is important to seek natural substitutes that are environmentally friendly. The field efficacy of the venom from the jellyfish Nemopilema nomurai against P. citri was assayed in a citrus garden. The frozen N. nomurai tentacles were sonicated in different buffers to isolate the venom. The venom isolated by PBS buffer (10 mM, pH 6.0) had the strongest acaricidal activity of the four samples, and the corrected field efficacy 7 days after treatment was up to 95.21%. This study demonstrated that jellyfish has potential use in agriculture.

Use of a Specific Phage Cocktail for Soft Rot Control on Ware Potatoes: A Case Study.

Using bacteriophages (bacterial viruses) to control pathogenic bacteria is a promising approach in horticulture. However, the application of this strategy in real conditions requires compliance with particular technological and environmental restraints. The presented paper concerns the process of phage selection to create a cocktail that is efficient against the circulating causal agents of potato soft rot. The resulting phage cocktail causes a complete lysis of a mixture of circulating pectobacterial strains in vitro. In the context of being used to treat ware potatoes during off-season storage, the protocol of phage application via the humidity maintenance system was designed. The phage cocktail was shown to reduce the population of Pectobacterium spp. 10-12-fold, achieving a population that was below a symptomatic threshold.

The pest kill rate of thirteen natural enemies as aggregate evaluation criterion of their biological control potential of Tuta absoluta.

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.

Exploration of microbiome of medicinally important plants as biocontrol agents against Phytophthora parasitica.

In a preliminary plant-based microbiome study, diverse bacterial taxa were identified from different medicinal plants using 16S rRNA gene sequencing. Based on initial antimicrobial screening, eight (8) bacterial endophytes in six (6) different genera, Streptomyces, Pseudomonas, Enterobacter, Bacillus, Arthrobacter, and Delftia, from four important medicinal plants Dodonaea viscosa, Fagonia indica, Caralluma tuberculata, and Calendula arvensis were selected for further analyses. Antimicrobial assays revealed that Pseudomonas taiwanensis MOSEL-RD23 has strong anti-Phytophthora activity. Volatiles produced by P. taiwanensis MOSEL-RD23and Bacillus flexus MOSEL-MIC5 inhibited the growth of Phytophthora parasitica by more than 80%. Ethyl acetate extracts of Streptomyces alboniger MOSEL-RD3, P. taiwanensis MOSEL-RD23, Enterobacter hormaechei MOSEL-FLS1, and Bacillus tequilensis MOSEL-FLS3, and Delftia lacustris MB322 displayed high potency against P. parasitica. All these bacterial extracts showed strong inhibition of more than 80% inhibition in vitro against P. parasitica at different concentrations (4-400 µg/mL). Bacterial extracts showing strong antimicrobial activity were selected for bioactivity-driven fractionation and showed anti-Phytophthoral activity in multiple fractions and different peaks observed in UV-Vis spectroscopy. In the detached-leaf assay against P. parasitica on tobacco, 1% ethyl acetate bacterial extract of S. alboniger MOSEL-RD3, P. taiwanensis MOSEL-RD23, E. hormaechei MOSEL-FLS1, B. tequilensis MOSEL-FLS3, and D. lacustris MB322 reduced lesion sizes and lesion frequencies caused by P. parasitica by 68 to 81%. Overall, P. taiwanensis MOSEL-RD23 showed positive activities for all the assays. Analyzing the potential of bacterial endophytes as biological control agents can potentially lead to the formulation of broad-spectrum biopesticides for the sustainable production of crops.

An LCI-like protein APC2 protects ginseng root from Fusarium solani infection.

AIMS: We aimed to purify an antimicrobial protein from Bacillus amyloliquefaciens FS6 culture supernatant, verify its antimicrobial activity against Fusarium solani and evaluate its biocontrol potential for ginseng root rot. METHODS AND RESULTS: The antimicrobial protein was purified from FS6 culture supernatant using ammonium sulphate precipitation, anion exchange and gel chromatography. Based on mass spectrometry results, the purified protein was identified as an antimicrobial protein of the LCI family and was designated APC2 . The APC2 recombinant protein expressed in Escherichia coli (BL21) significantly inhibited F. solani and decreased the infection and spread of F. solani in ginseng root. An overexpressing APC2 strain FS6-APC2 was constructed and shown to have enhanced antimicrobial activity compared to the wild-type strain FS6. CONCLUSIONS: The APC2 protein shows strong antimicrobial activity against F. solani, reduces the incidence and severity of ginseng root rot caused by F. solani and exhibits a great biocontrol potential. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the inhibitory activity of APC2 protein (LCI family) against F. solani and its protective efficacy on ginseng root rot. These findings provide a scientific basis for future research on the biocontrol mechanism, as well as the development and application of FS6.

Combined application of antagonistic Wickerhamomyces anomalus BS91 strain and Cynara cardunculus L. leaf extracts for the control of postharvest decay of citrus fruit.

Combined use of biocontrol agents and plant extracts can be considered a viable and promising strategy for protecting plant tissues with different synergistic mechanisms of action that improve the antimicrobial activity of the mixtures. Treatments of citrus fruits with Wickerhamomyces anomalus BS91 have been previously reported as effective measures to reduce the incidence of green mold caused by Penicillium digitatum. On the opposite, the knowledge of the antifungal activity of cultivated cardoon (Cynara cardunculus L. var. altilis DC.) leaf extract, vegetable widespread in some Mediterranean areas, is still very limited. In this study, experimental trials were conducted to evaluate the effectiveness in vitro of leaf aqueous, methanolic and ethanolic extracts of C. cardunculus against seven fungal pathogens responsible for considerable food losses in the postharvest stage. In addition, biocontrol yeast W. anomalus BS91 and the three C. cardunculus extracts were tested in vivo both as a single treatment and in mixture, against Penicillium digitatum on 'Tarocco' oranges and 'Femminello' lemons. The combination of W. anomalus BS91 and leaf ethanolic extract reduced with the highest efficacy the incidence and severity of green mold on orange and lemon fruits with respect to the control, and was more effective than treatment with antagonistic yeast or leaf extracts applied alone. Incidence and severity of citrus decay were more consistently reduced when mixtures were applied 24 h before the inoculation of the pathogen, thus suggesting the relevance of preventive treatments. The mixtures of antagonistic W. anomalus BS91 and ethanolic leaf extract were more effective in controlling green mold decay on oranges than on lemons. These results indicate that biocontrol agents and leaf extracts, used in appropriate combination, can provide a stronger protection than when used singularly. However, compatibility between microbial antagonist and antimicrobial extract should be preliminary verified.

Impregnation of pectin-cedarwood essential oil nanocapsules onto mini cotton bag improves larvicidal performances.

The use pesticide is one of the indispensable means to combat mosquito borne diseases. However, the repeated use of synthetic pesticides has induced resistance in the vector pest along with undesirable impact on the environment. The biodegradability, non-persistent and user's safety are the root cause to prefer plant-derived pesticides to synthetic ones. The botanical based pesticides tend to degrade rapidly under the influence of several environmental factors. For the feasible application as pesticides, the plant products are formulated either as liquid or as purely solid. Despite well-established formulation technology in pesticide delivery, their handling trouble is being ignored. There is difficulty in liquid formulation of pesticide products, as they are prone to splashing and spillage, resulting in contamination, wastage and direct exposure to skin; whereas a solid formulation tends to produce dust. In the present work, cedarwood (Cedrus deodara) essential oil embedded pectin nanocapsules were produced. The nanocapsules were characterized according to their morphology, size, encapsulation efficiency and thermal stability. Furthermore, the nanocapsules were impregnated onto mini cotton tea bags to be employed as RTU (ready to use) formulation for treating the breeding sites of mosquitoes. The larvicidal activity of the bags treated with pectin-cedar wood nanocapsules was assessed against malaria vector, Anopheles culicifacies and 98% mortality was recorded till 4 weeks, this suggests its potential and hassle free applications in controlling mosquito vector.