Greenhouse biofumigation with <Emphasis Type="Italic">Melia azedarach</Emphasis> controls <Emphasis Type="Italic">Meloidogyne</Emphasis> spp. and enhances soil biological activity

The need for environmentally friendly agricultural practices has led to the development of plant-based nematicides for root-knot nematode control. The efficacy of these botanicals has been tested primarily under laboratory and rarely under actual field conditions. Moreover, any side effects on non-target soil organisms that support soil biological activity are usually ignored. Herein, we evaluate the efficacy of Melia azedarach preparations against Meloidogyne spp. in a tomato greenhouse, by root gall examination and soil J2 enumeration. We also assessed side effects on soil microbes through PLFA analysis and microbivorous nematodes, and we quantified several plant growth parameters (e.g., fruit number and weight, root weight). Different treatments within the greenhouse included M. azedarach ripe fruit powder (MFP), ripe fruit water extract (MWE) and furfural, one of the principal active ingredients of M. azedarach and previously known to exhibit fumigant nematicidal activity. Results were compared to those obtained with the commercial nematicide oxamyl (Vydate^® 10 SL) and an untreated control. All treatments were repeated every 20 days throughout the cultivation period. MFP and MWE suppressed Meloidogyne spp. often at the same levels obtained by furfural and oxamyl treatments and enhanced soil biological activity, as indicated by the proliferation of soil microbes and microbial feeding nematodes. Furfural and oxamyl adversely affected the soil community, especially the free-living nematodes. Moreover, furfural was phytotoxic to tomato plants in spite of its natural origin.     

The relationship between the chemical composition of three essential oils and their insecticidal activity against Acanthoscelides obtectus (Say)

The chemical composition of the essential oils isolated from various parts of three Greek aromatic plants (Lavandula hybrida Rev, Rosmarinus officinalis L and Eucalyptus globulus Labill) collected at different seasons was determined by GC/MS analysis. The insecticidal action of these oils and of their main constituents on Acanthoscelides obtectus (Say) adults was evaluated and their LC50 values were estimated. All essential oils tested exhibited strong activity against A. obtectus adults, with varying LC50 values depending on insect sex and the composition of the essential oils. A correlation between total oxygenated monoterpenoid content and activity was observed, with oxygenated compounds exhibiting higher activity than hydrocarbons. Among the main constituents, only linalyl and terpinyl acetate were not active against A. obtectus, while all the others exhibited insecticidal activity against both male and female adults, with LC50 values ranging from 0.8 to 47.1 mg litre(-1) air. An attempt to correlate the insecticidal activity to the monoterpenoid's structure is presented, and the difference in sensitivity between male and female individuals is also explored.     

Synergistic and antagonistic interactions of terpenes against Meloidogyne incognita and the nematicidal activity of essential oils from seven plants indigenous to Greece

BACKGROUND: Biorational means for phytonematode control were studied within the context of an increasingly ecofriendly pest management global approach. The nematicidal activity and the chemical composition of essential oils (EOs) isolated from seven plants grown in Greece and ten selected compounds extracted from them against second‐stage juveniles (J2) of Meloidogyne incognita (Kof. & White) Chitwood were evaluated using juvenile paralysis experiments. Additionally, synergistic and antagonistic interactions between nematicidal terpenes were studied using an effect addition model, with the comparison made at one concentration level. RESULTS: The 96 h EC₅₀ values of Foeniculum vulgare Mill., Pimpinella anisum L., Eucalyptus meliodora A Cunn ex Schauer and Pistacia terebinthus L. were 231, 269, 807 and 1116 µg mL^?1, respectively, in an immersion bioassay. Benzaldehyde (9 µg mL^?1) was the most toxic compound, followed by γ‐eudesmol (50 µg mL^?1) and estragole (180 µg mL^?1), based on 96 h EC₅₀ values. The most potent terpene pairs between which synergistic actions were found, in decreasing order, were: trans‐anethole/geraniol, trans‐anethole/eugenol, carvacrol/eugenol and geraniol/carvacrol. CONCLUSION: This is the first report on the activity of F. vulgare, P. anisum, E. meliodora and P. terebinthus, and additionally on synergistic/antagonistic nematicidal terpene interactions, against M. incognita, providing alternative methods for nematode control. Copyright © 2010 Society of Chemical Industry     

Effect of geraniol,a plant-based alcohol monoterpene oil,against <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

In continuing previous work on the role of AAA+ proteins of the apple proliferation agent ‘Candidatus Phytoplasma mali’ in phytoplasma virulence and suppression of virulence, 147 full-length deduced protein sequences of AAA+ ATPase AP460 of single- and multiple-strain accessions were examined. This approach revealed that in this protein two regions can be distinguished. Region 1, located in the N-terminal part, is characterized by 14 highly conserved substitutions associated with the suppression of virulence. However, these substitutions were not present in all attenuated strains. In the more diverse region 2, located in the C-terminal part, highly conserved substitutions associated with two groups of virulence were identified. In addition to the virulence-related residues, three other groups of conserved substitutions are present in strains with attenuated virulence with or without the presence of suppression-associated substitutions in region 1. In one of these groups, substitutions next to key residues of the ATPase motifs sensor 1 and 2 and arginine finger do occur that seem to affect ATPase function. This group of substitution was present in all attenuated strains. From these findings it can be concluded that two different mechanisms of suppression exist of which the supposed effect on ATPase function seems to be more important than the suppression-associated substitutions in region 1. The presence of virulent, avirulent and suppressive strains in a tree leads in the commonly occurring multiple infections to interactions in which the resulting virulence is determined by the numerical relation of virulent and attenuated strains.     

Integrative diagnosis and molecular phylogeny of dagger and needle nematodes of olives and grapevines in the island of Crete,Greece, with description of Xiphinema cretense n. sp. (Nematoda,Longidoridae)

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated and wild olive and grapevine in Crete Island were investigated. Morphological and morphometrical studies identified five Longidorus and six Xiphinema species, with frequencies of prevalence (for wild and cultivated olives and grapevines, respectively) as follows: Longidorus closelongatus (2.0–13.3 %), L. cretensis (1.0–6.7 %), L. moesicus (13.3 % only in grapevines), L. orientalis (3.3 % only in grapevines), L. pseudoelongatus (7.0 % only in olives), Xiphinema cretense n. sp. (3.0 % only in olives), X. index (3.0–23.3 %), X. israeliae (6.3 % only in olives), X. italiae (3.3–10.0 %), X. pachtaicum (26.7–42 %) and X. simile (3.3 % only in grapevines). Xiphinema cretense n. sp. is characterized by a body size 3,872–6,135 μm long, lip region anteriorly rounded, separated from the rest of the body by a depression, odontostyle and odontophore 140.6 and 80.3 μm long respectively, vulva position at 46.0–50.5 %, female tail 31.0–38.0 μm long, nearly hemispherical with curvature essentially dorsal and with a tip completely rounded or presenting a very short bulge, c ratio (119.1–186.9), c’ ratio (0.7–0.8). Molecular characterisation using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids.     

Influence of soil physicochemical and biological properties on the degradation and adsorption of the nematicide fosthiazate

The degradation and adsorption of the organophosphorus nematicide fosthiazate were investigated in nine soils with various physicochemical and biological characteristics. Fosthiazate was more persistent in acidic soils (pH <6), with half-life (t1/2) values ranging from 53.3 to 57.7 days, compared to soils with higher pH (pH >7), with t1/2 ranging from 14.1 to 20.7 days. Application of antibacterial and antifungal antibiotics to soil samples resulted in a significant inhibition of fosthiazate degradation only in two of the three acidic soils. In contrast, soil autoclaving resulted in doubling the t1/2 of fosthiazate in all studied soils, suggesting that both microbial and abiotic processes contribute to fosthiazate degradation. Statistical analysis indicated a significant negative correlation (P < 0.01) between soil pH and t1/2. Fosthiazate was generally weakly adsorbed with Freundlich adsorption coefficient (Kf) values ranging from 1.23 to 2.74 mL/g. Fosthiazate concentration was strongly correlated with soil organic matter content with higher Kf values in soils with higher organic matter content (P < 0.01). The mean t1/2 and Kf values derived from the laboratory studies were used to parametrize the FOCUS groundwater (GW) models PRZM, PELMO, PEARL, and MACRO for nematicide application in potato and tomato crops. Predicted environmental concentrations produced by the models PEARL and MACRO suggested a potential risk for GW in several scenarios, unlike PELMO and PRZM, which predicted low risk for GW. These findings suggest that the environmental fate of fosthiazate is strongly influenced by soil characteristics and that this nematicide should be used with care in acidic, light soils with low organic matter content.     

The potential use of carvacrol for the control of <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

The protein kinase HOG1 in high osmolarity glycerol pathway is one of mitogen-activated protein kinases and plays an important role in fungi cellular responses to external stress and stimuli. In this study, AlHOG1, a yeast HOG1 homolog, was isolated from tobacco pathogenic fungus Alternaria longipes and its disrupted mutants were generated using homologous recombination. Compared with the wild-type strain, the mutants grew slower under Minimal-Media, produced less conidia on Potato-Dextrose-Agar and showed a retardation phenotype in conidial germination rate. Colony color observation indicated that AlHOG1 most likely negatively regulated melanin production under nutrients-deficient or osmotic stress. The same action mode of AlHOG1 on hyphal growth in response to certain carbon or nitrogen sources was speculated. Additionally, the disrupted strains were sensitive to osmotic and oxidative stress, and showed increased resistance to dicarboximide, phenylpyrrole and carbendazim fungicides. Meanwhile, results from the comparative analysis between the AlHOG1- and the AlHK1 (encoding a group III histidine kinase)-disrupted mutants showed that AlHOG1 mediated-pathway was partly regulated by the upstream AlHK1 kinase for cellular resistance to salts osmotic stress and that AlHK1 was a primary regulator for cellular sugar and fungicides stress adaptation and likely modulated both AlHOG1-mediated pathway and other unknown mechanisms. Noteworthily, AlHOG1 but not AlHK1 was confirmed to be associated with high concentration glycerol adaptation, which is the first report to our knowledge. Overall, our results indicated that AlHOG1 was involved in multiple physiological processes in A. longipes, including mycelial and conidial development, melanin production, nutrients sensing and multi-stress responses.     

Residue distribution of the acaricide coumaphos in honey following application of a new slow-release formulation

Acaricide used in beehives for the control of varroa often leaves residues in bee products. The behaviour and distribution of the acaricide coumaphos in honey following the application of a new slow-release strip formulation (CheckMite+) was assessed. The bee colonies were allowed to build new combs without foundation, and two strips were hung in the brood chamber of each colony for a period of 42 days. The distribution of coumaphos residues in honey in relation to the position of the frame and the duration of treatment was examined by collecting samples from each comb at various time intervals up to 145 days after treatment. In the brood chamber, coumaphos was incorporated into honey from the first day of application, and residues accumulated mainly in combs placed next to strips. In the adjacent combs, residues remained at low concentrations with slight variations. In the honey chamber, residue concentrations on the day of strip removal ranged between 0.006 and 0.020 mg kg(-1), while 79 days after application the concentration of coumaphos residues was below 0.020 mg kg(-1). Residues above the EC fixed maximum residue limit (MRL) of 0.1 mg kg(-1) were measured only in brood chamber honey obtained from those combs placed next to strips. In these samples, 0.060-0.111 mg kg(-1) of coumaphos was detected up to 103 days after strip removal. Coumaphos residues in honey extracted from combs that were placed at the edge of the brood chamber were found below the MRL value, even during the 42 day period of CheckMite+ strip treatment.