Efficacy of Azadirachtin in the Integrated Management of the Root Knot Nematode Meloidogyne incognita on Short- and Long-Cycle Crops.

Activity of azadirachtin on phytoparasitic nematodes has been documented for some decades, but the relationship between its nematicidal efficacy and crop cycle length has not yet been clarified. This study aimed to assess the efficacy of an azadirachtin-based nematicide, for controlling the infestation of the root-knot nematode Meloidogyne incognita, on the short- and long-cycle crops, lettuce and tomato, respectively. Experiments on lettuce and tomato were carried out in a greenhouse infested by M. incognita, including non-treated soil, or treated with the nematicide fluopyram, as controls. In the experiment on the short-cycle lettuce crop, the azadirachtin product effectively suppressed M. incognita infestation and increased crop yield, without significant differences from fluopyram. In the tomato crop, both azadirachtin and fluopyram were not able to control nematode infestation, but resulted in significantly higher yields. Data from this study indicated that azadirachtin can be a valid alternative to fluopyram and other nematicides, for root-knot nematode control in short-cycle crops. Integration of azadirachtin with a synthetic nematicide or nematode-suppressive agronomical techniques, should be more suitable to long-cycle crops.

Nematicidal Activity of a Garlic Extract Formulation against the Grapevine Nematode Xiphinema index.

The nematicidal activity of garlic extracts is known on root-knot nematodes but never investigated on the grapevine nematode Xiphinema index. In this study, the nematicidal activity of a commercial garlic extract formulate (GEF) was assessed on X. index, both in vitro and in a pot assay. In the in vitro assays, mixed specimens of X. index were exposed to a 0-4 mL L-1 range of GEF concentrations, checking nematode immotility and mortality after 2, 4 or 8 h. In the experiments on potted grapevines, plants cultivated in soil infested by X. index were irrigated twice at a 15-day interval with 0.05, 0.2 and 0.5 mL L-1 solutions of GEF, including nontreated soil as a control. An almost complete mortality of X. index specimens occurred after a 2 h exposure to a 2 mL L-1 GEF concentration, while an 8 h exposure to even the 0.0312 and 0.0156 mL L-1 solutions resulted in about 50% and 30% mortality, respectively. Soil treatment with a 0.5 mL L-1 GEF solution significantly reduced the population of X. index and increased the grapevine root growth compared to nontreated soil or soil treated with the lower dosages. Results of this study indicated that garlic-based nematicides could be an effective tool for X. index management in organic and integrated vineyards.

Synergistic Effects of Trichoderma harzianum, 1,3 Dichloropropene and Organic Matter in Controlling the Root-Knot Nematode Meloidogyne incognita on Tomato.

Environmental concerns raised by synthetic nematicides are encouraging integrated management strategies based on their combination with non-chemical control tools, such as biocontrol agents and/or organic amendments. In this study, the combination of the fumigant 1,3-dichloropropene (1,3-D) with a commercial formulation of the biocontrol agent Trichoderma harzianum (TH) and an organic fertilizer (OF) was investigated in two consecutive tomato crops for its effect on the root-knot nematode Meloidogyne incognita and plant growth and yield. The application of 1,3-D was only performed on the first crop, while TH and OF were provided to both crops. Almost all treatments significantly reduced nematode infestation in both crops, though the greatest nematicidal effect was caused by a combination of the three products. The treatment with 1,3-D limited its nematicidal efficacy to the first crop only. Fumigant integration with TH and OF also resulted in the greatest increases of plant growth and yield. Therefore, the integrated management of root-knot nematodes with a soil fumigant, a bionematicide as T. harzianum and a source of organic matter demonstrated effective nematode suppression though limiting the number of chemical applications.

Nematicidal Potential of Sulla (Hedysarum coronarium L.) against the Root-Knot Nematode Meloidogyne incognita.

The content of nematicidal metabolites such as saponins, flavonoids and tannins in sulla (Hedysarum coronarium L.) suggests its potential nematicidal activity. In this study, the biocidal activity of 62.5-1000 µg mL-1 concentrations of flavonoid and tannin fractions from sulla was assessed in in vitro assays on the infective juveniles (J2) of the root-knot nematode (RKN) Meloidogyne incognita, while the suppressive effects of soil amendments with 10-40 g kg-1 soil rates of sulla biomass were investigated on potted tomato infested by M. incognita. The content of total nitrogen, carbon, flavonoids, tannins and saponins of sulla experimental material was also determined. After a 96-h exposure, more than 80% of the M. incognita J2 were killed even by a 125 µg mL-1 concentration of the flavonoid extract, while mortality peaked at 89% only at the 1000 µg mL-1 concentration of the tannin solution. Soil incorporation with sulla biomass significantly reduced the M. incognita densities both on tomato roots and in the soil, compared to either the non-treated control and chemical treatment with Fluopyram. The data confirmed the nematicidal potential of sulla, mainly due to its content of flavonoids and tannins, suggesting its suitability as green manure or a soil amendment for sustainable RKN management.

Antibiotic and Nematocidal Metabolites from Two Lichen Species Collected on the Island of Lampedusa (Sicily).

The antibiotic and nematocidal activities of extracts from two coastal lichen species collected on Lampedusa Island (Sicily), Ramalina implexa Nyl. and Roccella phycopsis Ach., were tested. Methyl orsellinate, orcinol, (+)-montagnetol, and for the first time 4-chlororcinol were isolated from Roccella phycopsis. (+)-Usnic acid was obtained from Ramalina implexa. The crude organic extract of both lichen species showed strong antibiotic activity against some bacterial species and nematocidal activity. Among all the pure metabolites tested against the infective juveniles (J2) of the root-knot nematode (RKN) Meloydogine incognita, (+)-usnic acid, orcinol, and (+)-montagnetol had significant nematocidal activity, comparable with that of the commercial nematocide Velum® Prime, and thus they showed potential application in agriculture as a biopesticide. On the contrary, methyl orsellinate and 4-chlororcinol had no nematocidal effect. These results suggest that the substituent pattern at ortho-para-position in respect to both hydroxyl groups of resorcine moiety, which is present in all metabolites, seems very important for nematocidal activity. The organic extracts of both lichens were also tested against some Gram-positive and Gram-negative bacteria. Both extracts were active against Gram-positive species. The extract of Ramalina implexa showed, among Gram-negative species, activity against Escherichia coli and Acinetobacter baumannii, while that from Roccella phycopsis was effective towards all test strains, with the exception of Pseudomonas aeruginosa. The antimicrobial activity of (+)-usnic acid, methyl orsellinate, and (+)-montagnetol is already known, so tests were focused on orcinol and 4-chlororcinol. The former showed antibacterial activity against all Gram positive and Gram-negative test strains, with the exception of A. baumannii and K. pneumoniae, while the latter exhibited a potent antibacterial activity against Gram-positive test strains and among Gram-negative strains, was effective against A. baumannii and K. pneumonia. These results suggest, for orcinol and 4-chlororcinol, an interesting antibiotic potential against both Gram-positive and Gram-negative bacterial strains.

Nematicidal Activity of Essential Oil from Lavandin (Lavandula × intermedia Emeric ex Loisel.) as Related to Chemical Profile.

Essential oils (EOs) from lavandin are known for a large spectrum of biological properties but poorly and contrastingly documented for their activity against phytoparasitic nematodes. This study investigated the toxicity of EOs from three different lavandin cultivars, Abrialis, Rinaldi Cerioni, and Sumiens, either to juveniles (J2) and eggs of the root-knot nematode Meloidogyne incognita and to infective stages of the lesion nematode Pratylenchus vulnus. The suppressive activity of treatments with EOs from the three lavandin cultivars in soil infested by M. incognita was also investigated in a greenhouse experiment on potted tomato. The compositional profiles of tested EOs were also analyzed by GC-FID and GC-MS. Linalool was the major component of all the three EOs, as accounting for about 66%, 48%, and 40% of total EO from cv Rinaldi Cerioni, Sumiens, and Abrialis, respectively. Linalool acetate was the second most abundant compound in the EOs from cv Abrialis (18.3%) and Sumiens (14.9%), while significant amounts of camphor (11.5%) and 1,8-cineole (12.1%) were detected in cv Rinaldi Cerioni and Sumiens EOs, respectively. The mortality of M. incognita J2 peaked 82.0%, 95.8%, and 89.8% after a 24 h treatment with 100 mg·mL-1 solutions of cv Abrialis, Rinaldi Cerioni, and Sumiens EOs, respectively. Infective specimens of P. vulnus were largely more sensitive than M. incognita J2, as there were peak mortality rates of 65.5%, 67.7%, and 75.7% after 4 h of exposure to Abrialis, Rinaldi Cerioni, and Sumiens EO, respectively. All three lavandin EOs significantly affected also M. incognita egg hatchability, which reduced to 43.6% after a 48 h egg mass exposure to a 100 µg·mL-1 solution of cv Rinaldi Cerioni EO. Soil treatments with the three lavandin EOs strongly reduced, according to a dose-effect relationship, density of M. incognita eggs, and J2 both on tomato roots and in soil, as well as significantly reduced gall formation on tomato roots. Finally, almost all soil treatments with the lavandin EOs also resulted in a positive impact on tomato plant growth.

Selection of Bacterial Strains for Control of Root-Knot Disease Caused by Meloidogyne incognita.

Root-knot disease caused by Meloidogyne incognita leads to significant crop yield losses that may be aggravated by the association with pathogenic fungi and bacteria. Biological agents can be effectively used against the complex disease of root-knot nematode and pathogenic fungi. In this study, 35 bacterial strains were analyzed for their in vitro nematicidal, antagonistic and growth stimulation activities. Based on results from the in vitro assays, grow-box experiments on tomato and cucumber were carried out with the strain BZR 86 of Bacillus velezensis applied at different concentrations. Effects of B. velezensis BZR 86 on the development of root-knot disease were evaluated by recording root gall index, number of galls and number of eggs in egg masses. Application of B. velezensis BZR 86 noticeably decreased the development of root-knot disease on tomato and cucumber plants, as well as significantly increased growth and biomass of cucumber plants in accordance with bacterial concentration. This study seems to demonstrate that strain B. velezensis BZR 86 could be an additional tool for an environmentally safe control of root-knot disease on horticultural crops.

Chemical Composition and Nematicidal Properties of Sixteen Essential Oils-A Review.

Essential oils (EOs) can be a large source of new food-safe and healthy nematicidal products, due to their strong activity on crop pathogens and pests, including phytoparasitic nematodes, as well as to their low environmental persistence. This review summarizes the results from our 10-year studies on chemical features and nematicidal properties of 16 EOs with different botanical origins and compositions, i.e., the EOs from Artemisia herba-alba Asso (Asteraceae), Cinnamomum camphora (L.) J. Presl. and Cinnamomum verum J. Presl. (Lauraceae), Citrus aurantium L., Cinnamomum. sinensis L. Osbeck and Ruta graveolens L. (Rutaceae), Eucalyptus citriodora Hook, Eucalyptus globulus Labill. and Syzygium aromaticum (L.) Marry et Perry (Myrtaceae), Mentha piperita L., Monarda didyma L., Monarda. fistulosa L., Rosmarinus officinalis L. and Thymus satureioides Cosson (Lamiaceae), Pelargonium asperum Ehrh ex Willd (Geraniaceae) and Schinus molle L. (Anacardiaceae). All these EOs were chemically characterized and tested in vitro and/or in vivo for their activity against the phytoparasitic species Meloidogyne incognita Kofoid et White (Chitw.), Pratylenchus vulnus Allen et Jensen and Xiphinema index Thorne et Allen. Toxicity bioassays were conducted by exposing 2nd stage juveniles (J2) of M. incognita, mixed-age specimens of P. vulnus and adult females of X. index to 2-100 µg mL-1 concentrations of EOs or EO's major constituents for 4-96 h and checking mortality effect after a further 24-72 h permanence in water. Egg hatchability bioassays consisted in exposing (24-48 h) M. incognita egg masses to 500-1000 mg mL-1 EO solutions followed by a 5-week hatching test in water. The in vivo experiments were undertaken in sandy soil strongly infested by M. incognita and treated with different doses of EOs, applied either in water solution or by fumigation. The effects of the treatments on nematode infestation on tomato and in soil were checked at the end of each experiment. Structure-activity relationships, as suggested by the different chemical compositions of tested EOs, were also highlighted. In agreement with literature data, our studies indicated that most of the tested EOs are highly suitable for the formulation of new safe nematicides, though still retarded by the lack of efficient stabilization processes and standardized EOs' components and extraction techniques.

Chemical Identification of Specialized Metabolites from Sulla (Hedysarum coronarium L.) Collected in Southern Italy.

Sulla (Hedysarum coronarium L.) is a biennal forage legume originated from the Mediterranean basin and used for animal feeding due to its high forage quality and palatability. Several species of Hedysarum have been considered for their nutritional, pharmaceutical, and biological properties, and different applications have been reported, both for human consumption and animal nutrition. Although a systematic investigation of the chemical constituents of Hedysarum spp. has been performed in order to provide chemotaxonomic evidences for the genus and to support the pharmacological application of several species within the genus, few data are available on the chemical constituents of H. coronarium, and only the content of condensed tannins and flavonoids in leaves has been previously reported. In the present paper, results from a detailed chemical analysis of the extracts from the leaves and flowers of H. coronarium grown wild in southern Italy are presented. Identification of the main specialized metabolites within the chemical classes of flavonoids, proanthocyanidins and saponins, is described, including considerations on their content in the two plant organs. Information acquired from this study expands the knowledge on H. coronarium as a source of valuable phytochemicals for different applications in human and animal health and nutrition.

Short-Time Impact of Soil Amendments with Medicago Plant Materials on Soil Nematofauna.

Soil amendments with plant materials from Medicago species are widely acknowledged for a suppressive effect on plant-parasitic nematodes but their impact on beneficial components of soil nematofauna is still unknown. A study on potted tomato was carried out to investigate the short-time effects on the overall nematofauna of dry biomasses from six different Medicago species, i.e., M. sativa, M. heyniana, M. hybrida, M. lupulina, M. murex and M. truncatula, incorporated to natural soil at 10, 20, or 40 g kg-1 soil rates. All amendments resulted in a significant decrease of the total nematofauna biomass, whereas total abundance was significantly reduced only by M. heyniana, M. hybrida, and M. lupulina biomasses. Almost all the Medicago amendments significantly reduced the relative abundance of plant-parasites and root fungal feeders. All amendments significantly increased the abundance of bacterivores, whereas fungivores significantly increased only in soil amended with M. heyniana, M. lupulina and M. sativa plant materials. Mesorhabditis and Rhabditis were the most abundant genera of bacterivores, whereas Aphelenchoides and Aphelenchus prevailed among the fungivores. Predators were poorly influenced by all the tested Medicago biomasses, whereas the abundance of omnivores was negatively affected by M. heyniana and M. lupulina. Values of the Maturity Index and Sum Maturity Index were reduced by treatments with M. heyniana, M. hybrida, M. lupulina and M. sativa plant materials, whereas most of the tested amendments decreased values of the Channel Index while increasing those of the Enrichment Index. Enrichment and bacterivore footprints raised following soil addition with Medicago biomasses, whereas composite and fungivore footprints were significantly reduced. According to their overall positive effects on soil nematofauna, amendments with Medicago plant materials or their formulated derivatives could represent an additional tool for a sustainable management of plant-parasitic nematodes.

Relationship between Chemical Composition and Nematicidal Activity of Different Essential Oils.

In this study, the relationship between nematicidal activity and chemical composition of ten essential oils (EOs) from different plant species was investigated both in in vitro assays on juveniles (J2) and eggs of the root-knot nematode Meloidogyne incognita and in experiments on tomato in soil infested by M. incognita. Nematode J2 were exposed for 4, 8 or 24 h to 0.78-100 µg mL-1 concentrations of each EO, whereas 24, 48 or 96 h exposures to 250, 500 and 1000 µg mL-1 solutions were tested on M. incognita egg masses. Treatments with 50, 100 or 200 µg kg soil rates of each EO were applied in the experiment on potted tomato. The highest nematicidal potential resulted for the C. verum EO, as highly toxic to both M. incognitaJ2 and eggs and strongly suppressive on nematode multiplication on tomato roots. The infestation of M. incognita on tomato roots was also strongly reduced by the EOs from E. citriodora and S. aromaticum, both highly toxic to M. incognitaJ2 but less active on nematode eggs. Adversely, R. graveolens EO strongly inhibited the egg hatch but was limitedly toxic to the infective J2. Chemical composition of the EOs was determined by GC-FID and GC-MS. The ten EOs showed a very different chemical composition in terms of major phytochemicals, with one or two dominant components totally amounting up to 85%. The structure-activity relationship based on the main phytochemicals identified in the assayed EOs and their nematicidal effects on M. incognita was also discussed. Results from this study confirmed that the selection of suitable EO raw materials can lead to the formulation on new effective nematicidal products.

Activity of Saponins from Medicago Species against Phytoparasitic Nematodes.

Content of bioactive saponins of Medicago species suggests that they may also exert, as previously demonstrated on M. sativa, nematicidal properties exploitable for the formulation of new products for sustainable phytoparasitic nematode management. This study was addressed to highlight the bioactivity of saponins from five different Medicago species still poorly known for their biological efficacy, i.e., M. heyniana, M. hybrida, M. lupulina, M. murex and M. truncatula, against the plant parasitic nematodes Meloidogyne incognita, Xiphinema index and Globodera rostochiensis. The bioactivity of the extracts from the five Medicago species was assessed by in vitro assays on the juveniles (J2) and eggs of M. incognita and G. rostochiensis and the adult females of X. index. The suppressiveness to M. incognita of soil treatments with the Medicago plant biomasses was also investigated in a tomato experiment. The nematicidal activity of the five Medicago species was reported and discussed in relation to their phytochemical profile.

Correction to: Nematicidal potential of Taraxacum officinale.

The original publication of this paper contains a mistake. Data on Table 1 under TR and TL column have been interchanged: that is compounds 1-12 and their amounts refer to TL; compounds 1-8 and related amounts refer to TR (see Fig. 1).

Nematicidal potential of Taraxacum officinale.

This study was aimed to investigate the activity of the Asteraceae species Taraxacum officinale against the root-knot nematode Meloidogyne incognita. Leaf and root extracts of T. officinale were tested in vitro at a range of 62.5-1000 and 250-1000 µg mL-1 concentrations on nematode juveniles and eggs, respectively, whereas treatments with 10-40 g kg-1 soil rates of dry leaf and root T. officinale biomass were applied to soil infested by M. incognita in greenhouse experiments on potted tomato. Peak 36 and 50% juvenile mortality and 14.8 and 23.8% egg hatchability reduction were recorded at the maximum concentration of leaf and root extracts, respectively. Soil treatments with T. officinale leaf and root material strongly suppressed nematode multiplication and gall formation on tomato roots and significantly increased plant growth. Chicoric acid and 3-O- and 3,5-di-O-caffeoylquinic acid were found to be the main components of leaf and root extract, respectively, and proved, as the total hydroalcoholic extracts from T. officinale leaf and root material, for an antioxidant activity. Data from this study indicate the suitability of plant materials from T. officinale for a potential formulation of nematicidal products to include in sustainable nematode management strategies.

Effect of water, soil temperatures, and exposure times on the survival of the sugar beet cyst nematode, heterodera schachtii.

ABSTRACT The effect of different combinations of temperatures and exposure times on the mortality of Heterodera schachtii eggs was assessed in two different experiments under laboratory conditions. In the first experiment, cysts in water were exposed to 25, 35, 37.5, 40, 42.5, 45, 47.5, 50, or 52.5 degrees C for a maximum period of 2 h. In the second experiment, cysts in naturally infested soil were exposed to 25, 32.5, 35, 37.5, 40, 42.5, or 45 degrees C for a minimum period of 2 h to a maximum of 2,048 h. Viability of eggs in cysts was assessed by a hatching test in 3 mM zinc chloride solution. Viability in water was suppressed after 2-h exposure at 50 degrees C and inhibited after 1 to 2 h at 52.5 degrees C. Emergence of juveniles from cysts in soil was greater at the lower temperature x exposure time combinations and suppressed at higher combinations. Egg mortality started after exposure for 256 h at 40 degrees C, 32 h at 42.5 degrees C, and 16 h at 45 degrees C, and 81, 31, and 7 h of exposure were necessary to kill 50% of the nematode egg population at 40, 42.5, and 45 degrees C, respectively. The data fitted the models P(t) = P(0)10(-t/ (q + mT)) and P(T1) = P(T0)10(-T/(z - pt)) for m = -0.0111, q = 0.8238, z = 2.444, and p = -0.23.