Potential of different species of actinobacteria in the management of Meloidogyne javanica.

Root-knot nematodes (RKN) are one of the most harmful soil-borne plant pathogens in the world. Actinobacteria are known phytopathogen control agents. The aim of this study was to select soil actinobacteria with control potential against the RKN (Meloidogyne javanica) in tomato plants and to determine mechanisms of action. Ten isolates were tested and a significant reduction was observed in the number of M. javanica eggs, and galls 46 days after infestation with the nematode. The results could be explained by the combination of different mechanisms including parasitism and induction of plant defense response. The M. javanica eggs were parasited by all isolates tested. Some isolates reduced the penetration of juveniles into the roots. Other isolates using the split-root method were able to induce systemic defenses in tomato plants. The 4L isolate was selected for analysis of the expression of the plant defense genes TomLoxA, ACCO, PR1, and RBOH1. In plants treated with 4L isolate and M. javanica, there was a significant increase in the number of TomLoxA and ACCO gene transcripts. In plants treated only with M. javanica, only the expression of the RBOH1 and PR1 genes was induced in the first hours after infection. The isolates were identified using 16S rRNA gene sequencing as Streptomyces sp. (1A, 3F, 4L, 6O, 8S, 9T, and 10U), Kribbella sp. (5N), Kitasatospora sp. (2AE), and Lentzea sp. (7P). The efficacy of isolates from the Kitasatospora, Kribbella, and Lentzea genera was reported for the first time, and the efficacy of Streptomyces genus isolates for controlling M. javanica was confirmed. All the isolates tested in this study were efficient against RKN. This study provides the opportunity to investigate bacterial genera that have not yet been explored in the control of M. javanica in tomatoes and other crops.

Inoculation of Pochonia chlamydosporia triggers a defense response in tomato roots, affecting parasitism by Meloidogyne javanica.

Pochonia chlamydosporia is a soil-dwelling fungus and biological control agent of nematodes, active ingredient in commercial bionematicides. The fungus is also endophytically associated with the roots of several plant species, promoting their growth and inducing systemic resistance. In this study, different pathways and tomato defense metabolites were studied to identify mechanisms induced by P. chlamydosporia that contribute to the control of Meloidogyne javanica, at early and late developmental stages. Some defense responses activated by the fungus appeared related to the nematode life cycle. Among the evaluated biochemical analysis, root colonization of P. chlamydosporia showed an increase in the concentration of phenolic compounds, such as chlorogenic acid. In addition, the expression of some host plant genes was also modified. The interaction of the fungus with roots parasitized by M. javanica resulted in the highest expression of Phenylalanine Ammonia-Lyase (PAL), Chalcone synthase (LECHS 2), and Protease Inhibitor (PI1) genes at 24 days post-inoculation. At the second sampling time (44 days), there was an increase in the expression of the Respiratory Burst Oxidase Homolog (RBOH) gene. Fungus reduced the expression of the ACC-oxidase and Pathogenesis-Related Proteins 1 (PR-1) genes in roots. Moreover, P. chlamydosporia inoculation changed metabolites and phytohormone profiles of the gall formed by M. javanica. Plant defense response appeared to involve the jasmonic acid and phytosphingosine cascades. With this analysis, it was possible to propose new molecular mechanisms induced by the fungus that contribute to the control of M. javanica.

Attraction and toxicity: Ways volatile organic compounds released by Pochonia chlamydosporia affect Meloidogyne incognita.

The production of volatile organic compounds (VOCs) acting against plant-parasitic nematodes has been characterized in different fungi; however, the role of VOCs emitted by Pochonia chlamydosporia in its trophic interaction with Meloidogyne incognita is still unknown. The aim of this study was to determine the effects of VOCs emitted by P. chlamydosporia strain Pc-10 on different stages (eggs, juveniles and female) of the M. incognita life cycle. Exposure of M. incognita eggs to VOCs released by Pc-10 resulted in a reduction up to 88 % in the nematode egg hatching, when compared to the control treatments. The VOCs emitted by Pc-10 also attracted M. incognita second-stage juveniles (J2). Through gas chromatography-mass spectrometry (GC-MS), three molecules were identified from the volatiles of the strain Pc-10, with 1,4-dimethoxybenzene being the major compound. In tests performed in vitro, 1,4-dimethoxybenzene at a concentration of 1050 µg mL-1 inhibited M. incognita egg hatching by up to 78.7 % compared to the control (0 µg mL-1) and attracted M. incognita J2 in all concentrations evaluated (1, 10, 100, 1000, and 10000 µg mL-1). The 1,4-dimethoxybenzene also showed fumigant and non-fumigant nematicidal activity against M. incognita. This compound presented lethal concentration for 50 % (LC50) of M. incognita J2 ranged from 132 to 136 µg mL-1. Fumigation with 1,4-dimethoxybenzene (100 mg) reduced egg hatching by up to 89 % and killed up to 86 % of M. incognita J2 compared to the control (0 µg mL-1). In vivo, the VOCs produced by Pc-10, 1,4-dimethoxybenzene, and the combination of both (Pc-10 + 1,4-dimethoxybenzene) attracted the M. incognita J2, compared to the respective controls. To the best of our knowledge, this is the first report on the attraction of M. incognita J2 and the toxicity to eggs and J2 by VOCs from P. chlamydosporia in which 1,4-dimethoxybenzene is the main toxin and attractant.

Redução de inóculo de Aphelenchoides besseyi em sementes de Brachiaria brizantha tratadas com óleos essenciais / Inoculum reduction of Aphelenchoides besseyi in Brachiaria brizantha seeds treated with essential oils

Óleos essenciais foram testados com o objetivo de controlar Aphelenchoides besseyi em sementes de Brachiaria brizantha cv. 'Marandu'. Sementes naturalmente infestadas por A. besseyi foram tratadas com óleos essenciais de mostarda (Brassica nigra), canela (Cinnamomum zeylanicum), verbena (Lippia citriodora) e Bioprotector (r) (produto comercial composto por extratos e óleos essenciais derivados de plantas), nas concentrações de 0,5% e 1,0%. Avaliou-se também o efeito dos tratamentos após aplicação direta nos juvenis (ensaio in vitro). Todos os óleos tiveram sua efetividade reduzida no ensaio em semente (in vivo) em relação ao ensaio in vitro. Os tratamentos não erradicaram o nematoide das sementes, sendo que os óleos essenciais de canela e verbena, nas concentrações 0,5% e 1,0%, Bioprotector (r) a 1,0% e óleo de mostarda a 1,0% reduziram o nível populacional do nematoide em sementes contaminadas. O óleo de mostarda a 1,0% destacou-se dos demais por ter apresentado ação nematicida sem alterar a germinação das sementes. .

Essencial oils were tested to control Aphelenchoides besseyi in Brachiaria brizantha cv. 'Marandu' seeds. Seeds naturally infested with A. besseyi were treated with essential oils of mustard (Brassica nigra), cinnamon (Cinnamomum zeylanicum), verbena (Lippia citriodora) and Bioprotector (r) (a commercial product based on formulation of plant extracts and essential oils), in concentrations of 0.5% and 1.0%. The effects of the same treatments after direct application on juveniles nematodes were evaluated in vitro. All treatments were less effective to control the nematode in the seeds (in vivo) comparing to the in vitro test. The treatments did not eradicate the nematode from the seeds. However, the essential oils of cinnamon and verbena (in both concentrations), mustard (1.0%), as well as Bioprotector (r) (1.0%), reduced the seed population of A. besseyi. Mustard oil (1.0%) stood out for its nematicide action with no effect on seed germination. .