Control of phytonematodes is very hard and requires a combination of techniques to succeed. Alternative control through plant extracts may result in the discovery of nematicide substances. Research aimed at evaluating the effect of 33 plants submitted to aqueous extraction against Panagrellus redivivus in vitro. Concentrations were prepared at 1.25, 2.5, 5, 10, and 20%. Monitoring happened at 0, 6, 12, 24 and 30 hours after preparation. Counting considered dead nematodes subtracted from alive ones. Juveniles were also counted, and extract efficiency was expressed in percentage of control or stimuli. Data were submitted to variance analysis. Significant results got with the Scott-Knott test (5%), and multiple linear regression analysis. Extracts were observed acting as controllers, but also as stimulators to nematode reproduction. The best controlling performance was set by Carica papaya (-66% at 20%; -33.7% at 10%), Euphorbia milii (-37% at 20%), Psychotria carthagenensis (-25.5% at 2.5%), Clusia variegate (-22% at 20%), and Zamioculcas zamiifolia (-21.5% at 20%). Stimulator extracts were Mentha villosa at 10% (+148%) and 2.5% (+131.5%), followed by Aloe vera (+123% at 5%), Schinus molle (+112.5% at 10%), Schefflera arboricola (+93.5% at 5%), C. variegate (+89% at 5%), and S. molle (+88% at 5%). Some extracts kept population stable throughout the experiment, presenting lower control indexes. Besides an additive effect, there was an individual influence of concentration or time on control.
Biological control is a method of controlling pests through the use of other living organisms. The purposes of this study were to test Hohenbuehelia species as biological control agents against Panagrellus redivivus in vitro, evaluating nematodes influence on mycelia growth; establishing daily indexes for predation and growth and setting predation percentage. Five species previously identified as 436-Hohenbuehelia mastrucata (Nematoctonus hamatus), 528-H. bullulifera (not described so far), 581-H. paraguayensis (N. sp.), 582-H. sp. (N. sp.) and 631-H. portegna (N. campylosporus) were submitted to anamorphic purification directly from basidioma. Afterwards, 100 nematodes were added to each pure colony for predation test. Evaluation started right after 24 hours of nematode-fungus interaction. Immobilized and/or penetrated nematodes were counted and mycelia growth was measured. Results were subjected to variance analyses. Hohenbuehelia mastrucata had the best performance in growth speed, followed by H. portegna and H. paraguayensis; Nematodes multiplyied much but none specie grew more as an influence of their movement under mycelium, however all species formed trap devices and some of them produced adhesive or repelent substances. Trap devices were formed in control plates also. The plates of H. paraguayensis without nematodes grew more than treatments. Cumulative predation of H. portegna was the highest at 24 (195.5%) and 48 hours (235%). At the last evaluation day, H. paraguayensis preyed the same amount (185.75%) than H. portegna, followed by H. mastrucata (109.51%). Resulst of predation daily indexes displayed chronological activity for each isolate, where H. portegna was very reactive at first 24 hours, H. mastrucata raised its predacious activity in 48 hours being constant from this time on and H. paraguayensis pointed out itself at 72 hours. Other species presented low predation and growth indexes throughout experiment.
Sugarcane-associated nematodes (Saccharum spp.) can reduce productivity up to 50%. Through the survey, it was possible to identify the main nematodes that occur in a certain region as a tool for designing the best management and control strategies. The aim of this study was to characterize the population of nematodes associated with sugarcane in the North Central, North Pioneiro and Northwest mesoregion of the state of Paraná, Brazil, quantify the nematode genera associated with the crop and identify the species of Pratylenchus and Meloidogyne. A total amount of 89 soil and root composite samples were collected in nine municipalities. Nematodes were extracted and counted in a Peters counting chamber under an optical light microscope. Morphological description followed identification keys. Pratylenchus spp. were identified by morphological characteristics; Meloidogyne spp. were identified by morphological characteristics and isoenzyme electrophoresis. Twelve genera of nematodes associated with sugarcane were identified:
Swine wastewater (SWW) is a residue from pig farming that presents a high load of nutrients and organic matter. The appliance of organic matter in soil alters the microbial dynamic and may suppress soilborn phytopathogens. This study aimed at evaluating the inhibition on mycelial growth of Sclerotinia sclerotiorum and Sclerotium rolfsii in vitro under SWW doses. Hereupon, three kilograms of a soil classified as red dystroferric latosol was collected and sieved. Half of it was autoclaved. SWW was incorporated at doses of 0 mL, 2.5 mL, 5 mL, 10 mL and 20 mL in both soil conditions, autoclaved and not autoclaved. Afterwards, 130 grams of each soil was separately put into Petri plates above what a thin layer (≅ 5 mL) of Water-Agar (2%) medium was carefully spread over. Above this agar layer, one disk (6 mm diameter) of pure mycelium from each fungal grown in Potato Dextrose Agar medium was individually placed on the center of each plate. Daily evaluations on mycelial growth measuring were taken and ended when mycelium in control plates (without SWW addition) reached plate borders. Results indicated that in autoclaved soil condition, the inhibition was proportional to the dose, what is to say that the higher the dose the less the mycelial growth. In not autoclaved soil there was no significant difference among treatments, suggesting stimuli on suppression effect for both pathogens caused by SWW. In addition, the confirmed potential of SWW as a suppressor of S. sclerotiorum and S. rolfsii leads to promising investigations on other phytopathogens hard to control.
This research aimed to evaluate the nematophagous ability of 4077-Verticillium chlamydosporium var. chlamydosporium and 4466-Hirsutella thompsonii isolates and relate mycelia growth to the influence provoked by movement of nematodes. Each fungus grew in PDA (potato, dextrose, agar) medium end up to pure colonization. Then, ten mycelia plugs of 8 mm diameter were removed from colony borders and transferred to the center of ten Petri plates containing water-agar 2% medium. These plates were previously divided into four quadrants that received a number of 25 individuals of free-living nematodes (Panagrellus redivivus), composing a total of 100 nematodes per plate. Evaluations started after 24 hours of interaction, considering predation percentage and mycelia growth as stimuli of nematodes presence. Results showed growing predation performance to both isolates, being higher for V. chlamydosporium var. chlamydosporium since from first evaluation time, controlling more than 50% of nematode population initially added. Its predation potential was 39.2%, 38.4% and 48.35% higher than H. thompsonii at first, second and third evaluation day, respectively. Generally, nematodes did not stimulate mycelia growth, unless for H. thompsonii at 72 hours of interaction compared to control plates (without nematodes). Stress resulting from isolates transference from PDA to water-agar 2% resulted in sparse mycelia growth and it could have affected the predation performance of H. thompsonii that controlled nematodes in low levels throughout experiment. Independently of predation level, pictures revealed that both isolates has ability to control P. redivivus through hyphae penetration.
Root-knotting nematodes are found in vineyards and are associated with grapevine decline. This study aimed to evaluate the resistance of four new hybrids, obtained by crossing Vits labruscana and Vits rotundifolia (IBRE 421, IBBT 481, IBRK 504 and IBMG 631), to Meloidogyne javanica. This study also evaluated three rootstocks (IAC 766, Paulsen 1103, VR 043- 43), widely used and showing resistance levels to M. javanica. The plants were obtained from cuttings, then grown in pots, and inoculated with a population of M. javanica. One year after inoculation, the plants presented no galls on their fine roots. The root nematodes were extracted and quantified. The reproduction factor of M. javanica in all genotypes was close to zero. The tested hybrids are not good hosts for the multiplication of M. javanica. Thus, these new hybrids can be used as a geneticcontrol strategy of M. javanica.
Impatiens walleriana (Balsaminaceae), popularly known as Impatiens, is an African succulent and a popular ornamental plant worldwide (GBIF, 2019). In Brazil it is broadly grown indoors and outdoors, including in public parks of Curitiba, State of Paraná (Viezzer et al. 2018). In September 2018, I. walleriana plants showing typical downy mildew symptoms were observed in wastelands and gardens in Curitiba. The symptoms included adaxial chlorotic leaf spots with abundant white sporulation on abaxial side (Supplementary figure 1). The disease led to severe defoliation of the plants and the incidence of the plant disease varied from 20 to 80% of plants in an area ranging from 400 to 40,000 m2. A representative sample was deposited in herbarium of the Museu Botânico Municipal de Curitiba (MBM 331601). The following morphology was observed: Sporangiophores (n = 30), hyaline, thin walled, emerging through stomata, 407.3 to 551.1 μm long, slightly swollen base, first branch at 165.8 to 324.7 μm from base, end branches 5.1 to 13.1 μm long, sporangia (n = 50) hyaline, thin-walled subglobose to ovoid, from 12.8 to 21.9 μm x 12.5 to 17.9 μm, slightly papillate. Due to morphological and genetic variations within the species Plasmopara obducens, Görg et al. (2017) proposed the new species P. velutina and P. destructor. The morphology of the Curitiba specimen was equivalent to that described for P. destructor (Görg et al. 2017). DNA was extracted from LEMIDPRTf-19-02 isolate and the ITS1 and cox2 regions were PCR amplified as described in Görg et al. (2017). The resulting sequences were deposited in GenBank (ITS1, MT680628; cox2, MT952335). A BLASTn analysis of the sequences revealed 100% homology with ITS (MF372742) and cox2 (MF372728) sequences of type strain of P. destructor (GLM-F107554). A Bayesian phylogenetic analysis was performed to compare the sequences from this study with reference sequences for P. obducens, P. destructor and P. velutina (Görg et al. 2017; Salgado-Salazar et al. 2018). The oomycete from Curitiba grouped in a reliable clade with P. destructor (Supplementary figure 2). Pathogenicity was carried out by ex vivo and in vivo tests. For ex vivo, stems with approximately four healthy leaves of I. walleriana (n = 10) were embedded in aluminum grid inside of gerbox with the stem bases immersed in distilled water. The inoculation of five stems was carried out by spraying a suspension with 6 x 104 sporangia mL-1 on the abaxial side of the leaves. Five stems with leaves inoculated with sterile water were used as controls. They were incubated in a growth chamber in the dark for 48 h at 20 °C and another 12 days in a 12 h light photoperiod. The confirmation of pathogenicity in plants (in vivo) was obtained with the inoculation of I. walleriana seedlings (one-month old) grown in 2 dm3 aluminum pots. The inoculation methodology and number of plants were the same as the stems test. After the inoculation, plants were incubated in a growth chamber for 48 h in the dark at 20 °C with 100% RH with nebulization, and another 10 days at a photoperiod of 12 hours of light. For both tests, abundant sporulation was observedwith morphology equivalent to Plasmopara destructor described by Görg et al. (2017). No disease developed on control plants. To our knowledge, this is the first report of P. destructor on I. walleriana in Brazil (Farr and Rossman 2019, Silva et al. 2019) representing a potential loss to flower production and a reduction in flowering period in public gardens and parks.
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