First report of Colletotrichum siamense causing anthracnose in Oenocarpus bacaba in Brazil.

Bacaba (Oenocarpus bacaba Mart.) is a native palm tree from Brazilian Amazon and Cerrado biomes. This tree produces a small, rounded fruit with dark skin and approximately 1.5 mm thick pulp, extensively utilized for palm heart extraction, juices, and jellies (De Cól et al. 2021). However, several diseases can adversely impact fruit yield and quality. During the 2021 growing season, anthracnose symptoms were observed in Bacaba fruits, with a disease incidence of 58% in fruits collected from the Abreulândia (9°37'15″ S, 49°9'3″ W) and Gurupi (12°25'46" S; 49°16'42" W) municipalities in Tocantins state, Brazil. A total of 198 fruits exhibiting anthracnose symptoms, characterized by deep necrotic spots, were collected. In the laboratory, symptomatic fruits had their external surfaces sterilized for 30 seconds in 70% ethanol, 1 min in 1.5% NaOCl, and then rinsed with sterile distilled water. Sterilized pieces of the fruit tissue were transferred to PDA medium and incubated for 7 days at 28 ºC with a 12 h photoperiod. After this period, two isolates were obtained from the colonies and were identified both macroscopically and microscopically as Colletotrichum sp. The colonies grown at PDA showed a white to grey cottony mycelia, with straight and fusiform conidia, ranging from 14.0 to 21.0 (mean value of 15.8 ± 1.8) µm in length and 4.0 to 7.0 (mean value of 5.5 ± 0.7) µm in width, (n = 50). For species identification, the intergenic spacer between DNA lyase, mating-type locus MAT1-2-1 (APN2/MAT-IGS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glutamine synthetase (GS), and ß-tubulin (TUB) loci were amplified and sequenced. Resulting sequences were deposited in GenBank (OR333843, OR333844, OR333845 and OR333846). BLAST analysis of the partial APN2/MAT-IGS (99%), GAPDH (99,48%), GS (99,32%) and TUB (99,48%) sequences showed highly similarity to C. siamense isolates (IIFT223 and CBS130147). Maximum likelihood multilocus analysis placed the isolate UFTC16 within the C. siamense clade with 98% bootstrap support, clearly assigning the isolate to this species. Morphological features were consistent with the description of C. siamense (Prihastuti et al., 2009). Inoculation of Bacaba fruits and seedlings was conducted to confirm pathogenicity. The surface of uninjured Bacaba fruits was inoculated with two drops (20 µL) of conidial suspension (106 conidia mL-1). The same methodology was adopted to placed healthy leaves of 35-day-old seedlings grown in plastic tubes. Two drops of sterile distilled water were inoculated on nonwounded healthy fruits and seedlings as a negative control. The fruits and seedlings were incubated for five days in a controlled chamber at 28 °C, 70-80% humidity and a "12-h photoperiod". The experiment was conducted with five replicates (five fruits and five seedlings inoculated per isolate) and repeated once. Typical symptoms of anthracnose were observed in the fruits and leaves of Bacaba seedlings five days after inoculation. No symptoms were observed in the negative control. The pathogen was reisolated from symptomatic fruits and leaves, showing similar morphological characteristics as the original isolate, fulfilling Koch's postulates. The identification of C. siamense as the causal agent of Bacaba anthracnose helps in the diagnosis and disease control strategies of the disease. Colletotrichum siamense is a cosmopolitan species and easily found in cultivated and non-cultivated species (Batista et al. 2023). However, to the best of our knowledge, this is the first report of C. siamense causing anthracnose on Bacaba.

Contributions of γ-Aminobutyric Acid (GABA) Receptors for the Activities of Pectis brevipedunculata Essential Oil against Drosophila suzukii and Pollinator Bees.

The γ-aminobutyric acid (GABA) receptors play pivotal roles in the transmission of neuronal information in the nervous system of insects, which has led these proteins to be targeted by synthetic and natural products. Here, we assessed the insecticidal potential of the essential oil of Pectis brevipedunculata (Gardner) Sch. Bip., a neotropical Asteraceae plant used in traditional medicine, for controlling Drosophila suzukii (Matsumura) adults by feeding exposure. By using in silico approaches, we disentangle the contribution of GABA receptors and other potential neuronal targets (e.g., acetylcholinesterase, glutathione-S-transferases) in insects that may explain the essential oil differential activities against D. suzukii and two essential pollinator bees (Apis mellifera Linnaeus and Partamona helleri Friese). Neral (26.7%) and geranial (33.9%) were the main essential oil components which killed D. suzukii with an estimated median lethal concentration (LC50) of 2.25 µL/mL. Both pollinator forager bee species, which would likely contact this compound in the field, were more tolerant to the essential oil and did not have their diet consumptions affected by the essential oil. Based on the molecular predictions for the three potential targets and the essential oil main components, a higher affinity of interaction with the GABA receptors of D. suzukii (geranial -6.2 kcal/mol; neral -5.8 kcal/mol) in relation to A. mellifera (geranial -5.2 kcal/mol; neral -4.9 kcal/mol) would contribute to explaining the difference in toxicities observed in the bioassays. Collectively, our findings indicated the involvement of GABA receptors in the potential of P. brevipedunculata essential oil as an alternative tool for controlling D. suzukii.

The Potential of Plant-Based Biorational Products for the Drosophila suzukii Control: Current Status, Opportunities, and Limitations.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is an invasive species that causes serious damage to soft-skinned fruits. The use of plant-based biorational insecticides (plant extracts and essential oils) to control this pest has grown extensively. We conducted a systematic review and meta-analysis to examine the current status, trends, and perspectives of these studies, with a focus on the plant families and major compounds used as insecticides to control D. suzukii. The first article in this research field was published in 2015, and there has been exponential growth in subsequent years. Thirty-six botanical families were studied in these articles, with a prevalent interest in Myrtaceae and Lamiaceae plant species. The major constituents of these plant-based biorational molecules belong to monoterpenoids, followed by monoterpenes, benzene derivatives, and others. Geranial was the most frequent major constituent of these plant-based compounds. Our analysis revealed a few crucial consequences of the bias provided by the investigations using plant-based biorational insecticides for controlling D. suzukii. Firstly, there is a major focus on the pest species, with little or no attention paid to undesired effects on non-target beneficial organisms (e.g., pollinator bees, predators; parasitoids) and non-target pests. Secondly, the poor knowledge of how these plant-based biorational insecticides act on target and non-target organisms. Finally, there is a need to assess the efficacy of these substances under field conditions. Thus, attention is needed to address these gaps so that plant-based biorational insecticides can become a viable pest management tool for controlling D. suzukii.

Genomic-proteomic analysis of a novel Bacillus thuringiensis strain: toxicity against two lepidopteran pests, abundance of Cry1Ac5 toxin, and presence of InhA1 virulence factor.

Bacillus thuringiensis (Bt) is a biological alternative to the indiscriminate use of chemical insecticides in agriculture. Due to resistance development on insect pests to Bt crops, isolating novel Bt strains is a strategy for screening new pesticidal proteins or strains containing toxin profile variety that can delay resistance. Besides, the combined genomic and proteomic approaches allow identifying pesticidal proteins and virulence factors accurately. Here, the genome of a novel Bt strain (Bt TOL651) was sequenced, and the proteins from the spore-crystal mixture were identified by proteomic analysis. Toxicity bioassays with the spore-crystal mixture against larvae of Diatraea saccharalis and Anticarsia gemmatalis, key pests of sugarcane and soybean, respectively, were performed. The toxicity of Bt TOL651 varies with the insect; A. gemmatalis (LC50 = 1.45 ng cm-2) is more susceptible than D. saccharalis (LC50 = 73.77 ng cm-2). Phylogenetic analysis of the gyrB gene indicates that TOL651 is related to Bt kenyae strains. The genomic analysis revealed the presence of cry1Aa18, cry1Ac5, cry1Ia44, and cry2Aa9 pesticidal genes. Virulence factor genes such as phospholipases (plcA, piplc), metalloproteases (inhA), hemolysins (cytK, hlyIII, hblA, hblC, hblD), and enterotoxins (nheA, nheB, nheC) were also identified. The combined use of the genomic and proteomic data indicated the expression of Cry1Aa18, Cry1Ac5, and Cry2Aa9 proteins, with Cry1Ac5 being the most abundant. InhA1 also was expressed and may contribute to Bt TOL651 pathogenicity. These results provide Bt TOL651 as a new tool for the biocontrol of lepidopteran pests.

Pseudomonas aeruginosa and Bacillus cereus Isolated from Brazilian Cerrado Soil Act as Phosphate-Solubilizing Bacteria.

The phosphate-solubilizing microorganism is essential for soil quality and plant development and can serve as an alternative to reduce such Brazilian needs for importing phosphate overseas. Here, we isolated and selected bacteria from Brazilian Cerrado soils capable of solubilize phosphate. We obtained 53 bacteria isolates, of which 23 could solubilize phosphate at a pH of 7.0, 17 could solubilize phosphate at a pH of 6.0, and 8 could solubilize at a pH of 5.5. Using 16S rRNA gene sequences, we identified nine bacteria species clustered in four groups: Bacillus sp., Pseudomonas sp., Priestia sp., and Klebsiella sp. Our results revealed that the UFT01 (P. aeruginosa) and UFT42 (B. cereus) isolates exhibited the best phosphate solubilization performance at all tested pH values. We further recorded higher levels of solubilization and phosphate availability six days after the soil inoculation with P. aeruginosa, and enzymatic analysis of the soil samples revealed that the P. aeruginosa-inoculated samples resulted in four-fold higher enzymatic activities when compared to non-inoculated soils. The B. cereus soil inoculation increased ß-glucosidase activities and resulted in reduced the activities of arylsulfatase. Altogether, our findings demonstrated that P. aeruginosa and B. cereus isolated from Cerrado soils showed high phosphate solubilization potential.

Acaricide activity of extract and an isolated compound of Lithraea brasiliensis on Rhipicephalus microplus and selectivity actions against a non-target organism.

Rhipicephalus microplus, known as the cattle tick, is a cause of great economic losses for dairy cattle farming because of its high frequency of occurrence and the difficulty in controlling it. This research characterized the chemical profile and evaluated the in vitro toxicity of crude Lithraea brasiliensis extract and its isolated compound against acaricide-resistant and acaricide-susceptible R. microplus strains. Acaricidal activity was evaluated using a larval immersion test and the selectivity against non-target organisms was assessed on Artemia salina assay. The chemical investigation by high-performance liquid chromatography coupled with mass spectrometry (i.e., HPLC-MS) analysis showed the presence of hydrolysable tannins as well as urushiol derivatives. Column chromatography (CC) was carried out on the extract to obtain fractions and an isolated compound. The extract exhibited significant activity against acaricide-resistant (LC50 0.64 mg/mL) and acaricide-susceptible (LC50 0.76 mg/mL) strains of R. microplus larvae. The isolated compound from the extract (urushiol II), exhibited LC50 of 1.11 mg/mL for acaricide-resistant larvae. For acute toxicity in A. salina, the extract showed LC50>100 µg/mL. Thus, our findings represent the first effort to demonstrate the potential of L. brasiliensis extract and urushiol II as potential natural acaricides to replace or to be integrated into the conventional control of R. microplus larvae.

Host Potential and Adaptive Responses of Drosophila suzukii (Diptera: Drosophilidae) to Barbados Cherries.

Biological invasions are a global threat to agricultural crops worldwide. In the Neotropical region, the spotted-wing Drosophila [Drosophila suzukii (Matsumura)] has rapidly expanded its geographical range spreading throughout South America in recent years. Besides climatic factors, the remarkable success of its establishment and subsequent distribution in this region is closely dependent on the diversity and availability of host plants. We evaluated the host potential (e.g., as food and oviposition sources) of fruits of jabuticaba [Plinia cauliflora (Mart.) Kausel (Myrtales: Myrtaceae)], Barbados cherry (Malpighia emarginata DC) (Malpighiales: Malpighiaceae), bonnet pepper (Capsicum chinense Jacq.) (Solanales : Solanaceae), and coffee (Coffea arabica L.) (Gentianales : Rubiaceae) and their effects on the biological and physiological traits of D. suzukii. For the fruit types where fly emergence occurred, we assessed the biological and physiological performance of the flies and compared these parameters with those recorded for flies reared on strawberries (Fragaria × ananassa Duchesne) (Rosales : Rosaceae) and an artificial diet. Our results revealed that oviposition into fruits and completion of the life cycle occurred on Barbados cherries only. Furthermore, field surveys revealed a higher emergence rate of D. suzukii on undamaged ripe Barbados cherries than damaged ones. Moreover, flies developing on Barbados cherries and an artificial diet presented earlier emergence, shorter developmental time, lower number of adults per female, and a female-biased sex ratio compared to flies developing on strawberries. Overall, our findings demonstrated suitability of Barbados cherry as a host for D. suzukii, which renders management of D. suzukii in Neotropical region an even more challenging task.

The lacewing Ceraeochrysa caligata as a potential biological agent for controlling the red palm mite Raoiella indica.

BACKGROUND: Compared to chemical control, the use of naturally occurring biological agents to control invasive pests is less threatening to the environment and human health. OBJECTIVES: Here, we assessed the ability of immature stages of the lacewing Ceraeochrysa caligata (Neuroptera: Chrysopidae) to prey upon different developmental stages of the red palm mite Raoiella indica (Acari: Tenuipalpidae), one of the most destructive invasive pests of palm trees in Neotropical regions. METHODS: Increasing densities of three stages of R. indica (eggs, immature stages, and adult females) were offered to C. caligata in coconut leaf arenas. The immature stages of C. caligata were less than 24 h old and were starved before being transferring to the arenas. The amount of prey consumed was recorded 6 h after releasing the C. caligata. RESULTS: Our results indicated that the ability of C. caligata to feed upon R. indica increased with the larval development of the predator. Higher feeding levels and shorter handling times were recorded for the first and second instars of C. caligata when preying upon the eggs and immature stages of R. indica. Furthermore, C. caligata individuals of different stages exhibited differential functional responses according to prey type (i.e., eggs, immatures, or adult females of R. indica). Ceraeochrysa caligata second instar individuals exhibited a sigmoid increase in consumption rate with increasing prey availability (i.e., a type III functional response) when preying upon immature stages of R. indica. However, when preying upon R. indica adult females, C. caligata second instar individuals exhibited a type II functional response (i.e., an increase in consumption rate with increasing prey availability, before reaching a plateau). Predator individuals of the first and third instar stages exhibited a type II functional response for all prey types. CONCLUSIONS: Collectively, our findings demonstrate that C. caligata, especially at the second instar stage, has potential as a tool for ecological management of the red palm mite.

Fumonisin Production by Fusarium verticillioides in Maize Genotypes Cultivated in Different Environments.

Fumonisins are mycotoxins (MTs) produced mainly by the fungus Fusarium verticillioides, the main pathogens of maize which cause ear rot. The aim of this work was to evaluate some factors that may lead to high fumonisin production by F. verticillioides in maize grains, correlating the pathogen inoculation method with different genotypes grown in four Brazilian states. Experiments were conducted in 2015-2016 in maize crops from experimental maize fields located in four distinct states of Brazil. Results showed that contamination by fumonisin mycotoxins occurred even on symptomatic or asymptomatic grains. In all municipalities, the samples showed levels of fumonisin B1 that were higher than would be tolerable for the human consumption of corn products (the current tolerance limit for fumonisin is 1.5 µg g-1). High severity of grains infected with F. verticillioides does not always show high concentrations of fumonisins. Environments with higher temperatures may influence the production of high concentrations of fumonisin in maize hybrids. Spray inoculation methods and inoculation at the center of spikes did not influence fumonisin concentrations. Results showed that the hybrids P3630H, P32R48 and P3250 presented higher disease severity, as well as higher mycotoxin levels in the studied locations with higher temperatures.

Sublethal dose of deltamethrin damage the midgut cells of the mayfly Callibaetis radiatus (Ephemeroptera: Baetidae).

In insects, the midgut performs multiple physiologic functions (e.g., digestion and nutrients absorption) and serves as a physical/chemical barrier against pathogens and chemical stressors such as deltamethrin, a pyrethroid insecticide, commonly used in insect control that are agricultural pests and human disease vectors. Here, we described the midgut cell ultrastructure of Callibaetis radiatus nymphs, which are bioindicators of water quality and the ultrastructural alterations in midgut under sublethal exposure to deltamethrin at three different periods (1, 12, 24 h). The digestive cells of deltamethrin-unexposed nymphs had long microvilli, many mitochondria in the apical cytoplasm, a rough endoplasmic reticulum, a basal labyrinth with openings for hemocele, and the midgut peritrophic matrix which is classified as type I. Nymphs exposed to deltamethrin exhibited digestive cells rich in autophagic vacuoles, basal labyrinth loss, and microvilli disorganization since the first hour of contact with deltamethrin. However, these midgut tissues underwent to autophagic cellular recovery along the 24 h of exposure to deltamethrin. Thus, the sublethal exposure to deltamethrin is sufficient to disturb the ultrastructure of C. radiatus midgut, which might reduce the abilities of these insects to survive in aquatic environments contaminated by pyrethroids.