First Report of Vascular Wilt on Peanut in Mexico Caused by Fusarium incarnatum.

During the fall of 2020, wilt symptoms were presented in a commercial peanut field (Arachis hypogaea L.), variety 'CHAMPS' in Buenavista de Benito Juárez, México (18.460501 N, 98.627100 W). A peanut field was scout 80 days after planting, and plants presenting symptoms of root and crown rot, vascular chlorosis, and tissue death, were sampled. Disease incidence was estimated at 55% of the field. Isolations of the pathogen were made from stem and root tissues. These samples were disinfected by immersion in 1% sodium hypochlorite (NaClO) for 3 min and immersion in 70% ethanol for 1 min with 3 rinses with sterile distilled water. Subsequently, 0.5 cm fragments were removed and placed on media of potato dextrose agar (PDA) and Rose Bengal. Petri dishes were incubated in complete darkness at 26 °C for 7 days. Abundant aerial white mycelium was observed, which turned tan to brown and showed a slightly orange color on the back of the plate. Finally, pure cultures were obtained by single sporing (Aslam et al. 2020). Colonies identified as Fusarium spp. (Leslie and Summerell, 2006) were sub cultured on PDA agar media and Spezieller Nährstoffarmer (Pérez-Vázquez et al. 2022) to observe microscopic characteristics of ten isolates. Colonies of a representative strain (MA-PET-03) produced hyaline septate hyphae, macroconidia dorsoventrally curved, tapering towards both ends of 51-57 × 4.6-5.4 µm (n = 80) with most having 7 septa. Microconidia were unicellular, nonseptate, hyaline, and ovoid, 12.4-20.6 × 3.6-4.1 µm (n = 80). Chlamydospores were abundant and globose 5-11 µm diam (n = 80), intercalary, and solitary in short chains (Figure 1). The observed microscopic characteristics correspond to the description of Fusarium incarnatum (Khoa et al. 2006; Leslie and Summerell, 2006; Xia et al. 2019). The molecular analyses were done with genomic DNA extracted as previously reported by Pérez et al. (2022). A region from the translation elongation factor gene was PCR amplified using EF688/EF1251 primers (Alves et al. 2008) and from the calmodulin (CMDA) gene, using CALDF1 and CALDR1 primers (Noel et al. 2022). The corresponding PCR products were purified with the Gen Elute™ PCR Clean-Up Kit from Sigma-Aldrich Co. (St. Louis Mo. USA) and sequenced at Macrogen Inc. (Seoul, South Korea). The phylogenetic analysis was inferred using the Bayesian Inference method with 1 million generations, final standard deviation was 0.008516. The nucleotide substitution model for Calmodulin (CMDA) was GTR + G and for TEF1 GTR + I + G. This analysis showed that strain MA-PET-03 shared 100% identity (Figure 2) with F. incarnatum ex-type strain CBS 132.73 (CMDA: MN170342; TEF1: JMN1704761) from Pointed gourd (Trichosanthes dioica) in Malawi Africa. The sequences of strain MA-PET-03 were deposited in GenBank (CMDA: OQ679820; TEF1: OQ679821). The pathogenicity tests were carried out with a total of 20 peanut plants, variety 'CHAMPS', 18 days after having been sown in groups of five seeds in 250 g plastic pots, containing a sterilized mixture of Peatmoss and Agrellite (1:1 v./v), with four repetitions. The seeds were inoculated by immersion in 20 mL of spore suspension (106 conidia/mL) isolated from F. incarnatum for 10 min. The plants were maintained in a greenhouse (70% relative humidity and 28 °C) until the appearance of disease symptoms of. Likewise, 10 control plants were inoculated with sterile water. The experiment was repeated twice. The symptoms developed 15 days after inoculation, the plants presented symptoms of chlorosis, wilting of leaves, stems, and roots, a manifestation similar to that observed in the field, while the control plants remained healthy. F. incarnatum was consistently reisolated from inoculated stems and roots and identified by the microscopic characteristics described above. Peanut leaf blight and wilt disease caused by F. incarnatum has been reported in India (Thirumalaisamy et al. 2019). This first report emphasizes that this phytopathogen is a new threat for peanut producers in Mexico, which is why our finding suggests the need to seek new strategies for its control.

Effiiency of the Essential Oil of Porophyllum linaria (Asteraceae) a Mexican Endemic Plant Against Sitophilus zeamais (Coleoptera: Curculionidae).

The insecticidal and repellent effect of essential oil isolated from fresh leaves of Porophyllum linaria on maize weevil was evaluated, as well as the effect on the grain germination after treated. In total, 28 constituents were identified by gas chromatography coupled with mass spectrometry accounting for 99.86% of whole essential oil. The main majority compounds were ß-myrcene (41.94%), D-limonene (20.29%), and estragole (20.03%). Contact toxicity significantly increased with dose and time after treatment. With the 800 ppm (highest concentration), the mortality (%) obtained for the tenth and fifteenth day was 43 and 82%, respectively, whereas with 50 ppm (lowest concentration) 30% mortality was obtained at the end of the experiment (fifteenth day). At 15 d (end of the experiment), the LC50 y LC90 were obtained with values of 329.01 ± 44.35 y 1058.86 ± 117.76 ppm, respectively. For a concentration of 800 ppm, a selection index of zero was obtained, indicating the preference of the pest to the untreated maize (control). The maize grains germination test showed a significant reduction both in the length of hypocotyl and radicle of maize grain. So, in the highest dose, the hypocotyl and radicle length was 1.40 ± 0.34 and 9.14 ± 0.55 cm, respectively, whereas the control group registered 3.28 ± 0.39 and 13.02 ± 0.97 cm, respectively. This finding is promising since as it could result in the identification of botanical substances capable of suppressing maize weevil, Sitophilus zeamais development.

Volatiles released by Beauveria bassiana induce oviposition behavior in the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae).

Microbial volatile organic compounds may act as semiochemicals, inciting different behavioral responses in insects. Beauveria bassiana is an entomopathogenic fungus, and physiological and environmental factors are positively related to fungal virulence. In this study, we examined the volatile profiles produced by eight B. bassiana strains, isolated from soil plots and mycosed insect cadavers, with different speeds of kill and determined if these compounds induce oviposition behavior in Spodoptera frugiperda. Fungal volatilome analysis revealed differences between the isolates. Isolates from mycosed insects showed higher virulence, larger egg mass area and length, and a higher number of eggs by mass, than those obtained from soil. Furthermore, a dilution of the fungal odoriferous compounds increased the insect response, suggesting that S. frugiperda is highly susceptible to the fungal compound's fingerprint. Otherwise, the insect response to the natural blend of volatiles released by the fungus was different from that obtained with 3-methylbutanol, which was the most abundant compound in all isolates. The ability of an entomopathogen to produce volatiles that can induce olfactory stimulation of egg-laying behavior could represent an ecological adaptive advantage in which the entomopathogen stimulates the insect population growth.

Biological Control of Charcoal Rot in Peanut Crop through Strains of Trichoderma spp., in Puebla, Mexico.

Charcoal rot is an emerging disease for peanut crops caused by the fungus Macrophomina phaseolina. In Mexico, peanut crop represents an important productive activity for various rural areas; however, charcoal rot affects producers economically. The objectives of this research were: (a) to identify and morphologically characterize the strain "PUE 4.0" associated with charcoal rot of peanut crops from Buenavista de Benito Juárez, belonging to the municipality of Chietla in Puebla, Mexico; (b) determine the in vitro and in vivo antagonist activity of five Trichoderma species on M. phaseolina, and (c) determine the effect of the incidence of the disease on peanut production in the field. Vegetable tissue samples were collected from peanut crops in Puebla, Mexico with the presence of symptoms of charcoal rot at the stem and root level. The "PUE 4.0" strain presented 100% identity with M. phaseolina, the cause of charcoal rot in peanut crops from Buenavista de Benito Juárez. T. koningiopsis (T-K11) showed the highest development rate, the best growth speed, and the highest percentage of radial growth inhibition (PIRG) over M. phaseolina (71.11%) under in vitro conditions, in addition, T. koningiopsis (T-K11) showed higher production (1.60 ± 0.01 t/ha-1) and lower incidence of charcoal rot under field conditions. The lowest production with the highest incidence of the disease occurred in plants inoculated only with M. phaseolina (0.67 ± 0.01 t/ha-1) where elongated reddish-brown lesions were observed that covered 40% of the total surface of the main root.

First Report of Macrophomina phaseolina Causing Charcoal Rot of Peanut (Arachis hypogaea L.) in Mexico.

Peanut (Arachis hypogaea L.) is the third most important oilseed crop in the world. The cultivated area in Mexico is currently 52,046 ha with a production of 91,109 ton in 2018 (FAO, 2020). Puebla state ranks third in the national production with 9,313 ton (SIAP, 2020). In September 2019, typical symptoms of charcoal rot (Macrophomina phaseolina (Tassi) Goid.) were observed in about 50% of cultivar Virginia Champs peanuts, and it affecting 1.5 ha located in Chietla (18° 27' 39" N; 98° 37' 11" W), Puebla, Mexico. Diseased plants showed brown discoloration in stem and root rot, with chlorotic foliage, dark microsclerotia were observed on the stem and premature dying. To isolate the causal agent of these symptoms, 20 infected plants were recovered and processed in the laboratory. Ten pieces of stem and root tissue were selected from each plant, cut into small pieces 5-mm in length, superficially disinfested with 1% sodium hypochlorite for 3 min, followed by three rinses with sterile distilled water. Later, dried on sterile paper and placed on Petri plates containing potato dextrose agar (PDA) medium, which were kept at 28°C for 7 days (12 h light and 12 h dark). Four colonies were purified via hyphal tip culture, fungus was consistently isolated from the analyzed tissues; additional microcultures were prepared to observe phenotypic characteristics. Colonies showed dense growth, with a gray initial mycelium, becoming black after 7 days. Microesclerotia with spherical to oblong in shape were observed after 5 days on PDA, with a black coloration, measuring an average of 74 µm width × 110 µm length (n=40). Phylogenetic analysis was conducted by amplification and sequencing of the internal transcribed spacer (ITS) region with the ITS5 and ITS4 primers (White et al. 1990). The obtained sequences were deposited in GenBank database under accession numbers: MW585378, MW585379, MW585380, and MW585381 containing approximately 601 bp of the ITS1-5.8S-ITS2 region (complete sequence); they were 99% identical with the reference sequence of Macrophomina phaseolina (GenBank accession KF951698) isolated in Phaseolus vulgaris from Mexico. Based on the symptoms in the field, colony morphology, color, and shape of the microsclerotia, and molecular identification, the fungus was identified as M. phaseolina (Tassi) Goid. The pathogenicity test was performed on peanut plants cultivar Virginia Champs grown on plastic pots with an autoclaved peat/soil mixture under greenhouse conditions (70% relative humidity and 28°C). Fifty two-month-old peanut plants were inoculated using the toothpick method. The toothpicks were previously sterilized and then placed in Petri plates with each of the four colonies of M. phaseolina until colonization. Small wounds were made with those toothpicks in the roots, and a sterile toothpick was used in the control plants, the assays were performed twice. After three weeks, the inoculated plants exhibited symptoms of wilting chlorosis on the leaves and brown to dark brown discoloration of the vascular ring, while control plants remained healthy. M. phaseolina was re-isolated from symptomatic root tissues and identified by phylogenetic approach, fulfilling Koch's postulates. To date, this fungus affects at least 372 hosts globally causing yield losses. Although in Mexico this fungus has been documented in Glycine max, Ipomoea batatas, Phaseolus vulgaris, Physalis ixocarpa, Saccharum officinarum, Sesamum indicum, Solanum melongena, S. tuberosum, and Sorghum bicolor (Farr and Rossman 2021). However, there are no reports of M. phaseolina as a potential pathogen on peanut; therefore, according to our knowledge, this is the first report of this fungus affecting A. hypogaea in Mexico.

Toxicity and Residual Activity of Insecticides Against Tamarixia triozae (Hymenoptera: Eulophidae), a Parasitoid of Bactericera cockerelli (Hemiptera: Triozidae).

Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) is one of the most economically important pests of potato, tomato, and peppers in Central America, Mexico, the United States, and New Zealand. Its control is based on the use of insecticides; however, recently, the potential of the eulophid parasitoid Tamarixia triozae (Burks) (Hymenoptera: Eulophidae) for population regulation has been studied. Because T. triozae is likely to be exposed to insecticides on crops, the objective of this study was to explore the compatibility of eight insecticides with this parasitoid. The toxicity and residual activity (persistence) of spirotetramat, spiromesifen, beta-cyfluthrin, pymetrozine, azadirachtin, imidacloprid, abamectin, and spinosad against T. triozae adults were assessed using a method based on the residual contact activity of each insecticide on tomato leaf discs collected from treated plants growing under greenhouse conditions. All eight insecticides were toxic to T. triozae. Following the classification of the International Organization of Biological Control, the most toxic were abamectin and spinosad, which could be placed in toxicity categories 3 and 4, respectively. The least toxic were azadirachtin, pymetrozine, spirotetramat, spiromesifen, imidacloprid, and beta-cyfluthrin, which could be placed in toxicity category 2. In terms of persistence, by day 5, 6, 9, 11, 13, 24, and 41 after application, spirotetramat, azadirachtin, spiromesifen, pymetrozine, imidacloprid, beta-cyfluthrin, abamectin, and spinosad could be considered harmless, that is, placed in toxicity category 1 (<25% mortality of adults). The toxicity and residual activity of some of these insecticides allow them to be considered within integrated pest management programs that include T. triozae.