Long-Lasting Silver Nanoparticles Synthesized with Tagetes erecta and Their Antibacterial Activity against Erwinia amylovora, a Serious Rosaceous Pathogen.

A rapid, eco-friendly, and simple method for the synthesis of long-lasting (2 years) silver nanoparticles (AgNPs) is reported using aqueous leaf and petal extracts of Tagetes erecta L. The particles were characterized using UV-Visible spectrophotometry and the analytical and crystallographic techniques of transmission electron microscopy (TEM). The longevity of the AgNPs was studied using UV-Vis and high-resolution TEM. The antibacterial activity of the particles against Erwinia amylovora was evaluated using the Kirby-Bauer disk diffusion method. The results were analyzed using ANOVA and Tukey's test (p ≤ 0.05). Both the leaf and petal extracts produced AgNPs, but the leaf extract (1 mL) was long-lasting and quasi-spherical (17.64 ± 8.87 nm), with an absorbance of UV-Vis λmax 433 and a crystalline structure (fcc, 111). Phenols, flavonoids, tannins, and terpenoids which are associated with -OH, C=O, and C=C were identified in the extracts and could act as reducing and stabilizing agents. The best antibacterial activity was obtained with a nanoparticle concentration of 50 mg AgNPs L-1. The main contribution of the present research is to present a sustainable method for producing nanoparticles which are stable for 2 years and with antibacterial activity against E. amylovora, one of most threatening pathogens to pear and apple productions.

[Identification and antifungal activity of halophilic bacteria isolated from saline soils in Campeche, México]. / Identificación y actividad antifúngica de bacterias halófilas aisladas de suelos salinos en Campeche, México.

Phytopathogenic fungi Alternaria alternata and Colletotrichum gloeosporioides cause diseases in plant tissues as well as significant postharvest losses. The use of chemical fungicides for their control has negative effects on health and the environment. Secondary metabolites from halophilic bacteria are a promising alternative for new antifungal compounds. In the present study, halophilic bacteria were isolated and characterized from two sites with saline soils called branquizales in Campeche, Mexico. A total of 64 bacteria were isolated. Agrobacterium, Bacillus, Inquilinus, Gracilibacillus, Metabacillus, Neobacillus, Paenibacillus, Priestia, Staphylococcus, Streptomyces and Virgibacillus were among the identified genera. The antifungal potential of the culture supernatant (CS) of 39 halophilic bacteria was investigated against C. gloeosporioides and A. alternata. The bacteria showing the greatest inhibition of mycelial growth corresponded to Bacillus subtilis CPO 4292, Metabacillus sp. CPO 4266, Bacillus sp. CPO 4295 and Bacillus sp. CPO 4279. The CS of Bacillus sp. CPO 4279 exhibited the highest activity and its ethyl acetate extract (AcOEt) inhibited the germination of C. gloeosporioides, with IC50 values of 8,630µg/ml and IC90 of 10,720µg/ml. The organic partition of the AcOEt extract led to three fractions, with acetonitrile (FAcB9) showing the highest antifungal activity, with values exceeding 66%. Halophilic bacteria from 'blanquizales' soils of the genus Bacillus sp. produce metabolites with antifungal properties that inhibit the phytopathogenic fungus C. gloeosporioides.

Preventive and Curative Effects of Treatments to Manage Strawberry Root and Crown Rot Caused by Neopestalotiopsis rosae.

Strawberry root and crown rot caused by the fungus Neopestalotiopsis rosae is an emerging disease that has caused yield losses reaching 70% in Mexico and other regions worldwide. This research evaluated the effects of biological and chemical fungicides applied as preventive and curative applications for controlling root and crown rot caused by N. rosae in strawberries under greenhouse conditions. Treatments included these chemical fungicides: prochloraz, prochloraz+thiram, cyprodinil+fludioxonil, difenoconazole+azoxystrobin, iprodione, captan, thiram, pydiflumetofen+fludioxonil, fluxapyroxad+pyraclostrobin, and hymexazol; each applied at commercial doses. Also tested were biological treatments based on Trichoderma koningiopsis, Trichoderma asperellum, Streptomyces sp., and Bacillus amyloliquefaciens strain D747 (B. velezensis). Disease incidence, severity, plant mortality, root length, and dry weight were determined. Results showed that overall, preventive applications of the fungicides pydiflumetofen+fludioxonil, cyprodinil+fludioxonil, and prochloraz resulted in the smallest area under the disease progress curve, and lowest final disease incidence, severity, and plant mortality. An intermediate group of effective treatments entailed hymexazol, iprodione, T. asperellum, and T. koningiopsis (50-75% efficacy). Treatments with greater efficacy (99 to 100%), prochloraz pydiflumetofen+fludioxonil, cyprodinil+fludioxonil, and prochloraz, also had maximal total plant biomass vis-à-vis the untreated control. In contrast, each treatment's efficacy was significantly reduced when applied curatively (0 to 37% treatment efficacy). These results suggest that certain treatments are useful for controlling strawberry root and crown rot caused by N. rosae, when applied preventively (as root dipping). These results will contribute to design more effective management programs of root rot and crown rot caused by N. rosae on strawberry.

[Ecological and biological benefits of the cosmopolitan fungus Trichoderma spp. in agriculture: A perspective in the Mexican countryside]. / Beneficios ecológicos y biológicos del hongo cosmopolita Trichoderma spp. en la agricultura: una perspectiva en el campo mexicano.

There is currently an extensive record of scientific studies on the general characteristics of filamentous fungus Trichoderma spp., which demonstrates its wide range of interrelation in ecosystems and its fungal activity that benefits the agricultural sector and agroindustry, as well as its importance in the preservation and restoration of the soil microbiota. The success of the biological and ecological benefits of Trichoderma is due to its reproductive capacity, as well as its efficiency in the use of soil nutrients; the efficacy of the genus has been reported against a variety of phytopathogenic fungi, as well as the potential to synthesize and release enzymes (cellulases, xylanases, and chitinases) that have been implemented in agroindustrial bioprocesses. It has also been reported that various species of Trichoderma spp. can produce auxins and gibberellin-type growth regulators, reported as growth promoters of some agricultural crops; however, their most relevant fact is their ability to prevail at certain doses of 'agrotoxic' active ingredients and contribute studies regarding processes for obtaining biofuel and bioremediation of the agricultural soil. In this overview, a general description of the current and relevant studies of the different subspecies of Trichoderma and their contribution in agriculture is made, presenting results obtained in vitro, in greenhouses and in the field. This analysis will serve as a starting point for future research in Mexico, specifically on the genus Trichoderma and its benefits for the Mexican countryside.

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.

Tomato Plant Microbiota under Conventional and Organic Fertilization Regimes in a Soilless Culture System.

Tomato is the main vegetable cultivated under soilless culture systems (SCSs); production of organic tomato under SCSs has increased due to consumer demands for healthier and environmentally friendly vegetables. However, organic tomato production under SCSs has been associated with low crop performance and fruit quality defects. These agricultural deficiencies could be linked to alterations in tomato plant microbiota; nonetheless, this issue has not been sufficiently addressed. Thus, the main goal of the present study was to characterize the rhizosphere and phyllosphere of tomato plants cultivated under conventional and organic SCSs. To accomplish this goal, tomato plants grown in commercial greenhouses under conventional or organic SCSs were tested at 8, 26, and 44 weeks after seedling transplantation. Substrate (n = 24), root (n = 24), and fruit (n = 24) composite samples were subjected to DNA extraction and high-throughput 16S rRNA gene sequencing. The present study revealed that the tomato core microbiota was predominantly constituted by Proteobacteria, Actinobacteria, and Firmicutes. Remarkably, six bacterial families, Bacillaceae, Microbacteriaceae, Nocardioidaceae, Pseudomonadaceae, Rhodobacteraceae, and Sphingomonadaceae, were shared among all substrate, rhizosphere, and fruit samples. Importantly, it was shown that plants under organic SCSs undergo a dysbiosis characterized by significant changes in the relative abundance of Bradyrhizobiaceae, Caulobacteraceae, Chitinophagaceae, Enterobacteriaceae, Erythrobacteraceae, Flavobacteriaceae, Nocardioidaceae, Rhodobacteraceae, and Streptomycetaceae. These results suggest that microbial alterations in substrates, roots, and fruits could be potential factors in contributing to the crop performance and fruit quality deficiencies observed in organic SCSs.

Novel Harzia ixtarensis Fungus on Annona cherimola Fruit in Mexico and Its Synergistic Relationship with Colletotrichum fragariae.

Since 2005 in Íxtaro, Michoacán, symptoms of Harzia infection have been observed on immature Annona cherimola fruit with Colletotrichum fragariae-induced anthracnose lesions and mummified fruit. This study aimed to identify the Harzia sp. and evaluate its pathogenicity. Four isolates were obtained from fruit exhibiting symptoms, cultured in four types of agar under various conditions, and characterized based on concatenated internal transcribes spacer (ITS) + large subunit and ITS + small subunit sequences. Additionally, the isolates were compared with two CBS species (two-type strains and two isolates) of Harzia patula and H. tenella under the same conditions as the Harzia isolates, and all known Harzia spp. in culture were included in two phylogenetic analyses. H. ixtarensis sp. nov. was proposed. Compared with H. patula CBS isolate 121524 which was the most closely phylogenetically related species, H. ixtarensis was characterized by slower colony growth (white to salmonish-beige), different percentages of two forms of conidia (elongated and globose; unicellular and hyaline to subhyaline), and smaller conidia. The conidia mainly germinated with two hyaline tubes without an appressorium. In situ inoculations (1 × 106 ml-1 conidia suspension) of fruit showed that fruit with wounds developed larger lesions than those without wounds. Harzia inoculation on anthracnose lesions (induced by prior inoculation with C. fragariae) produced larger anthracnose lesions than C. fragariae alone. When C. fragariae or H. ixtarensis was inoculated alone, the lesion size was 51 and 99% smaller, respectively, indicating synergy between C. fragariae and H. ixtarensis. Thus, H. ixtarensis may have a parasitic-synergistic and necrotrophic lifestyle, and exhibited symptoms on anthracnose lesions.

Health-Related Quality of Life in Primary Caregivers of People Receiving Palliative Home Care.

Primary caregivers face constant challenges because of changes in the clinical situation of the person receiving palliative home care. These changes can alter the health-related quality of life and all its dimensions. This study aimed to describe the health-related quality of life of 137 primary caregivers of people enrolled in a palliative home care program in Bogotá, Colombia, applying a quantitative, descriptive, and cross-sectional research design. The Caregiver's Quality of Life Instrument, initially developed by Ferrell, and the sociodemographic characteristics form for caregivers of people with chronic disease, both previously validated in the Colombian population, were used. The results showed that the primary caregivers have a good and adequate overall health-related quality of life; however, they presented some alterations in the physical, psychological, and social dimensions. Therefore, nursing and interdisciplinary palliative care teams should aim their interventions not only at patients but also at primary caregivers during palliative home care.

Identification based on morphological and molecular characters of Phassus huebneri (Geyer) (Lepidoptera: Hepialidae), a ghost moth feeding on blackberry in Michoacán, Mexico: its biology and parasitoids.

Larvae of an unidentified Lepidopteran hepialid were found boring stems and crown roots of blackberry (Rubus sp.) in Michoacán, Mexico. In this study, the morphological and molecular identification of larvae and adults of this insect was performed. Preliminary aspects of biology, including information on parasitoids associated to larval stage, are also recorded. A total of 109 larvae of various instars were collected and three were reared to adult. The two females and one male were identified as Phassus huebneri (Geyer) (Lepidoptera: Hepialidae) following morphological characters. This identification was corroborated by comparing the cytochrome oxidase I (COI) barcode of two field-collected larvae (IIAFL1 and IIAFL2) with sequences of Phassus spp. obtained from GenBank. A Neighbor-Joining similarity analysis yielded a phenogram with two subclades. The first subclade grouped the IIAFL1 and IIAFL2 sequences with two other Mexican sequences categorized as P. huebneri, whereas in the second subclade other species belonging to the genus Phassus from Guatemala and Costa Rica were grouped together. Larval development of nine instars took about 14 months. Larvae reached a maximum length of 10 cm. The pupal stage lasted 39-41 days. Each of the two females laid over 1,000 unfertilized eggs within a period of 48 h. Larvae were parasitized by Dinera grisescens Fallen (Diptera: Tachinidae) and another unidentified Dinera sp. This study provides additional evidences on the distribution pattern of P. huebneri in western Mexico and presents the first record of this species feeding on one of the most economically important crops in Mexico.

Comparative Performance of Fungicides and Biocontrol Products in the Management of Fusarium Wilt of Blackberry.

Fusarium wilt of blackberry (FWB) is an emerging disease caused by a Fusarium oxysporum species complex. More than 3,000 ha of blackberry (Rubus spp.) crops have been lost in Mexico since 2011. The objectives of this research were: to evaluate the sensitivity of pathogenic F. oxysporum isolates recovered from symptomatic blackberry plants to fungicides with different modes of action; to assess the potential of these fungicides and plant resistance inducers against FWB in the greenhouse; and to determine the effects of commercial biofungicides and two indigenous strains of Trichoderma spp. on the incidence of FWB. The EC50 values of the fungicides prochloraz, thiabendazole, azoxystrobin, thiophanate-methyl, difenoconazole, triflumizole, and potassium phosphite for six pathogenic F. oxysporum isolates were determined. In a separate experiment, the fungicides acibenzolar-s-methyl (ASM), potassium phosphite, and commercial biofungicides, as well as two soil microbial inoculants and two indigenous Trichoderma strains, were tested for protection against wilt development in blackberry plants in the greenhouse. Prochloraz showed an average sensitivity for EC50 of 0.01 µg ml-1 for the tested F. oxysporum isolates, followed by difenoconazole and thiabendazole. Prochloraz and ASM proved to be the most effective treatments in the greenhouse. In contrast, potassium phosphite was ineffective in both the in vitro and in vivo experiments. The soil bioinoculants MicroSoil, Baktillis, T. koningiopsis, and T. asperellum significantly reduced the incidence of disease in the greenhouse. These results provide evidence for the potential of the various tools as useful components of integrated FWB management in the field.

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.

A methodology for the technical-economic analysis of municipal solid waste systems based on social cost-benefit analysis with a valuation of externalities.

Countries face a serious problem due to the generation and management of higher volumes of waste. Large-scale production of waste has promoted the establishment of various operations (collection, transport, treatment and disposal) for its management. When a MSW management system is implemented, it can generate different impacts or consequences (internal or external impacts). Generally, external impacts (social and environmental impacts) are not reflected in MSW economic analysis or taken into consideration in decision-making processes in regard to MSW management options. For this reason, the objective of this paper is present a methodology with which is viable to conduct the technical-economic analysis of municipal solid waste management projects based on social cost-benefit analysis (sCBA) as it considers internal and external impacts. Its main objectives are to determine the total benefits (the difference between revenues and costs) generated by a project and to reduce uncertainty and risk of investing in particular MSW management system. Finally, a case study was carried out to verify the validity of the methodology through analysis and valuation of different impacts of a light packaging waste and bulky waste facility. Through the application of the methodology, it has been possible to visualize that this facility is viable operationally (BP = 42.94 €/ton) as economically (BT = 87.73 €/ton).

A Comprehensive Evaluation of Enterobacteriaceae Primer Sets for Analysis of Host-Associated Microbiota.

Enterobacteriaceae is one of the most important bacterial groups within the Proteobacteria phylum. This bacterial group includes pathogens, commensal and beneficial populations. Numerous 16S rRNA gene PCR-based assays have been designed to analyze Enterobacteriaceae diversity and relative abundance, and, to the best of our knowledge, 16 primer pairs have been validated, published and used since 2003. Nonetheless, a comprehensive performance analysis of these primer sets has not yet been carried out. This information is of particular importance due to the recent taxonomic restructuration of Enterobacteriaceae into seven bacterial families. To overcome this lack of information, the identified collection of primer pairs (n = 16) was subjected to primer performance analysis using multiple bioinformatics tools. Herein it was revealed that, based on specificity and coverage of the 16S rRNA gene, these 16 primer sets could be divided into different categories: Enterobacterales-, multi-family-, multi-genus- and Enterobacteriaceae-specific primers. These results highlight the impact of taxonomy changes on performance of molecular assays and data interpretation. Moreover, they underline the urgent need to revise and update the molecular tools used for molecular microbial analyses.

Endophytic Trichoderma Species Isolated from Persea americana and Cinnamomum verum Roots Reduce Symptoms Caused by Phytophthora cinnamomi in Avocado.

Avocado root rot caused by the oomycete Phytophthora cinnamomi is a severe disease that affects avocado production in Mexico and worldwide. The use of biological control agents such as Trichoderma species isolated from places where the disease is always present, represents an efficient alternative to reduce losses. Thus, the objective of this research was to evaluate the biocontrol ability of 10 endophytic Trichoderma spp. strains against P. cinnamomi tested both in vitro and in the greenhouse. The endophytic Trichoderma spp. were recovered from Persea americana and Cinnamomum verum roots, isolated and purified on potato-dextrose-agar medium. Ten strains were identified by phylogenetic reconstruction of the internal transcribed spacer region of rDNA sequences as T. asperellum (T-AS1, T-AS2, T-AS6, and T-AS7), T. harzianum (T-H3, T-H4, and T-H5), T. hamatum (T-A12), T. koningiopsis (T-K8 and T-K11), and P. cinnamomi (CPO-PCU). In vitro dual-culture assay, the percentage of inhibition of radial growth (PIRG) between Trichoderma spp. and P. cinnamomi strains was measured according to the Bell's scale. PIRG results indicated that T-AS2 reached the highest value of 78.32%, and T-H5 reached the lowest value of 38.66%. In the greenhouse, the infection was evaluated according to the percentage of disease incidence. Plants with the lowest incidence of dead by avocado root rot were those whose seedlings were inoculated with T-AS2 and T-AS7, resulting in only 5% death by root rot caused by P. cinnamomi. The disease incidence of seedlings with wilt symptoms and death decreased more than 50% in the presence of Trichoderma spp. Relying on the results, we conclude that T. asperellum and T. harzianum contribute to the biocontrol of soil-borne pathogenic oomycete P. cinnamomi.

An Emerging Strawberry Fungal Disease Associated with Root Rot, Crown Rot and Leaf Spot Caused by Neopestalotiopsis rosae in Mexico.

In the 2017 strawberry season, several transplant losses reaching 50% were observed in Zamora, Michoacán Valley, Mexico, due to a new fungal disease associated with root rot, crown rot, and leaf spot. In this year the disease appeared consistently and increased in the following seasons, becoming a concern among strawberry growers. Thus, the aim of this research was to determine the etiology of the disease and to determine the in vitro effect of fungicides on mycelial growth of the pathogen. Fungal isolates were obtained from symptomatic strawberry plants of the cultivars 'Albion' and 'Festival' and were processed to obtain monoconidial isolates. Detailed morphological analysis was conducted. Concatenated phylogenetic reconstruction was conducted by amplifying and sequencing the translation elongation factor 1 α, ß-tubulin partial gene, and the internal transcribed spacer region of rDNA. Pathogenicity tests involving inoculation of leaves and crowns reproduced the same symptoms as those observed in the field, fulfilling Koch's postulates. Morphology and phylogenetic reconstruction indicated that the causal agent of the described symptoms was Neopestalotiopsis rosae, marking the first report anywhere in the world of this species infecting strawberry. N. rosae was sensitive to cyprodinil + fludioxonil, captan, iprodione, difenoconazole, and prochloraz.

A Molecular Tool for Rapid Detection and Traceability of Cyclospora cayetanensis in Fresh Berries and Berry Farm Soils.

Due to recent outbreaks of cyclosporiasis associated with consumption of fresh berries, producers are demanding modern microbiological tools for the rapid and accurate identification of the human pathogen Cyclospora cayetanensis in berries and environmental samples. The aim of the present work was to develop a molecular tool based on a PCR approach for the rapid and accurate detection of C. cayetanensis. A nested PCR assay was validated for the amplification of a 294 bp size region of the 18S rRNA gene from C. cayetanensis. The limit of detection for the nested PCR assay was validated using 48 berry samples spiked with ~0, 10, 100, and 1000 oocyst per gram of sample. With this assay, it was possible to detect as few as 1 oocyst per gram of berry, in a 50 g sample. Sanger DNA sequencing and phylogenetic analysis were carried out to confirm the presence of C. cayetanensis in berry (n = 17) and soil (n = 5) samples. The phylogenetic analysis revealed that the C. cayetanensis sequences obtained from Mexico clustered within a group recovered from China, Peru, Guatemala-Haiti, and Japan. The PCR protocol designed in the present study could be an important tool for the rapid and accurate detection of this human pathogen in environmental and food samples.

Conventional and qPCR reveals the presence of 'Candidatus Liberibacter solanacearum' haplotypes A, and B in Physalis philadelphica plant, seed, and Βactericera cockerelli psyllids, with the assignment of a new haplotype H in Convolvulaceae.

The husk tomato (Physalis philadelphica Lam.) is an important Solanaceae native to Mesoamerica that is grown for its green fruit used as an important ingredient in domestic and international cuisine. Nevertheless, husk tomato plants with symptoms resembling those caused by 'Candidatus Liberibacter solanacearum' (CLso) have been observed during the last decade in plantations located in the State of Mexico, Michoacan and Sinaloa in Mexico. These areas are located near other solanaceous crops where Bactericera cockerelli the well-known psyllid transmitter of CLso is frequently present. Thus, the goal of this study was to determine if CLso haplotypes are present in husk tomato varieties in commercial fields in Mexico. From 2015 to 2016, plants and fruit showing evident symptoms of CLso infection, as well as psyllids were collected in these states and assayed by PCR for CLso using primer sets OA2/OI2c and LpFrag 1-25F/427R. Phylogenetic reconstruction was performed with Bayesian analysis and maximum likelihood methods using amplicon sequences obtained in this work along with those deposited in the GenBank database corresponding to the CLso detected in Solanaceae, Apiaceae, and Convolvulaceae host families. In addition, all the sequences were subjected to haplotype determination through an analysis of DNA polymorphisms using the DnaSP software. Furthermore, quantitative PCR (qPCR) was performed using CLso-specific primers and probes. Phylogenetic reconstruction and qPCR confirmed the presence of CLso in plants, seeds and insect-vectors, and CLso sequences from plants and seeds completely matched haplotype B, whereas CLso haplotypes A and B were detected in B. cockerelli psyllids. Polymorphism analysis identified a novel Convolvulaceae-associated CLso haplotype, which was named haplotype H. The results of this study will enable the dissemination of infected seeds to new husk tomato production areas to be avoided.

Respuesta del simbiosistema frijol (Phaseolus vulgaris L.) y Rhizobium tropici CIAT899 ante el efecto alelopático de Ipomoea purpurea L. Roth / Responses of the common bean (Phaseolus vulgaris L.) and Rhizobium tropici CIAT899 symbiosystem to induced allelopathy by Ipomoea purpurea L. Roth

La alelopatía es un fenómeno que involucra la producción de metabolitos secundarios que influyen en el crecimiento de las plantas, pero este potencial alelopático ha sido poco estudiado en la simbiosis rizobio-leguminosa. Esta investigación tuvo los siguientes objetivos: 1) evaluar el potencial alelopático de lixiviados acuosos de Ipomoea purpurea L. Roth en la germinación de semillas y en el crecimiento radical de plántulas de frijol (Phaseolus vulgaris L.); 2) determinar el efecto de estos lixiviados en el crecimiento in vitro de Rhizobium tropici CIAT899, y 3) evaluar el potencial alelopático de I. purpurea en el crecimiento, la fisiología y la nodulación de frijol en simbiosis con R. tropici. Tanto el lixiviado acuoso de raíz como el de la parte aérea de I. purpurea estimularon la germinación de semillas de frijol y la elongación radical. El crecimiento in vitro de R. tropici fue inhibido al aplicar los 2 tipos de lixiviado. La presencia de I. purpurea tuvo un efecto negativo en el crecimiento y en las respuestas fisiológicas de las plantas de frijol, que fue atenuado cuando las plantas fueron inoculadas con Rhizobium tropici; no obstante, la nodulación asociada a esta bacteria fue afectada en presencia de la planta alelopática. Los resultados indican que la simbiosis de rizobios en las raíces de frijol es un elemento importante en la atenuación de los danos producidos por la planta alelopática I. purpurea.

Allelopathy is a phenomenon that involves the production of secondary metabolites that influence the growth of plants and microorganisms; however, this alellopathic effect has been scarcely studied on the rhizobia-legume symbiosis. The aims of this research were 1) to assess the allelopathic potential of aqueous extracts of Ipomoea purpurea L. Roth on seed germination and root length of common bean seedlings (Phaseolus vulgaris L.), 2) to determine its effects on the in vitro growth of Rhizobium tropici CIAT899, and 3) to evaluate the allelopathic potential of I. purpurea on the growth, nodulation and physiology of common bean plants inoculated with R. tropici. After 48 h, 15% of the aqueous root extract of I. purpurea stimulated seed germination, whereas 4% of the aqueous shoot extracts stimulated such germination. Both the root or shoot extracts stimulated seed germination and e root length. In vitro growth of R. tropici was inhibited as a result of the application of both aqueous extracts. The presence of I. purpurea negatively affected both the growth and physiological responses of common bean plants, and this effect was attenuated after the inoculation of R. tropici; nevertheless, this allelopathic plant affected root nodulation. Our results suggest that the symbiosis of rhizobia and roots of common bean plants is an important element for attenuating the negative effects caused by the allelopathic plant.

[Responses of the common bean (Phaseolus vulgaris L.) and Rhizobium tropici CIAT899 symbiosystem to induced allelopathy by Ipomoea purpurea L. Roth]. / Respuesta del simbiosistema frijol (Phaseolus vulgaris L.) y Rhizobium tropici CIAT899 ante el efecto alelopático de Ipomoea purpurea L. Roth.

Allelopathy is a phenomenon that involves the production of secondary metabolites that influence the growth of plants and microorganisms; however, this alellopathic effect has been scarcely studied on the rhizobia-legume symbiosis. The aims of this research were 1) to assess the allelopathic potential of aqueous extracts of Ipomoea purpurea L. Roth on seed germination and root length of common bean seedlings (Phaseolus vulgaris L.), 2) to determine its effects on the in vitro growth of Rhizobium tropici CIAT899, and 3) to evaluate the allelopathic potential of I. purpurea on the growth, nodulation and physiology of common bean plants inoculated with R. tropici. After 48h, 15% of the aqueous root extract of I. purpurea stimulated seed germination, whereas 4% of the aqueous shoot extracts stimulated such germination. Both the root or shoot extracts stimulated seed germination and e root length. In vitro growth of R. tropici was inhibited as a result of the application of both aqueous extracts. The presence of I. purpurea negatively affected both the growth and physiological responses of common bean plants, and this effect was attenuated after the inoculation of R. tropici; nevertheless, this allelopathic plant affected root nodulation. Our results suggest that the symbiosis of rhizobia and roots of common bean plants is an important element for attenuating the negative effects caused by the allelopathic plant.

Colletotrichum fructicola, a Member of Colletotrichum gloeosporioides sensu lato, is the Causal Agent of Anthracnose and Soft Rot in Avocado Fruits cv. "Hass".

The filamentous Ascomycota Colletotrichum gloeosporioides sensu lato is a fungus that has been reported worldwide as a causal agent of anthracnose disease in avocado and other crops. In Mexico, this species affects fruits from an early stage of development in the orchard until the post-harvest stage. Although fungicides are continuously applied to control Colletotrichum species, pericarp cankers and soft rot mesocarp in fruits are still frequently observed. Considering the lack of a precise description of the causative agent, the aim of the current study was to determine the pathogens involved in this symptomatology. Twenty-four isolates were consistently obtained from the pericarp of avocado fruits cv. "Hass" collected in the central avocado-producing area of Mexico. Morphological features such as colony growth, conidia size, and mycelial appressorium were assessed. Bayesian multilocus phylogenetic analyses were performed using amplified sequences of the internal transcribed spacer region of the nuclear ribosomal DNA; actin, chitin synthase, glyceraldehyde-3-phosphate dehydrogenase partial genes; and APn2-Mat1-2 intergenic spacer and mating type Mat1-2 partial gene from the nine selected isolates. In addition, fruits were inoculated with a conidial suspension and reproducible symptoms confirmed the presence of Colletotrichum fructicola in this area. This pathogenic species can now be added to those previously reported in the country, such as C. acutatum, C. boninense, C. godetiae, C. gloeosporioides, and C. karstii. Disease management programs to reduce the incidence of anthracnose should include C. fructicola to determine its response to fungicides that are routinely applied, considering that the appearance of new species is affecting the commercial quality of the fruits and shifting the original population structure.