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.

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.

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.

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.

In a belowground multitrophic interaction, Trichoderma harzianum induces maize root herbivore tolerance against Phyllophaga vetula.

BACKGROUND: Trichoderma spp. are soil fungi that interact with plant roots and associated biota such as other microorganisms and soil fauna. However, information about their interactions with root-feeding insects is limited. Here, interactions between Trichoderma harzianum and the root-feeding insect Phyllophaga vetula, a common insect pest in maize agroecosystems, were examined. RESULTS: Applications of T. harzianum and P. vetula to the root system increased and decreased maize growth, respectively. Induced tolerance against herbivore attack was provided by T. harzianum maintaining a robust and functional root system as evidenced by the increased uptake of Cu, Ca, Mg, Na and K. Herbivore tolerance also coincided with changes in the emission of root volatile terpenes known to induce indirect defense responses and attract natural enemies of the herbivore. More importantly, T. harzianum induced de novo emission of several sesquiterpenes such as ß-caryophyllene and δ-cadinene. In addition, single and combined applications of T. harzianum and P. vetula altered the sucrose content of the roots. Finally, T. harzianum produced 6-pentyl-2H-pyran-2-one (6-PP) a volatile compound that may act as an antifeedant-signaling compound mitigating root herbivory by P. vetula. CONCLUSION: Our results provide novel information about belowground multitrophic plant-microbe-arthropod interactions between T. harzianum and P. vetula in the maize rhizosphere resulting in alterations in maize phenotypic plant responses, inducing root herbivore tolerance.

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.

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.

Insecticide resistance of adults and nymphs of Asian citrus psyllid populations from Apatzingán Valley, Mexico.

BACKGROUND: Control of the Asian citrus psyllid Diaphorina citri Kuwayama, the most important pest of citrus worldwide, is based on the use of insecticides, though unsatisfactory results have recently been reported. In this study, insecticide resistance of D. citri to three insecticides (bifenthrin, malathion, and chlorpyrifos) was examined. RESULTS: Three populations (designated Dci-CParácuaro, Dci-El Junco, and Dci-Antúnez) of both adults and fourth-instar D. citri individuals were collected in 2014 at two different times and on one occasion, respectively, from three locations (Crucero de Parácuaro, El Junco, and Antúnez). These locations represent the major commercial Mexican lemon production areas in the Apatzingán Valley in the state of Michoacán, Mexico. The three populations of D. citri adults and fourth-instar nymphs at the different collection times showed low levels of resistance (≤7-fold) to bifenthrin, but were very resistant to malathion (≤345- and ≤432-fold for adults and fourth instars, respectively) and chlorpyrifos (≤2435- and ≤1424-fold for adults and fourth instars, respectively). CONCLUSION: Resistance levels to the tested insecticides were highly variable but homogeneous among seasons and localities. Resistance management programmes that include crop sanitation, use of biological and cultural control practices, and rotation of insecticide classes should be established, particularly in areas where D. citri has developed resistance to malathion and chlorpyrifos. © 2017 Society of Chemical Industry.