Exploring interactions between Beauveria and Metarhizium strains through co-inoculation and responses of perennial ryegrass in a one-year trial.

Perennial ryegrass (Lolium perenne L.) possesses a high level of nutritional quality and is widely used as a forage species to establish permanent pastures in southern Chile. However, the productivity of most such pastures is limited by various environmental agents, such as insect pests and drought. In this context, our work stresses the need for elucidating the ability of fungal endophytes to establish interactions with plants, and to understand how these processes contribute to plant performance and fitness. Therefore, we evaluated the colonization and impact of two native strains of the endophytic insect-pathogenic fungus (EIPF) group isolated from permanent ryegrass pastures in southern Chile. Roots and seeds of ryegrass and scarabaeid larvae were collected from nine different ryegrass pastures in the Los Ríos region of southern Chile to specifically isolate EIPFs belonging to the genera Beauveria and Metarhizium. Fungal isolations were made on 2% water agar with antibiotics, and strains were identified by analyzing the entire internal transcribed spacer (ITS) 1-5.8S-ITS2 ribosomal DNA region. Four strains of Beauveria and 33 strains of Metarhizium were isolated only in scarabaeid larvae from ryegrass pastures across four sites. Experimental mini-pastures that were either not inoculated (control) or co-inoculated with conidia of the strains Beauveria vermiconia NRRL B-67993 (P55_1) and Metarhizium aff. lepidiotae NRRL B-67994 (M25_2) under two soil humidity levels were used. Ryegrass plants were randomly collected from the mini-pastures to characterize EIPF colonization in the roots by real-time PCR and fluorescence microscopy. Aboveground biomass was measured to analyze the putative impact of colonization on the mini-pastures' aboveground phenotypic traits with R software using a linear mixed-effects model and the ANOVA statistical test. Seasonal variation in the relative abundance of EIPFs was observed, which was similar between both strains from autumn to spring, but different in summer. In summer, the relative abundance of both EIPFs decreased under normal moisture conditions, but it did not differ significantly under water stress. The aboveground biomass of ryegrass also increased from autumn to spring and decreased in summer in both the inoculated and control mini-pastures. Although differences were observed between moisture levels, they were not significant between the control and inoculated mini-pastures, except in July (fresh weight and leaf area) and October (dry weight). Our findings indicate that native strains of B. vermiconia NRRL B-67993 (P55_1) and M. aff. lepidiotae NRRL B-67994 (M25_2) colonize and co-exist in the roots of ryegrass, and these had little or no effect on the mini-pastures' aboveground biomass; however, they could have other functions, such as protection against root herbivory by insect pests.

Comparison of New Kairomone-Based Lures for Cydia pomonella (Lepidoptera: Tortricidae) in Italy and USA.

Studies were conducted during the period 2019/2020 to evaluate the effectiveness of four lures for codling moth (Cydia pomonella L.) in pome fruits in Italy and the USA. Multi-component blends of sex pheromone ((E,E)-8,10-dodecadien-1-ol, PH), pear ester ((E,Z)-2,4-ethyl decadienoate, PE), (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), and pyranoid linalool oxide (6-ethenyl-2,2,6-trimethyloxan-3-ol, LOX) were loaded in either a halobutyl elastomer septum or a PVC matrix and always used in combination with acetic acid (AA) loaded in a closed membrane co-lure. Total moth capture was significantly greater with the PVC than the septum lure loaded with PH/PE + AA in both countries. Female capture in the USA study was significantly greater for 8 weeks in traps baited with the PE/DMNT/LOX blend + AA co-lure than with other lures and adding PH to this blend in a PVC lure significantly reduced female capture. In contrast, female capture in Italy did not differ among lures and counts were similar in both apple and pear crops treated with or without mating disruption. These results suggest that the effectiveness of 'female removal' strategies to manage codling moth may be geographically limited and further comparisons are needed in other production regions and in walnut.

Survey of conspecific herbivore-induced volatiles from apple as possible attractants for Pandemis pyrusana (Lepidoptera: Tortricidae).

BACKGROUND: Studies were conducted to identify volatiles released by apple foliage untreated or sprayed with a yeast and from untreated and sprayed foliage with actively feeding larvae of Pandemis pyrusana Kearfott. Field studies then evaluated various combinations of these volatiles when paired with acetic acid as possible adult attractants. RESULTS: The most abundant volatiles released following herbivore feeding were four green leaf volatiles (GLVs) and acetic acid. Nineteen volatiles were found to be released in significantly higher amounts from foliage with herbivore damage than from intact leaves. The combination of yeast followed by herbivore injury increased the levels of methyl salicylate and phenylacetonitrile compared with herbivory alone. Levels of acetic acid released were not significantly different among the four treatments. Only phenylacetonitrile and 2-phenylethanol with acetic acid caught similar and significantly more total and female moths than acetic acid alone. Moth catches with 12 other volatiles plus acetic acid were not significantly higher than with acetic acid alone, and were lower than with acetic acid and 2-phenylethanol. CONCLUSION: These data show that herbivore injury does not create a unique chemical signal for adults to locate oviposition or rendezvous sites. Instead, moths may cue to the aromatic-acetic acid combination as a nutritional cue to locate sugary resources. © 2017 Society of Chemical Industry.

Adding yeasts with sugar to increase the number of effective insecticide classes to manage Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) in cherry.

BACKGROUND: Drosophila suzukii is a major pest of cherry in the western United States. We evaluated whether the addition of sugary baits could improve the efficacy of two classes of insecticides not considered to be sufficiently effective for this pest, diamides and spinosyns, in laboratory and field trials in cherry. RESULTS: Adding cane sugar alone or in combination with the yeasts Saccharomyces cerevisiae or Aureobasidium pullulans significantly improved insecticide efficacy. However, the significance of adding yeasts to the sugar plus insecticide on fly mortality varied with respect to both the insecticide and yeast species. The addition of S. cerevisiae to sugar also did not significantly reduce egg densities in fruit compared with sugar alone. The addition of a yeast plus sugar significantly reduced egg densities in three field trials with cyantraniliprole and in two out of three trials with spinosad. CONCLUSION: The addition of cane sugar with or without yeast can improve the effectiveness of diamide and spinosyn insecticides for D. suzukii in cherry. Inclusion of these two insecticides in D. suzukii management programs may alleviate the strong selection pressure currently being imposed on a few mode-of-action insecticide classes used by growers to maintain fly suppression over long continuous harvest periods of mixed cultivars. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

Improving the Performance of the Granulosis Virus of Codling Moth (Lepidoptera: Tortricidae) by Adding the Yeast Saccharomyces cerevisiae with Sugar.

Studies were conducted with the codling moth granulosis virus (CpGV) to evaluate whether adding the yeast Saccharomyces cerevisiae Meyen ex E. C. Hansen with brown cane sugar could improve larval control of Cydia pomonella (L.). Larval mortalities in dipped-apple bioassays with S. cerevisiae or sugar alone were not significantly different from the water control. The addition of S. cerevisiae but not sugar with CpGV significantly increased larval mortality compared with CpGV alone. The combination of S. cerevisiae and sugar with CpGV significantly increased larval mortality compared with CpGV plus either additive alone. The addition of S. cerevisiae improved the efficacy of CpGV similarly to the use of the yeast Metschnikowia pulcherrima (isolated from field-collected larvae). The proportion of uninjured fruit in field trials was significantly increased with the addition of S. cerevisiae and sugar to CpGV compared with CpGV alone only in year 1, and from the controls in both years. In comparison, larval mortality was significantly increased in both years with the addition of S. cerevisiae and sugar with CpGV compared with CpGV alone or from the controls. The numbers of overwintering larvae on trees was significantly reduced from the control following a seasonal program of CpGV plus S. cerevisiae and sugar. The addition of a microencapsulated formulation of pear ester did not improve the performance of CpGV or CpGV plus S. cerevisiae and sugar. These data suggest that yeasts can enhance the effectiveness of the biological control agent CpGV, in managing and maintaining codling moth at low densities.

Measuring local genetic variability in populations of codling moth (Lepidoptera: Tortricidae) across an unmanaged and commercial orchard interface.

The genetic structure of adult codling moth, Cydia pomonella (L.), populations was characterized both inside a managed apple, Malus domestica Borkdhausen, orchard and in surrounding unmanaged hosts and nonhost trees in central Chile during 2006-2007. Adult males were collected using an array of sex pheromone-baited traps. Five microsatellite genetic markers were used to study the population genetic structure across both spatial (1-100 ha) and temporal (generations within a season) gradients. Analysis of molecular variance (AMOVA) found a significant, but weak, association in both the spatial and temporal genetic structures. Discriminant analysis also found significant differentiation between the first and second generation for traps located either inside or outside the managed orchard. The Bayesian assignment test detected three genetic clusters during each of the two generations, which corresponded to different areas within the unmanaged and managed apple orchard interface. The lack of a strong spatial structure at a local scale was hypothesized to be because of active adult movement between the managed and unmanaged hosts and the asymmetry in the insecticide selection pressure inside and outside the managed habitats. These data highlight the importance of developing area-wide management programs that incorporate management tactics effective at the landscape level for successful codling moth control.