Resistance inducers applied alone or in association with fungicide for the management of leaf rust and brown eye spot of coffee under field conditions

The use of resistance inducers is a promising development in the management of plant diseases, owing to their ability to control a broad spectrum of pathogens and improve the efficacy of fungicides. This study evaluates different sources of phosphonates (potassium, manganese, copper), a formulation prepared from the by‐products of the coffee industry (Greenforce CuCa), as well as the effects of their application, alone or in association with fungicide, in the management of two important coffee fungal diseases in Brazil: leaf rust and brown eye spot, caused by Hemileia vastatrix and Cercospora coffeicola, respectively. The effect of these products on defoliation, productivity and chemical composition of coffee beans (content of trigonelline, chlorogenic acid, caffeine and total soluble solids) was evaluated. Among all the alternative products tested individually, potassium phosphonate (P₂O₅—33.6% + K₂O—29%) stood out, particularly for rust control, which was similar to the results of fungicide treatments. Treatments with fungicide, Greenforce CuCa and cuprous oxide, individually, caused less plant defoliation. Regarding the chemical composition of the coffee beans, the manganese phosphonate treatment showed the highest values for trigonelline, chlorogenic acid, caffeine and total soluble solid content. The results of this study show that resistance inducers can be useful in disease management, may come to eventually replace traditional fungicides and can also contribute to the beverage quality.     

Effectiveness of acibenzolar-S-methyl, fungicides and antibiotics for the control of brown eye spot, bacterial blight, brown leaf spot and coffee rust in coffee

This study was carried out to evaluate the potential of acibenzolar‐S‐methyl (ASM), combined or not combined with fungicides and antibiotics for the control of brown eye spot (Cercospora coffeicola) and bacterial blight (Pseudomonas syringae pv. garcae) in coffee seedlings, and ASM combined with conventional fungicide application schedules for the control of coffee rust (Hemileia vastatrix) and brown leaf spot (Phoma costarricencis) under field conditions in two coffee crops in the State of São Paulo, Brazil. ASM protected coffee seedlings against C. coffeicola when applied at the rates of 2.5 and 5 g of active ingredient per hectolitre of water (g a.i. hL^?1), providing 34–55% of disease control, and against bacterial blight, when applied at the rates of 2.5, 10 and 20 g a.i. hL^?1, with 38–57% of disease control. Tebuconazole (100 g a.i. hL^?1) and azoxystrobin (10 g a.i. hL^?1) showed the best results for brown eye spot control. Oxytetracycline + streptomycin, kasugamycin hydrochloride, oxytetracycline + metallic copper, copper oxychloride and mancozeb + copper oxychloride also controlled bacterial blight in levels similar to those shown by ASM. In the field experiments, all fungicide application schedules tested, cyproconazole (December, February, April), epoxiconazole (December, March), tetraconazole (December, February, April), cyproconazole (December, February) and azoxystrobin (January, March) were effective for coffee rust control and provided partial control of brown leaf spot. The results also showed that for all experiments, there was no synergistic effect of the combination of ASM with azoxystrobin, cyproconazole or cupric fungicides.     

Foliar fertilizers for the management of phoma leaf spot on coffee seedlings

The growing demand for alternative strategies for plant disease management has sought a reduction in the use of fungicides via the employment of resistance inducers and foliar fertilizers, among others. Therefore, this study aimed at evaluating the following foliar fertilizers for the management of phoma leaf spot: a foliar fertilizer based on macro‐ and micronutrients (Fmm: 10% N, 4% S, B, 5% Fe and 5% Zn); one based on cobalt and molybdenum (Fcm: 2% Co and 3% Mo); manganese phosphite (FMn: 30% P₂O₅ and 9% Mn); and the FMn+Fmm, Fcm+Fmm and FMn+Fcm+Fmm associations compared to a boscalid fungicide and a control with no treatment. The disease severity, the chlorophyll a and b contents, the net photosynthetic rate (LPR), the phenylalanine ammonia lyase (PAL) activity and the lignin content in leaves were assessment. Based on the severity, the area under the disease severity progress curve (AUSPC) and the efficiency of disease control in relation to untreated plants were calculated. The experimental design was in randomized blocks with eight treatments and three replications. Treatments FMn+Fmm and Fcm were the most effective in reducing the AUPSC in comparison with the control and promoted an increased activity of PAL. FMn was the treatment that promoted the highest increase in the LPR. There were no effects of the treatments on the lignin content compared to the control.     

Suppression of Rust and Brown Eye Spot Diseases on Coffee by Phosphites and By‐products of Coffee and Citrus Industries

The rust and brown eye spot, caused by Hemileia vastatrix and Cercospora coffeicola, respectively, are the most important fungal diseases on coffee in South America. Their management is mainly by chemical treatment, and there is no genetic resistance to brown eye spot known so far. Considering the need for developing alternative products for their control, the goal of this work was to evaluate the effects of phosphites and by‐products of coffee and citrus industries on rust and brown eye spot. Formulations of coffee and citrus industry by‐products, phosphites and their combination with fungicide were evaluated in field experiments, and their effect on fungal urediniospores and conidia was evaluated in vitro. In the field, treatments were applied individually or in combination and the in vitro assays were performed with manganese phosphite (Reforce Mn), potassium phosphite and citrus industry by‐product (Fortaleza), copper phosphite and coffee industry by‐product (Fitoforce Full), and fungicide. The severity and incidence of rust and brown eye spot on coffee leaves, yield, and leaf retention were evaluated in the field. Percentage of spore germination was evaluated in vitro for both fungi, whereas mycelial growth was evaluated for C. coffeicola only. The treatments Fortaleza, Reforce Mn and Fitoforce Full suppressed both diseases with a reduction in defoliation. In the year 2012, the plants treated with Reforce Mn and Reforce Mn + Fortaleza showed a yield increase of 72 and 88%, respectively, which was similar to the results shown by the fungicide treatment. In vitro inhibition of germination of H. vastatrix urediniospores and of C. coffeicola conidia was observed and suggests that the products exert some toxic effects to both fungi. Finally, the results observed indicate that the combined use of by‐products of plant‐processing industries and phosphites is an alternative and can be added efficiently to the management of coffee diseases.     

Investigating variability and behaviour of Colletotrichum gloeosporioides strains from lesions of coffee blister spot

Coffee blister spot has been associated with species from the Colletotrichum genus, but there is no information on the variability of isolates present on leaf lesions. This study evaluated a population of Colletotrichum gloeosporioides strains from blister spot lesions in Coffea arabica. Colletotrichum spp. isolates were collected from blister spot lesions on leaves of coffee trees from Catuaí and Topázio cultivars (Coffea arabica). Monosporic cultures were obtained from colonies with sporulation. A pathogenicity test was carried out by inoculation of pathogens on the leaves of young coffee plants. C. gloeosporioides strains were characterized by morphologial, cytological and physiological analyses. The molecular analysis was carried out using Inter‐Retrotransposon Amplified Polymorphism (IRAP) markers. C. gloeosporioides strains showed no pathogenicity on coffee plants and presented a wide variability in all traits evaluated. The presence of sexual strains, formation of CATs (conidial anastomosis tubes) among conidial strains and high mycelial compatibility among strains observed suggest the occurrence of sexual and asexual recombination. The role of these C. gloeosporioides strains on the lesions of coffee plant leaves is unclear.     

Population genetic analysis of a parasitic mycovirus to infer the invasion history of its fungal host

Hymenoscyphus fraxineus mitovirus 1 (HfMV1) occurs in the fungus Hymenoscyphus fraxineus, an introduced plant pathogen responsible for the devastating ash dieback epidemic in Europe. Here, we explored the prevalence and genetic structure of HfMV1 to elucidate the invasion history of both the virus and the fungal host. A total of 1298 H. fraxineus isolates (181 from Japan and 1117 from Europe) were screened for the presence of this RNA virus and 301 virus‐positive isolates subjected to partial sequence analysis of the viral RNA polymerase gene. Our results indicate a high mean prevalence (78.7%) of HfMV1 across European H. fraxineus isolates, which is supported by the observed high transmission rate (average 83.8%) of the mitovirus into sexual spores of its host. In accordance with an expected founder effect in the introduced population in Europe, only 1.1% of the Japanese isolates were tested virus positive. In Europe, HfMV1 shows low nucleotide diversity but a high number of haplotypes, which seem to be subject to strong purifying selection. Phylogenetic and clustering analysis detected two genetically distinct HfMV1 groups, both present throughout Europe. This pattern supports the hypothesis that only two (mitovirus‐carrying) H. fraxineus individuals were introduced into Europe as previously suggested from the bi‐allelic nature of the fungus. Moreover, our data points to reciprocal mating events between the two introduced individuals, which presumably initiated the ash dieback epidemic in Europe.     

Temporal Dynamics of Powdery Mildew (Oidium neolycopersici) and its Effects on the Host Growth Dynamics of Tomato

Controlled glasshouse experiments were conducted to investigate the temporal progress of powdery mildew and its effects on host dynamics of tomato, without and with one fungicide application. Healthy tomato transplants (5‐ to 6‐week old) were artificially inoculated with powdery mildew, and disease progress as well as host growth were monitored in both fungicide sprayed and unsprayed treatments and compared with non‐inoculated plants. Actual disease severity on a plant basis increased in unsprayed plants reaching maximum severity in the proportionate range of 0.53–0.83. One fungicide spray significantly reduced the maximum disease severity by two‐ to fourfolds. Despite adjustments for defoliation, declines in the proportion of disease severity between successive assessments were evident. Whereas the estimated growth rates of diseased plants were significantly lower than that of healthy plants, no significant differences were observed in the maximum leaf area formed of inoculated and non‐inoculated plants. A considerable effect of the powdery mildew epidemics was manifested through hastened shrivelling and defoliation of diseased leaves within the tomato canopy. An average of 18–29% and 40–52% of leaves had abscised from the plant canopy at the last date of assessment in sprayed and non‐sprayed plants, respectively. Accordingly, defoliation accounted for 14–33.3% and 58.3–63.1% losses in leaf area of sprayed and non‐sprayed plants, respectively. Duration of healthy leaf area and yield of inoculated plants were also significantly reduced by powdery mildew epidemics.     

Temporal response and attraction of Diaphorina citri to visual stimuli

As the vector of the global disease of citrus greening or huanglongbing, Asian citrus pysllids, Diaphorina citri Kuwayama (Hemiptera: Liviidae), are the greatest threat to the worldwide citrus industry. Critical to management of D. citri and huanglongbing is optimization of surveillance methodologies. Although phytophagous insects may find host plants by multimodal cues, some appear to primarily use visual cues. In this study, we examined the behavior of Asian citrus psyllids toward light from light‐emitting diodes (LEDs) in the insect visible spectrum. The periodicity of attraction of psyllids to visual cues was evaluated in the field (yellow sticky traps) and laboratory (multi‐colored LEDs) with a strong peak of activity during the afternoon in both the field and the laboratory (both 14:00 to 18:00 hours). In laboratory evaluations of psyllids to differently colored LEDs, strongest attraction was to LEDs emitting ultraviolet (390 nm), green (525 nm), and yellow (590 nm) light. Male and female psyllids did not differ significantly in their responses to visual cues. These findings provide the basis for formulating better traps that reflect UV and yellow light and potentially incorporate UV LEDs for monitoring psyllids and a better understanding of Asian citrus psyllid visual behavior.     

Combining hyperspectral imaging and chemometrics to assess and interpret the effects of environmental stressors on zebrafish eye images at tissue level

Changes on an organism by the exposure to environmental stressors may be characterized by hyperspectral images (HSI), which preserve the morphology of biological samples, and suitable chemometric tools. The approach proposed allows assessing and interpreting the effect of contaminant exposure on heterogeneous biological samples monitored by HSI at specific tissue levels. In this work, the model example used consists of the study of the effect of the exposure of chlorpyrifos‐oxon on zebrafish tissues. To assess this effect, unmixing of the biological sample images followed by tissue‐specific classification models based on the unmixed spectral signatures is proposed. Unmixing and classification are performed by multivariate curve resolution‐alternating least squares (MCR‐ALS) and partial least squares‐discriminant analysis (PLS‐DA), respectively. Crucial aspects of the approach are: (1) the simultaneous MCR‐ALS analysis of all images from 1 population to take into account biological variability and provide reliable tissue spectral signatures, and (2) the use of resolved spectral signatures from control and exposed populations obtained from resampling of pixel subsets analyzed by MCR‐ALS multiset analysis as information for the tissue‐specific PLS‐DA classification models. Classification results diagnose the presence of a significant effect and identify the spectral regions at a tissue level responsible for the biological change.      

Nectar yeasts enhance the interaction between Clematis akebioides and its bumblebee pollinator

• It has been hypothesised that intense metabolism of nectar‐inhabiting yeasts (NIY) may change nectar chemistry, including volatile profile, which may affect pollinator foraging behaviours and consequently plant fitness. However, empirical evidence for the plant–microbe–pollinator interactions remains little known.
• To test this hypothesis, we use a bumblebee‐pollinated vine Clematis akebioides endemic to southwest China as an experimental model plant. To quantify the incidence and density of Metschnikowia reukaufii, a cosmopolitan NIY in floral nectar, a combination of yeast cultivation and microscopic cell‐counting method was used. To examine the effects of NIY on plant–pollinator interactions, we used real flowers filled with artificial nectar with or without yeast cells. Then the volatile metabolites produced in the yeast‐inoculated nectar were analysed with coupled gas chromatography and mass spectrometry (GC‐MS).
• On average 79.3% of the C. akebioides flowers harboured M. reukaufii, and cell density of NIY was high to 7.4 × 10⁴ cells mm^?3. In the field population, the presence of NIY in flowers of C. akebioides increased bumblebee (Bombus friseanus) pollinator visitation rate and consequently seed set per flower. A variety of fatty acid derivatives produced by M. reukaufii may be responsible for the above beneficial interactions.
• The volatiles produced by the metabolism of M. reukaufii may serve as an honest signal to attract bumblebee pollinators and indirectly promote the female reproductive fitness of C. akebioides, forming a potentially tripartite plant–microbe–pollinator mutualism.

     

A preliminary assessment of the response of a native reptile assemblage to spot‐spraying invasive Bitou Bush with glyphosate herbicide

During recent work examining the effects of Bitou Bush (Chrysanthemoides monilifera ssp. rotundata) invasion on native reptile assemblages in coastal heathland vegetation in Eastern Australia, unplanned spot‐spraying of glyphosate occurred at some of our experimental sites invaded by Bitou Bush. We used this unexpected herbicide application as an opportunity to provide a preliminary assessment of the short‐term impacts on reptiles of glyphosate spot‐spraying of Bitou Bush. Using an M‐BARCI design, we compared reptile assemblages among uninvaded (reference) sites, invaded (control) sites and invaded and sprayed (impact) sites before and after spraying. We found no significant short‐term (7 – 10 months) differences in reptile abundance, species richness or assemblage composition among invaded, uninvaded and sprayed sites before and after glyphosate application. We cautiously interpret our results to generate a preliminary finding that spot‐spraying of Bitou Bush with glyphosate appears not to have a deleterious effect on reptile assemblages at seven and ten months following herbicide application. While we would not recommend basing management decisions on the outcomes of our study alone, we suggest that our findings can be used to assist in the development of strategic analyses of glyphosate impacts on native flora and fauna.