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.     

Silicon and Triadimenol for the Management of Coffee Leaf Rust

This study documents an experiment that was undertaken in the 2006/2007, 2007/2008 and 2008/2009 growing seasons on Coffea arabica cv. ‘Catuaí Vermelho IAC 144’ that sought to evaluate the effects of various calcium silicate rates combined with the fungicide triadimenol on the incidence of coffee leaf rust. The experimental design was a randomized complete block in a split plot with five treatments (with varied calcium silicate rates and with or without triadimenol) and four replications. Each experimental unit (split plot) consisted of seven coffee plants (14 m²), which were the central five plants used for the evaluations. Calcium silicate (CS) and lime (L) were used according to the following mixtures (M): M1: 0% CS and 100% L; M2: 25% CS and 75% L; M3: 50% CS and 50% L; M4: 75% CS and 25% L; and M5: 100% CS and 0% L. The leaf Si concentration did not increase as CS rates increased in the soil. There was no reduction in the area under rust progress curve (AURPC) as the rates of CS increased in the soil. During the growing seasons 2006/2007, 2007/2008 and 2008/2009, rust incidence reached 94, 96 and 92% on plants that did not receive triadimenol, respectively, whereas the incidence did not exceed 6, 38 and 16%, respectively, for those plants that did. For yield, no interaction was observed between the calcium silicate rates and with or without triadimenol. The yield increased by 117% for plants receiving triadimenol compared with those that did not. The 3‐year experiments indicated that soil amendment with calcium silicate had no effect on either reducing coffee leaf rust incidence or increasing yield. Conversely, as expected, coffee leaf rust symptoms were dramatically reduced on plants sprayed with triadimenol, and this was accompanied by a significant gain in yield.     

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.     

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.     

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.     

Diagrammatic Scale for Assessment of Bacterial Blight in Coffee Leaves

Due to the lack of a standardized visual method for assessing bacterial blight (Pseudomonas syringae pv. garcae) in coffee leaves, a diagrammatic scale was developed and validated to quantify the disease. Leaves were collected in crops and nursery with different intensity of symptoms, and the true severity was determined electronically. Based on the frequency distribution of severity values and according to the Weber–Fechner's law of visual stimulus, the minimum and maximum limits and the intermediate levels in the scale were determined. Validation was performed by ten evaluators who estimated the severity of 50 leaves with different intensity of symptoms. One evaluation was performed without diagrammatic scale and two evaluations with the scale at 7‐day intervals. The accuracy, precision, repeatability and reproducibility of the estimates were evaluated. The scale had nine levels: 0 (0%), 1 (0.1–0.99%), 2 (1–2%), 3 (2.01–4%), 4 (4.01–8%), 5 (8.01–16%), 6 (16.01–25%), 7 (25.01–45%) and 8 (≥45.1%). Using the scale, the evaluators were able to improve accuracy, precision, reproducibility and repeatability of estimates, compared to evaluators without scale. The scale was appropriate to visual estimation of severity of bacterial blight in coffee leaves.     

Diagrammatic Scales for Assessing Brown Eye Spot (Cercospora coffeicola) in Red and Yellow Coffee Cherries

Brazil is the world's largest coffee producer. Brown eye spot (BES) (Cercospora coffeicola) is a major crop disease that can cause 15–30% production losses and decrease the coffee beverage quality. Although the influence of this disease on beverage quality has been studied, diagrammatic scales for assessment of the disease severity in berries are still unexplored. Thus, this study has developed and validated diagrammatic scales to assess the severity of BES. Two diagrammatic scales were designed to assess the disease in berries, which can be yellow or red depending on the cultivar. The scale of yellow berries had nine grades: grade 0: 0%; 1: 0.1–5.0%; 2: 5.1–10.0%; 3: 10.1–15.0%; 4: 15.1–20.0%; 5: 20.1–30.0%; 6: 30.1–40.0%; 7: 40.1–60.0%; and 8: higher than 60.0%, while the scale of red berries had eight grades: grade 0: 0%; 1: 0.1–2.5%; 2: 2.6–5.0%; 3: 5.1–10.0%; 4: 10.1–20.0%; 5: 20.1–30.0%; 6: 30.1–50.0%; and 7: higher than 50.0% severity. Using scales improved accuracy and precision with R² = 0.99 and provided good repeatability and reproducibility of assessments of disease severity. Both scales can be used because the different berry colour influenced the accuracy and precision between the scales.     

Genome rearrangements derived from homoeologous recombination following allopolyploidy speciation in coffee

Allopolyploidization is widespread and has played a major role in flowering plant diversification. Genomic changes are common consequences of allopolyploidization, but their mechanisms of occurrence and dynamics over time are still poorly understood. Coffea arabica, a recently formed allotetraploid, was chosen as a model to investigate genetic changes in allopolyploid using an approach that exploits next‐generation sequencing technologies. Genes affected by putative homoeolog loss were inferred by comparing the numbers of single‐nucleotide polymorphisms detected using RNA‐seq in individual accessions of C. arabica, and between accessions of its two diploid progenitor species for common sequence positions. Their physical locations were investigated and clusters of genes exhibiting homoeolog loss were identified. To validate these results, genome sequencing data were generated from one accession of C. arabica and further analyzed. Genomic rearrangements involving homoeologous exchanges appear to occur in C. arabica and to be a major source of genetic diversity. At least 5% of the C. arabica genes were inferred to have undergone homoeolog loss. The detection of a large number of homoeologous exchange events (HEEs) shared by all accessions of C. arabica strongly reinforces the assumption of a single allopolyploidization event. Furthermore, HEEs were specific to one or a few accessions, suggesting that HEE accumulates gradually. Our results provide evidence for the important role of HEE in allopolyploid genome evolution.     

Identification of attractant and repellent plants to coffee berry borer,Hypothenemus hampei

Colombia is one of the world's largest producers of coffee [Coffea arabica L. (Rubiaceae)]. The coffee berry borer (CBB), Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae), is the main pest of coffee. This insect is controlled through an integrated pest management program that includes cultural, biological, and chemical control strategies. Despite research seeking CBB attractants and repellents, these potential management tools have not been successfully incorporated into control programs. This work proposes the use of plant functional diversity for CBB management, for which a number of plants related to coffee and weeds were selected. CBB preference to these plants was determined by olfactometry and volatile compounds emitted by them were identified. Field trials were performed to test CBB preference under field conditions. These trials determined the olfactory preference of CBB to coffee berries accompanied by material of the plants Crotalaria micans Link (Fabaceae), Lantana camara L. (Verbenaceae), Nicotiana tabacum L. (Solanaceae), Artemisia vulgaris L., Calendula officinalis L., Stevia rebaudiana (Bertoni) Bertoni, and Emilia sonchifolia (L.) DC. (all four Asteraceae). Under laboratory conditions N. tabacum, L. camara, and C. officinalis were identified as repellents for CBB in olfactometer assays, whereas E. sonchifolia functioned as attractant. Controlled field trials corroborated CBB repellency of N. tabacum and L. camara; both release volatile sesquiterpenes. Selected candidate attractants included E. sonchifolia plants, for showing attraction in the laboratory. The potential use of these plants in agroecological management of coffee plantations is discussed.     

Relationships between phoma leaf spot and development of stem canker (Leptosphaeria maculans) on winter oilseed rape (Brassica napus) in southern England

Models were constructed to describe the relationships between incidence of phoma leaf spot at different growth stages in autumn/winter or early spring and incidence of stem canker (basal canker or stem lesions) in summer on winter oilseed rape in southern England. Model 1, describing the phoma leaf spot/basal canker relationship, was y₁=β₀+β₁x₁+β₂(x₂ ‐ x₁) if x₂ > x₁, and y₁=β₀+β₁x₁ if x₂≤x₁, in which y₁ was the incidence (% plants affected) of basal canker at harvest, x₁ was the maximum incidence of phoma leaf spot during the period from sowing to growth stage (G.S.) 1,6‐1,7 (about 100 days after sowing) and x₂ was the maximum incidence of phoma leaf spot between G.S. 1,7 and G.S. 2,0 (start of stem extension). Model 2, describing the phoma leaf spot/stem lesion relationship, was y₂=α₀+α₁x₃+α₂x₄, in which y₂ was the incidence of stem lesions at harvest, x₃ was the incidence of phoma leaf spot at G.S. 3,3–3,5 (flower buds visible) and x₄ was the incidence of phoma leaf spot at G.S. 4,5–5,5 (flower buds opening). Data from field experiments with four winter oilseed rape cultivars at Boxworth or Rothamsted in the 1992/93, 1993/94, 1996/97, 1997/98 or 1998/99 seasons were used to test the models. The values of R² for the regression equations testing model 1 for the phoma leaf spot/basal canker relationship were 0.75, 0.93, 0.91 and 0.89 for cvs Apex, Bristol, Capitol and Envol, respectively. The values of R² for the regression equations testing model 2 for the phoma leaf spot/stem lesion relationship were 0.58, 0.57, 0.54 and 0.71 for cvs Apex, Bristol, Capitol and Envol, respectively. The phoma leaf spot/basal canker relationship (model 1) could also be fitted to the combined data set for all four cultivars (R²= 0.65), whereas the phoma leaf spot/stem lesion relationship (model 2) could not to be fitted to the combined data set for the four cultivars. The relationships between incidence and severity of stem canker were examined and the values of R² for the regressions of severity on incidence were 0.91 for basal canker and 0.89 for stem lesions.     

First report of Bipolaris bicolor causing a leaf spot disease on rubber tree

The rubber tree (Hevea brasiliensis) is the only resource for commercial natural rubber production and thus has economic importance in Southeast Asia. A spot disease on the leaves of a rubber tree was first discovered in 2017 in Hainan, China. In this study, the fungal isolate MA1 from the infected tissues was determined to be a pathogen of the spot disease by satisfying Koch's postulates. The isolate MA1 was identified as Bipolaris bicolor based on the morphological characteristics and multigene phylogenetic analysis. Among fungicides, prochloraz, iprodione and pyraclostrobin significantly inhibited hyphal growth of B. bicolor under in vitro conditions. This study constitutes the first report on the association of B. bicolor with leaf spot disease of rubber trees worldwide.     

夜间低温对两种咖啡光合作用的影响

为了探讨夜间低温对滇南地区主要栽培的小粒种咖啡(Coffea arabica)和大粒种咖啡(Coffea liberica)光合作用的影响,于西双版纳雾凉季对盆栽幼苗进行了连续3d的4℃夜间低温处理,并于原地恢复3d,测定了叶片的一些生理生态指标。结果表明,经4℃夜间低温处理3d后,虽然两种咖啡非辐射耗散能力增强,胡萝卜素／叶绿素(Car／Chl)升高,但净光合速率(Pn)、气孔导度(Gs)和黎明原初光化学效率(Fv／Fm)降低,初始荧光(Fo)升高,以及过氧化氢(H2O2)和丙二醛(MDA)的积累,表明夜间低温对两种咖啡光合机构均产生了明显的破坏,导致净光合速率下降的气孔因素和非气孔因素同时存在。虽然正常条件下两种咖啡净光合速率没有显著差异,但与大粒种咖啡相比,低温处理后,小粒种咖啡的Pn,Fv／Fm下降的比例小,MDA积累的量少,且在3d的恢复后基本可以恢复到原初值,表明小粒种咖啡相对比较耐寒,这与其较强的非辐射耗散能力,较大的Car／Chl及较强的恢复能力有关。     

Temporal analysis of brown eye spot of coffee and its response to the interaction of irrigation with phosphorous levels

Brown eye spot (BES—Cercospora coffeicola) is a major disease of coffee, and its occurrence is affected by water supply and nutritional balance. Little is known about the effect of phosphorous (P) on coffee fields under irrigation. Thus, this study evaluated the effect of the interaction between different water application levels and phosphorus levels (5 irrigation levels × 4 phosphorus levels) on the intensity of this disease. The area under the incidence progress curve (AUIPC) was calculated and subjected to analysis of variance. The progress curve of the average incidence of BES varied in both evaluation years. In Year 1 (November 2011 to December 2012), the incidence peaked on August 12, 2012 (22.45%), while in Year 2 (January 2013 to January 2014), the incidence reached its highest level on September 12, 2013 (16.29%). The exponential nonlinear model was adjusted for the two years. There was an interaction between irrigation and phosphorus levels on October 07, 2012. The incidence interacted significantly with the harvest dates. On the first evaluation date, an increase in phosphorus levels at shallower irrigation depths and an absence of phosphate fertilizer at higher irrigation levels caused higher incidences.     

First report of Alternaria leaf spot caused by Alternaria alternata (Fries.) Kiessler on Sonchus oleraceus L. and Convolvulus arvensis L. in Algeria

Alternaria leaf spot caused by Alternaria alternata (Fries.) Kiessler was found on sow thistle (Sonchus oleraceus L.) and field bindweed (Convolvulus arvensis L.) in the experimental station of ENSA (Ecole Nationale Supérieure d'Agronomie) in Algiers, Algeria, during the winter of 2016. Necrotic spots in the form of concentric circles were observed on the leaves of both weeds with disease incidence of approximately 70% and disease severity ranging from 50% to 70%. Fungi were isolated from the infected leaves and identified as A. alternata, based on morphological and molecular analyses (using genetic marker internal transcribed spacer, ITS of rDNA). Pathogenicity tests confirmed that A. alternata is the causing agent of leaf spot disease of sow thistle and field bindweed in accordance with the original symptoms. To the best of our knowledge, this is the first report of sow thistle and field bindweed naturally infected by A. alternata in Algeria.     

Interactions Between Rhizoctonia Root Rot and the Foliar Common Bean Diseases Anthracnose and Rust

The effects of co‐inoculation of Rhizoctonia solani and Colletotrichum lindemuthianum or Uromyces appendiculatus at different inoculum levels were studied on the disease dynamics and on the growth of bean plants under greenhouse conditions. Bean seeds were sown in R. solani‐infested soil. Additional experiments in which seedlings were transplanted to infested soil were also carried out. Conidial suspensions of C. lindemuthianum or uredospores of U. appendiculatus were inoculated onto leaves at plant developmental stages V2 and V3, respectively. Interactions between root rot and the aerial diseases were observed depending on the inoculum levels and on the timing of R. solani inoculation. Anthracnose severity tended to be higher on R. solani‐infected plants. Conversely, R. solani infection significantly reduced diameter of pustules and rust severity. When seedlings were transplanted to soil infested with low levels of R. solani, root rot severity and density of R. solani in the soil were magnified at high levels of C. lindemuthianum or U. appendiculatus. In these experiments, a synergistic interaction between root rot and anthracnose was observed to affect the plant dry weight. Antagonistic effects on the plant dry weight were found for the combination root rot/rust only when seeds were sown in infested soil.     

Assessment of genetic diversity of coffee leaf rust pathogen Hemileia vastatrix using SRAP markers

Coffee leaf rust caused by the fungus Hemileia vastatrix (Berk and Br.) is a major disease occurring in coffee plantations. Although the rust fungus exists in different physiological races, the genetic difference between them is meagrely understood. In this study, genetic diversity of 14 identified and two unidentified leaf rust races was determined by sequence‐related amplified polymorphism (SRAP) markers. Of 48 SRAP primer pairs tested, 35 primers are polymorphic and generated 347 distinct scorable fragments. The number of fragments ranged from 4 to 18 with a mean of 9.97 fragments per primer combination. Of the total 347 amplified fragments, 185 fragments (53.31%) are polymorphic with an average of 5.41 fragments per primer combination. The average resolving power (Rp) and the average polymorphism information content (PIC) of the 35 SRAP primer combinations were 13.60 and 0.356, respectively. Of 35 SRAP primer pairs, 15 primer pairs were more informative and generated 25 unique fragments, which are useful for race discrimination. The study demonstrated the existence of genetic variability among various leaf rust races and this information will be helpful in coffee breeding programmes.     

Characterization of Colletotrichum orchidophilum,the agent of black spot disease of vanilla

Black spot symptoms were reported on vanilla plants in Reunion Island in 2011. They have repeatedly reduced annual pod production by 10% to 30%. The disease is characterized by dark spots that appear in slight depressions on flowers, pods, leaves and stems. The spots then develop into broad clearly depressed necrotic plaques. Morphological and molecular analyses, as well as pathogenicity tests, identified the fungus Colletotrichum orchidophilum (Ascomycota) as the causal agent of the disease. Inoculation tests in controlled conditions confirmed that the two C. orchidophilum strains isolated from fruit lesions are pathogenic on the leaves and fruits of Vanilla planifolia (accessions CR0001 and CR0020). However, these strains induced symptoms only when the epidermis of leaves and fruits had been punctured by a needle. In the absence of injury, no symptom appeared. Colletotrichum arxii and Fusarium proliferatum (Ascomycota) are fungal species that are also frequently isolated from black spot lesions. However, they are not pathogenic to vanilla. This is the first report of C. orchidophilum in Reunion Island. It is also the first demonstration of C. orchidophilum's pathogenicity to an orchid. Simple preventive control measures were proposed to reduce the incidence of black spot disease in vanilla plots.