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.     

Reduction of Winter Wheat Yield Losses Caused by Stripe Rust through Fungicide Management

Stripe rust of winter bread wheat (Triticum aestivum L.) causes substantial grain yield loss in Central Asia. This study involved two replicated field experiments undertaken in 2009–2010 and 2010–2011 winter wheat crop seasons. The first experiment was conducted to determine grain yield reductions on susceptible winter wheat cultivars using single and two sprays of fungicide at Zadoks growth stages Z61–Z69 in two farmers’ fields in Tajikistan and one farmer's field in Uzbekistan. In the second experiment, four different fungicides at two concentrations were evaluated at Zadoks growth stage Z69. These included three products from BASF – Opus (0.5 l/ha and 1.0 l/ha), Platoon (0.5 l/ha and 1.0 l/ha) and Opera (0.75 l/ha and 1.5 l/ha) – and locally used fungicide Titul 390 (0.5 l/ha and 1.0 l/ha). One and two sprays of fungicides did not differ significantly (P > 0.05) in increasing grain yield. Stripe rust reduced grain yield and 1000‐kernel weight (TKW) from 24 to 39% and from 16 to 24%, respectively. The benefits from the two concentrations of the same fungicide did not consistently resulted in significantly higher grain yield, suggesting that the lower concentrations could be more cost effective. Our study provides important information about the selection of fungicides, spray concentrations and number of spray to control stripe rust and increase grain yield. The findings could play an important role in developing stripe rust management approaches such as fungicide rotation and strategic fungicide applications in Central Asian countries.     

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.     

Conversion of nicotinic acid to trigonelline is catalyzed by N-methyltransferase belonged to motif B′ methyltransferase family in Coffea arabica

Trigonelline (N-methylnicotinate), a member of the pyridine alkaloids, accumulates in coffee beans along with caffeine. The biosynthetic pathway of trigonelline is not fully elucidated. While it is quite likely that the production of trigonelline from nicotinate is catalyzed by N-methyltransferase, as is caffeine synthase (CS), the enzyme(s) and gene(s) involved in N-methylation have not yet been characterized. It should be noted that, similar to caffeine, trigonelline accumulation is initiated during the development of coffee fruits. Interestingly, the expression profiles for two genes homologous to caffeine synthases were similar to the accumulation profile of trigonelline. We presumed that these two CS-homologous genes encoded trigonelline synthases. These genes were then expressed in Escherichiacoli, and the resulting recombinant enzymes that were obtained were characterized. Consequently, using the N-methyltransferase assay with S-adenosyl[methyl-¹⁴C]methionine, it was confirmed that these recombinant enzymes catalyzed the conversion of nicotinate to trigonelline, coffee trigonelline synthases (termed CTgS1 and CTgS2) were highly identical (over 95% identity) to each other. The sequence homology between the CTgSs and coffee CCS1 was 82%. The pH-dependent activity curve of CTgS1 and CTgS2 revealed optimum activity at pH 7.5. Nicotinate was the specific methyl acceptor for CTgSs, and no activity was detected with any other nicotinate derivatives, or with any of the typical substrates of B′-MTs. It was concluded that CTgSs have strict substrate specificity. The K_m values of CTgS1 and CTgS2 were 121 and 184 μM with nicotinic acid as a substrate, and 68 and 120 μM with S-adenosyl-l-methionine as a substrate, respectively.     

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.     

Trigonelline inheritance in the interspecific Coffea pseudozanguebariae×C. liberica var. dewevrei cross

Trigonelline alkaloid is present in coffee beans, and during roasting it gives rise to the major coffee aroma compounds (several alkyl-pyridines and pyrroles). In this study we investigated the genetic inheritance of trigonelline accumulation in green beans in an interspecific cross between a wild east African species, Coffea pseudozanguebariae (PSE) and the west African species C. liberica var. dewevrei (DEW). Trigonelline content was measured by HPLC in both parental species, F₁ hybrids and the reciprocal backcross hybrids (BCDEW and BCPSE). The results showed that, on average, PSE accumulated twice as much trigonelline as DEW. No year effect or interaction (genotype×year) was recorded. Trigonelline showed high heritability (71%), which meant that the genotypic value could be easily estimated from the phenotypic value. However, the fact that this trait was not additive suggested the possibility of nucleo-cytoplasmic inheritance. This hypothesis was confirmed by: (1) similar levels of trigonelline content in the PSE, F₁, BCPSE and BCDEW groups, all having the same maternal cytoplasm, and (2) the location of one nuclear QTL on the G linkage group. Received: 2 May 2000 / Accepted: 19 June 2000     

Activity of Ten Fungicides against Phytophthora capsici Isolates Resistant to Metalaxyl

Pepper Phytophthora blight (PPB), caused by Phytophthora capsici, is an important disease of pepper in China. The extensive application of metalaxyl has resulted in widespread resistance to this fungicide in field. This study has evaluated the activities of several fungicides against the mycelial growth and sporangium germination of metalaxyl‐sensitive and metalaxyl‐resistant P. capsici isolates by determination of EC₅₀ values. The results showed that the novel carboxylic acid amide (CAA) fungicide mandipropamid exhibited excellent inhibitory activity against PPB both in vitro and in vivo, with averagely EC₅₀ values of 0.075 and 0.004 μg/ml in mycelial growth and sporangium germination, respectively, and over 88% efficacy in controlling PPB. The other three CAA fungicides also provided over 70% efficacy in controlling PPB. The mycelial growth was less sensitive to quinone outside inhibitor (QoI) fungicides azoxystrobin and trifloxystrobin than that of sporangium germination in P. capsici isolates. However, azoxystrobin and trifloxystrobin provided over 80% efficacy in controlling PPB. It was noted that propamocarb and cymoxanil did not exhibit activity against the mycelial growth or sporangium germination of P. capsici isolates in the in vitro tests, with over 70% efficacy in controlling PPB. The new fungicide mixture 62.5 g/l fluopicolide + 625 g/l propamocarb (trade name infinito, 687.5 g/l suspension concentrate (SC)) produced over 88% efficacy in controlling PPB caused by both metalaxyl‐sensitive and metalaxyl‐resistant isolates. The data of this study also proved that there was obviously no cross‐resistance between metalaxyl and the other tested fungicides. Therefore, these fungicides should be good alternatives to metalaxyl for the control of PPB and management of metalaxyl resistance.     

Combined changes of process conditions improved aromatic properties of Vietnamese Robusta

The flavor and taste of coffee are affected by roasting conditions and extraction temperature. This study assessed changes in the flavors and tastes of coffee extracted from Vietnamese Robusta with different roasting times and temperatures, as well as different extraction temperatures. Vietnamese Robusta green beans were roasted for different times ranging from 5 to 20 min and at different temperatures ranging from 100 to 250°C. The roasted coffee was then extracted at five different temperatures ranging from 90 to 120°C. The coffee flavor was evaluated in terms of 5 key odorants: guaiacol; 4-ethylguaiacol; 2-ethyl-3,5-dimethylpyrazine; 2,3-diethyl-5-methylpyrazine, and 2-furfuryl. The taste of coffee was evaluated in terms of 6 main compounds that confer bitterness and sourness: caffeine, trigonelline, chlorogenic acid and citric acid, acetic acid, formic acid. The optimized roasting and extracting conditions were identified that 200 g of VN Robusta green beans, roasting at 230°C for 18 min, and then extracted with Espresso Machine at 110°C.     

On the hunt for the alternate host of Hemileia vastatrix

Coffee leaf rust (CLR), caused by the fungal pathogen Hemileia vastatrix, has plagued coffee production worldwide for over 150 years. Hemileia vastatrix produces urediniospores, teliospores, and the sexual basidiospores. Infection of coffee by basidiospores of H. vastatrix has never been reported and thus far, no alternate host, capable of supporting an aecial stage in the disease cycle, has been found. Due to this, some argue that an alternate host of H. vastatrix does not exist. Yet, to date, the plant pathology community has been puzzled by the ability of H. vastatrix to overcome resistance in coffee cultivars despite the apparent lack of sexual reproduction and an aecidial stage. The purpose of this study was to introduce a new method to search for the alternate host(s) of H. vastatrix. To do this, we present the novel hypothetical alternate host ranking (HAHR) method and an automated text mining (ATM) procedure, utilizing comprehensive biogeographical botanical data from the designated sites of interests (Ethiopia, Kenya and Sri Lanka) and plant pathology insights. With the HAHR/ATM methods, we produced prioritized lists of potential alternate hosts plant of coffee leaf rust. This is a first attempt to seek out an alternate plant host of a pathogenic fungus in this manner. The HAHR method showed the highest‐ranking probable alternate host as Psychotria mahonii, Rubus apetalus, and Rhamnus prinoides. The cross‐referenced results by the two methods suggest that plant genera of interest are Croton, Euphorbia, and Rubus. The HAHR and ATM methods may also be applied to other plant–rust interactions that include an unknown alternate host or any other biological system, which rely on data mining of published data.     

Variation in Aggressiveness Components in the Hemileia vastatrix Population in Brazil

The Pucciniomycete fungus Hemileia vastatrix causes leaf rust on coffee trees. The pathogen is responsible for considerable yield losses in susceptible coffee cultivars if appropriate management strategies are not implemented. Rapid spread and epidemics of rust fungi are usually associated with the emergence of new races of the pathogen that overcome resistance or with the emergence of more aggressive populations of the pathogen. In Brazil, coffee production is dominated by susceptible cultivars of Coffea arabica and Coffea canephora. We assessed aggressiveness in 46 populations of H. vastatrix from Minas Gerais and Espírito Santo, two of the most important coffee‐producing states in Brazil. We observed a significant difference in the incubation period between the populations from Minas Gerais and Espírito Santo when 183 single‐pustule isolates were inoculated onto Catuaí Vermelho IAC 44, a susceptible C. arabica cultivar. Variation in aggressiveness components was observed between and within localities. Isolates with longer incubation periods also tended to have longer latent periods, although there was only a low correlation between these two aggressiveness components (r² = 0.34, P = 2.2 × 10^?16). Low‐sporulating isolates also had significantly longer incubation and latent periods. The H. vastatrix population from Minas Gerais and Espírito Santo is structured by the formation of groups of individuals with differential level of aggressiveness. Our results indicate that the variation in aggressiveness of the Brazilian H. vastatrix population may be associated with the geographic coffee‐producing areas.     

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.     

Trunk injection of fosetyl‐aluminium controls the root disease caused by Phytophthora cinnamomi on Quercus ilex woodlands

In Spain, Quercus open woodlands are animal ranching systems of organic production seriously threatened by the exotic pathogen Phytophthora cinnamomi. The root disease it causes kills thousands of oaks annually. Effective disease management needs to integrate different techniques, and the use of a resistance inducer such as fosetyl‐Al can play a key role, because the use of potassium phosphite is prohibited in Spain. In a woodland where the pathogen recently arrived, 60 holm oaks in three different defoliation classes (asymptomatic, slight and moderate defoliation) were selected for trunk injection with pressurised capsules containing 4% of commercial fosetyl‐Al or water (controls). Holm oaks were checked periodically for defoliation and presence of the pathogen in roots and rhizosphere soil. Three years after treatments, defoliation was significantly lower in oaks treated with fosetyl‐Al, which even increased canopy cover, in comparison with control oaks, independent of the initial defoliation class considered. Chlamydospore density in rhizosphere soil, as well as the presence of the pathogen into the roots, was not significantly influenced by fosetyl‐Al treatments, although a trend to a lower presence of P. cinnamomi in roots was observed in treated oaks at every soil inoculum density detected. This study has shown that fosetyl‐Al, a phosphonate registered as a fungicide in the European Union, provides protection to holm oaks against P. cinnamomi, even exhibiting a therapeutic effect on pre‐existing infections. Consequently, this effective measure should be considered as part of the integrated approach to control this highly destructive pathogen in holm oak woodlands.     

Interspecies gene transfer provides soybean resistance to a fungal pathogen

Fungal pathogens pose a major challenge to global crop production. Crop varieties that resist disease present the best defence and offer an alternative to chemical fungicides. Exploiting durable nonhost resistance (NHR) for crop protection often requires identification and transfer of NHR‐linked genes to the target crop. Here, we identify genes associated with NHR of Arabidopsis thaliana to Phakopsora pachyrhizi, the causative agent of the devastating fungal disease called Asian soybean rust. We transfer selected Arabidopsis NHR‐linked genes to the soybean host and discover enhanced resistance to rust disease in some transgenic soybean lines in the greenhouse. Interspecies NHR gene transfer thus presents a promising strategy for genetically engineered control of crop diseases.     

Inheritance of caffeine and heteroside contents in an interspecific cross between a cultivated coffee species Coffea liberica var dewevrei and a wild species caffeine-free C. pseudozanguebariae

 Coffee species originating from Africa, in particular the two major cultivated species C. arabica and C. canephora, usually contain caffeine in their beans, whereas almost all Malagasy coffee species are caffeine-free. However, one wild coffee species C. pseudozanguebariae, collected near the coast in south Kenya, is also caffeine-free. Beans of this species contain a specific heteroside diterpene (hereinafter referred to simply as heteroside) and give a bitter coffee beverage. We have investigated the inheritance of the caffeine and heteroside contents in the first and second generations of an interspecific cross between C. pseudozanguebariae and C. liberica var. dewevrei, for which the caffeine content is about 1% dmb (dry matter basis). The caffeine content of F₁ hybrids (0.2% dmb) was lower than the parental average (0.47% dmb). Caffeine and heteroside contents appeared to be under polygenic control with a strong genetic effect. Nevertheless, one major gene with two alleles seemed to be involved in the control of both compounds. Absence of caffeine was apparently controlled by one recessive gene. Heteroside content seemed to be controlled by one co-dominant gene, heterozygotes being intermediate between the two different groups of homozygotes. Received: 15 September 1997 / Accepted: 6 October 1997     

Leaf gas exchange and chlorophyll a fluorescence in soybean leaves infected by Phakopsora pachyrhizi

Asian soybean rust (ASR), caused by Phakopsora pachyrhizi, is one of the most important foliar diseases affecting soybean production worldwide. This study aimed to investigate the photosynthetic performance (leaf gas exchange, chlorophyll (Chl) a fluorescence images and photosynthetic pigment pools) of soybean plants sprayed with Acibenzolar‐S‐Methyl (ASM) and the fungicide epoxiconazole + pyraclostrobin (Epo+Pyr) and further inoculated with P. pachyrhizi. The ASR symptoms progressed much faster on the leaves of plants from the control treatment (water spray) in comparison with the ASM and Epo+Pyr treatments. In general, the values for the leaf gas exchange parameters net carbon assimilation rate (A), stomatal conductance to water vapour (g_s), internal CO₂ concentration (C_i) and transpiration rate (E) increased for the infected plants sprayed with ASM or Epo+Pyr in comparison with plants from the control treatment. The values for the initial fluorescence (F_o), maximal fluorescence (F_m), maximal photosystem II quantum efficiency (F_v/F_m), effective photosystem II quantum yield (Y(II)) and quantum yield of regulated energy dissipation (Y(NPQ)) were consistently higher for the ASM and Epo+Pyr treatments in comparison with the control treatment at advanced stages of fungal infection. By contrast, the values for quantum yield of non‐regulated energy dissipation (Y(NO) were significantly lower for the ASM and Epo+Pyr treatments. The concentrations of total Chl a+b and carotenoids significantly increased for infected plants sprayed with ASM and Epo+Pyr in comparison with plants from the control treatment. The results of this study demonstrated that the spray of soybean plants with either ASM or Epo+Pyr contributed to reduce the negative effect of ASR on the photosynthesis of soybean plants.     

Characterization of Pythium spp. associated with root rot of tobacco seedlings produced using the float tray system in Zimbabwe

The study was undertaken to identify and characterize Pythium isolates associated with root rot disease of tobacco seedlings as a first step towards developing management strategies for the pathogen. A total of 85 Pythium isolates were collected from diseased tobacco seedlings during 2015–2016 tobacco growing season. The isolates were identified to species level using sequencing of the internal transcribed spacer region. Thereafter, a subset of the isolates was tested for sensitivity to the commonly used fungicides, metalaxyl, azoxystrobin and a combination of fenamidone/propamocarbby growing isolates on Potato Dextrose Agar plates amended with the fungicides. The sequence analysis of the ITS‐rDNA identified Pythium myriotylum as the dominant Pythium species associated with the root rot of tobacco seedlings in Zimbabwe. Pythium aphanidermatum and P. insidiosum were also identified albeit at lower frequencies. Phylogenetic analyses of the ITS region of the P. myriotylum isolates showed little sequence diversity giving rise to one distinct clade. The fungicide sensitivity tests showed that metalaxyl provided the best control of P. myriotylum in vitro, as compared to other fungicides. To the best of our knowledge, this is the first comprehensive study to determine and characterize Pythium species associated with root rot of tobacco in the float seedling production system in Zimbabwe.