Effects of three esca-associated fungi on Vitis vinifera L.: I. Characterization of secondary metabolites in culture media and host responses to the pathogens in calli

Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) produced scytalone, isosclerone and pullulans in liquid cultures, as well as in calli. Secondary metabolites and host defense compounds were shown to occur in calli of Vitis vinifera cv. Italia and cv. Matilde infected by the esca-associated fungi Pch, Tmi and Fomitiporia mediterranea (Fme). Calli of both cvs. were grown as dual cultures with Pch, Tmi and Fme. The fungi grew well in the presence of calli of both cvs., but callus growth was reduced. Accumulation and changes of total phenolics and recurring phenolics, and of two phytotoxic pentaketides and pullulans were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that concentrations of phenolics are strongly related to the cv. The paper discusses the possible relationship between melanin biosynthesis in Pch and Tmi, which utilize pentaketide metabolites as intermediates and their pathogenicity related to phytotoxity of scytalone and isosclerone.     

Effects of three esca-associated fungi on Vitis vinifera L.: II. Characterization of biomolecules in xylem sap and leaves of healthy and diseased vines

Secondary metabolites and host defense compounds were shown to occur in xylem sap, and leaves of Vitis vinifera cv. Italia and cv. Matilde naturally infected by the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme). Samples of xylem sap and leaves were collected from healthy vines and from vines showing severe symptoms of brown wood-streaking caused by Pch and Tmi, or from vines with symptoms of both brown wood-streaking and white rot caused by Fme. Xylem sap collection was carried out during the early spring of 2003 and 2004, corresponding to the phenological phases: (A) cotton bud; (B) green tip; (C) leaves out; (D) stretched out leaves; and (E) visible clusters. In the present work we have studied the accumulation of biomolecules (pentaketides and α-glucans), host defense compounds (benzaldehydes, benzoic acid and cinnamic acid derivatives, flavonols, flavanols, flavan-3-ol derivatives and stilbenes) at different stages of grapevine development. Accumulation and changes in total phenolics and recurring phenolics, and of three phytotoxic secondary metabolites (scytalone, isosclerone and pullulan) were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that phenolic concentrations are strongly related to the cv.     

Effects of three esca-associated fungi on Vitis vinifera L.: II. Characterization of biomolecules in xylem sap and leaves of healthy and diseased vines

Secondary metabolites and host defense compounds were shown to occur in xylem sap, and leaves of Vitis vinifera cv. Italia and cv. Matilde naturally infected by the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme). Samples of xylem sap and leaves were collected from healthy vines and from vines showing severe symptoms of brown wood-streaking caused by Pch and Tmi, or from vines with symptoms of both brown wood-streaking and white rot caused by Fme. Xylem sap collection was carried out during the early spring of 2003 and 2004, corresponding to the phenological phases: (A) cotton bud; (B) green tip; (C) leaves out; (D) stretched out leaves; and (E) visible clusters. In the present work we have studied the accumulation of biomolecules (pentaketides and α-glucans), host defense compounds (benzaldehydes, benzoic acid and cinnamic acid derivatives, flavonols, flavanols, flavan-3-ol derivatives and stilbenes) at different stages of grapevine development. Accumulation and changes in total phenolics and recurring phenolics, and of three phytotoxic secondary metabolites (scytalone, isosclerone and pullulan) were analyzed by HPLC. On comparing results for cv. Italia and cv. Matilde, it can be seen that phenolic concentrations are strongly related to the cv.     

Effects of three esca-associated fungi on Vitis vinifera L.: V. Changes in the chemical and biological profile of xylem sap from diseased cv. Sangiovese vines

A vineyard of Vitis vinifera cv. Sangiovese was surveyed for incidence of esca and xylem sap collection. Sap samples were collected from healthy vines and from those with dual infection by Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) or triple infection by Pch, Tmi and Fomitiporia mediterranea (Fme), during each early spring in a 3-year period (2001–2003). In order to analyse the possible trends in the climatic data, temperature and rainfall were assessed. At sap harvesting, aliquots of sap were assayed for phytotoxicity and extracted with ethyl acetate for phytotoxin recovery. Moreover, the exopolysaccharide (EPS) content was evaluated on several sap samples during the bleeding period. Conidia of Pch and Tmi, mycelium of Fme and their secondary metabolites were found in the sap of the esca-affected vines, indicating that the pathogens and their by-products together with some defence substances were accumulated and then translocated. Bioactivity tests showed toxicity of the sap from esca-affected vines to healthy detached leaves of cv. Sangiovese. The daily amount of sap, the pH, and the volume (J_v) and solute (J_s) fluxes were analysed as a function of the infecting fungi. Pullulan, glucogalactomannan(s) and arabinogalactan(s) are the main EPS in the esca-infected vines, whereas in the sap of healthy vines no traces of pullulan were found. Scytalone and isosclerone usually produced in vitro by Pch and Tmi were also detected in the sap of vines infected by Pch and Tmi or by Pch, Tmi and Fme. The endogenous phytohormone content of healthy vines evaluated by the cutting bioassay was different from that of infected vines. Four phenolics belonging to three classes e.g., benzoic acid derivatives, stilbenes and flavonol-glycosides were separated and identified by HPLC.     

Effects of three esca-associated fungi on Vitis vinifera L.: III. Enzymes produced by the pathogens and their role in fungus-to-plant or in fungus-to-fungus interactions

When the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme) were grown in liquid stationary cultures, it was seen that they were able to live in media containing resveratrol (RES) or tannic acid (TA) as the sole carbon source and that the fungi were able to convert both compounds. Particular attention is paid here to detecting RES and TA conversion. Pch, Tmi and Fme were partially inhibited by RES or TA. Pch, Tmi and Fme produced extracellular tannase, laccase and peroxidase enzymes in liquid or agarized cultures, whether glucose was present or not. When colonies of Pch, Tmi and Fme were confronted, they showed spatially and temporally heterogeneous patterns of laccase and peroxidase activity. The results indicate the non-synergistic, competitive association of Pch and Tmi and the inhibition of Fme growth. Muconic acid, a well-known intermediate in a large number of lignin and phenol oxidative processes, can partly or completely inhibit the lignolytic agent Fme, but is tolerated by Pch and Tmi. An explanation for wood pigmentation patterns by Pch, Tmi and Fme is given.     

Effects of three esca-associated fungi on Vitis vinifera L.: V. Changes in the chemical and biological profile of xylem sap from diseased cv. Sangiovese vines

A vineyard of Vitis vinifera cv. Sangiovese was surveyed for incidence of esca and xylem sap collection. Sap samples were collected from healthy vines and from those with dual infection by Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi) or triple infection by Pch, Tmi and Fomitiporia mediterranea (Fme), during each early spring in a 3-year period (2001–2003). In order to analyse the possible trends in the climatic data, temperature and rainfall were assessed. At sap harvesting, aliquots of sap were assayed for phytotoxicity and extracted with ethyl acetate for phytotoxin recovery. Moreover, the exopolysaccharide (EPS) content was evaluated on several sap samples during the bleeding period. Conidia of Pch and Tmi, mycelium of Fme and their secondary metabolites were found in the sap of the esca-affected vines, indicating that the pathogens and their by-products together with some defence substances were accumulated and then translocated. Bioactivity tests showed toxicity of the sap from esca-affected vines to healthy detached leaves of cv. Sangiovese. The daily amount of sap, the pH, and the volume (J_v) and solute (J_s) fluxes were analysed as a function of the infecting fungi. Pullulan, glucogalactomannan(s) and arabinogalactan(s) are the main EPS in the esca-infected vines, whereas in the sap of healthy vines no traces of pullulan were found. Scytalone and isosclerone usually produced in vitro by Pch and Tmi were also detected in the sap of vines infected by Pch and Tmi or by Pch, Tmi and Fme. The endogenous phytohormone content of healthy vines evaluated by the cutting bioassay was different from that of infected vines. Four phenolics belonging to three classes e.g., benzoic acid derivatives, stilbenes and flavonol-glycosides were separated and identified by HPLC.     

Effects of three esca-associated fungi on Vitis vinifera L.: III. Enzymes produced by the pathogens and their role in fungus-to-plant or in fungus-to-fungus interactions

When the esca-associated fungi Phaeomoniella chlamydospora (Pch), Togninia minima (Tmi) and Fomitiporia mediterranea (Fme) were grown in liquid stationary cultures, it was seen that they were able to live in media containing resveratrol (RES) or tannic acid (TA) as the sole carbon source and that the fungi were able to convert both compounds. Particular attention is paid here to detecting RES and TA conversion. Pch, Tmi and Fme were partially inhibited by RES or TA. Pch, Tmi and Fme produced extracellular tannase, laccase and peroxidase enzymes in liquid or agarized cultures, whether glucose was present or not. When colonies of Pch, Tmi and Fme were confronted, they showed spatially and temporally heterogeneous patterns of laccase and peroxidase activity. The results indicate the non-synergistic, competitive association of Pch and Tmi and the inhibition of Fme growth. Muconic acid, a well-known intermediate in a large number of lignin and phenol oxidative processes, can partly or completely inhibit the lignolytic agent Fme, but is tolerated by Pch and Tmi. An explanation for wood pigmentation patterns by Pch, Tmi and Fme is given.     

Effects of three esca-associated fungi on Vitis vinifera L.: IV. Diffusion through the xylem of metabolites produced by two tracheiphilous fungi in the woody tissue of grapevine leads to esca-like symptoms on leaves and berries

Two naphthalenone pentaketides (scytalone and isosclerone) and α-glucans (pullulan) are produced in vitro and in planta by Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi), two tracheiphilous fungi associated with the ‘esca’ disease of grapevines. The possible role of such fungal metabolites in inducing symptoms on leaves and berries was studied in a vineyard of Vitis vinifera cv. Italia located in southern Italy. During early spring, pruning off two to four branches per vine allowed samples of xylem sap to be collected. Vine bleeding, assessed as ml day⁻¹ vine⁻¹, reached its maximum at bud burst and stopped within 28 days. The total amount of sap collected from healthy vines was about a quarter of that from esca-affected vines. During the same period, the leaf water potential of diseased vines increased progressively (i.e. showed less negative values), indicating a dysfunction in water and nutrient supply to the new growth. Both fungi were isolated from the xylem sap and from the woody tissue of branches and the trunk of diseased vines. Conidia isolated from the sap showed a high germination rate (>90%). Bioactive concentrations of the two pentaketides were detected in xylem sap, leaves and berries at various stages of seasonal growth. Exopolysaccharides, including pullulan, were found in the xylem sap. Absorption of culture filtrates of Pch and Tmi, as well as weak solutions of purified preparations of scytalone, isosclerone or pullulan, on detached leaves and berries caused symptoms similar to those shown by the esca-affected vines in the field.     

Esca in young and mature vineyards,and molecular diagnosis of the associated fungi

Esca is one of the most important grapevine trunk diseases, and it can induce severe decline. In the past, the disease occurred mostly on mature vines, but today it is also present in young vineyards. The aim of this study was to investigate the incidence of esca in young (< 7 years old) and mature (> 11 years old) vineyards on cvs Montepulciano, Sangiovese, Verdicchio and Passerina located in the main viticultural areas of the Marche Region, central-eastern Italy. The average incidence of diseased plants was higher in mature (32.6%) than young (5.2%) vineyards, and Verdicchio and Passerina appeared to be the most sensitive among the cultivars considered, followed by Sangiovese and Montepulciano. The analysis of the spatial spread of esca carried out in two mature vineyards on cv. Verdicchio and a young vineyard on cv. Sangiovese showed a fluctuation in the numbers of infected plants over the three years of observation. The fungi associated with symptomatic plants were detected by classical and molecular tools. Isolation on agar media yielded colonies of Phaeomoniella chlamydospora (Pch), Botryosphaeria spp. (Bot), Fomitiporia mediterranea (Fomed) and, sporadically, Phaeoacremonium aleophilum (Pal). In samples from young plants, Bot and Pch were recurrent, while Pch and Fomed were found in mature vines and old rootstocks. Molecular detection with specific primer pairs for Pch, Pal, Fomed, and B. dothidea confirmed the data obtained using classical tools, and in some cases it was more sensitive. This study thus provides a further contribution to the association between causal agents and esca symptoms, and it confirms the importance of molecular tools for a sensitive detection of associated pathogens, which can also be present in propagative materials.     

Melanin biosynthesis in Pyricularia oryzae: Site of tricyclazole inhibition and pathogenicity of melanin-deficient mutants

Tricyclazole (5-methyl-1,2,4-triazolo[3,4-b]benzothiazole) inhibits melanin synthesis in Pyricularia oryzae at concentrations less than 0.01 μg/ml. The primary site of inhibition in the biosynthetic pathway occurs between scytalone and vermelone. Accumulation of several metabolites derived from melanin precursors along branch pathways is associated with inhibition of melanin biosynthesis. At low tricyclazole concentrations (0.01–1 μg/ml), predominant accumulation of 2-hydroxyjuglone and 3,4-dihydro-3,4,8-trihydroxy-1-(2H)-naphthalenone (3,4,8-DTN) occurs as a result of the primary block between 1,3,8-trihydroxynaphthalene and vermelone. As the concentration of tricyclazole is increased from 1 to 10 μg/ml, flaviolin accumulation is markedly enhanced, whereas that of 3,4,8-DTN and 3,4-dihydro-4,8-dihydroxy-1-(2H)-naphthalenone is depressed, indicating possible secondary sites of inhibition in the main and branch pathways. Five melanin-deficient mutants of P. oryzae that phenotypically resemble the tricyclazole-treated wild-type strain were nonpathogenic or rarely infected two rice varieties. Three of the mutants studied were genetically defective in the melanin biosynthetic pathway at the site blocked by tricyclazole in the wild type. The wild-type strain converted both scytalone and vermelone to melanin; whereas the three mutants and the tricyclazole-treated wild type converted only vermelone to melanin. The data suggest a relationship between melanin biosynthesis and pathogenicity in P. oryzae.     

Biochemical factors involved in vetch resistance toOrobanche aegyptiaca

Experiments were performed to elucidate the biochemical nature of the differential response of resistantVicia atropurpureacv. Popany and susceptibleVicia sativacv. Yovel to the parasitic weedOrobanche aegyptiaca. Plant root material was obtained by growing vetch plants in association withOrobancheseeds on glass microfibre filter sheets inserted in polyethylene bags replenished by nutrient solution. Higher concentrations of bound phenolics, free phenolics and lignin, and higher activity of peroxidase were observed in cv. Popany roots as a result ofO. aegyptiacainfection than were found in cv. Yovel vetch. No differences in hydroxyproline-rich glycoproteins levels were detected between infected and non-infected roots of either vetch cultivar. These results suggest that the resistant vetch defense mechanism involves elevated induction of the phenylproponoid pathway uponOrobancheinfection, conferring mechanical and chemical barriers confronting the invading parasite.     

Biodegradation kinetics of dicofol by selected microorganisms

The capacity of some plant growth promoters (seven species of bacteria namely, Azospirillium barasilense, Azotobacter chroococcum, Klebsilense pneumoneae, Pseudomonas cepacia, Bacillus subtilis, Pseudomonas fluorescens and Bacillus polymyxa) and some control agents for plant pathogens (two species of fungi namely, Trichoderma viride and Trichoderma harzianum) to degrade dicofol pesticide (DCF) in liquid culture media was investigated. The recovered amount of DCF was extracted based on the solid phase extraction (SPE) with methylene chloride and then analyzed by HPLC. Contrary to published reports, no intermediate or final degradation metabolites of DCF could be observed. About 26-33% of DCF degradation was observed after 3 days of incubation with the tested bacteria which increased to 61-80%, 74-85%, 77-87% and 75-94% after 7, 14, 21 and 28 days of incubation, respectively. On the other hand, the tested fungi removed roughly 35% of available DCF within the first three days of incubation which increased to 84-87%, 89-95%, 91-95% and 92-96% after 7, 14, 21 and 28 days of incubation, respectively. A biphasic model was assumed to explore the disappearance of DCF from media enriched with either bacteria or fungi, where the rate of disappearance in the first phase was faster than the second. This is clearly reflected in the half-life (t_1/2) for the first and second phases, where the t_1/2 values of DCF ranged from 2.82-4.42 to 19.32-29.73 days, respectively. The results have implications for the development of a bioremediation strategy.     

Combined effects of salt stress and cucurbit downy mildew (Pseudoperospora cubensis Berk. and Curt. Rostov.) infection on growth,physiological traits and antioxidant activity in cucumber (Cucumis sativus L.) seedlings

Abiotic and biotic stresses both decrease quality and quantity of cultivated plants. We compared responses of Cucumis sativus L. cvs Sardes and Beith alpha which were treated with 50 or 100 mM NaCl and/or infected by Pseudoperospora cubensis Berk. and Curt. Rostov.. Both cultivars were affected remarkably when fungus infection was combined with salt stress however; cv Sardes was found out to be relatively tolerant with sustainable growth performance, better leaf water status and lower lipid peroxidation levels under this condition. This relative tolerance could be related to the fine tuning of the activities of the antioxidative enzymes and proline levels.     

Host genotype- and temperature-dependent colonizationof In vitro carnation callus cultures byFusarium oxysporum f.sp.dianthi race 2

Differentin vivo resistance/susceptibility levels of 14 carnation cultivars toFusarium oxysporum f.sp.dianthi race 2, the causal agent of Fusarium wilt disease of carnation, were also expressed in anin vitro system and assayed as the degree of fungal colonization of callus cultures at 20° C. Temperature influenced thein vitro expression of carnation resistance. An incubation temperature of 27° C increased the colonization of calli derived from both the susceptible (‘Corrida’ and ‘Ambra’) and the resistant (‘Pulcino’ and ‘Pallas’) cultivars. At 15°C, the colonization of calli derived from Pulcino and Pallas diminished significantly more than for Ambra and Corrida. Inhibition of fungal growth on resistant calli was correlated to retardation in hyphal development. Both scanning electron microscopy and light microscopy observations showed that hyphae did not penetrate into carnation cells.     

Histological and proteomics analysis of apple defense responses to the development of Colletotrichum gloeosporioides on leaves

Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. is an important fungal pathogen known to cause glomerella leaf spot (GLS). The objective of this study was to analyze the infection of C. gloeosporioides on leaves of apple cv. Fuji (resistant) and cv. Gala (susceptible) and apply proteomics techniques to study the apple defense responses 48 h after inoculation (h.a.i.). On both of cultivars, C. gloeosporioides started to germinate at 3 h.a.i. on adaxial surface and produced appressoria adhering to epidermal cell juxtapositions. Histological analysis showed more stratified parenchyma in leaves of cv. Fuji than cv. Gala associated with differences in the chemical composition of cell walls. Total and unique proteins expressed by cvs. Fuji and Gala at 3, 12, 24 and 48 h.a.i. were detected by comparative proteomes analysis. A total of 42 unique proteins expressed at 24 and 48 h.a.i. were identified by MALDI/TOF mass spectrometry, and most of these proteins were identified as directly involved in defense responses.     

Symptom development, pathogen isolation and Real-Time QPCR quantification as factors for evaluating the resistance of olive cultivars to Verticillium pathotypes

Verticillium wilt is the most serious olive disease in the Mediterranean countries and worldwide. The most effective control strategy is the use of resistant cultivars. However, limited information is available about the level and source of resistance in most of the olive cultivars and there are no published data using microsclerotia, the resting structures of Verticillium dahliae, as the infective inoculum. In the present study, we correlated symptomatology and the presence of the fungus along with the DNA relative amount (molecules μl⁻¹) of a defoliating (D) and a non-defoliating (ND) V. dahliae strain in the susceptible cv. Amfissis and the tolerant cvs Kalamon and Koroneiki, as quantified by the Real-Time QPCR technology. The viability of the pathogen in the plant tissues was confirmed by isolating the fungus on PDA plates, while symptom assessment proved the correlation between the DNA relative amount of V. dahliae in plant tissues and cultivar susceptibility. It was further demonstrated that the D and ND strains were present at a significantly higher level in cv. Amfissis than in cvs Kalamon and Koroneiki. It was finally observed that the relative amount of the pathogen in roots was lower than in stems and shoots and declined in plant tissues over time. These data constitute a valuable contribution in evaluating resistance of olive cultivars or olive root-stocks to V. dahliae pathotypes.     

Host status of cultivated plants to <Emphasis Type="Italic">Meloidogyne hispanica</Emphasis>

The reproduction of a Meloidogyne hispanica isolate from Portugal was evaluated in 63 plant species/cultivars, in pot assays at 25?±?2.0°C, on the basis of root gall index (GI) and reproduction factor (Rf?=?final/initial egg density) at 60 days after inoculation. Cultivars of aubergine, bean, beetroot, broccoli, carnation, corn, cucumber, French garlic, lettuce, melon, onion, parsley, pea, potato, spinach, and tobacco and two of cabbage were susceptible (3?≤?GI?≤?5; 1.15?≤?Rf?≤?262.86). Cabbage cv. Bacalan, cauliflower cv. Temporão and pepper cv. Zafiro R2 were hypersusceptible or poor hosts (Rf??2) and pepper cvs. Aurelio and Solero were resistant (0.0?≤?GI?≤?0.4; 0.00?≤?Rf?≤?0.03). The response of the pepper cultivars and the Mi-1 resistant tomato cv. Rossol was also conducted in pots using two inoculum levels and four temperatures, three growth chamber (25?±?2.7°C, 29.3?±?1.8°C and 33.6?±?1.2°C) and one outdoors (24.4?±?8.2°C). At 24.4?±?8.2°C and 25?±?2.7°C, the reproduction on the resistant tomato was significantly lower compared to the susceptible cv. Easypeel. At all temperatures, resistance was evident for the pepper cultivars, despite the fact they were not found to contain any of the Me1, Me3, Me7 and N genes. The eggs obtained on cv. Aurelio at 33.6?±?1.2°C were used to get a selected resistance breaking isolate of M. hispanica that was able to reproduce on the three pepper cultivars. Our results suggest that the initial M. hispanica isolate is a mixture of virulent and avirulent individuals. The pepper cultivars tested, have potential to reduce M. hispanica populations in agro-ecosystems under certain conditions, but they should be used as a part of an integrated management strategy in order to prevent the development of virulent populations.     

Characterization of <Emphasis Type="Italic">Phaeoacremonium</Emphasis> isolates associated with Petri disease of table grape in Northeastern Brazil,with description of <Emphasis Type="Italic">Phaeoacremonium nordesticola</Emphasis> sp. nov.

Severe outbreaks of Alternaria leaf blotch and fruit spot were recently observed in cv. Pink Lady apples in northern Israel, especially on fruit. Such severe outbreaks have not been reported from other countries. Symptoms involved cracks and rot around the calyx and external rot of the fruit body. Up to 80 % of the fruit in some orchards were affected by the disease. Microscopic examinations, fulfillment of Koch’s postulates and molecular (genetic) analyses confirmed the causal agent as Alternaria alternata f. sp. mali. The incidence of Alternaria increased as the degree of calyx cracking increased, or if fruit were both cracked and rotted. Injecting spore suspensions into the fruit produced typical rot symptoms. Injection assays of detached fruit of eight apple cultivars showed that cvs. Pink Lady and Golden Delicious were susceptible whereas cv. Jonathan was resistant. Pink Lady and Golden Delicious produced more fruit rot as the inoculum concentration increased. Rot in all three cultivars was moderate close to the skin but more severe close to the seed locule. Aqueous extracts taken from Jonathan fruit peel inhibited germ tube elongation of A. alternata f. sp. mali in vitro. This is the first report on heavy infection of Pink Lady fruit in Israel caused by A. alternata f. sp. mali.     

Preference and infestation level of Anastrepha fraterculus (Wiedemann, 1830) on fruits of some Psidium guajava L. cultivars and relation to their physicochemical parameters

This study aimed to verify the preference and infestation level of Anastrepha fraterculus (Diptera: Tephritidae) [South American fruit fly] in fruits of guava cultivars and to correlate them to variables such as peel coloration, soluble solids and pH of fruit cultivars. The following cultivars were used: Pedro Sato, Paluma, Cascão and Século XXI. The infestation was evaluated in cages, considering two scenarios: no-choice and multiple choice. In both tests, evaluations of the fruit attraction to insects were conducted for a period of 1′, 3′, 5′, 10′, 20′, 30′, 1 h, 2 h, 6 h, 12 h and 24 h. The visit of A. fraterculus on the assayed cultivars in relation to the time was studied by logistic regression. After 10 days, the number of larvae in each fruit was recorded. In the multiple choice test, the visit proportions were significantly higher in the fruits of cvs. Século XXI and Pedro Sato than in those of cvs. Paluma and Cascão. In the no-choice test, the visit proportions were significantly lower in the Paluma fruits. In both tests, the rate of fruit infestation by A. fraterculus did not differ among cvs. Pedro Sato, Paluma and Cascão, whereas the fruits of cv. Século XXI were more infested. The indexes of pH did not interfere with the infestation of A. fraterculus, whereas a high rate of soluble solids and low color angle appear to be crucial for discriminating the fruits of the most susceptible cultivars. Infestation rate of Século XXI fruits displayed significant correlations with: °Brix (r= 0.7078) and color angle (h) (r= –0.9499) of guava fruits under the multiple choice conditions.