Grapes In Chile Research Articles

A Conservative Mutant Version of the Mrr1 Transcription Factor Correlates with Reduced Sensitivity to Fludioxonil in Botrytis cinerea.

Fludioxonil is a highly effective phenylpyrrole fungicide for controlling Botrytis cinerea. Although the field efficacy of fludioxonil remains high, Botrytis cinerea isolates with reduced sensitivity have been reported globally. The molecular target of fludioxonil still remains unknown; however, a mechanism of reduced sensitivity to fludioxonil underlies the overexpression of the ATP binding cassette (ABC) transporter AtrB in a dependent pathway of the Mrr1 transcription factor. Fludioxonil is a key player in controlling B. cinerea infection in table grapes in Chile. However, some isolates with a reduced sensitivity to fludioxonil were detected. This study observed endogenous atrB overexpression in Chilean isolates with reduced sensitivity to fludioxonil (n = 22) compared to the sensitive isolates (n = 10). All isolates increased the expression of atrB in a growth medium supplemented with fludioxonil (0.05 μg/mL). However, sensitive isolates showed lower atrB expression than those with reduced fludioxonil sensitivity. Remarkably, a mutant version of the transcription factor Mrr1 carrying 21 amino acid modifications was identified in all isolates with reduced sensitivity to fludioxonil. These changes alter the protein's transcription factor domain and the C-terminal portion of the protein but not the Zn (2)-C6 fungal-type DNA-binding domain. These results suggest a direct relationship between the conserved and divergent mutant version of mrr1 and sensitivity to fludioxonil. This study provides a new target for developing molecular diagnostic strategies to monitor B. cinerea's sensitivity to fludioxonil in the field.

Indigenous Yeasts for the Biocontrol of Botrytis cinerea on Table Grapes in Chile.

One hundred twenty-five yeast strains isolated from table grapes and apples were evaluated for the control Botrytis cinerea of in vitro and in vivo. Ten strains were selected for their ability to inhibit mycelial growth of B. cinerea in vitro. In the in vivo assays, these yeasts were tested at 20 °C on 'Thompson Seedless' berries for 7 days; only three were selected (m11, me99 and ca80) because they significantly reduced the incidence of gray mold. These three yeast strains were then evaluated at different concentrations (1 × 107, 1 × 108 and 1 × 109 cells mL-1) on 'Thompson Seedless' grape berries at 20 °C. The strains m11, me99 and ca80 reduced the incidence of B. cinerea to 11.9, 26.1 and 32.1%, respectively, when the berries were submerged in a yeast suspension at a concentration of 1 × 109 cells mL-1 24 h before inoculation with B. cinerea. The most favorable pH for antifungal activity was 4.6 in the three isolates. The three yeast strains secreted the hydrolytic enzymes chitinase and β-1-glucanase, and two strains (me99 and ca80) produced siderophores. The three yeast strains exhibited low oxidative stress tolerance and only strain m11 had the ability to produce biofilms. The strains were identified using 5.8S-ITS rDNA PCR-RFLP and correspond to the Meyerozyma guilliermondii (m11) and Aureobasidium pullulans (me99 and ca80) species.

Lobesia botrana: A Biological Control Approach with a Biopesticide Based on Entomopathogenic Fungi in the Winter Season in Chile

Simple SummaryLobesia botrana, also known as the European grapevine moth, is one of the main pests that affect grapes. In Chile, this type of moth is classified as a quarantine pest, which requires fumigating the fruit with methyl bromide to prevent the immature stages of the pest from being strained and reaching the export-destination countries. In the fields, the larvae of this moth feed on grapes, which can introduce diseases such as Botrytis cinerea, thereby increasing the costs of managing the crop. One way to control this pest is to use entomopathogenic fungi on the winter pupae to reduce moth populations in the spring. In the present study, six fungi were characterized, formulated, and evaluated. The selected strains RGM 2184 and RGM 678 were evaluated in two regions of Chile during two seasons. These strains reached maximum efficiencies of 80% and 88%, respectively. Therefore, the use of entomopathogenic fungi is an environmentally friendly alternative to control L. botrana and reduce the use of chemical pesticides.Lobesia botrana (Denis and Shiffermüller) (Lepidoptera: Tortricidae) is one of the main pests that affect the production and export of table grapes in Chile. Because this pest has quarantine status, the fruit must be fumigated with methyl bromide, which reduces the fruit’s export competitiveness in the destination market. In the present study, to help resolve this issue, six native entomopathogenic fungi were identified through multilocus analysis, including three Beauveria pseudobassiana and three Metarhizium robertsii. These fungi were evaluated in the laboratory to control L. botrana in its pupal stage in a silk cocoon and compared against a biological control product. Formulations with additional carbon sources improved the performance of the fungi. The treatments with outstanding performance contained the fungal strains B. pseudobassiana RGM 2184 and M. robertsii RGM 678. These strains were evaluated in the field during the winter season in two different regions of the country; the strains reached maximum efficacies of 80% and 88%, respectively, at 21 days post first application. Therefore, entomopathogenic fungi can contribute to reducing pupal populations in winter, thereby decreasing the moth population in spring–summer.

LC-MS-Based Metabolomics Discriminates Premium from Standard Chilean cv. Cabernet Sauvignon Wines from Different Valleys.

Cabernet Sauvignon grapes in Chile, mainly grown between the 30° S and 36° S, account for more than 30% of Chilean wine production, and yield wines with different characteristics which influence their quality. The aim of this study was to apply a liquid chromatography – mass spectrometry (LC–MS)-based metabolomic protocol to investigate the quality differentiation in a sample set of monovarietal wines from eight valleys covering 679 km of the north-south extension. All samples were produced using a standardized red winemaking process and classified according to a company categorization in two major groups: premium and standard, and each group in two subcategories. The results pointed out that N-containing metabolites (mainly small peptides) are promising biomarkers for quality differentiation. Moreover, the premium wines were characterized by higher amounts of anthocyanins and other glycosylated and acetylated flavonoids, as well as phenolic acids; standard quality wines, on the other hand, presented stilbenoids and sulfonated catabolites of tryptophan and flavanols.

Spray drift with several nozzles under wind breeze in a vineyard nursery

The area planted with grapes in Chile has grown nearly 50 % in the last 12 years, causing a drastic increase in use of agrichemicals. In particular, herbicides need to be applied more precisely to reach the desired targets. Weeds are a limiting factor in nurseries, mostly because of the reduced distance between rows. The spray nozzles are of vital importance because they distribute the mixture over the target. A comparative study between conventional extended rage flat spray (XR) and drift reducing nozzles (TT, DG and AI) was conducted in Vitis vinifera L. cv. Thompson Seedless plants self-rooted in the nursery, studying the drift detection and quantification, of a 5.8 km.h-1 wind breeze, using the food tracer Brilliant Blue FD & C-1. Under this breeze condition, at the plants level, the drops produced with the XR nozzle suffered a trajectory deviation. A strong decrease in the spray deposits occurred while the spray area distance increased. At ground level, it was appreciated that with wind conditions, there is an increase in all the nozzles of the tanks before the plant, next to it, and after cultivation, but these did not become statistically significant at this wind level

Phenotyping tools for genetic improvement of table grapes in Chile

Although Chile is the world leader for table-grape export by volume, the renewal of table-grape cultivars has just recently taken off. Among these, 'Maylen' ('Iniagrape-One'), a mid-season black cultivar from the Chilean Breeding Program at INIA, stands out for its flavor and excellent postharvest life. In order to accelerate the development of new table-grape cultivars under local conditions, we have developed two software tools to improve the phenotyping of fruit quality characteristics either in the laboratory or in the field. The first of these tools, Vinetracker, is a smartphone application that replaces the use of field blocks: it combines a cloud-located database of the breeding program with the mobile network capacity to allow simultaneous evaluation of vine or fruit phenotypes, resulting in the reduction of cost, time and involuntary errors. It also allows the user to filter and consult data in real time. The second of these tools, Berry Analyzer, is a digital image-analysis tool for fruit phenotyping that aims at fast retrieval of information from berries, such as the polar and equatorial diameters, shape and color, in a small fraction of the time needed in comparison with standard tools or image software that requires human intervention.

First report of Rhizopus stolonifer causing Rhizopus bunch rot on grapes in Pakistan

During June 2016, a postharvest survey of table grapes was carried out in the main fruit markets of Attock (33°46'07.9"N 72°21'43.0"E) and Jehlum (32°56'22.3"N 73°43'31.4"E) districts of Punjab Province. At the time of sampling, two cultivars (King’s Ruby and Perlette) were sampled at five different locations of both districts. Disease incidence % of bunches averaged 3 to 4 %. Some infected fruit appeared water-soaked, light brown and covered by fluffy mycelium consisting of erect sporangiophores with black sporangia on the top (Figure 1). Symptomatic tissue pieces were surface-sterilized with 0.1% NaOCl for 30 sec, rinsed three times with sterile distilled water, dried on filter paper for 45 sec and incubated on potato dextrose agar (PDA) at 25°C. After 1 day, mycelium on PDA was transferred to a fresh PDA plate and incubated at 25°C with a 12-h photoperiod. Within 3 days, white to yellow colonies with black aerial sporangia were formed (Figure 2). A total of 64 isolates were examined morphologically. Sporangiophores were erect, light brown and 623 to 3800 µm long. One to three rhizoids were observed opposite to each sporangiophore. Sporangia were black, globose to sub-globose, 91 to 124 μm in diameter. Columellae were conical to cylindrical and 86 to 187 μm long × 72 to 205 μm wide. Sporangiospores were hyaline to light dark grey, globose, ellipsoidal in shape and 6.8 to 12.4 × 3.6 to 12.5 μm (Figure 3). These features were identical to the description of Rhizopus stolonifer (Ehrenb.) Vuill (Liou et al., 2007). For molecular identification, the ITS1-5.8S-ITS2 region of two representative isolates (Rizo 05 and Rizo 07) was amplified with primers ITS1/ITS4 (White et al., 1990). Sequence comparison of two isolates Rizo05 and Rizo 07 (Accession no. MH348205 and MH356272) revealed 100% identity with previously reported isolates of Rhizopus stolonifer (Accession no. MG865992, KU729185, HM051076, and MF374842). To complete Koch’s postulates, 10-µl aliquots of spore suspensions (106 spores/ml) of Rizo 05 and Rizo 07 were pipetted onto three non-wounded and four wounded asymptomatic grape berries (seven berries per isolate), Sterile distilled water was applied to asymptomatic berries to serve as a negative control. Berries were incubated at 25 ± 2°C in sterile moisture chambers, and the experiment was conducted twice (Ghuffar et al., 2018). Black to light brown, fluffy mycelium with the original symptoms was observed on both wounded and non-wounded inoculated berries after 3 days, whereas no symptoms were recorded on the negative control (Figure 4). The morphology of the fungus that was re-isolated from each of the inoculated berries was identical to that of the original cultures. Previously, Rhizopus stolonifer has been reported as a pathogen on grapes in Chile (Latorre et al., 2002). To our knowledge, this is the first report of Rhizopus stolonifer causing bunch rot of grapes in Pakistan. This finding will help to plan effective disease management strategies against Rhizopus rot of grapes in Pakistan.

Frequency of fungicide-resistant Botrytis cinerea populations isolated from ‘Thompson Seedless’ table grapes in the Central Valley of Chile

During the 2013 and 2014 growth seasons, 526 single spore isolates of Botrytis cinerea were collected from naturally infected ‘Thompson Seedless’ table grape flowers from fifteen orchards in three regions of the Central Valley of Chile. The isolates were tested for resistance to azoxystrobin, boscalid, fenhexamid, fludioxonil and pyrimethanil. Among the 526 isolates, 106 (20.15%) were sensitive to all fungicides tested; from north to south, the frequency of sensitive isolates in the regions of Valparaiso, Metropolitana and O’Higgins ranged from 48.15% to 21.1% and 5.88%, respectively. Four hundred and twenty isolates (79%) showed resistance to single or multiple fungicides, 134 (25.4%) were simultaneously resistant to azoxystrobin and pyrimethanil. No fludioxonil-resistant isolates were found, indicating that fludioxonil has great potential for gray mold control in table grapes in Chile. From sixty randomly selected B. cinerea isolates, only the azoxystrobin-resistant isolates carried the G143A point mutation; according to the cytochrome b ( cyt b ) gene structure, the third intron Bcbi-143/144 was only detected in the azoxystrobin-sensitive isolates. The H272R and H272Y point mutations in the succinate dehydrogenase subunit B ( sdhB ) gene were associated only with the boscalid-resistant isolates. The F412S and F412V point mutations were found in the sequenced erg27 gene of randomly selected fenhexamid-resistant isolates. These results contribute to the knowledge of B. cinerea fungicide resistance for table grape vine crops in Central Chile, particularly for the development of multiple-resistance and the associated resistance mechanisms of azoxystrobin, boscalid and fenhexamid-resistant isolate populations. Antiresistance strategies are discussed in a general manner.

Response of Table Grape to Irrigation Water in the Aconcagua Valley, Chile

The irrigation water available for agriculture will be scarce in the future due to increased competition for water with other sectors, and the issue may become more serious due to climate change. In Chile, the table grape is only cultivated under irrigation. A five-year research program (2007–2012) was carried out in the Aconcagua Valley, the central area of grapes in Chile, to evaluate the response of table grape vines (Vitis vinifera L., cv Thompson Seedless) to different volumes of irrigation water. Four irrigation treatments were applied: 60, 88, 120 and 157% of crop evapotranspiration (ETc) during the first four years, and 40, 54, 92 and 108% of ETc in the last year. Irrigation over 90%–100% of ETc did not increase fruit yield, whereas the application of water below 90% ETc decreased exportable yield, berry size and pruning weight. For example, 60% ETc applied water reduced exportable yield by 20%, and only 40% of the berries were in the extra and large category size, while pruning weight was 30% lower in comparison to the treatment receiving more water.

Botryosphaeriaceae species affecting table grape vineyards in Chile and cultivar susceptibility

Morales, B.A. Latorre, E. Piontelli, and X. Besoain. 2012. Botryosphaeriaceae species affecting table grape vineyards in Chile and cultivar susceptibility. Cien. Inv. Agr. 39(3): 445-458. Several Botryosphaeriaceae species have been identified as the causes of cankers and dieback of Vitis vinifera in several grape-growing regions around the world. This research was conducted to further study the species of Botryosphaeriaceae associated with table grapes in Chile, to estimate the prevalence and severity of the disease as a function of vineyard age, to study the susceptibility of table grape cultivars to infection by Botryosphaeriaceae species, and to evaluate the effect of tissue age on the infection caused by Botryosphaeriaceae species. Symptoms were characterized by the presence of the partial or total death of the grapevine cordons and distorted leaves. Brown V-shaped or U-shaped cankers and black spots were observed in cross-sections, while brown vascular streaks were observed in longitudinal sections of the cordons and trunks. Pathogenic isolates of Diplodia seriata , D. mutila and Spencermartinsia viticola were consistently obtained from wood cankers and/or vascular streaking; D. seriata was the most common (83.3%) Botryosphaeriaceae species. In 11- to 20-year-old vineyards, the disease incidence varied between 22.0 and 69.0%, and the severity varied between 6.0 and 21.3%. The table grape cultivars ‘Thompson Seedless’, ‘Redglobe’ and ‘Flame Seedless’ were equally susceptible to infection by . The age of the inoculated tissue had no significant effect on the development of the vascular necrosis. This is the first report of D. mutila and S. viticola infections of grapevines in Chile.

Low Occurrence of Patulin-Producing Strains of Penicillium in Grapes and Patulin Degradation during Winemaking in Chile

<i>Penicillium expansum</i> has emerged as the cause of storage decay of table grapes (<i>Vitis vinifera</i>) and has been frequently isolated from apparently healthy clusters of grapes in Chile. The objectives of this study were to identify patulin-producing strains of <i>Penicillium</i> associated with winegrapes and wineries in Chile and to determine the potential presence of patulin in wines made with grapes infected with <i>P. expansum</i>. In this study, <i>P. brevicompactum, P. expansum,</i> and <i>P. glabrum</i> were identified on apparently healthy grape clusters and in the air of vineyards and wineries. Of 132 <i>Penicillium</i> isolates, 4 <i>P. brevicompactum</i> and 11 <i>P. expansum</i> strains were patulin-producing, determined by HPLC-UV/DAD. Patulin was also detected in Cabernet Sauvignon musts produced with grapes contaminated with a patulin-producing strain of <i>P. expansum.</i> However, patulin concentrations decreased during fermentation by 67.3 to 83.3%. Overall, the frequency of <i>P. expansum</i> isolation from grapes was relatively low; thus, considering the rapid degradation of patulin produced during fermentation, the risk of patulin contamination of bottled wine appears to be low.

Outbreaks of Cladosporium Rot Associated with Delayed Harvest Wine Grapes in Chile.

Severe outbreaks of Cladosporium rot have occurred on berries in clusters of late harvest wine grapes (Vitis vinifera L.) during the 2003 to 2006 growing seasons. This disease was especially prevalent on Cabernet Sauvignon (CS) vineyards in central Chile where disease incidence commonly comprised 50 to 100% of the clusters at harvest. Symptoms appeared on mature grapes (total soluble solids [TSS] >22%) and were characterized by berry dehydration, a firm decay affecting a small portion of the berry and a superficial olive-green mold. Isolations made on acidified potato dextrose agar (APDA) consistently yielded olive-green colonies after 7 days at 20°C. On the basis of colony morphology and morphological characteristics of conidiophores and conidia, Cladosporium herbarum (Pers.:Fr) Link and C. cladosporioides (Fres.) de Vries were identified (1). These species were primarily separated by the presence of rough and smooth conidial surfaces, respectively. Koch's postulates were completed by inoculating 80 wounded mature (20% TSS) CS berries and incubating them at 10, 20, and 30°C. An equal number of wounded but noninoculated berries were left as controls. Berries were surface sterilized (75% ethanol for 30 s and 0.5% sodium hypochlorite for 60 s), inoculated with 10 μl of a 106 conidial suspension per ml applied to the wounds made with a sterile hypodermic needle. Dark, necrotic lesions, 1 to 6 mm in diameter, and a dark mycelial colony appeared on the surface after 7 days in chambers with relative humidity of >95%. Disease incidence was significantly (P < 0.05) influenced by temperature, with 90, 100, and 49% of inoculated berries becoming infected when incubated at 10, 20, or 30°C, respectively. C. herbarum and C. cladosporioides appeared to be equally pathogenic, producing symptoms similar to naturally infected CS berries, and were reisolated (100%) on APDA. These same isolates were pathogenic when tested on mature (TSS >16%) Thompson Seedless (TS) berries. Fungicide sensitivity tests were performed on detached TS berries challenged by placing 10 μl of a 106 conidial suspension per ml of C. herbarum on injured berries. Boscalid (0.5 mg/ml, Cantus WG; BASF, Santiago, Chile), iprodione (0.5 mg/ml, Rovral WP; Bayer Crop Science, Santiago, Chile), pyraclostrobin (0.085 mg/ml, Comet SC; BASF), and pyraclostrobin (0.009 mg/ml) mixed with boscalid at 0.017 mg/ml (Bellis WG; BASF) provided a significant control (P < 0.05) with efficacy between 84.3 and 95.9%. Azoxystrobin (0.188 mg/ml, Quadris SC; Syngenta Crop Protection, Santiago, Chile), kresoxim methyl (0.067 mg/ml, Stroby SC; BASF), and trifloxystrobin (0.06 mg/ml, Flint WG; Bayer Crop Science) provided partial control with efficacy between 23.1 and 42.1%. Cladosporium spp. have been previously reported (2). However, severe outbreaks of Cladosporium rot occur when berries become partially senescent because of a considerable delay in harvest (3). This appears to favor the development of these pathogens.

Canopy structure and photosynthesis modelling of grapevines (Vitis vinifera L. cv. Sultana) grown on an overhead (parronal) trellis system in Chile

Most table grapes in Chile are produced on an overhead (parronal) trellis system with typically high values for leaf area index (LAI). An apparent lack of sunlight inside such canopies often raises questions as to the photosynthetic effectiveness of shaded leaves, and the comparative importance of successive leaf layers (top to bottom) for the total canopy carbon balance. In particular, are some leaves parasitic with respect to daily carbon balance? Accordingly, we undertook a study of canopy architecture, light climate and photosynthetic physiology to address those issues. Canopy photosynthesis was inferred from a process-based model using measured values for: the vertical distribution of (1) leaf angle, (2) LAI, (3) the fraction of sunlit leaves and (4) single-leaf photosynthetic capacity. Photon flux densities (PFD) and measured values of CO2 assimilation by leaves located on different parts of the canopy were obtained at flowering (mid-October) and during active berry growth (mid-December). Results of leaf angle revealed that Sultana canopies on this system behave in an erectophile manner with an average leaf inclination of 60o above horizontal. Modelling the contribution made by different leaf layers to overall canopy photosynthesis demonstrated that leaf layers contributing most were those under a shade plus sunfleck regime. They were located between 20 and 40 cm above the horizontal trellis structure itself. Indeed, some photoinhibition was observed in leaves under continuous direct radiation towards the top of the canopy, and located above 40 cm layer. However, such leaves represented only a small proportion of total LAI, and thus compromised canopy photosynthesis to only a limited extent. Outcomes from our simulation studies imply that LAI values commonly reached by canopies managed according to the parronal system of grape trellising do not result in parasitic leaves.

Severe Outbreaks of Bunch Rots Caused by Rhizopus stolonifer and Aspergillus niger on Table Grapes in Chile.

Severe outbreaks of bunch rots (BR) have occurred recently during harvest of table grapes (Vitis vinifera L.) in Chile. Previously, BR was almost exclusively associated with Botrytis cinerea Pers.:Fr. (2,3); however, in 2000 to 2002, BR symptoms were associated with black molds and possibly nonfilamentous yeasts and bacteria. Cvs. Thompson Seedless, Flame Seedless, Ruby Seedless, and Red Globe were severely affected. Symptoms start at the pedicels as soft, watery rots that partially or completely decay infected berries. Longitudinal cracks are produced, a black mold usually develops along the crack fissures, and the skin of the berry turns light gray. Isolations on potato dextrose agar acidified with 1 N lactic acid (APDA) at 0.5 ml/liter, consistently yielded Rhizopus stolonifer (Ehrenb. ex Fr.) Vuillemin and Aspergillus niger Tiegh. R. stolonifer on APDA produced a white-to-gray aerial and nonseptate mycelium, black and globose sporangia with an elliptical collumela, one-celled, globose to oval, striated, almost hyaline sporangiospores, rhizoids, and stolons. A. niger produced septate mycelium. Single-celled, black, rough walled, globose conidia developed on short chains on the second phialides at the tip of globose, upright conidiophores. Mature (soluble solids >16%) detached berries of cv. Thompson Seedless were inoculated with sporangiospores (≈107 spores per ml) of R. stolonifer isolates RS6, RS52, RS73, and RS79 and conidia (≈108 conidia per ml) of A. niger isolates AN12, AN69, and AN75. When berries were aseptically punctured with a sterile hypodermic syringe prior to inoculation, 60 to 86.7% and 42.5 to 100% of berries were infected with R. stolonifer and A. niger, respectively, and both developed BR symptoms (significantly different from control berries) after 48 h in humid chambers at 23°C. Injuries were needed for infection since no infection or only 23.3% of noninjured berries were infected with R. stolonifer and A. niger, respectively. For both pathogens, there was a significant (P < 0.043) interaction between isolates and the presence or absence of injuries. Both pathogens were successfully reisolated on APDA. Fungicide sensitivity tests were performed on detached cv. Thompson Seedless berries challenged by placing an ≈6 μl-drop of inoculum suspension (106 or 107 spores per ml of R. stolonifer isolate RS52 and A. niger isolate AN12, respectively) on injured berries. Pyraclostrobin (0.067 mg/ml) mixed with nicobifen at 0.134 mg/ml (BAS 516 01 F at 0.201 mg a.i./ml, BASF) and copper oxide at 1.2 mg/ml (Cuprodul 60 WP, Quimetal Chile) significantly (P < 0.01) inhibited infection (100% control) by R. stolonifer and A. niger. R. stolonifer was completely controlled by dicloran at 1.88 mg/ml (Botran 75 WP) and partially controlled by captan at 1.6 mg/ml (Captan 80 WP), but A. niger was not controlled by either fungicide. To our knowledge this is the first report of R. stolonifer causing BR of table grape in Chile (4). The severe outbreaks may be associated with warm weather conditions during harvest and injuries caused by birds, insects, or cultural practices. Infection caused by R. stolonifer or A. niger may be followed by sour rot organisms (yeasts or bacteria), as has been suggested elsewhere (1,2).

Occurrence of resistant strains of Botrytis cinerea to anilinopyrimidine fungicides in table grapes in Chile

Effort to control gray mold caused by Botrytis cinerea failed in a table grapevine ( Vitis vinifera) vineyard near Santiago, Chile where cyprodinil (Vangard 50 WP), a new fungicide of the anilinopyrimidine group, had been applied alone up to four times per year during two growing seasons. A relatively high frequency (38.5%) of resistant isolates of B. cinerea (EC 50 for mycelial growth inhibition varied from 2.9 to 4.84 μg ml −1) may explain the partial loss of field control efficacy obtained. Resistance was correlated with a complete loss of in vivo sensitivity to cyprodinil. Resistant isolates of B. cinerea showed cross resistance to the anilinopyrimidines fungicides mepanipyrim and pyrimethanil. Cyprodinil partially impaired conidia germination and differentially affected conidial germination of resistant and sensitive isolates. Significant differences ( p<0.05) in growth rate, sclerotia production and osmotic sensitivity were found among isolates of B. cinerea, but no correlation could be drawn between these biological differences and resistance or sensitivity. This indicates a disruptive selection characteristic of monogenic resistance. Thus, strategies were implemented to avoid the further development and spread of resistance in B. cinerea to the anilinopyrimidine fungicides. To our knowledge this constitutes the first mention of resistance in B. cinerea populations to anilinopyrimidine fungicides in South America.

Characterization of Botrytis cinerea from Table Grapes in Chile Using RAPD-PCR.

Random amplified polymorphic DNA (RAPD) analysis was performed on 29 isolates of Botrytis cinerea Pers. ex Fr. isolated from table grapes (Vitis vinifera L.) and other crops in Chile with 29 decaprimers. No single primer was found to differentiate either the host or the geographical origin of each of the B. cinerea isolates tested. The DNA profiles obtained, particularly with primers OPA4 and OPA11, distinguished isolates of B. cinerea from other epiphyte fungi found on table grapes, including Alternaria alternata, Aspergillus niger, Cladosporium herbarum, Epiccocum nigrum, Rhizopus stolonifer, a Penicillium sp., and yeasts (Cryptococcus laurentii, Rhodotorula glutinis, and Saccharomyces cerevisiae). Regardless of host origin, primers OPA4 and OPA11 amplified a strong fragment of 1.2 kilobases (kb) and two fragments of 1.10 and 0.7 kb, respectively. These DNA fragments were obtained even when only one conidium of B. cinerea was in the test sample. Three main groups were clearly defined based on the genetic similarities found in additional RAPD analysis with 19 arbitrary decaprimers and 15 selected isolates of B. cinerea. The overall similarity coefficients (SC) between the groups obtained ranged from 0.326 to 0.891. Interestingly, all isolates from table grapes were included in group I (SC: 0.761 to 0.826), isolates from apple and tomato were in group II (SC: 0.739 to 0.848), while isolates from blueberry were either in group I (SC: 0.804) or III (SC: 0.673). Consequently, the genetic variability determined by RAPD analysis among these B. cinerea isolates suggested a possible host:pathogen relationship. However, further research is needed to clarify its pathological significance.

Crown and root rots of table grapes caused by Phytophthora spp. in Chile

Phytophthora crown and root rot of grapevine ( Vitis vinifera L.) occurs frequently in Chile, where grapes are cultivated on their own roots. P. cinnamomi, P. cryptogea , and P. drechsleri were isolated from diseased root or crown tissue of the table grape cvs Flame Seedless, Italia, Red Globe, and Thompson Seedless with P. cryptogea being the most frequently isolated species. These pathogens were identified on the basis of standard morphological and cultural features and were compared by SDS-PAGE profiles of mycelial proteins. All isolates were pathogenic on 1-year-old cane segments and 1-year-old rooted grapevines of cvs Thompson Seedless and Red Globe. The latter cultivar was more susceptible than Thompson Seedless. P. cinnamomi and P. cryptogea have been associated with root rot of grapevines elsewhere, but this is the first report of P. drechsleri as a pathogen of grapevines and the first report of Phytophthora spp. affecting table grapes in Chile.