The Friend Within: Endophytic Bacteria as a Tool for Sustainability in Strawberry Crops.

Strawberry (Fragaria x ananassa, Duch.) is an important crop worldwide. However, since it is a highly demanding crop in terms of the chemical conditions of the substrate, a large part of strawberry production implies the application of large amounts of fertilizers in the production fields. This practice can cause environmental problems, in addition to increases in the fruit's production costs. In this context, applying plant growth-promoting bacteria in production fields can be an essential strategy, especially thanks to their ability to stimulate plant growth via different mechanisms. Therefore, this study aimed to test in vitro and in vivo the potential of bacteria isolated from strawberry leaves and roots to directly promote plant growth. The isolates were tested in vitro for their ability to produce auxins, solubilize phosphate and fix nitrogen. Isolates selected in vitro were tested on strawberry plants to promote plant growth and increase the accumulation of nitrogen and phosphorus in the leaves. The tested isolates showed an effect on plant growth according to biometric parameters. Among the tested isolates, more expressive results for the studied variables were observed with the inoculation of the isolate MET12M2, belonging to the species Brevibacillus fluminis. In general, bacterial inoculation induced strain-dependent effects on strawberry growth. In vitro and in vivo assays showed the potential use of the B. fluminis MET12M2 isolate as a growth promoter for strawberries.

Impact of diseases and pests on premature fruit drop in sweet orange orchards in São Paulo state citrus belt, Brazil.

BACKGROUND: Despite technical improvements in the citrus chain and leadership in orange production achieved in the past decades, premature fruit drop remains a major component of crop loss in São Paulo state citrus belt, the largest sweet orange production area in the world. The present study aimed to determine, during five consecutive seasons, the impact of the diseases and pests on premature fruit drop in the orange belt. RESULTS: Fruit drop due to the main diseases and pests averaged approximately 11.0%, which corresponded to approximately 63% of the annual fruit drop. The average fruit drop rate due to fruit borer and fruit flies combined was 4.0%, Huanglongbing (HLB) 3.3%, black spot 2.6%, leprosis 1.0% and citrus canker 0.3%. The average amount of fruit drop (million 40.8 kg boxes) and value of crop losses (million US$ dollars), in five seasons, were 12.7 and 66.2 for fruit borer/fruit flies, 11.0 and 57.9 for HLB, 8.1 and 42.2 for black spot, 3.1 and 15.6 for leprosis, and 0.9 and 4.9 for citrus canker, respectively. CONCLUSION: Fruit borer and fruit flies (combined), HLB, black spot, leprosis and citrus canker are, in this order, the main diseases and pests in the orange belt of São Paulo state. All of these causes significantly increased the overall fruit drop rate in the evaluated seasons. The results will contribute to the development of the Brazilian citrus industry, while showing to other citrus-growing regions the potential that diseases and pests have to jeopardize production. © 2022 Society of Chemical Industry.

Fusarium graminearum Species Complex: A Bibliographic Analysis and Web-Accessible Database for Global Mapping of Species and Trichothecene Toxin Chemotypes.

Fusarium graminearum is ranked among the five most destructive fungal pathogens that affect agroecosystems. It causes floral diseases in small grain cereals including wheat, barley, and oats, as well as maize and rice. We conducted a systematic review of peer-reviewed studies reporting species within the F. graminearum species complex (FGSC) and created two main data tables. The first contained summarized data from the articles including bibliographic, geographic, methodological (ID methods), host of origin and species, while the second data table contains information about the described strains such as publication, isolate code(s), host/substrate, year of isolation, geographical coordinates, species and trichothecene genotype. Analyses of the bibliographic data obtained from 123 publications from 2000 to 2021 by 498 unique authors and published in 40 journals are summarized. We describe the frequency of species and chemotypes for 16,274 strains for which geographical information was available, either provided as raw data or extracted from the publications, and sampled across six continents and 32 countries. The database and interactive interface are publicly available, allowing for searches, summarization, and mapping of strains according to several criteria including article, country, host, species and trichothecene genotype. The database will be updated as new articles are published and should be useful for guiding future surveys and exploring factors associated with species distribution such as climate and land use. Authors are encouraged to submit data at the strain level to the database, which is accessible at https://fgsc.netlify.app.

Aggressiveness and Mycotoxin Production by Fusarium meridionale Compared with F. graminearum on Maize Ears and Stalks in the Field.

Fusarium meridionale and F. graminearum both cause Gibberella ear rot (GER) and Gibberella stalk rot (GSR) of maize in Brazil, but the former is much more common. Recent work with two isolates of each from maize suggested this dominance could be caused by greater aggressiveness and competitiveness of F. meridionale on maize. We evaluated pathogenicity and toxigenicity of 16 isolates of F. graminearum and 24 isolates of F. meridionale recovered from both wheat and maize. Strains were individually inoculated into ears of four maize hybrids in field trials. GER severity varied significantly between isolates within each species. Although ranges overlapped, the average GER severity induced by F. meridionale (25.2%) was two times as high overall as that induced by F. graminearum (12.8%) for isolates obtained from maize but was similar for those isolated from wheat (19.9 and 21.4%, respectively). In contrast, severity of GSR was slightly higher for F. graminearum (22.2%) than for F. meridionale (19.8%), with no effect of the host of origin. Deoxynivalenol and its acetylated form 15ADON were the main mycotoxins produced by F. graminearum (7/16 strains), and nivalenol toxin was produced by F. meridionale (17/24 strains). Six isolates of F. graminearum and three of F. meridionale also produced zearalenone. Results confirmed that F. meridionale from maize is, on average, more aggressive on maize but also suggested greater complexity related to diversity among the isolates within each species and their interactions with different hybrids. Further studies involving other components of the disease cycle are needed to more fully explain observed patterns of host dominance.

Endophytic bacteria from strawberry plants control gray mold in fruits via production of antifungal compounds against Botrytis cinerea L.

Botrytis cinerea causes the gray mold disease in a wide range of plant hosts, especially in post-harvest periods. The control of this phytopathogen has been accomplished through the application of fungicides. However, this practice can cause environmental problems and increase fruit production costs. In addition, this fungus species has developed resistance to conventional synthetic fungicides. In this context, plant growth-promoting bacteria have shown potential for application in agricultural production because they are able to stimulate plant growth through different mechanisms, including the biological control of phytopathogens (indirect growth promotion mechanism). The aim of this work was to evaluate in vitro and in fruits the potential for indirect plant growth-promotion of bacteria isolated from strawberry leaves and roots. Dual plate method and inverted plate method were used to verify the ability of controlling in vitro the growth of Botrytis cinerea via the production of diffusible and volatile antifungal compounds, respectively. The effect of six bacterial isolates that showed greater potential for biological control in vitro was evaluated by scanning electron microscopy. Antifungal compounds produced by these bacterial isolates were identified by liquid chromatography coupled with mass spectrometry. Six bacterial strains were tested on strawberry pseudofruits. Five selected strains belong to the genus Bacillus and one to the genus Pantoea sp. Selected strains were able to inhibit more than 80 % of the mycelial growth of B. cinerea by the production of diffusible compounds and 90 % by volatile antifungal compounds production. Scanning electron microscopy showed the intense degradation of fungal hyphae caused by the presence of all bacterial strains. Bioactive compounds (salycilamide, maculosin, herniarin, lauroyl diethanolamide, baptifoline, undecanedioic acid, botrydial, 8 3-butylidene-7-hydroxyphthalide and N-(3-oxo-henoyl)-homoserine lactone) were obtained from liquid culture of these strains and extraction with ethyl acetate. All six isolates tested in vivo reduced the incidence of gray mold in strawberry pseudofruits in postharvest. It is concluded that isolates 26, 29, 65, 69, 132 (Bacillus sp.) and MQT16M1 (Pantoea sp.) have potential application for the biological control of Botrytis cinerea in strawberry via the production of diffusible and volatile antifungal compounds.

The Dominance of Fusarium meridionale Over F. graminearum Causing Gibberella Ear Rot in Brazil May Be Due to Increased Aggressiveness and Competitiveness.

In Brazil, Gibberella ear rot (GER) of maize is caused mainly by Fusarium meridionale, whereas F. graminearum is a minor contributor. To test the hypothesis that F. meridionale is more aggressive than F. graminearum on maize, six experiments were conducted in the south (summer) and one in the central-south (winter), totaling seven conditions (year × location × hybrid). Treatments consisted of F. graminearum or F. meridionale (two isolates of each) inoculated once 4 days after silk, inoculated sequentially and alternately (F. graminearum → F. meridionale or F. meridionale → F. graminearum) 6 days apart, or (in the central-south) inoculated sequentially without alternating species (F. meridionale → F. meridionale or F. graminearum → F. graminearum). Overall, severity was two times greater in the south (37.0%), where summer temperatures were warmer (20 to 25°C) than in central-south. In the south, severity was greatest in F. meridionale treatments (67.8%); followed by F. meridionale → F. graminearum (41.1%), then F. graminearum → F. meridionale (19.4%), and lowest in F. graminearum (2.1%), suggesting an antagonistic relationship. In the central-south (15 to 20°C), severity was generally higher in the sequential nonalternating inoculation treatments (F. meridionale → F. meridionale or F. graminearum → F. graminearum) than when either species was inoculated only once. Only nivalenol (NIV) or deoxynivalenol was detected when F. meridionale or F. graminearum, respectively, was inoculated singly, or sequentially with no alternation. Both toxins were found in grains harvested from the F. meridionale → F. graminearum treatment, whereas only NIV was found in kernels from the F. graminearum → F. meridionale treatment, suggesting that F. meridionale was more competitive than F. graminearum in coinoculations. The dominance of F. meridionale as a cause of GER in Brazil may be due in part to its higher aggressiveness and competitiveness compared with F. graminearum.

Diversity and Cross-Infection Potential of Colletotrichum Causing Fruit Rots in Mixed-Fruit Orchards in Kentucky.

Fungi in the genus Colletotrichum cause apple, blueberry, and strawberry fruit rots, which can result in significant losses. Accurate identification is important because species differ in aggressiveness, fungicide sensitivity, and other factors affecting management. Multiple Colletotrichum species can cause similar symptoms on the same host, and more than one fruit type can be infected by a single Colletotrichum species. Mixed-fruit orchards may facilitate cross-infection, with significant management implications. Colletotrichum isolates from small fruits in Kentucky orchards were characterized and compared with apple isolates via a combination of morphotyping, sequencing of voucher loci and whole genomes, and cross-inoculation assays. Seven morphotypes representing two species complexes (C. acutatum and C. gloeosporioides) were identified. Morphotypes corresponded with phylogenetic species C. fioriniae, C. fructicola, C. nymphaeae, and C. siamense, identified by TUB2 or GAPDH barcodes. Phylogenetic trees built from nine single-gene sequences matched barcoding results with one exception, later determined to belong to an undescribed species. Comparison of single-gene trees with representative whole genome sequences revealed that CHS and ApMat were the most informative for diagnosis of fruit rot species and individual morphotypes within the C. acutatum or C. gloeosporioides complexes, respectively. All blueberry isolates belonged to C. fioriniae, and most strawberry isolates were C. nymphaeae, with a few C. siamense and C. fioriniae also recovered. All three species cause fruit rot on apples in Kentucky. Cross-inoculation assays on detached apple, blueberry, and strawberry fruits showed that all species were pathogenic on all three hosts but with species-specific differences in aggressiveness.

Are Demethylation Inhibitor Plus Quinone Outside Inhibitor Fungicide Premixes During Flowering Worthwhile for Fusarium Head Blight Control in Wheat? A Meta-Analysis.

Fusarium head blight (FHB), caused mainly by Fusarium graminearum, is best controlled with demethylation inhibitor (DMI) fungicides during flowering. However, the use of premixes of DMI and quinone outside inhibitor (QoI) fungicides to control FHB has increased in Brazil. Data on FHB severity and wheat yields measured in field experiments conducted in Brazil were gathered from both peer- and nonpeer-reviewed sources published from 2000 to 2018. After selection criteria were applied, 73 field trials from 35 bibliographic sources were identified, among which 50% of the data were obtained from cooperative network trials conducted after 2011. To be included in the analysis, DMI plus QoI premixes or tebuconazole were tested in at least 14 trials and 3 years. Four premixes met the criteria. Estimates of percent control (and respective 95% confidence intervals) by a network model fitted to the log of the treatment means ranged from 44.1% (pyraclostrobin plus metconazole applied once; 32.4 to 53.7) to 64.3% (pyraclostrobin plus metconazole; 58.4 to 69.3); the latter did not differ from tebuconazole (59.9%; 53.6 to 65.3). Yield response was statistically similar for pyraclostrobin plus metconazole (532.1 kg/ha; 441 to 623) and trifloxystrobin plus prothioconazole (494.9 kg/ha; 385 to 551), and both differed statistically from a group composed of tebuconazole (448.2 kg/ha; 342 to 554), trifloxystrobin plus tebuconazole (468.2 kg/ha; 385 to 551), azoxystrobin plus tebuconazole (462.4 kg/ha; 366 to 558), and pyraclostrobin plus metconazole applied once (413.7 kg/ha; 308 to 518). The two categories of FHB index (7% cutoff) and yield (3,000 kg/ha cutoff), both in the nontreated check, did not explain the heterogeneity in the estimates. Considering only the fungicide effects on yield, two sequential sprays of tebuconazole or one spray of pyraclostrobin plus metconazole as management choices are likely more profitable than DMI plus QoI premixes sprayed twice during flowering.

Modeling Yield Losses and Fungicide Profitability for Managing Fusarium Head Blight in Brazilian Spring Wheat.

Fusarium head blight (FHB) and wheat yield data were gathered from fungicide trials to explore their relationship. Thirty-seven studies over 9 years and 11 locations met the criteria for inclusion in the analysis: FHB index in the untreated check ≥ 5% and the range of index in a trial ≥ 4 percentage points. These studies were grouped into two baseline yields, low (Yl ≤ 3,631 kg ha-1) or high (Yh > 3,631 kg ha-1), defined based on the median of maximum yields across trials. Attainable (disease-free) yields and FHB index were predicted using a wheat crop and a disease model, respectively, in 280 simulated trials (10 planting dates in a 28-year period, 1980 to 2007) for the Passo Fundo location. The damage coefficient was then used to calculate FHB-induced yield loss (penalizing attainable yield) for each experiment. Losses were compared between periods defined as before and after FHB resurge during the early 1990s. Disease reduction from the use of one or two sprays of a triazole fungicide (tebuconazole) was also simulated, based on previous meta-analytic estimates, and the response in yield was used in a profitability analysis. Population-average intercepts but not the slopes differed significantly between Yl (2,883.6 kg ha-1) and Yh (4,419.5 kg ha-1) baseline yields and the damage coefficients were 1.60%-1 and 1.05%-1, respectively. The magnitudes and trends of simulated yield losses were in general agreement with literature reports. The risk of not offsetting the costs of one or two fungicide sprays was generally higher (>0.75) prior to FHB resurgence but fungicide profitability tended to increase in recent years, depending on the year. Our simulations allowed us to reproduce trends in historical losses, and may be further adjusted to test the effect and profitability of different control measures (host resistance, other fungicides, etc.) on quality parameters such as test weight and mycotoxin contamination, should the information become available.

Fitness Traits of Deoxynivalenol and Nivalenol-Producing Fusarium graminearum Species Complex Strains from Wheat.

Fusarium graminearum of the 15-acetyl-deoxynivalenol (15-ADON) chemotype is the main cause of Fusarium head blight (FHB) of wheat in southern Brazil. However, 3-ADON and nivalenol (NIV) chemotypes have been found in other members of the species complex causing FHB in wheat. To improve our understanding of the pathogen biology and ecology, we assessed a range of fitness-related traits in a sample of 30 strains representatives of 15-ADON (F. graminearum), 3-ADON (F. cortaderiae and F. austroamericanum), and NIV (F. meridionale and F. cortaderiae). These included perithecia formation on three cereal-based substrates, mycelial growth at two suboptimal temperatures, sporulation and germination, pathogenicity toward a susceptible and a moderately resistant cultivar, and sensitivity to tebuconazole. The most important trait favoring F. graminearum was a two times higher sexual fertility (>40% perithecial production index [PPI]) than the other species (<30% PPI); PPI varied among substrates (maize > rice > wheat). In addition, sensitivity to tebuconazole appeared lower in F. graminearum, which had the only strain with effective fungicide concentration to reduce 50% of mycelial growth >1 ppm. In the pathogenicity assays, the deoxynivalenol producers were generally more aggressive (1.5 to 2× higher final severity) toward the two cultivars, with 3-ADON or 15-ADON leading to higher area under the severity curve than the NIV strains in the susceptible and moderately resistant cultivars, respectively. There was significant variation among strains of the same species with regards asexual fertility (mycelial growth, macroconidia production, and germination), which suggested a strain- rather than a species-specific difference. These results contribute new knowledge to improve our understanding of the pathogen-related traits that may explain the dominance of certain members of the species complex in specific wheat agroecosystems.

Standard Area Diagrams for Aiding Severity Estimation: Scientometrics, Pathosystems, and Methodological Trends in the Last 25 Years.

Standard area diagrams (SAD) have long been used as a tool to aid the estimation of plant disease severity, an essential variable in phytopathometry. Formal validation of SAD was not considered prior to the early 1990s, when considerable effort began to be invested developing SAD and assessing their value for improving accuracy of estimates of disease severity in many pathosystems. Peer-reviewed literature post-1990 was identified, selected, and cataloged in bibliographic software for further scrutiny and extraction of scientometric, pathosystem-related, and methodological-related data. In total, 105 studies (127 SAD) were found and authored by 327 researchers from 10 countries, mainly from Brazil. The six most prolific authors published at least seven studies. The scientific impact of a SAD article, based on annual citations after publication year, was affected by disease significance, the journal's impact factor, and methodological innovation. The reviewed SAD encompassed 48 crops and 103 unique diseases across a range of plant organs. Severity was quantified largely by image analysis software such as QUANT, APS-Assess, or a LI-COR leaf area meter. The most typical SAD comprised five to eight black-and-white drawings of leaf diagrams, with severity increasing nonlinearly. However, there was a trend toward using true-color photographs or stylized representations in a range of color combinations and more linear (equally spaced) increments of severity. A two-step SAD validation approach was used in 78 of 105 studies for which linear regression was the preferred method but a trend toward using Lin's correlation concordance analysis and hypothesis tests to detect the effect of SAD on accuracy was apparent. Reliability measures, when obtained, mainly considered variation among rather than within raters. The implications of the findings and knowledge gaps are discussed. A list of best practices for designing and implementing SAD and a website called SADBank for hosting SAD research data are proposed.

Quantitative Review of the Effects of Triazole and Benzimidazole Fungicides on Fusarium Head Blight and Wheat Yield in Brazil.

Triazole and benzimidazole fungicides have been used for controlling Fusarium head blight (FHB) in wheat for over two decades. In Brazil, it was only during the last five years that uniform fungicide trials for FHB control have been established yearly, thus contributing to a new large body of fungicide efficacy data for this country. A systematic review of both peer- and non-peer-reviewed studies on chemical control conducted since 2000 in Brazil was performed. Fungicides of interest were the triazoles tebuconazole (TEBU1x and TEBU2x) and propiconazole (PROP2x), and the benzimidazole carbendazim (CARB2x). Most fungicides were applied twice, the first at early to mid-flowering and the second 7 to 10 days later. Only TEBU was tested as one or two applications, and thus four treatments were evaluated. For these fungicides, there were 35 trials reporting FHB index and 48 reporting mean yield. Network meta-analytic models were fitted to the data of the log of the means of FHB index for each fungicide and for the nontreated check. The meta-analytic estimates were used to obtain control efficacy ( C¯ ), or percent disease reduction relative to the nontreated check. The absolute mean difference ( D¯ ) in yield (kg/ha) between the fungicide-treated and the nontreated check plots was also estimated. Yield response relative to the nontreated check ( Y¯ ) was also calculated based on the difference in the logs of the means of yield between fungicide-treated and nontreated check. The TEBU1x, TEBU2x, and CARB2x treatments performed similarly with regards to control efficacy (= 59%, 53% and 55%, respectively), and although better than PROP2x (47%), the difference was marginally significant. Yield response ( D¯ ) was highest for TEBU2x, ( D¯ = 558 kg/ha, Y¯ = 19.2%) followed by PROP2x (497 kg/ha, 16.0%), TEBU1x (457 kg/ha, 17.3%), and CARB2x (456 kg/ha, 12.8%). For an average 2016 scenario of fungicide plus application costs (FC = $18 U.S./ha) and average wheat price (WP = $215 U.S./MT), the probability of breaking even on the financial investment in the four treatments ranged from 59 to 63%. For 140 scenarios (four fungicides) created based on the combination of five WP ($133 to 266 U.S./MT) and seven FC ($5 to 35 U.S./ha), the probability of breaking even was >50% for all but two scenarios. The information may serve as a guide for planning future trials and provides a baseline and first step toward optimizing FHB management in Brazil.