- Author: Surendra K. Dara
- Author: Dave Peck, Manzanita Berry Farms
Botrytis cinerea infection appears as wilted flowers and a layer of spores on ripe fruit. Photo by Surendra Dara
Botrytis fruit rot or gray mold caused by Botrytis cinerea is an important disease of strawberry and other crops damaging flowers and fruits. Pathogen survives in the plant debris and soil and can be present in the plant tissues before flowers form. Infection is common on developing or ripe fruits as brown lesions. Lesions typically appear under the calyxes but can be seen on other areas of the fruit. As the disease progresses, a layer of gray spores forms on the infected surface. Severe infection in flowers results in the failure of fruit development. Cool and moist conditions favor botrytis fruit rot development. Sprinkler irrigation, rains, or certain agricultural practices can contribute to the dispersal of fungal spores.
Although removal of infected plant material and debris can reduce the source of inoculum in the field, regular fungicide applications are typically necessary for managing botrytis fruit rot. Since fruiting occurs continuously for several months and fungicides are regularly applied, botrytis resistance to fungicides is not uncommon. Applying fungicides only when necessary, avoiding continuous use of fungicides from the same mode of action group (check FRAC mode of action groups), exploring the potential of biological fungicides to reduce the risk of resistance development are some of the strategies for effective botrytis fruit rot management. In addition to several synthetic fungicides, several biological fungicides continue to be introduced into the market offering various options for the growers. Earlier field studies evaluated the potential of various biological fungicides and strategies for using them with synthetic fungicides against botrytis and other fruit rots in strawberry (Dara, 2019; Dara, 2020). This study was conducted to evaluate some new and soon to be released fungicides in fall-planted strawberry to support the growers, ag input industry, and to promote sustainable disease management through biological and synthetic pesticides.
Methodology
This study was conducted at the Manzanita Berry Farms, Santa Maria in strawberry variety 3024 planted in October, 2020. While Captan and Switch were used as synthetic standards, a variety of biological fungicides of microbial, botanical, and animal sources were included at various rates and different combinations and rotations. Products and active ingredients evaluated in this study included Captan Gold 4L (captan) from Adama, Switch 62.5 WG (cyprodinil 37.5% + fludioxinil 25%) from Syngenta, NSTKI-14 (potassium carbonate 58.04% + thyme oil 1.75%) from NovoSource, A22613 [A] (botanical extract) from Syngenta, Regalia (giant knotweed extract 5%) from Marrone Bio Innovations, EXP14 (protein 15-20%) from Biotalys, Gargoil (cinnamon oil 15% + garlic oil 20%) and Dart (caprylic acid 41.7% + capric acid 28.3%) from Westbridge, Howler (Pseudomonas chlororaphis strain AFS009), Theia (Bacillus subtilis strain AFS032321), and Esendo (P. chlororaphis strain AFS009 44.5% + azoxystrobin 5.75%) from AgBiome, ProBlad Verde (Banda de Lupinus albus doce – BLAD, a polypeptide from sweet lupine) from Sym-Agro with Kiplant VS-04 (chitosan 2.3%) or Nu-Film-P spreader/sticker, AS-EXP Thyme (thyme oil) from AgroSpheres, and AgriCell FunThyme (thyme oil) provided by AgroSpheres.
Table 1. List of treatments color coded according to the kind of fungicide (light blue=synthetic fungicide; dark blue=synthetic+biological fungicide active ingredient; peach=synthetic and biological fungicides alternated; green=biological fungicides)
Excluding the untreated control, rest of the 24 treatments can be divided into synthetic fungicides, a fungicide with synthetic + biological active ingredients (a formulation with two application rates), synthetic fungicides alternated with biological fungicides, and various kinds of biological fungicides (Table 1). Treatments were applied at a 7-10 day interval between 22 April and 17 May, 2021. Berries for pre-treatment disease evaluation were harvested on 19 April, 2021. Each treatment had a 5.67'X15' plot replicated four times in a randomized complete block design. Strawberries were harvested 3 days before the first treatment and 3-4 days after each treatment for disease evaluation. On each sampling date, marketable-quality berries were harvested from random plants within each plot during a 30-sec period and incubated in paper bags at outdoor temperatures under shade. Number of berries with botrytis infection were counted on 3 and 5 days after harvest (DAH) and percent infection was calculated. This is a different protocol than previous years' studies where disease rating was made on a 0 to 4 scale. Treatments were applied with a backpack sprayer equipped with Teejet Conejet TXVK-6 nozzle using 90 gpa spray volume at 45 PSI. Water was sprayed in the untreated control plots. Dyne-Amic surfactant at 0.125% was used for treatments that contained Howler, Theia, Esendo, AgriCell FunThyme, AS-EXP Thyme, and EXP 14. Research authorization was obtained for some products and crop destruction was implemented for products that did not have California registration.
Percent infection data were arcsine-transformed before subjecting to the analysis of variance using Statistix software. Significant means were separated using the least significant difference test.
Results
Pre-treatment infection was very low and occurred only in some treatments with no statistical difference (P > 0.05). Infection levels increased for the rest of the study period. There was no statistically significant difference (P > 0.05) among treatments for disease levels 3 or 5 days after the first spray application. Differences were significant (P = 0.0131) in disease 5 DAH after the second spray application where 13 treatments from all categories had significantly lower infection than the untreated control. After the third spray application, infection levels were significantly lower in eight treatments in 3 DAH observations (P = 0.0395) and 10 treatments in 5 DAH observations (P = 0.0005) compared to untreated control. There were no statistical differences (P > 0.05) among treatments for observations after fourth spray application or for the average of four applications. However, there were numerical differences where infection levels were lower in several treatments than the untreated control plots.
In general, the efficacy of both synthetic and biological fungicides varied throughout the study period among the treatments. When the average for post-treatment observations was considered, infection was numerically lower in all treatments regardless of the fungicide category. Multiple biological fungicide treatments either alone or in rotation with synthetic fungicides appeared to be as effective as synthetic fungicides.
Conclusions
Botanical and microbial fungicides can be effective against either for using alone or in rotation with synthetic fungicides for suppressing botrytis fruit rot in strawberry. Additional studies can help optimize the application rates and use strategies for those fungicides that were not as effective as others. Sanitation practices and use of synthetic and biological fungicides help manage botrytis fruit rot.
Acknowledgements: Thanks to AgBiome, AgroSpheres, Biotalys, NovaSource, Sym-Agro, Syngenta, and Westbridge for funding and Chris Martinez for his technical assistance.
References
Dara, S. K. 2019. Five shades of gray mold control in strawberry: evaluating chemical, organic oil, botanical, bacterial, and fungal active ingredients. UCANR eJournal of Entomology and Biologicals. https://ucanr.edu/blogs/blogcore/postdetail.cfm?postnum=30729
Dara, S. K. 2020. Evaluating biological fungicides against botrytis and other fruit rots in strawberry. UCANR eJournal of Entomology and Biologicals. https://ucanr.edu/blogs/blogcore/postdetail.cfm?postnum=43633
- Author: Surendra K. Dara
Botrytis fruit rot or gray mold, caused by Botrytis cinerea, is common fruit disease in California strawberries (Koike et al. 2018). Botrytis cinerea has a wide host range infecting several commercially important crops including blueberry (Saito et al. 2016), grapes (Saito et al., 2019), and tomato (Breeze, 2019). Fungal infection can cause flower or fruit rot. Fruit can be infected directly or through a latent infection in the flowers. Moist and cool conditions favor fungal infections and increased sugar content in the ripening fruit can also contribute to the disease development. Initial symptoms of infection appear as brown lesions and a thick mat of gray conidia is characteristic symptom in the later stages of infection. As chemical fungicides are primarily used for gray mold control, fungicide resistance is a common problem around the world (Panebianco et al., 2015; Liu et al., 2016; Stockwell et al., 2018; Weber and Hahn, 2019). In strawberry, cultural control options such as removing diseased plant material or using cultivars with traits that can reduce gray mold infections may not be practical when the disease is widespread in the field or cultivar choice is made based on other factors. Non-chemical control options are necessary to help reduce the risk of chemical fungicide resistance, prolong the life of available chemical fungicides, achieve desired disease control, and to maintain environmental health. Although there are several botanical and microbial fungicides available for gray mold control, limited information is available on their efficacy in California strawberries. A study was conducted in the spring of 2019 to evaluate the efficacy of several chemical, botanical, and microbial fungicides in certain combinations and rotations to help identify effective options for an integrated disease management strategy.
Methodology
Strawberry cultivar San Andreas was planted late November, 2018 and the study was conducted in April and May, 2019. Each treatment had a 20' long strawberry plot with two rows of plants replicated in a randomized complete block design. Plots were maintained without any fungicidal applications until the study was initiated. Table 1 contains the list of treatments, application rates and dates of application, and Table 2 contains the type of fungicide used and their mode of action. Beauveria bassiana and Metarhizium anisopliae s.l. are California isolates of entomopathogenic fungi, isolated from an insect and a soil sample, respectively. These fungi are pathogenic to a variety of arthropods and some strains are formulated as biopesticides for arthropod control. However, earlier studies in California demonstrated that these fungi are also known to antagonize plant pathogens such as Fusarium oxysporum f.sp. vasinfectum Race 4 (Dara et al., 2016) and Macrophomina phaseolina (Dara et al., 2018) and reduce the disease severity. To further evaluate their efficacy against B. cinerea, these two fungi were also included in this study alternating with two chemical fungicides.
Treatments were applied with a CO2-pressurized backpack sprayer using 66.5 gpa spray volume. Five days before the first spray application and 3 days after each application, all ripe fruit were harvested from each plot and incubated at the room temperature in vented plastic containers. The level of gray mold on fruit from each plot was rated using a 0 to 4 scale (where 0=no disease, 1=1-25% fruit with fungal infection, 2=26-50% infection, 3=51-75%, and 4=76-100%) 3 and 5 days after each harvest (DAH). Due to the rains, fruit could not be harvested after the 3rd spray application for disease rating, but was harvested and discarded after the rains to avoid cross infection for the following week's harvest. Data were analyzed using analysis of variance using Statistix software and significant means were separated using Least Significant Difference separation test.
Results
Gray mold occurred at low to moderate levels during the study period. Along with B. cinerea, there were a few instances of minor fungal infections from Rhizopus spp. (Rhizopus fruit rot) and Mucor spp. (Mucor fruit rot). Pre-treatment disease ratings were statistically not significant (P = 0.6197 and 0.5741) 3 and 5 DAH. While the chemical standard treatment with the rotation of Captan, Merivon, Switch, and Pristine (treatment 2) appeared to result in the lowest disease rating throughout the observation period, treatments 3 and 5 after the 1st spray application, treatments 5 and 11 along with 3, 4 and 6 after the 2nd spray application, and treatments 3 and 5 along with 11 after the 4th spray application also had similar disease control at 3 DAH. When disease at 5 DAH was compared, the lowest rating was seen in treatment 2 after the 1st and 2nd spray applications, and treatments 2, 3, and 11 after the 4th application. Several other treatments also provided statistically similar control during these days.
When the average disease rating for the three post-treatment observation events was considered, treatment 2, 3, 5, and 11 had the lowest disease at both 3 and 5 DAH. Treatments 4 and 12 at 3 DAH also had a statistically similar level of disease control to treatment 2.
In general, most of the treatments provided moderate to high control compared to the disease in untreated control when the post-treatment averages were considered. Only treatment 7 and 13 had lower control at 3 DAH.
Discussion
This study compared a variety of registered and developmental products along with two entomopathogenic fungi in managing B. cinerea. Considering the fungicide resistance problem in B. cinerea in multiple crops, having multiple non-chemical control options is very important to achieve desirable control with integrated disease management strategies. Since the active ingredients in the botanical and bacterial fungicides used in this study are not public, discuss will be limited on their modes of action and efficacy at this point. Similarly, the active ingredient of WXF-17001 is also not known, however, an earlier study by Calvo-Garrido et al. (2014) demonstrated that a fatty acid-based natural product reduced B. cinerea conidial germination by 54% and disease severity in grapes by 96% compared to untreated control. The product used by Calvo-Garrido et al. (2014) is thought to be fungistatic and reduce the postharvest respiratory activity and ethylene production in fruits.
While chemical fungicides have a specific mode of action, biological and other products act in multiple manners either directly antagonizing the plant pathogen or by triggering the plant defenses. For example, amending the potting medium with biochar resulted in induced systemic resistance in tomato and reduced B. cinerea severity by 50% (Mehari et al., 2015). Luna et al. (2016) also showed that application of β-aminobutyric acid and jasmonic acid promoted seed germination and long-term resistance to B. cinerea in tomato. Burkholderia phytofirmans, beneficial endophytic bacterium, offered protection against B. cinerea in grapes by mobilizing carbon resources (callose deposition), triggering plant immune system (hydrogen peroxide production and priming of defense genese), and through antifungal activity (Miotto-Vilanova et al. 2016). Similarly, entomopathogenic fungi such as B. bassiana are also known to induce systemic resistance against plant pathogens (Griffin et al. 2006). Compared to other options evaluated in the study, entomopathogenic fungi have an advantage of controlling both arthropod pests and diseases, while also having plant growth promoting effect (Dara et al. 2017).
Rotating fungicides with different mode of actions reduces the risk of resistance development and using some combinations will also maintain control efficacy. This study provided the efficacy of multiple control options and their combinations and rotations for B. cinerea. This is also the first study demonstrating the efficacy of entomopathogenic fungi against B. cinerea in strawberry.
Acknowledgements: Thanks to Sipcam Agro and Westbridge for funding the study, technical assistance of Hamza Khairi for data collection, and the field staff at the Shafter Research Station for the crop maintenance.
References
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