Progress 09/01/23 to 08/31/24
Outputs
Target Audience:Mushroom Growers, mushroom industry and general public, graduate and undegraduate students Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Students and researchers presented their research findings at the national conferences How have the results been disseminated to communities of interest?1. Students, researchers and investigators presneted at extension meetings as well as demonstrattions of research during farmer field days. 2. Researchers interacted with muhroom industry. What do you plan to do during the next reporting period to accomplish the goals?We are testing casing layers in combination with different bioactive compounds for their effectivness against mushroom fungi and flies.
Impacts
What was accomplished under these goals?
1.We identified a class of flavonoids present in a sorghum extract and produced through boiling in an acidic medium and a liquid-liquid extraction with the organic solvent isoamyl alcohol. The 3-deoxy flavonoids, along with several possible methylated derivatives were identified. We also explored how the polarity and water content of solvents may affect the extraction of flavonoids from the same sorghum tissue.A 75% concentration of acidified ethanol or acetone in water yielded greater concentrations of anthocyanins compared with our original isoamyl alcohol method. 2.UHPLC-MS profiling of the sorghum flavonoid extracts was performed.UHPLC, with a pump that can go up to 19,000 psi, was used along with a 1.7 µm C18 column with a dimension of 2.1 x 100 mm. Commercially available flavonoids were used as standards for identification of compounds present in the total flavonoid extract. 3. Acetone and Ethanol Gradients for Solvent Comparison. Dried sorghum leaveswereadded to an increasing gradient of acidified acetone or ethanol in dH2O (25,50,75,100 v/v with 0.1% HCl). A ratio of 10 ml of solvent to 1g of fresh plant material was used. The solutions underwent ultrasonic-assisted extraction at 60 Hz for 1 hour using a FS28 Ultrasonic Cleaner (Thermo Fisher Scientific, Waltham, MA, USA). The maceration was then shaken at 100 rpm for 24 hours and the supernatant was filtered off. The filtered supernatant was evaporated using a BUCHI Rotavapor®- R (Brinkmann Instruments, Westbury, NY, USA). The residue was resuspended with a minimal amount of acidified methanol and stored at 4°C for later spectral analysis using a Cytation3 microplate reader (BioTek, Winooski, VT, USA).Absorbance values were analyzed at 520 nm for anthocyanins and 550 nm for flavan-4-ols. Anthocyanins were expressed as µg cyanidin-3-glucoside per gram of dry tissue (MW= 449.2 g mol-1 ; ε extinction coefficient= 26,900). Flavan-4-ols were expressed as absorbance per gram of dry tissue.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2024
Citation:
Lesko T., Wolfin M., Chopra S. 2024. Can Sorghum Flavonoids Help Combat Mushroom Flies? Presented at PSU Graduate School Exhibition. March 27, 2024, State College, PA
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Chatterjee, D., Lesko, T., Peiffer, M., Felton, G., Chopra, S. 2023. Sorghum and maize flavonoids affect the growth and survival of fall armyworm. Poster presentation. In Plant Biology. American Society of Plant Biologist, Savannah, Georgia, Aug 5-9, 2023.
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Progress 09/01/22 to 08/31/23
Outputs
Target Audience:Mushroom scientists,biologist, chemists, and mushroom extension specialists were consulted to discuss the scope of issue of mushroom flies and fungal diseases of mushrooms. We are also interacting with mushroom growers and folks from mushroom industry during the mushroom short course in Kenneth Square, PA and at the International Mushroom Conferences. Graduate and Undergraduate students, technical staff and Post-doctoral scholar are also integral part of this project Changes/Problems:It was noted that spots/residue of flavonoid extract formed at the bottom of several arenas that resulted in death when adult flies contacted them. We are attempting to improve dissolving of sorghum flavonoids in water or ethanol to have homogenous nature of the solution. We are also using less harsh solvents to extract flavonoids. What opportunities for training and professional development has the project provided?Students and researchers are receiving training in plant genetics, flavonoid extractions, mushroom diseases and insect pests. One of the specific areas they are focusing on is to understand the molecular mechanisms that are needed to transcribe flavonoid genes in sorghum. How have the results been disseminated to communities of interest?Students and postdoctoral researchers are able to attend scientific conferences and present their research results. Students have also been interacting with mushroom industry researchers and getting awareness of the issues in mushroom crops. What do you plan to do during the next reporting period to accomplish the goals?Through another project in the lab, we have identified new sources of sorghum flavonoids. We are planning to perform extractions from this new sorghum germplasm. Further, we will be performing LC-MS based identification of compounds and profiling them to check abundances of different flavonoid species.
Impacts
What was accomplished under these goals?
I. We established flavonoid extraction methods from the medium scale biomass from dried leaf tissue of a sorghum line carrying a functional flavonoid pathway genes. The ground tissue was boiled in 2N HCl at a ratio of 1:10 tissue:solvent for 1 hour. After cooling, the solution was centrifuged at 20,000 xg for 15 min, and the supernatant was collected. A liquid-liquid extraction was performed twice using alcoholic solvents and supernatant. was evaporated from the extract using a BUCHI Rotavapor®- R (Brinkmann Instruments, Westbury, NY, USA). The residue was resuspended in a minimal amount of acidified methanol (100%MeOH with 0.1% HCl). Commercial standards of different flavonoids were used to measure the concentrations of different flavonoid species using a UVmini-1240 UV-VIS spectrophotometer (Shimadzu, Kyoto, Japan) at 480 nm. All flavonoid extracts were stored at -20°C until further use. II. The sorghum flavonoid extracts were tested against Lecanicillium fungicola. Sorghum flavonoids reduced bubble count and did not affect yield for infected or uninfected mushroom crops. Treatments receiving sorghum extract negatively affected with bubble counts reduced by half after the third flush. Uninfected treatments that received sorghum extract produced similar yields compared with the control, suggesting resistance by the A. bisporus. III. Contact with flavonoid residue increased mortality rates of mushroom fly pests. The sorghum flavonoid extract's effectiveness was tested against adult flies. Results showed that the extracts killed one-third to over half of the insects tested. During this work, if a sciarid landed on an accumulated spot of residue, it was less likely to break free than its phorid counterparts.A reduction in mortality was observed when the concentration was increased for both the mushroom flies- phorids and sciarids.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
Lesko, T., Chatterjee, D., Paley, K., Beyer, D., and Chopra, S. 2021. Effectiveness of Sorghum Compounds as Biofungicides in Mushroom Cultivation. Poster presented at the International Society for Mushroom Science (ISMS) September 14-17 (online)
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
McCracken, T., Lesko, T., Chopra, S., and Wolfin, M. 2023. Controlling two species of mushroom fly with sorghum Flavonoids. Presented at Gamma Sigma Delta Research Expo. March 17, 2023. Penn State University, University Park, PA.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Effectiveness of Sorghum and Maize Flavonoids Against Pests. Master of Science thesis in Agricultural and Environmental Plant Science. February 2023. Penn State University, University Park, PA.
https://etda.libraries.psu.edu/catalog/29553tkl5215
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