Evaluation of Plant Defense Inducers and Plant Growth Regulators for Fire Blight Management Using Transcriptome Studies and Field Assessments.

Fire blight, caused by Erwinia amylovora, is a destructive disease of pome fruit trees. In the United States, apple and pear growers rely on applications of copper and antibiotics during bloom to control fire blight, but such methods have already led to regional instances of resistance. In this study, we used transcriptome analyses and field trials to evaluate the effectiveness of three commercially available plant defense elicitors and one plant growth regulator for fire blight management. Our data indicated that foliar applications of acibenzolar-S-methyl (ASM; Actigard 50WG) triggered a strong defense-related response in apple leaves, whereas applications of Bacillus mycoides isolate J (LifeGard WG) or Reynoutria sachalinensis extract (Regalia) did not. Genes upregulated by ASM were enriched in the biological processes associated with plant immunity, such as defense response and protein phosphorylation. The expression of several pathogenesis-related (PR) genes was induced by ASM as well. Surprisingly, many differentially expressed genes in ASM-treated apple leaves overlapped with those induced by treatment with prohexadione-calcium (ProCa; Apogee), a plant growth regulator that suppresses shoot elongation. Further analysis suggested that ProCa likely acts similarly to ASM to stimulate plant immunity because genes involved in plant defense were shared and significantly upregulated (more than twofold) by both treatments. Our field trials agreed with the transcriptome study, demonstrating that ASM and ProCa exhibit the best control performance relative to the other biopesticides. Taken together, these data are pivotal for the understanding of plant response and shed light on future improvements of strategies for fire blight management.

Reynoutria Rhizomes as a Natural Source of SARS-CoV-2 Mpro Inhibitors-Molecular Docking and In Vitro Study.

More than a year has passed since the world began to fight the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) responsible for the Coronavirus disease 2019 (COVID-19) pandemic, and still it spreads around the world, mutating at the same time. One of the sources of compounds with potential antiviral activity is Traditional Chinese Medicinal (TCM) plants used in China in the supportive treatment of COVID-19. Reynoutria japonica is important part of the Shu Feng Jie Du Granule/Capsule-TCM herbal formula, recommended by China Food and Drug Administration (CFDA) for treatment of patients with H1N1- and H5N9-induced acute lung injury and is also used in China to treat COVID-19, mainly combined with other remedies. In our study, 25 compounds from rhizomes of R. japonica and Reynoutria sachalinensis (related species), were docked into the binding site of SARS-CoV-2 main protease. Next, 11 of them (vanicoside A, vanicoside B, resveratrol, piceid, emodin, epicatechin, epicatechin gallate, epigallocatechin gallate, procyanidin B2, procyanidin C1, procyanidin B2 3,3'-di-O-gallate) as well as extracts and fractions from rhizomes of R. japonica and R. sachalinensis were tested in vitro using a fluorescent peptide substrate. Among the tested phytochemicals the best results were achieved for vanicoside A and vanicoside B with moderate inhibition of SARS-CoV-2 Mpro, IC50 = 23.10 µM and 43.59 µM, respectively. The butanol fractions of plants showed the strongest inhibition of SARS-CoV-2 Mpro (IC50 = 4.031 µg/mL for R. sachalinensis and IC50 = 7.877 µg/mL for R. japonica). As the main constituents of butanol fractions, besides the phenylpropanoid disaccharide esters (e.g., vanicosides), are highly polymerized procyanidins, we suppose that they could be responsible for their strong inhibitory properties. As inhibition of SARS-CoV-2 main protease could prevent the replication of the virus our research provides data that may explain the beneficial effects of R. japonica on COVID-19 and identify the most active compounds worthy of more extensive research.

Reducing Chlorothalonil Use in Fungicide Spray Programs for Powdery Mildew, Anthracnose, and Gummy Stem Blight in Melons.

Fungicides are applied to nearly 80% of U.S. melon acreage to manage the numerous foliar and fruit diseases that threaten yield. Chlorothalonil is the most widely used fungicide but has been associated with negative effects on human and bee health. We designed alternative fungicide programs to examine the impact of reducing chlorothalonil use (Bravo Weather Stik) on watermelon, cantaloupe, and honeydew melon in 2016, 2017, and 2018 in Maryland. Chlorothalonil was replaced in the tank mix of weekly sprays of targeted fungicides with either polyoxin D zinc salt (Oso) or an extract of Reynoutria sachalinensis (Regalia). Powdery mildew (PM; Podosphaera xanthii), gummy stem blight (GSB; Stagonosporopsis spp.), and anthracnose (Colletotrichum orbiculare) were the most prevalent diseases to occur in the 3 years. Replacing chlorothalonil with the biopesticides as the tank-mix component of the fungicide spray program was successful in reducing GSB and PM severity in cantaloupe, honeydew melon, and watermelon compared with the untreated control, with the exception of GSB in 2017 in cantaloupe, and similar to the program including chlorothalonil in all cases, except anthracnose in watermelon. Anthracnose disease severity was not significantly reduced compared with the untreated control when chlorothalonil was replaced with the biopesticides and yields were not improved over the chlorothalonil-alone treatment in any of the trials. Therefore, replacement of chlorothalonil may not fully address its loss as a fungicide resistance management tool but efficacy can be maintained when polyoxin D is alternated with R. sachalinensis as a tank mix with targeted fungicides to manage PM and GSB.

Simultaneous Determination of Four Compounds, Campesterol, Emodin8-O-ß-D-Glucopyranoside, Quercetin, and Isoquercitrin in Reynoutria sachalinensis by High-performance Liquid Chromatography-Diode Array Detector.

BACKGROUND: Reynoutria sachalinensis is a well-known and used herbal medicine to treatment of arthralgia, jaundice, amenorrhea, coughs, carbuncles, and sores. OBJECTIVE: We have developed high-performance liquid chromatography analysis method for simultaneous determination of isolated four compounds, campesterol, emodin8-O-ß-D-glucopyranoside, quercetin, and isoquercitrin from R. sachalinensis is. MATERIALS AND METHODS: The four compounds were separated on Shiseido C18 column (S-5 µm, 4.6 mm I.D. ×250 mm) at a column temperature of 25°C. The mobile phase composed of water and methanol with gradient elution system, and flow rate is 1.0 ml/min. The detection wavelength was set at 205 nm. RESULTS: Validation of this analytical method was evaluated by linearity, precision, and accuracy test. This established method had good linearity (R2 > 0.997). The relative standard deviation values of intra- and inter-day testing were indicated that <2%, and accuracy is 91.66%-103.31% at intraday and 91.69%-103.31% at intraday. The results of recovery test were 92.60%-108.99%. CONCLUSION: In these results, developed method was accurate and reliable to the quality evaluation of campesterol, emodin 8-O-ß-D-glucopyranoside, quercetin, and isoquercitrin isolated from R. sachalinensis. SUMMARY: We have developed high-performance liquid analysis method for simultaneous determination of 4 compounds of Reynoutria sachalinensis. Abbreviations used: HPLC: High-performance liquid chromatography, DAD: Diode array detector, LOD: Limit of detection, LOQ: Limit of quantitation, ICH: International Conference on Harmonisation.