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Anti-Rice Pathogenic Microbial Activity of Persicaria Sp. Extracts

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Anti-Rice Pathogenic Microbial Activity of Persicaria Sp. Extracts P-ISSN 2586-9000 E-ISSN 2586-9027 Homepage : https://tci-thaijo.org/index.php/SciTechAsia Science & Technology Asia Vol. 23 No.4 October - December 2018 Page: [ 32-41 ] Original research article Anti-Rice Pathogenic Microbial Activity of Persicaria sp. Extracts Pragatsawat Chanprapai1,3, Isao Kubo2 and Warinthorn Chavasiri3,* 1Biotechnology Program, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand 2Department of Environmental Science, University of California, Berkley, CA 94720-3112, USA 3Center of Excellence in Natural Products Chemistry, Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand Received 18 July 2018; Received in revised form 5 September 2018 Accepted 14 September 2018; Available online 19 December 2018 ABSTRACT The dichloromethane and methanol extracts, and the essential oil of Persicaria sp. were subjected to in vitro anti-rice pathogenic microbial activity tests. The essential oil displayed the most potential antimicrobial activity. GC-MS analysis revealed thirteen main compounds as dodecanal (54%), decanal (15%), trans-caryophyllene (8%), cyclododecane (7%) and α- humulene (5%). Strong antimicrobial activities of the oil and dodecanal were found against Rhizoctonia solani (IC50 of 0.066 and 0.851 mg/mL) and Xanthomonas oryzae pv. oryzicola (MIC/MBC of 0.78/12.50 and 0.78/25.00 mg/mL), and potent activities against Bipolaris oryzae (IC50 of 3.047 and 3.341 mg/mL) and X. oryzae pv. oryzae (MIC/MBC of 3.12/12.50 and 3.12/25.00 mg/mL). In terms of structure-activity relationship, 2-dodecanone and 2-dodecanol displayed significantly anti-fungal activity, while 1- and 2-dodecanols expressed potent anti- bacterial activity. The essential oil might be used for new microcides controlling rice pathogenic bacteria and fungi. Keywords: Antimicrobial activity; Essential oil; Persicaria sp.; Rice pathogens; Structure activity relationship 1. Introduction edible vegetable Persicaria odorata (Lour) The antimicrobial activities of the Sojak (synonym: Polygonum odoratum Lour; essential oils and the extracts from various Vietnamese coriander) and P. pulchra plants have long been utilized and explored (Blume) Sojak (synonym: Polygonum towards the discovery of innovative tomentosum Lour) were chosen for this study. antimicrobial compounds to control There were main reasons of our study pathogenic microorganisms. The essential oil choosing the plants for the experiment on and the extracts from a Thai cultivar of the antimicrobial activity. Some of them were the *Corresponding author: [email protected] doi: 10.14456/scitechasia.2018.30 P. Chanprapai et al. | Science & Technology Asia | Vol.23 No.4 October – December 2018 major constituents of the essential oils from usually used fungicides and bactericides to P. odorata in cultivars from Vietnam, suppress rice diseases. Although the Singapore, Australia and Thailand reported to chemicals successfully control rice be decanal, dodecanal, 1-decanol, β- pathogenic microorganisms, those also harm caryophyllene and caryophyllene oxide [1-9]. to human and animal health, and The antimicrobial activity of the oil was environment. Therefore, it is a great need to mostly affected on bacteria such as search for sustainable approaches, particularly Salmonella choleraesuis subsp. choleraesuis, natural agents from plants to control rice S. typhi, Methicillin-resistant Staphylococcus pathogenic bacteria and fungi. Many studies aureus (MRSA), Streptococcus pneumoniae, have described that plant essential oil and S. pyogenes, Escherichia coli, Pseudomonas extracts provided the proper answer for this aeruginosa and Bacillus subtilis [5-7, 9-12]. posed problem. Meanwhile, the phytochemicals of the whole In this work, the antimicrobial plant extracts from P. pulchra were reported activities of the essential oil and the extracts as glycosides, tannins, flavonoids, saponins, from two Thai cultivars of P. odorata and P. sterols, terpenes and alkaloids that could only pulchra were evaluated against two fungal combat bacteria including multi-drug resistant species (R. solani and B. oryzae) and two Klebsiella sp. [13, 14]. Therefore, the extracts pathovars of the bacterial species X. oryzae from the plants should be able to control the (Xoc and Xoo) that are significant rice growth of rice diseases. pathogens in Thailand. This is also the first Thailand was previously the largest rice report to indicate the ability of the extracts to exporting country globally and has been decrease rice pathogenic microorganisms. remained in the top three countries. Although rice export is performed entirely by a cartel of 2. Materials and Methods private companies, the government subsidizes 2.1 Plant materials rice and provides reduced control over rice The whole plants of P. odorata were exports because of the considerable role that purchased from Pak Klong Talad Market, rice plays in the welfare of a large sector of Bangkok, Thailand in 2014. The plant was the country’s poorest people [15]. labeled, identified and kept at the herbarium Nevertheless, rice production and quality of Kasin Suvathabandhu Herbarium, Faculty absolutely depended on the environmental of Science, Chulalongkorn University (BCU- factors including weeds, soil problems, Herbarium: BCU 016034, Collector number: environmental stresses, insect pests, Pragatsawat Chanprapai 1). P. pulchra was biological agents, climate changes, relative collected from Phranakorn Sri Ayutthaya humidity, rainfall, leaf wetness effects and province in 2014. The plant was also pathogen have been reported [16-20]. All identified by the herbarium of Kasin factors are thus likely that rice yields in Suvathabandhu Herbarium. Thailand shall be enlarging speedily. From 2.2 Rice pathogens the factors influencing rice production, the Rice pathogenic fungi (R. solani, most crucial factor is rice pathogens caused DOAC1406 and B. oryzae, DOAC1760) and by vires, nematodes, bacteria and fungi [21, bacteria (Xoo, TB0006 and Xoc, TS8203) 22]. Those induced by fungi including sheath were supplied and purchased from the Plant blight (Rhizoctonia solani) and brown spot Pathology Research Group, Plant Protection (Bipolaris oryzae); concurrently, bacterial Research and Development Office, Bangkok, leaf blight (Xanthomonas oryzae pv. oryzae, Thailand. The fungal stock cultures were Xoo) and bacterial leaf streak (Xanthomonas maintained on potato dextrose agar (PDA) at oryzae pv. oryzicola, Xoc) have been the room temperature (25–32oC), while the bacterial diseases recorded [23]. Rice farmers 33 P. Chanprapai et al. | Science & Technology Asia | Vol.23 No.4 October – December 2018 bacteria were maintained on nutrient agar screen for the antibacterial activity of plant (NA) at 28oC. essential oil and its principal components. 2.3 Compounds The two bacterial pathovars (X. oryzae, Xoo Dodecanal (≥95%, main component), and Xoc) were cultivated in NA at 37oC for 1-dodecanol (reagent grade), 1-dodecene 18–24 h. Then four or five single colonies of (≥99%) and 2-dodecanol (99%) were each tested bacteria were selected and purchased from Fluka-Chemika Co cultivated in nutrient broth (NB) at 37oC for (Steinheim, Switzerland). 2-Dodecanone 2–5 h. These bacterial suspensions in NB (≥97%) and 1,2-dodecene oxide (90%) were were then standardized to a cell density of obtained from Sigma-Aldrich, respectively. 1.5x108 colony forming units (CFU)/mL Nativo 750 WG as the positive control was (McFarland No. 0.5) using sterile 0.85% procured from Bayer Co., Bangkok, Thailand. (w/v) NaCl solution. The adjusted bacterial 2.4 Solvent extraction suspension (1 mL) was then mixed into NA Ten kilograms of the whole plants of P. media (19 mL) and poured into Petri plates odorata and P. pulchra were air dried, milled and allowed to set. Wells of 6 mm diameter to fine powder and extracted with were created with a cork border and 40 µL of the dissolved extract in DMSO (or DMSO dichloromethane (CH2Cl2) and then methanol only for the solvent control) was added into (CH3OH) by Soxhlet extraction for three times. Each extract was filtered, evaporated these wells (in duplicate plates) and incubated o and kept at room temperature for at 37 C overnight. The bacterial lawn antimicrobial activities. The percentage yield (opaque) was then observed and the diameter of extracts was presented in Table 1. of the zones of inhibition (clear zone) was measured. 2.5 Hydrodistillation The minimum inhibitory concentration The essential oil was derived as (MIC) was measured using the macro-dilution reported [24]. Briefly, the fresh whole plants broth susceptibility assay. Stock solutions of of P. odorata (1.2 kg) was cut into small the test essential oil and each of the principal pieces and hydrodistilled using a Dean-Stark components were prepared in DMSO and apparatus for 3 h [25-27]. The distillate was then ten-fold serial diluted from 100 mg/mL dried over anhydrous sodium sulfate and to 0.08 mg/mL with sterile NB. Each serial concentrated using rotatory evaporator to dilution (1 mL) was pipetted with 1 mL of leave the essential oil. The pale yellow test bacteria to a concentration of 1.5x108 essential oil was obtained (0.03% yield CFU/mL and incubated at 37oC for 24 h. The (w/w), Table 1). The oil was protected from lowest concentration of the tested compound oxidation and photo-degradation of light by that showed no visible bacterial growth was bottle wrapping with aluminum foil and taken as the MIC and the mean MIC value stored at 4oC until further used. was calculated
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