Aquino, S.O.

Botelho, D.M.S.

Guerra-Guimarães, L.

Moreira, S.I.

Pozza, E.A.

Resende, M.L.V.

Santiago, W.D.

Teixeira, A.R.

English

UTL Repository

www.alphavisa.com/asic/2021Posters  - Session 2 - Posters Session 2: Plant pathology & protection22428th Conference of Association for the Science and Information on Coffee28 June - 1 July 2021 Montpellier, FranceBook of abstractsS2-P-13Cercosporin quantification in Cercospora coffeicola isolates by spectrophotometry and high-performance liquid chromatography: a comparative analysisBotelho Deila Magna S1 (deilamagna@hotmail.com), Resende Mário Lúcio V1,  Teixeira Alexandre Rezende1, Santiago Wilder D1, Pozza Edson Ampélio1, Moreira Silvino Intra2, Aquino Sinara Oliveira1, Guerra-Guimarães Leonor*31 Universidade Federal de Lavras, Lavras, Minas Gerais, Brazil ; 2 Universidade Estadual Paulista, Ilha Solteira, São Paulo, Brazil ; 3 CIFC, Instituto Superior de Agronomia, Universidade de Lisboa, Lisboa, Portugal*Corresponding author: leonorguimaraes@edu.ulisboa.ptRATIONALECoffee brown eye spot (BES) is an important disease, caused by the agent Cercospora coffeicola. BES affects all coffee growing stages, from seedlings in nurseries to adult plants, leading to losses up to 30%. Cercosporin is activated by light toxin produced by Cercospora and has been detected in some Pseudocercosporella and Colletotrichum species. Cercosporin has been considered a possible component of aggressiveness in coffee plants. Sensitive and accessible methods for cercosporin detection and quantification are required for better understanding its role in plant pathogenesis. A comparative analysis of cercosporin quantification by spectrophotometry (SPEC) and high-performance liquid chromatography (HPLC) was performed in Cercospora coffeicola isolatesMETHODSTeen monoconidial isolates C. coffeicola were cultured in 9 cm diameter Petri dishes containing 9 mL PDA (potato dextrose agar) culture media and were maintained in BOD incubator (Bio-Oxygen Demand) under 12 h photoperiod at 25 0C for 12 days.Four mycelial plugs (6 mm diameter) removed from each petri dish colony were immersed in 8 mL of 5N KOH and maintained in the dark for 4 hours. Cercosporin determination by SPEC was performed at 480 ηm (Jenns et al. 1989, Yamazaki and Ogawa 1972) and by HPLC was performed based on Gunasinghe et al. (2016).RESULTSThe cercosporin production varied among the C. coffeicola isolates, ranging from 0.01 to 34.52 µM. When comparing cercosporin quantification obtained by SPEC and HPLC for each isolate, although SPEC quantification was always higher than HPLC no significantly differences were obtained (Tukey test, p < 0.05). Furthermore, the Spearman correlation showed a significant linear association between cercosporin quantification values obtained by SPEC and HPLC methods (r=0.94).CONCLUSIONS & PERSPECTIVESBoth methods are equally valid for the cercosporin evaluation from C. coffeicola grown in vitro. The HPLC is an analytical procedure more sensitive, accurate and precise. On the other hand, SPEC is a fast and simple technique involving ordinary lab equipment; the choice of technique depending on lab facilities available.ACKNOWLEDGEMENTSThis work was supported by INCT-Café (Instituto Nacional de Ciência e Tecnologia do Café), Fundação de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG). L Guerra-Guimarães acknowledge CAPES PVE grant.References:• Gunasinghe et al. 2016 Plant Disease 100: 1521-1531 (doi: 10.1094/PDIS-10-15-1192-RE)• Jenns et al. 1989 Phytopathology 79: 213–219• Yamazaki, Ogawa 1972 Agricultural and Biological Chemistry 36:1707–1718

Cercosporin quantification in Cercospora coffeicola isolates by spectrophotometry and high performance liquid chromatography: a comparative analysis

https://www.repository.utl.pt/bitstream/10400.5/23335/1/Botelho%20et%20al_Asic2021.pdf

Cercosporin quantification in Cercospora coffeicola isolates by spectrophotometry and high performance liquid chromatography: a comparative analysis

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