Phenolic compounds involved in pine wilt disease: HPLC-based method development and validation for their quantification
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ORIGINAL ARTICLE
Phenolic compounds involved in pine wilt disease: HPLC-based method development and validation for their quantification Sara Canas1,2
•
Caˆndida Sofia Trindade3
•
Baoshan Sun1,4
•
Pedro Naves3,5
Received: 8 April 2020 / Accepted: 30 August 2020 Ó Society for Plant Biochemistry and Biotechnology 2020
Abstract Pine Wilt Disease, caused by the pine wood nematode, is a severe and unsolved sanitary problem of pine forests in Europe and Eastern Asia. However, not all Pinus species are affected. Differences in their phenolic pattern may imply different resistance/susceptibility to the nematode, but this topic has not yet been investigated in detail. Therefore, a thorough analysis of phenolic compounds in different Pinus species is crucial to assess their role as markers of resistance/susceptibility to the nematode in order to understand the defense mechanisms against it, and to develop innovative strategies to manage and control wilt disease. A HPLC-based method for the simultaneous quantification of gallic acid, ferulic acid, epicatechin, taxifolin, rutin, resveratrol and secoisolariciresinol in Pinus tissues was developed and validated. The method exhibits good sensitivity, low limits of detection and quantification, good precision, and high specificity and selectivity. The method was successfully applied to the quantification of the above-mentioned phenolic compounds in thirty-six extracts of Pinus halepensis and Pinus sylvestris tissues, before and after nematode inoculation. The results showed that the studied compounds can act as potential interspecific markers of resistance/susceptibility to the pathogen. Hence, the method lends itself readily for application in scientific studies and routine analysis on this sanitary issue. Keywords Phenolic compounds Pine wilt disease Pinus spp. HPLC based-method Abbreviations PWD PWN HPLC
UPLC Pine wilt disease Pine wood nematode High-performance liquid chromatography
& Sara Canas [email protected] 1
Instituto Nacional de Investigac¸a˜o Agra´ria e Veterina´ria, INIAV-Dois Portos, Quinta da Almoinha, 2565-191 Dois Portos, Portugal
2
MED – Mediterranean Institute for Agriculture, Environment and Development, Instituto de Formac¸a˜o Avanc¸ada, Universidade de E´vora, Po´lo da Mitra, Ap. 94, 7006-554 E´vora, Portugal
3
Instituto Nacional de Investigac¸a˜o Agra´ria e Veterina´ria, INIAV-Oeiras, Quinta do Marqueˆs, 2780-157 Oeiras, Portugal
4
School of Functional Food and Wine, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang 110016, China
5
GREEN-IT Bioresources for Sustainability, ITQB NOVA, Av. da Repu´blica, 2780-157 Oeiras, Portugal
GC LC-ESI-FT-ICR-MS
Epicat Ferul Gall Resv Rut Secoi Taxif
Ultra-performance liquid chromatography Gas chromatography Liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry (-)-Epicatechin Ferulic acid Gallic acid Resveratrol Rutin (?)-Secoisolariciresinol Taxifolin
Introduction Pine Wilt Disease (PWD) is caused by the pine wood nematode (PWN), Bursaphelenchus xylo
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