caa a22 4500 606208836 CHVBK 20210128101025.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00425-015-2257-9 doi (NATIONALLICENCE)springer-10.1007/s00425-015-2257-9 In vitro co-cultures of Pinus pinaster with Bursaphelenchus xylophilus [Elektronische Daten] a biotechnological approach to study pine wilt disease [Jorge Faria, Inês Sena, Inês Vieira da Silva, Bruno Ribeiro, Pedro Barbosa, Lia Ascensão, Richard Bennett, Manuel Mota, A. Figueiredo] Main conclusion : Co-cultures of Pinus pinaster with Bursaphelenchus xylophilus were established as a biotechnological tool to evaluate the effect of nematotoxics addition in a host/parasite culture system. The pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD), was detected for the first time in Europe in 1999 spreading throughout the pine forests in Portugal and recently in Spain. Plant in vitro cultures may be a useful experimental system to investigate the plant/nematode relationships in loco, thus avoiding the difficulties of field assays. In this study, Pinus pinaster in vitro cultures were established and compared to in vivo 1year-old plantlets by analyzing shoot structure and volatiles production. In vitro co-cultures were established with the PWN and the effect of the phytoparasite on in vitro shoot structure, water content and volatiles production was evaluated. In vitro shoots showed similar structure and volatiles production to in vivo maritime pine plantlets. The first macroscopic symptoms of PWD were observed about 4weeks after in vitro co-culture establishment. Nematode population in the culture medium increased and PWNs were detected in gaps of the callus tissue and in cavities developed from the degradation of cambial cells. In terms of volatiles main components, plantlets, P. pinaster cultures, and P. pinaster with B. xylophilus co-cultures were all β- and α-pinene rich. Co-cultures may be an easy-to-handle biotechnological approach to study this pathology, envisioning the understanding of and finding ways to restrain this highly devastating nematode. Springer-Verlag Berlin Heidelberg, 2015 Maritime pine nationallicence Monoxenic culture nationallicence Pinewood nematode nationallicence Relative water content nationallicence Shoots structure nationallicence Volatiles nationallicence BAP : 6-Benzylaminopurine nationallicence DAI : Days after inoculation nationallicence EPPO : European and Mediterranean Plant Protection Organization nationallicence GC : Gas chromatography nationallicence GC-MS : Gas chromatography coupled to mass spectrometry nationallicence IBA : Indole-3-butyric acid nationallicence LM : Light microscopy nationallicence PAS : Periodic acid-Schiff's reagent nationallicence PWD : Pine wilt disease nationallicence PWN : Pinewood nematode (Bursaphelenchus xylophilus) nationallicence RI : Retention index nationallicence RWC : Relative water content nationallicence SEM : Scanning electron microscopy nationallicence SH : Schenk and Hildebrandt medium nationallicence SHe : Schenk and Hildebrandt elongation medium nationallicence SHm : Schenk and Hildebrandt multiplication medium nationallicence t : Trace nationallicence Faria Jorge Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut Sena Inês Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut Vieira da Silva Inês Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut Ribeiro Bruno NemaLab/ICAAM, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra, Ap. 94, 7002-554, Évora, Portugal aut Barbosa Pedro NemaLab/ICAAM, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra, Ap. 94, 7002-554, Évora, Portugal aut Ascensão Lia Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut Bennett Richard Universidade de Trás-os-Montes e Alto Douro, Quinta dos Prados, Apartado 1013, 5000-911, Vila Real, Portugal aut Mota Manuel NemaLab/ICAAM, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra, Ap. 94, 7002-554, Évora, Portugal aut Figueiredo A. Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut Planta Springer Berlin Heidelberg 241/6(2015-06-01), 1325-1336 0032-0935 241:6<1325 2015 241 425 https://doi.org/10.1007/s00425-015-2257-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00425-015-2257-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Faria Jorge Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut NATIONALLICENCE 700 1- Sena Inês Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut NATIONALLICENCE 700 1- Vieira da Silva Inês Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut NATIONALLICENCE 700 1- Ribeiro Bruno NemaLab/ICAAM, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra, Ap. 94, 7002-554, Évora, Portugal aut NATIONALLICENCE 700 1- Barbosa Pedro NemaLab/ICAAM, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra, Ap. 94, 7002-554, Évora, Portugal aut NATIONALLICENCE 700 1- Ascensão Lia Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut NATIONALLICENCE 700 1- Bennett Richard Universidade de Trás-os-Montes e Alto Douro, Quinta dos Prados, Apartado 1013, 5000-911, Vila Real, Portugal aut NATIONALLICENCE 700 1- Mota Manuel NemaLab/ICAAM, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra, Ap. 94, 7002-554, Évora, Portugal aut NATIONALLICENCE 700 1- Figueiredo A. Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, CESAM, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016, Lisbon, Portugal aut NATIONALLICENCE 773 0- Planta Springer Berlin Heidelberg 241/6(2015-06-01), 1325-1336 0032-0935 241:6<1325 2015 241 425