Forging sustainability

PlantBioNem – Plant biotechnology in nematodes invasion mechanism elucidation
Coordinator - Ana Cristina da Silva Figueiredo
CESAM Responsible researcher - Ana Cristina da Silva Figueiredo
Programme - PTDC/AGR-CFL/117026/2010
Execution dates - 2012-01-02 - 2015-01-01 (36 Months)
Funding Entity - FCT - Fundação para a Ciência e a Tecnologia
Funding for CESAM - 135727 €
Total Funding - 176797 €
Proponent Institution - FCiências.ID
Participating Institutions
Universidade de Lisboa
Universidade de Évora

Phytoparasitic nematodes are, nowadays, an alarming, difficult to control, threat to national agriculture and forestry industries. Their life cycle and infection mechanism seems to be dependant on volatile cues, and several phytochemicals have been shown to affect their growth and development. Bursaphelenchus xylophilus (Pinewood Nematode, PWN) along with the members of the genus Globodera (Potato-Cyst Nematodes), Heterodera (Soybean-Cyst Nematodes) and Meloidogyne (Root-Knot Nematodes) and are among the most damaging, and economically devastating, phytoparasites, to agriculture and forestry. Pest control is often a complex process, which relies, generally, on traps and/or pesticides targeted to the insect vector or on synthetic nematicides that are unspecific, expensive and harmful to the environment. In this context, there is an urgent need to find solutions that are safer for the environment and public health, while less costly. The approaches commonly used to study nematode-caused diseases seldom evaluate the real time invasive mechanism. Additionally, the determination of potential phytonematicidal compounds relies mostly on phytonematicide-nematode direct-contact assays, that do not take into account the plant response nor do they assess the toxicity to the plant. Plant in vitro culture is a useful experimental system that can be controlled in terms of nutrition and environmental conditions, providing additional advantages, to conventional systems, in the evaluation of phytochemicals as nematicidals. Additionally, it is an ideal system to study biotransformation based decontamination mechanisms, as well as to investigate the plant-nematode relationships in loco without the need of field assays. In an attempt to clarify the invasive mechanism of different types of nematodes, and to identify the plant response to nematode and to potential nematicides, viewing at obtaining alternative strategies for nematode control, the current project proposes the use of in vitro plant-nematode co-cultures as testing models that simulate the in vivo infection conditions. Particularly, and using - two types of nematodes that adversely affect the national economy, B. xylophilus, responsible for pine wilt disease (PWD), by affecting the aerial parts of plants, and nematodes of the genus Meloidogyne, causative agents of the root galls that destroy the root system, and - in vitro cultures of micropropagated Pinus spp. and Solanum tuberosum hairy roots already established in the course of the PhD work of one of the team members (Jorge Faria, SFRH/BD/43738/2008) We propose: To use two types of in vitro co-culture model systems: co-culture of micropropagated Pinus spp./B. xylophilus and Solanum tuberosum hairy roots/M. chitwoodi, in order to: a) characterize the volatile metabolomic profile resulting from nematode infection and proliferation, b) follow the morpho-anatomical mechanisms of infection, c) elucidate the host and parasite responses to phytonematicide application, and d) evaluate the changes in gene expression as a result of nematode invasion and of phytonematicide application. To successfully carry out the research work, the involved research units will join their skills in the areas related to this study. The main research unit, Instituto de Biotecnologia e Bioengenharia, Centro de Biotecnologia Vegetal, Faculdade de Ciências da Universidade de Lisboa (IBB/CBV/FCUL), has a solid knowledge and know-how in the areas of phytochemistry, plant in vitro culture, morphological and anatomical characterization and molecular biology [1-5]. These abilities coupled with the long experience of the University of Évora (UE), in nematology and assessment of nematicidal activity against B. xylophilus [6-8] [a-e]. The main research unit relies on other institutions, as consultants, for certain stages / phases of the research work, namely, the IMAR, Departamento de Ciências da Vida, Universidade de Coimbra (Prof. Dr Isabel Abrantes) for her scientific knowledge on the genus Meloidogyne, and in whose lab the PhD student already performed a training period on the specific growth conditions of this nematode. The research team will also have the support of Laboratório de Biotecnologia Florestal of Instituto de Tecnologia Química e Biológica (Grupo Pinus/ITQB) (Dra Célia Miguel) currently involved in the transformation of embryogenic Pinus cultures and studying the role of genes potentially involved in response to the nematode. The results will complement a completed project (PTDC/AGR-AAM/74579/2006) in partnership with UTAD an UE, and will allow the successful conclusion of the PhD of one of the team researchers. Taking advantage of the knowledge acquired to date, the expected results will add more knowledge both to scientific community as well as to the public and will follow the national effort to develop strategies, and implement environmentally friendly control measures for the control of damaging phytoparasitic nematodes.

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