Projects / Programmes
The synergy of knowledge - Development of methods and procedures for the rapid detection and management of bacterial diseases caused by Xylella fastidiosa and its vectors
| Code |
Science |
Field |
Subfield |
| 4.03.05 |
Biotechnical sciences |
Plant production |
Phytomedicine |
| Code |
Science |
Field |
| B006 |
Biomedical sciences |
Agronomics |
| Code |
Science |
Field |
| 4.01 |
Agricultural and Veterinary Sciences |
Agriculture, Forestry and Fisheries |
Plant diseases, quarantine plant pathogens, disease vectors, on-site detection, safe food
Researchers (12)
Organisations (2)
Abstract
The project encompasses (i) optimizing laboratory detection of Xylella fastidiosa, (ii) elucidating occurance and bionomy of vectors (known and potential), (iii) collecting data to support decision making of plant health authorities (inventory of host plants, vectors, recommendations for olive producers and providing selected data for preparation of the national contingency plan) and (iv) raise awarness about the disease and associated riska among differen stakeholders and general public.
The project is designed by combining the special expertise of individual partners: knowledge of X. fastidiosa bacteria and development and use of diagnostic methods (NIB) with a continuous presence in practice (extension), regular contact with growers, knowledge to recognize disease symptoms, conduct of visual inspections and entomological knowledge (KGZSNG).
The two partners are involved in national and international research and technical spheres which will allow for efficient transfer of knowledge among project partners and from project to various stakeholders e.g. professionals and the general public. Already established systems of information and communication will be used for the effective implementation of the proposed project and end-users will be involved in the critical stages of development of recommendations and guidelines. The acquired knowledge will form the basis for making recommendations to growers and a basis of the national contingency plan.
Inventory of plants and vectors, and laboratory testing will focus on the areas of higher risk (Primorska region close to the seaside with higher average annual temperatures and Mediterranean climate) where 10 kilometer squeres will be selected for screening (UTM quadrants) as well as smaller locations selected for a more thorough analysis of vector populations. Dissemination and raising of awarness however, will be targeted to the whole area of Slovenia.
The work package LABORATORY DETECTION is intended to bring the diagnosis closer to the field, provide a fast and simple analysis of symptomatic samples and increase the capacity for laboratory analysis of samples.
Isothermal LAMP, adapted from Harper et al., Phytopath (2010) 100: 1282-1288), be optimized for fluorescent, real-time detection and applied to various plant and insect samples testing different sample preparation approaches. Transfer of method from turbidity detection to fluorescence based detection of the amplified product allows us to monitor amplification in real time using a portable apparatus, and the subsequent verification of the specificity of the resulting product by determining the melting temperature (Tm).
Samples used for verification will include both healthy plant material of various plants and healthy material to which a variety of known and specified amounts of bacteria X. fastidiosa will be added, as well as naturally contaminated material from infected areas in Europe. Vectors will be tested in a similar way. As a control bacteria Xylella fastidiosa CoDiRO strain will be used, which originates from an infected area in the south of Italy. In addition, we will test selected other bacteria, which occur on various host plants and are kept in working bacterial culture collection at NIB. The presence of bacteria in artificially contaminated and natural (not artificially contaminated) samples will be examined in parallel with the real-time PCR to determine the absolute and relative analytical specificity and sensitivity of the analytical method LAMP and most appropriate reaction conditions (temperature and time amplification). We will compare different methods of sample preparation, which will vary according to the complexity and amount of starting material, and choose the most suitable one for use in the field.
One specific aim is to test sample preparation as used for phytoplasma detection also for X. fastidiosa testing. For the determination of these harmful organisms similar plant material is collected (leaf v
Significance for science
The results of the proposed project will significantly contribute to the identification and application of optimal procedures for sample preparation in a variety of host plants and identify their effect of different matrices on the sensitivity of analytical methods to determine X. fastidiosa. The inventory of potential and known vectors will provide a completely new scientific information which is indispensable for the development of procedures and their management will be important to complement the knowledge base on vectors in the Mediterranean and Europe.
Inventory of the potential host plants will enable the review and possible update of the list of host plants that occur in our country and are relevant for visual inspection and/or testing. Also, data will enable a more focused and targeted approach to visual inspections and testing in general.
The data obtained will be useful for other countries with similar geographical conditions and will promote our cooperation in international circles.
Significance for the country
One of the objectives of the project integrates sample preparation for phytoplasma and X. fastidiosa testing. Most European researchers focuse on olive trees and other plants most relevant to the southern Mediterranean, which have already been confirmed as infected in Europe, while the methods that would be appropriate for certain host plants of high importance for Slovenia (eg. for fruit trees) remain unoptimized. With the transfer of knowledge from phytoplasmology we plan to increase the range of host plants in which we can reliably establish the presence of Xylella fastidiosa. In the event of a successful transfer of sample preparation to grapevines, one of the main hosts Xylella fastidiosa, we can significantly increase the number of samples analyzed through joint preparation of samples and simultaneous analysis thus making it economically cheaper and increasing capacity.
Over the course of the project we will build on the expertize on the recognition of host plants, possible symptoms and vector (entomological expertise), and an integrated view of the problems in individual host plants, which will significantly contribute to greater competence in the entire field of plant protection. Involvement of all stakeholders in the project will greatly expand the potential for rapid identification of pathogens by the producers, and consequently reduce the ecological and economic damage.
The results will be included in the documents that will form the basis for the preparation of contingency plans in the event of the entry of X. fastidiosa i.e. they will form the basis for strategic decision-making, policy formulation and / or transfer of knowledge into practice. By supporting the preparation of the contingency plan and the involvement of all actors at the stage of forming recommendations and advice we anticipate an increased their willingness to participate also in the execution part.
The results of the project will also strengthen the preparadness of Slovenia for the possible introduction of X. fastidiosa and thereby contribute to increasing the competitiveness of agriculture and forestry, environmental protection and the conservation of genetic resources. Also, data will enable a more targeted and resources efficient approach to sample analysis.
An effective system for the protection of plants also for X. fastidiosa and demonstration of our activities during the project will increase the confidence of trading partners in Slovenian crops and products. With a focus on the popularization of knowledge we will enhance cooperation between the partner institutions, government departments, producers and end-users of produce and upport the development of value chains and networks (olive trees, ornamental plants).
Most important scientific results
Final report
Most important socioeconomically and culturally relevant results
Annual report
2016,
2018,
final report
