Innovative technologies for biological protection of mediterranean crops from telluric pathogens and parasites

Place: Bari, italy, Europe
Sustainable management of natural resources Sustainable management of natural resources
Total Budget: € 190.575,00 | Period: From November 2009 To October 2013


Characterization and formulation of useful rhizosphere and soil microorganisms, active against phytoparasitic
nematodes (Heterodera, Meloidogyne, Globodera spp.) and fungi (Pyrenochaeta lycopersici, Phytophthora or
Armillaria spp.) to develop sustainable practices for biological control and management in organic agriculture.
1a) Collection and isolation of antagonists, divulgation
1b) Collection and biological and molecular characterization of antagonists
1c) Identification and evaluation of biological antagonists
1d) Development of models
2a) Formulation of substrates with soil organisms or other microbial matrices
2b) Experimental assays in greenhouse for evaluation of formulates
2c) Field experimental assays for evaluation of formulates against soil nematodes and fungi, divulgation


National Research Council, Institute for Sustainable Plant Protection - Italy

Lead applicant

The Italian National Research Council (Consiglio Nazionale delle Ricerche, CNR, http://www.cnr.it) is a public Agency whose role is to carry out, promote, disseminate, trasfer and add value to research activities in the main sectors of knowledge, to increase scientific, technological, economic and social growths. The Institute for Sustainable Plant Protection (IPSP) is part of the Agricultural and Food Department of CNR. IPSP is located in the CNR area of research in Sesto Fiorentino (Florence) with Organizational Support Units in Bari, Turin and Portici. The mission of IPP is the study of plants biotic and abiotic stress factors, in order to identify resistance mechanisms and methods of defense against stress and protection, to improve quality and quantity of agricultural food production, for a sustainable and environmentally friendly growth. The activities are divided into 5 IPSP research lines.

Italian Association for Organic Agriculture - Italy

Initiative partner

AIAB is an association of producers, engineers and citizen-consumers. It is the networking of the organic movement and represents primarily the interests of organic producers, through the promotion of organic agriculture as a sustainable development model, based on the principles of conservation and exploitation of resources, the environment, animal welfare and the health of the consumer. Organic farming is, in fact, a model of development for the Italian countryside alternative to '"industrial agriculture", capable of addressing the ecological sense in the behavior of market participants and general public and, in particular, an approach to the method of production and consumption. The interests of organic producers affect the interests of citizen-consumers and enhance the role and function of the technicians that specialize in organic farming.

National Researc Council, Institute for Mediterranean Agricultural and Forestry Systems - Italy

Initiative partner

ISAFOM research strategic objective is contributing to the sustainable development of agricultural and forestry systems as concerns the effects of climate changes through research lines integrating specific aspects of biological and ecological balances for optimal conservation of forest and agricultural resources, in regions with Mediterranean climate. Further researches concern: the enhancement and innovation of agri-food and forestry environments in Mediterranean regions, with particular attention to their overall quality; the cultivation and use of non-food products and by-products of food processing for energy purposes; the study on the effects of the atmosphere and climate on present and future productivity and on natural and human interactions with crop environments using aero-flow measurement platforms; the monitoring, advanced planning, inventory and evaluation of ecosystem services of forest resources. ISAFOM is central in the scientific debate on sustainable agricultural and forestry systems and transfer of research results.

Agriculture Research and Experiment Council, Plant Pathology Research Centre - Italy

Initiative partner

CRA PAV deals with the protection of agricultural and forestry plants by biotic agents (bacteria, fungi, viruses, viroids, phytoplasmas, etc..) and abiotic stress (physiological disorders, adverse climatic conditions). Studies are carried out on the interaction between plants and pathogens, on the epidemiology and characterization of populations of pathogenic organisms using traditional methods and molecular biology to control diseases. Researches focus on the genetic sources of resistance to pests and tolerance, to be applied to the development of varieties / lines resistant through the use of conventional methods and biotechnology. CRA PAV investigates crop protection methods in the context of conventional, integrated and organic practices. Attention is given to methods useful in the diagnosis of disease prevention, to techniques and methods of treatment which may minimize the occurrence of pesticide residues in food and the environment. Applied projects aim at remediation techniques for the production of propagation material certified free of pathogens.

The project aims at the development of innovative technologies for biological control of pests and soil pathogens (fungi and nematodes) of Mediterranean vegetable crops (carrot, potato and tomato in Puglia, Lazio and Sicily) and forest areas in the Apennines (walnut and chestnut in Tuscan-Emilian Apennines). Formulations are developed based on micro-organisms of the rhizosphere. The parasites are: Heterodera carotae, Meloidogyne and Globodera spp. for nematodes and Pyrenochaeta lycopersici, agent of tomato and vegetables corky root rot; Phytophthora and Armillaria spp. on trees. The antagonists considered wre: nematophagus bacteria and fungi, predatory nematodes (Diploasteridae); bacteria and Trichoderma spp. against soil-borne fungi; fungi and nematodes myco- or bacteriophagous against pathogenic fungi (Phytophthora, Armillaria). At the molecular characterization follows the production of formulates, with essays in the field of molecular probes and descriptive models.

Main technologies for soil-borne pathogens and pests like fungi or nematodes relied on
soil fumigants. We propose innovative organic approaches respectful of the environment,
as alternative to chemicals. At the end of the trial we have available: 1) new tools for plant
protection based on microorganisms; 2) data on the biology, sequencing and molecular
probes for monitoring of biological antagonists; 3) collections of micro-organisms; 4)
experimental data from biological control assays on nematodes and fungi; 5) prototype
formulations; 6) molecular probes and non-linear predictive or descriptive models.

Model describing the effect of the fungus Pochonia chlamydosporia against p Effect of the parameter sigma on parasitism and biocontrol efficacy. The parameter controls the fungus saprotrophic growth. Increasing the fungus density in soil as a saprotroph also increases nematode parasitism

The Italian Ministry of Agricultural and Forestry Policies supports research on organic farming and
food safety through projects based on the National Program for Research in Organic Agriculture.
Project BIOMED reflects the mentioned program, starting from principles such as
multifunctionality of agriculture, sustainability, reduced use of fossil fuels, reuse of organic waste
or secondary materials, improvement of cultivation techniques and protection of plant and natural
antagonists diversity. The research focuses on new techniques to produce scientific and technical
data, integrating lab and field assays. The connection between research and production includes
knowledge for biopesticides efficacy and safety, product quality, organic biomass recycling and
respect for the environment. BIOMED goal is the substitution of high environmental impact
technologies (fumigations), by others based on knowledge about the organisms and
environments involved.

Eggs of a plant parasitic nematode parasitized by a fungus Nematophagous fungi like Pochonia or Lecanicillium spp. are capable to digest the eggs of many invertebrate pests, including insects or nematodes parasitizing plants.

Products or deliverables available:
- Line 1: Microorganisms and methods for containment of soil parasites: isolation of fungal antagonists
of nematodes; of Pasteuria penetrans and Pasteuria spp. and Trichoderma spp. antagonistic of
Pyrenochaeta lycopersici, Phytophthora cinnamomi and Armillaria; collections of microorganisms;
biological and molecular characterization of antagonists and sequencing; descriptive models of
host-parasite relationships and derived data; selection and production of biological antagonists.
- Line 2: Evaluation of the ability of parasite-repressive organic formulates: data on formulations,
micro-organisms or antagonists; production methods of organic substrates for field trials; data from
greenhouse trials with organic and / or antagonists against nematodes and fungi; biological control of
nematodes, P. lycopersici or P. cinnamomi.

../file-system/small/pdf ../file-system/small/pdf Document Hajer R., Ciancio A., Horrigue R.N. & Rosso L. C. 2011. Detection and biocontrol potential of Verticillium leptobactrum parasitising Meloidogyne spp. World J. Appl. Microbiol. Biotechnol. 27: 1615-1623
../file-system/small/pdf ../file-system/small/pdf Document Colagiero M., Rosso L. C., Ciancio A., Murga-Gutierrez S.N.2011. Observations on the biology of a predatory nematode belonging to Diplogasteridae. Redia 94: 133-135.
../file-system/small/pdf ../file-system/small/pdf Document Infantino A., Di Giambattista G., De Felice S. 2011. Valutazione di isolati di Trichoderma spp. per il controllo della suberosi radicale del pomodoro. Atti I Congresso Nazionale della Rete Italiana per la Ricerca in Agricoltura Biologica - RIRAB. 7-8 novembre 2011, Catania (Italia): p. 104
../file-system/small/pdf ../file-system/small/pdf Document Ciancio A., Colagiero M., Rosso L. 2011. Modeling trophic interactions in the rhizosphere of horticultural crops for application of fungi and other biological antagonists of nematodes. Atti I Congresso Nazionale della Rete Italiana per la Ricerca in Agricoltura Biologica - RIRAB. 7-8 novembre 2011, Catania (Italia): p. 35
../file-system/small/pdf ../file-system/small/pdf Document Ciancio A., Colagiero M., Rosso L.C., Murga Gutierrez S.N., Grasso G. 2012. Molecular analysis and morphology of Hirsutella tunicata sp. nov. (Ophiocordycipitaceae), a novel mite parasite from Peru. Mycoscience 54: 378-386.
../file-system/small/pdf ../file-system/small/pdf Document Ciancio A. 2012. Modelling approaches in nematode ecology studies. Nematropica 42: 380
../file-system/small/pdf ../file-system/small/pdf Document Ciancio A., Miranda I. 2012. Density-independent nematode regulation mechanisms of soil fungi with saprotrophic-parasitic behaviours. Nematropica 42: 376
../file-system/small/pdf ../file-system/small/pdf Document Rosso L. C., Colagiero M., Salatino N., Ciancio A. 2012. Effect of nutritional conditions on gene expression of the nematode parasitic fungus Pochonia chlamydosporia. Nematropica 42: 401
../file-system/small/pdf ../file-system/small/pdf Document Rosso L. C., Colagiero M., Salatino N., Ciancio A. 2014. Effect of trophic conditions on gene expression of Pochonia chlamydosporia. Annals of Applied Biology, 164: 232-243.

Scientific benefits include new knowledge on the biology and epidemiology of soil pests and pathogens, of
their antagonists and on biological control in different Italian environments. Benefits concern the availability,
for producers and industries active in the field of biopesticides, of prototype formulations for biological
protection of carrot, potato, tomato, walnut and chestnut, and the technology transfer to industries. They also
include the development of organic technologies for protection of chestnut and walnut with amendments and
their diffusion, due to the impact of these crops on the economy of rural areas with low agricultural
intensification. Main social impact is linked to the development of organic farming and food productions.
Results dissemination is a second social impact, promoting biological technologies between farmers and
producers and a constructive exchange among researchers involved in different research lines and operators
or users (technicians, farmers).

Effect of Trichoderma sp. on Pyrenochaeta lycopersici To test for efficacy of biocontrol, isolates of Trichoderma sp. are tested in vitro against main fungal targets, in this case the agent of the tomato corky rot, Pyrenochaeta lycopersici.
Effect of chicken manure against isolates of Phytophthora Results from in vitro tests of chicken manure on radial growth of Phytophthora.
../file-system/small/pdf ../file-system/small/pdf Document Ciancio A. 2011. La lotta biologica contro nematodi e funghi del terreno. Bioagricoltura 125: 39-41

Unità Coordinatrice: CNR, Istituto per la Protezione Sostenibile delle Piante.
Coordinator: Aurelio Ciancio, IPP CNR, Via Amendola 122/D, 70126 Bari; E-mail: a.ciancio@ba.ipp.cnr.it
1) CNR, Istituto per la Protezione Sostenibile delle Piante, Bari and Florence. Researchers (5 people): A. Ciancio, Laura Rosso,
Mariantonietta Colagiero, Isabella Pentimone; Tullio Turchetti. Technician (1 people): Nicola Salatino.
2) CNR, Istituto per i Sistemi Agricoli e Forestali del Mediterraneo, Catania. Researchers (2 people) Ezio Riggi, M. Cementero.
Str. V. Lancia, Blocco Palma I, Zona Industriale, 95121 Catania; e-mail: ezio.riggi@cnr.it
3) AIAB, Associazione Italiana per l’Agricoltura Biologica, Roma (2 people). Cristina Micheloni, Livia Ortolani.
Via Piave 14, 00187 Roma, e-mail: c.micheloni@aiab.it
4) CRA Patologia Vegetale, Roma. Resp.: Alessandro Infantino (1 people)
CRA PAV, Via C. G. Bertero, 22, 00156 Roma; e-mail: alessandro.infantino@entecra.it

Field and greenhouse trials in Sicily Experimental assays for control of root-knot nematodes with organic products

Dealing with living organisms to produce a product offering a standard service for crop protection does
not always warrant an efficacy or stability of results. This is the case of predatory nematodes, that in
spite of their direct impact on a target nematode prey, appear difficult to rear in vitro for practical
exploitation, due to a susceptibility to their own pathogens and antagonists. Also the isolates of fungi or
bacteria often show differences as concerns their suitability as basic ingredients of biopesticides, due to
variable genetic effects or compatibility, and require in vitro testings for selection, that are time

Predatory nematode Koernersia sp. was tested for biocontrol applications. The nematode, however, is subject to its own bacterial antagonists affecting its production in vitro

Two fungi, Pochonia chlamydosporia and Trichoderma virens, with related species, are close to the
step of becoming the basic ingredients of industrial biopesticides or ammendants, due to their positive
impacts on protection of vegetables in Mediterranean horticulture. Pochonia furthermore is not only a
nematode killer, but also a plant growth promoter, due to the induction of resistance gene in plants that
it can colonize as an endophyte. Organic approaches to pest and disease management like this one
protect soil microbial biodiversity and save energy, relying on living rganisms. Similarly, the
management of tree crops like chestnut and walnut with organic matters, including chicken manure,
showed a positive impact for management of ink disease and other fungal based pathologies for which
no simple, cost effective management tools were previously available. It also extends and completes
the lifecycle of organic manure, offering a safe method for its recycling.

Table 1. Antagonistic activity of Trichoderma spp. against the agent of cor Results of in vitro tests for determining the antagonistic activity of Trichoderma spp. against the agent of corky root rot, P. licopersici.

There are two type of fallouts: the first is mainly scientific, related to the publications describing the
new knowledge produced, and to the information acquired, which is now integrated in further scientific
initiatives. The second fallout is technical, promoting new approaches for sustainable pest and
disease management, in the field. First fallouts will be durable in time given they are published in
international journals and are also the basis for new ongoing projects or proposals, dealing with
Pochonia or Trichoderma. The second fallout is expected to increase the demand of safer organic
tools for pest and disease management, possibily traducing the experience gained in field trials in new biopesticide industrial products.


Articles, dissemination papers and web pages.
Meetings: a first kick off meeting at CNR (Jan. 2010, Rome). An presentation by AIAB at MIPAAF organic farming conference (July
2010). A meeting organized by AIAB in Rome (July 2011) for programming and dissemination, including production of articles.
Data presented at:
I and II RIRAB Meetings, Catania (November 2011) and Rome (June 2013)
X Congress of the Italian Society of Nematology, Portici (2010);
XLIV and XLV Meetings of ONTA (Organization of Tropical Nematologista of America), Cancun (2012) and La Serena (2013);
6th International Nematology Congress, Cape Town (2013).

BIOMED was presented as lecture of series: "Impacto Ambiental y Conservación del Territorio y el Paisaje: el Mediterráneo", by Instituto para el Estudio multidisciplinar del medio ambiente "Ramon Margalef", Univ. of Alicante and CASA MEDITERRÁNEO, consortium of Spain Ministry of Foreign Affairs, Gener. Valenciana and Municipality of Xàbia, Benidorm and Alicante.

../file-system/small/docx ../file-system/small/docx Web links Web links