**5. Phytosanitary certification program: The Italian experience in the last twenty years**

The production of healthy, high-quality olive products depend to a large extent on the quality of the plant material used for olive cultivation. In general, the production of "healthy" plants for planting occurs through defined certification procedures by which a particular cultivated selection, whose health status and trueness to type have been officially attested, is propagated following specific requirements. In a typical certification scheme, the certified material is descended by a defined number of propagation steps from individual plants, found to be free from pathogens and pests, maintained and propagated under rigorous conditions that exclude recontamination.

To this end, appropriate regulations are necessary to ensure the production, marketing and movement of certified plant propagation material with high standards and free from harmful pathogens (Annex I). Over the past twenty years, through globalisation and the expansion of several crops in new areas, concerns were raised about new disease emergencies transmitted by infected propagating material. In an attempt to limit the potential impact of the spread of pests and pathogens through the movement of infected plants, specific laws have been issued at regional, national and European levels.

The Council Directive 2008/90/EEC of 29 September 2008 (recast version of Directive 92/34/EEC) on the marketing of fruit plant (including olive) propagating material and fruit plants intended for fruit production established a harmonized Community regime which ensures that growers throughout the Community receive propagating material and fruit plants which are healthy and of good quality. This applies to fruit plant propagating material and fruit plants of genera and species listed in Directive Annex I, which may only be marketed if they are either CAC (*Conformitas Agraria Communitatis*), Pre-basic, Basic or Certified material.

To be classified as such, material must comply with the criteria of quality, plant health, testing methods and procedures, propagation systems and varietal aspects and must have been recognised following official inspections. In addition, propagating material or fruit plants may, in most circumstances, only be marketed by accredited suppliers, whose production methods and facilities meet the requirements of the Directive. Propagating material and fruit plants from Countries outside of the European Union (EU) may only be marketed within the Community if they offer the same guarantees as materials produced in the Community complying with Council Directive 92/34/EEC. Each EU Member State adopts its own enforcement and implementation policies using the EU Directive as a guide.

120 Olive Germplasm – The Olive Cultivation, Table Olive and Olive Oil Industry in Italy

simultaneously with four other olive viruses (Bertolini et al., 2003).

rigorous conditions that exclude recontamination.

**twenty years** 

Certified material.

Diagnosis of *V. dahliae* is preliminarily performed through an accurate search for foliar symptoms and vascular browning. Foliar chlorosis and necrosis could be due to other causes such as root rot diseases, whereas browning on cross section of stems was sometimes not found. Conclusive detection is attempted by isolating the fungus on agar media from olive tissues and possibly using PCR or nested PCR with *V. dahliae* specific primers (Nigro et al., 2002; Mercado-Blanco et al., 2002) or by Real Time Scorpion PCR (Schena et al., 2004). Identification of *P. savastanoi* pv. *savastanoi* is very easy when the typical knots are present on plants. Nevertheless, its presence in latent and systemic form has been reported by Penyalver et al., (2006). The presence of *P. savastanoi* pv. *savastanoi* both as epiphytic and entophytic agent makes its control in the certified material absolutely compulsory; moreover, for a more sensitive and reliable diagnosis, molecular techniques are needed. Recently, molecular protocols of nested-PCR have been set up to obtain reliable diagnoses of latent infections (Bertolini et al., 2001b). This is also possible (and has been done)

**5. Phytosanitary certification program: The Italian experience in the last** 

The production of healthy, high-quality olive products depend to a large extent on the quality of the plant material used for olive cultivation. In general, the production of "healthy" plants for planting occurs through defined certification procedures by which a particular cultivated selection, whose health status and trueness to type have been officially attested, is propagated following specific requirements. In a typical certification scheme, the certified material is descended by a defined number of propagation steps from individual plants, found to be free from pathogens and pests, maintained and propagated under

To this end, appropriate regulations are necessary to ensure the production, marketing and movement of certified plant propagation material with high standards and free from harmful pathogens (Annex I). Over the past twenty years, through globalisation and the expansion of several crops in new areas, concerns were raised about new disease emergencies transmitted by infected propagating material. In an attempt to limit the potential impact of the spread of pests and pathogens through the movement of infected

The Council Directive 2008/90/EEC of 29 September 2008 (recast version of Directive 92/34/EEC) on the marketing of fruit plant (including olive) propagating material and fruit plants intended for fruit production established a harmonized Community regime which ensures that growers throughout the Community receive propagating material and fruit plants which are healthy and of good quality. This applies to fruit plant propagating material and fruit plants of genera and species listed in Directive Annex I, which may only be marketed if they are either CAC (*Conformitas Agraria Communitatis*), Pre-basic, Basic or

plants, specific laws have been issued at regional, national and European levels.

In Italy, it is compulsory that the production and marketing of olive propagating material fulfils the requirements established by the Italian Decree amended on 14/04/1997 in compliance with the aforementioned EU Directive. According to this law, assessment of true to type and certification of sanitary status of olive propagating materials are compulsory and plants are certified as CAC. In this kind of certification plants must be free from *Euzophera pinguis*, *Meloidogine* spp., *S. oleae*, *P. savastanoi* pv. *savastanoi*, *V. dahliae* and all known viruses (15 to date). However, several aspects concerning for example the procedures for inspections and controls are not well defined, leading to misinterpretations and heterogeneous application of the law in the different Italian regions by the regional Phytosanitary Services. Besides this compulsory system, which relies on the propagation of olive mother plants identified by the single nurseryman and found free from the target pathogens, a voluntary certification system has been activated since 1987 at the national level by the Ministry of Agriculture. During the last 10 years, the legislation has been revised, and in late 2006 the revision process was completed with the publication of 2 decrees concerning the revised organisation of the system (DM 24/7/2003, DM 4/5/2006) and 4 decrees concerning the updated official technical operations for the certification of pommes, stone fruit, olive and strawberry (DM 20/11/06).

The DM 20/11/06 provides detailed guidance on the production of olive trees and rootstocks. Plant material produced according to this certification scheme is derived from nuclear-stock plants (also identified as Primary source) officially recognised and registered in the database of certified accessions by the Ministry of Agriculture. The material deriving from the first multiplication of the nuclear-stock material enters in the certification process as Pre-basic material. Once the nuclear-stock has been registered, the breeders or Institutions or Research Centres responsible for its production and selection must keep the material under conditions that minimise recontamination risks.

The main outlines of the Italian certification scheme (Tab. 3 and Fig. 6) are the following:

a. *Registration of nuclear-stock material*: Breeders or researchers that intend to introduce a new accession for a specific variety in the certification system must provide detailed information about the trueness to type and sanitary controls performed by filling the official forms defined in the DM 20/11/06. The evaluation and eligibility of the registration request is carried out by a technical certification committee authorised by the Ministry of Agriculture.

	- b. *Maintenance and propagation of Pre-basic material*: Pre-basic olive material derives directly from the propagation of the nuclear-stock; the Pre-basic plants must be maintained in insect-proof green-houses (at the Conservation for Premultiplication Repository) with at least two replications. The plants are grown in *V. dahliae* and *X. diversicaudatum* -free soil mixture and periodically tested for viruses using molecular tools (10% of the plants each year, starting from the 5th year). Molecular tests should be also performed if, after visual inspection, plants show symptoms of *V. dahliae* or phytoplasmas. Cuttings and seeds collected from the Pre-basic material is used to produce, in the same facilities, the Basic plants for the establishment of the Premultiplication Repository.

Phytosanitary Certification 123

**Controls and certification released by** 

Regional Phytosanitary Services

**Current active Repository for olive certified material** 

**in Italy** 

houses Several Research Centers

Bari


Open field ConsorzioVivaisticoPugliese



Sperimentale "Improsta"


houses

Nurseries --

As aforementioned, the Italian voluntary certification program involves several woody crop species. In most cases, the main reason prompting for the certification of such accessions is the presence of pathogens that can cause detrimental effects on the affected plants (i.e. quarantine pests for stone fruit or citrus). Contrastingly, in the case of olive plants, the main aspect that promoted the adoption of this program has been the high level of genetic and phenotypic variability within each cultivar, which could results in heterogeneous plants and

The certification scheme adopted in Italy ensures trueness-to-type and uniformity, since the certified plants are obtained through subsequent clonal propagation steps from a single

**Table 3.** Organization of the Certification program for olive propagating material

**Steps Plant category Facilities** 

stock Nuclear-stock Screen-

Premultiplication Pre-basic Screen-

Premultiplication Basic Open field

Multiplication Certified

misidentification of such cultivars.

registered accession.

Selection of Nuclear-

Conservation for


In order to facilitate the certification and the availability of certified material for new olive cultivars or clones, the Premultiplication and the Multiplication blocks may be created directly using planting material deriving from the first multiplication of the nuclear-stock. In this way, the timeframe between the approval of a new accession in the system and the availability of certified plants in the nurseries is effectively reduced.

This certification program has been supported until now mainly by public funds that cover the costs for the management of the Conservation for the Premultiplication and Premultiplication repositories; starting in 2012 the program should shift to a self-sustaining system, in which taxes recovered on each released certification label will make up for the costs of the repository management.


Multiplication mother blocks.

at least once within a 30-year period.

single plant or seedling.

costs of the repository management.

b. *Maintenance and propagation of Pre-basic material*: Pre-basic olive material derives directly from the propagation of the nuclear-stock; the Pre-basic plants must be maintained in insect-proof green-houses (at the Conservation for Premultiplication Repository) with at least two replications. The plants are grown in *V. dahliae* and *X. diversicaudatum* -free soil mixture and periodically tested for viruses using molecular tools (10% of the plants each year, starting from the 5th year). Molecular tests should be also performed if, after visual inspection, plants show symptoms of *V. dahliae* or phytoplasmas. Cuttings and seeds collected from the Pre-basic material is used to produce, in the same facilities, the

c. *Maintenance and propagation of Basic material:* Basic olive planting material is the propagation material that is obtained from Pre-basic material, maintained in open field (Repository for the Premultiplication) in a variable replication number (2 minimum) depending on the importance of the cultivar. Premultiplication field plots must be tested and found free from *V. dahliae* and *X. diversicaudatum,* and have a 20 meters non-cultivated border. Basic plants must be periodically inspected and tested as defined for the Pre-basic material. Cuttings and seeds collected from the Basic material are used to produce the certified mother plants for the establishment of the

d. *Maintenance and propagation of Certified material:* Certified mother plants obtained from the propagation of the Basic material represent the source for nursery certified olive plant production. Mother plants are grown in open fields in variable replication numbers depending on the market of the specific cultivar. Plants are visually inspected at least once a year, while each plant must undergo laboratory tests for virus detection

e. *Certified nursery productions*: Production takes place in authorized nurseries that join the certification program. The nursery production must comply with the requirements established by the DM 20/11/06 in terms of: (i) soil mixture (free from *V. dahliae*, *X. diversicaudatum, Meloidogyne incognita, M. javanica and Pratylenchus vulnus*), (ii) location of the certifiable olive blocks; (iii) maintenance of a farm business registry. The regional phytosanitary service, following visual inspections and examination of the documentation, is in charge of releasing the official certification (blue label) for every

In order to facilitate the certification and the availability of certified material for new olive cultivars or clones, the Premultiplication and the Multiplication blocks may be created directly using planting material deriving from the first multiplication of the nuclear-stock. In this way, the timeframe between the approval of a new accession in the system and the

This certification program has been supported until now mainly by public funds that cover the costs for the management of the Conservation for the Premultiplication and Premultiplication repositories; starting in 2012 the program should shift to a self-sustaining system, in which taxes recovered on each released certification label will make up for the

availability of certified plants in the nurseries is effectively reduced.

Basic plants for the establishment of the Premultiplication Repository.

**Table 3.** Organization of the Certification program for olive propagating material

As aforementioned, the Italian voluntary certification program involves several woody crop species. In most cases, the main reason prompting for the certification of such accessions is the presence of pathogens that can cause detrimental effects on the affected plants (i.e. quarantine pests for stone fruit or citrus). Contrastingly, in the case of olive plants, the main aspect that promoted the adoption of this program has been the high level of genetic and phenotypic variability within each cultivar, which could results in heterogeneous plants and misidentification of such cultivars.

The certification scheme adopted in Italy ensures trueness-to-type and uniformity, since the certified plants are obtained through subsequent clonal propagation steps from a single registered accession.

**Figure 6.** Outline of the general certification steps and facilities

Phytosanitary Certification 125

*Annex I. – List of the reference European Union (EU) Directives and Italian national regulations* 

material and fruit plants intended for fruit production

fruit production, pursuant to Council Directive 92/34/EEC

materiale di propagazione vegetale delle piante da frutto

Gazzetta Ufficiale n. 141 del 20 giugno 2007)

*Council Directive 92/34/EEC* of 28 April 1992 on the marketing of fruit plant propagating

 *Council Directive 2008/90/EC* of 29 September 2008 (Recast version of Directive 92/34/EEC) on the marketing of fruit plant propagating material and fruit plants

 *Commission Directive 93/48/EEC* of 23 June 1993 setting out the schedule indicating the conditions to be met by fruit plant propagating material and fruit plants intended for

 *Commission Directive 93/64/EEC* of 5 July 1993 setting out the implementing measures concerning the supervision and monitoring of suppliers and establishments pursuant to Council Directive 92/34/EEC on the marketing of fruit plant propagating material and

 *Commission Directive 93/79/EEC* of 21 September 1993 setting out additional implementing provisions for lists of varieties of fruit plant propagating material and

 *DM 14 aprile 1997* Recepimento delle direttive delle Commissione n. 93/48/CEE del 23 giugno 1993, n. 93/64/CEE del 5 luglio 1993 e n. 93/79/CEE del 21 settembre 1993, relative alle norme tecniche sulla commercializzazione dei materiali di moltiplicazione

 *DM 4 maggio 2006* Disposizioni generali per la produzione di materiale di moltiplicazione delle specie arbustive ed arboree da frutto, nonché delle specie erbacee

 *DM 20 novembre 2006* norme tecniche per la produzione di materiali di moltiplicazione certificati di Agrumi, Fragola, Olivo, Pomoidee, Prunoidee (supplemento ordinario alla

 *Decreto Legislativo 25 giugno 2010, n.124* Attuazione della direttiva 2008/90 relativa alla commercializzazione dei materiali di moltiplicazione delle piante da frutto destinate alla produzione di frutti (refusione) (pubblicato nella Gazzetta Ufficiale n. 180 del 4

In this chapter an overview on the olive graft-transmissible pathogens and on the latest phytosanitary directives embodied by the EU and by the Italian Ministry of Agriculture is

delle piante da frutto e delle piante da frutto destinate alla produzione di frutto *DM 24 luglio 2003* Organizzazione del servizio nazionale di certificazione volontaria del

fruit plants, as kept by suppliers under Council Directive 92/34/EEC

Basic EU Directive

Basic Italian Regulations

a moltiplicazione agamica

agosto 2010)

**6. Conclusion** 

given.

intended for fruit production

Implementing measures of Directive 92/34/EEC :

fruit plants intended for fruit production

*Annex I. – List of the reference European Union (EU) Directives and Italian national regulations* 

#### Basic EU Directive

124 Olive Germplasm – The Olive Cultivation, Table Olive and Olive Oil Industry in Italy

**Figure 6.** Outline of the general certification steps and facilities


Implementing measures of Directive 92/34/EEC :


Basic Italian Regulations

