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Brome mosaic virus (BMV) and cocksfoot mottle virus (CfMV) are pathogens of grass species including all economically important cereals. Both viruses have been identified in Poland therefore they create a potential risk to cereal crops. In this study, a duplex—reverse transcription—polymerase chain reaction (duplex-RT-PCR) was developed and optimized for simultaneous detection and differentiation of BMV and CfMV as well as for confirmation of their co-infection. Selected primers CfMVdiag-F/CfMVdiag-R and BMV2-F/BMV2-R amplified 390 bp and 798 bp RT-PCR products within coat protein (CP) region of CfMV and replicase gene of BMV, respectively. Duplex-RT-PCR was successfully applied for the detection of CfMV-P1 and different Polish BMV isolates. Moreover, one sample was found to be co-infected with BMV-ML1 and CfMV-ML1 isolates. The specificity of generated RT-PCR products was verified by sequencing. Duplex-RT-PCR, like conventional RT-PCR, was able to detect two viruses occurring in plant tissues in very low concentration (as low as 4.5 pg/µL of total RNA). In contrast to existing methods, newly developed technique offers a significant time and cost-saving advantage. In conclusion, duplex-RT-PCR is a useful tool which can be implemented by phytosanitary services to rapid detection and differentiation of BMV and CfMV.

Agriculturists have been treating seeds to protect them from pathogens and pests for centuries, even before the nature of plant diseases was understood. Today, the use of seed treatments is a highly sophisticated strategy that has evolved into a very valuable, effective, and environmentally friendly component of agricultural production practices. Chemical seed treatments can be used to achieve a variety of benefits, including: improved emergence, through protection from seedborne pathogens and soilborne pathogens and insects; prevention of seed transmission of seedborne pathogens; protection of above-ground plant parts from infection by airborne pathogens or feeding by insect pests and disease vectors; improved vigor and uniformity of crop growth; deterrence of deterioration or insect feeding in storage; fulfillment of phytosanitary requirements and prevention of pathogen spread. These benefits all contribute to maximizing crop yield and quality while minimizing negative impacts through efficient use of crop protection chemicals. Seed treatment allows for highly targeted application of low, uniform doses of product, which is effective while reducing the risk of selection pressure for pathogen or pest resistance. Seed treatments are commercially available with contact, locally systemic, or fully systemic activity. Common active ingredients can be used for protection against Oomycetes, fungi, insects, and nematodes. There are numerous examples of improvements in stand establishment and yield as a result of seed treatment use in a wide range of crops. Combinations of active ingredients are becoming more common as products improve for efficacy against specific pathogen groups. In maize, seed treatment is nearly universal and standard practices may include a combination of four fungicides, an insecticide, and a nematicide. This provides a high level of protection across a wide pathogen spectrum as well as prevention of feeding damage to the seed and seedling. Seed treatments are playing an increasing role in the productivity of agriculture, as well as its sustainability and efficiency. Seed application of crop protection compounds provides unique benefits that make it a preferable approach compared to other tactics. It is a reliable technology that guarantees a uniform crop establishment in a variety of environments, soils and cultural practices; benefits provided by seed treatments cannot be duplicated because most of the target diseases and pests cannot be controlled after planting.

Trapping of tephritid fruit flies forms the backbone of many activites related to phytosanitary issues and regulation of trade of fresh fruits and vegetables between countries. Detection of incipient fruit fly populations can occur through a number of means, such as visual surveys, fruit cutting (to reveal the presence of eggs and larvae), collection and holding of fruits to determine if fruit flies emerge from the collected fruits, and perhaps the most commonly used method, trapping of adult flies using some combination of specific trap types, such as the McPhail, MultiLure, Jackson, etc., and semiochemical attractant, such as a food lure, pheromone, or male lure and kairomonal attractant.

Viral pathogens comprise approximately half of the emerging diseases in plants, and plant introductions (including the international movement of seed) are considered to be one of the most important contributing factors to the emergence of these pathogens. For the most part plant viruses are incapable of surviving outside of host tissue making their long-term propagation dependent on their hosts. Thus infected seeds are an effective strategy that not only allows for pathogen survival from one season to the next, but also for their dispersal. The Potyviridae, as the largest plant virus family, is often considered to be the most economically important and its members rank among the most successful plant pathogens. Seed transmission within the Potyviridae family is not uncommon, however the exact mechanism of viral entry into the germ line is currently unknown, and the genetic basis of seed transmission has yet to be completely elucidated. Seed transmission rates are influenced by complex interactions among a variety of factors including the host cultivar, the virus isolate, environmental conditions, the timing of infection, vector characteristics, and viral synergism. Seed transmission can have an enormous effect on the epidemiology of crop pathogens due in part to the ecology of plant viruses which are often secondarily disseminated via insect vectors with the effect that extremely low frequencies of seed transmission can result in devastating epidemics. This is compounded by the fact that vertically infected seedlings often do not exhibit symptoms of viral infection. Given the potential for seed transmitted viral pathogens to initiate epidemics, it is vital to understand how seed transmission rates translate into epidemics. In addition, as seed transmission is a means of dispersal for these viral pathogens, effective phytosanitary measures to control the spread of these pathogens are crucial.

Since a number of important pests are seed-borne or seed-transmissible, the movement of infested seed may pose a risk for the international spread of pests. One of the main roles of the European and Mediterranean Plant Protection Organization (EPPO) is to help its member countries to prevent entry or spread of dangerous pests. The Organization has therefore been given the task of identifying pests which may present a risk for the region (early warning), evaluating them and making proposals on the phytosanitary measures which can mitigate the risk. Once a pest has been identified as presenting a risk for the EPPO region, guidance on how to detect and identify it are developed (phytosanitary procedures for inspection and diagnostic protocols) as well as recommendations on how to eradicate and/or control it. To perform these activities, EPPO collects information and makes it available to its member countries. Different databases have been developed including PQR (Plant Quarantine data Retrieval system) and the EPPO database on Diagnostic expertise. In addition to pest-specific activities, EPPO also develops recommendations for quality assurance in laboratories, in order to harmonize procedures in the EPPO region and improve diagnostic quality. The different activities conducted in this framework are presented with a special focus on activities related to seed-borne pests.

The Scientific Panel on Plant Health (PLH) of the European Food Safety Authority was established in 2006 as the reference EU body for risk assessment in the plant health area in the frame of phytosanitary measures to prevent the introduction and further spread of organisms harmful (pests) to plants or plants products. Proper diagnosis and detection are critical for the categorization and assessment of pests that are or may qualify as quarantine pests, and for appropriate application of phytosanitary measures. In carrying out its responsibilities, the PLH Panel is frequently faced by the changes in taxonomy and nomenclature: these may often lead to a re-evaluation of previous pest records, concerns with published literature, and in some cases with the increased use of molecular taxonomic and identification tools. Pathogens that were previously considered strains/varieties of a single species, in some cases have now been elevated to specific status. In other cases pathogens previously recognised as species complexes have now been partitioned into named species, using molecular criteria rather than traditional methods. The problems that can arise are illustrated in this Chapter by examples representative of the main pathogen taxa.

Systematic deployment of attractant-baited traps has long been a mainstay of phytosanitary programs for tephritid fruit flies. Trapping arrays are used for detecting, delimiting, monitoring, and confirming eradication of tephritid populations, as well as for demonstrating minimal risk of infestation of commodities in support of trade. Designing and optimizing trapping systems to meet these various functions requires a basic understanding of the efficiency of the trapping system, including the influence of programmatic variables, such as trap density and distribution. Distance-capture functions, based on results of release-recapture tests, are discussed as one tool for assessing the sensitivity of trapping systems. As a demonstration of their use, these functions are incorporated into Monte Carlo simulations for estimating probabilities of detecting populations, including the effects of population size, time (in fly generations), and programmatic variables. Examples are provided for three key pest species: Ceratitis capitata (Wiedemann), Bactrocera dorsalis (Hendel), and B. cucurbitae (Coquillett). Results of these or comparable analyses can be used to predict potential ranges of population size at the time of detection or estimate the maximum expected size of a residual population after control measures are applied given a continued absence of captures. Optimal design of trapping systems for any given location and purpose requires consideration of a number of factors that may include, among other things, the frequency of introduction of the target pest(s), availability and feasibility of using various control methods (insecticides, sterile insect releases, etc.), potential of the pest population to spread, and/or costs of survey, eradication, and other program activities. In combination with this information, quantitative assessments of tephritid trapping systems can be used to ensure adequate and appropriate system sensitivity, identify opportunities to improve overall cost-effectiveness of phytosanitary programs, and provide scientific justification for program strategies and protocols.

Official attempts to prevent the introduction and/or spread of quarantine pests began in China in 1928, and acquired more complexity after 1980. China has seen accelerated economic development, especially during the past 10 years following membership in the World Trade Organisation (WTO). Simultaneously, China faces many challenges from the global market and international regulations involving trade of plants and plant products. During the past 15 years research and education related to Plant Biosecurity have been strengthened, especially within China’s National Academies and agricultural universities. According to requirements of the International Plant Protection Convention (IPPC) and Agreement on the Application of Sanitary and Phytosanitary Measures (SPS Agreement), the transparent and scientific phytosanitary measures in China have been adopted based on Pest Risk Assessment. Invasive plant pests (insects, pathogens and weeds) are detected and managed through quarantine inspection, identification and treatments. Here we introduce the regulations, bureaucracy, technology, education and research associated with plant quarantine and invasive species management in China. More progress is expected for the development of plant biosecurity in the process of preventing the invasion of plant pests, protecting the production of agriculture and forestry, and promoting international trade.

The International Plant Protection Convention (IPPC) is an international treaty that came into existence during 1952, superseding all previous international plant protection agreements. The IPPC is intended to secure common and effective action to prevent the introduction and spread of pests and diseases of plants and plant products and to promote measures for their control. In 2013, there were 179 contracting parties to the Convention. A Secretariat for the IPPC and a Committee of Experts on Phytosanitary Measures (CEPM) were established in 1993 to address the task of developing International Standards for Phytosanitary Measures (ISPMs). ISPMs provide a useful framework for National Plant Protection Organizations (NPPOs), particularly when pest management systems and regulatory decision-making have an important role in international trade. ISPMs in themselves are not regulatory instruments but come into force after countries establish phytosanitary measures within their national legislation. Countries may establish phytosanitary measures that provide a higher level of protection than ISPMs, as long as these national measures are science based and technically justified. Subject Matter Experts from member countries draft ISPMs, which have many levels of review and revision before their adoption. Thirty-six (36) International Standards for Phytosanitary Measures relating to all aspects of plant health have been adopted between 1995, when standard setting began, until 2013. Many more are in various stages of development.

Regulatory plant science has a unique evolution which distinguishes it from many disciplines with similar functions. The design and role of most National Plant Protection Organizations is closely linked to the International Plant Protection Convention and more recently to the World Trade Organization Agreement on the Application of Sanitary and Phytosanitary Measures. The international regulatory framework created by these agreements and the activities surrounding them represent a major adjustment for the phytosanitary community and is expected to be a significant factor in determining the direction of future efforts in regulatory plant science. It is clear that a focus on the application of phytosanitary measures in trade will continue to be crucial; complemented by substantial efforts in harmonization and capacity building to meet the anticipated demands of commerce. Other aspects of the Convention that may have had greater attention in the past are likely to fade in importance as new challenges occupy the efforts of the phytosanitary community on its journey into the digital era with a mix of old and new approaches and tools but with an eye towards flexibility and change. Developing analytical capacity will be critical to advancing defendable regulatory decisions and challenging others. The role of science and technology will be dramatically greater, especially in developing more sophisticated and cost-effective programs based on the continuum concept.

Research on antimicrobial peptides is continuously growing because of the possibilities of applications they offer in different domains including food safety, human medicine, and plant biocontrol (phytosanitary).

In recent years, invasive plant pathogens such as Phytophthora ramorum and Puccinia graminis (TTKS) have emerged as global threats that move rapidly across international boundaries. To counter such threats, the accurate monitoring of global plant health concerns necessitates the involvement of all agricultural sectors: governmental agencies, universities and the agricultural industry. Many members of industry conduct (i) scouting of research and seed production fields for pests, (ii) phytosanitary field inspections, and (iii) provide disease diagnostics testing for their customers. The Pest Information Platform (PIPE) proved to be an excellent vehicle for fostering collaborative efforts between the public and private sector in monitoring for Soybean Rust (Phakopsora pachyrhizi) in North America. The American Seed Trade Association’s Phytosanitary Committee has developed a draft proposal for an industry PIPE platform for a narrow list of new and emerging pests of corn, soybeans and watermelon. Combining private industry and public pest data into a central database can provide (i) useful pest distribution information for state extension specialists; (ii) improved monitoring for emerging and exotic pest threats; and (iii) improved pest distribution records for state and federal regulators to enhance their decision making. Expanding PIPE like systems beyond national borders has potential to provide additional information for agencies to prepare and respond to exotic pest threats. Many members of industry with international operations have the capacity to jump start the development of a global PIPE. However, a global PIPE would require a comprehensive data sharing policy that protects national, state and corporate interests.

Available strategies for the management of nematode vectors of plant viruses are reviewed, focusing on the nematode vector species, their associated viruses, as well as their geographic distribution and spread. Diagnostic procedures including morphological identification of virus vectors, plant tests and transmission assays as well as the application of molecular detection tools are reviewed, in the light of preventive and phytosanitary procedures. Management of GFLV on grapevine requires production of healthy plants for certification and marketing schemes, to be used in soils found free of its vector, Xiphinema index. In fields already infested, some integrated management options may be applied including, in order of importance, agronomic practices with long rotations (5-7 years) intercropping with poor or antagonistic hosts, chemical control, application of organic amendments and natural products, biofumigation, nematicidal plants and biological control agents. Given the risk that due to some nematodes’ parthenogenetic reproduction epidemics may arise even from a single individual vector, emphasis must be given to preventive and continuous field monitoring procedures.

The purpose of the International Plant Protection Convention (IPPC) is to secure cooperation and action to prevent the spread and introduction of pests of plants and plant products and to promote appropriate measures for their control. Under the IPPC, pests include: “any species, strain or biotype of plant, animal or pathogenic agent injurious to plants or plant products”. Although the first version of the IPPC came into force in 1952 it was the recognition of the IPPC by the Sanitary and Phytosanitary Agreement of the World Trade Organization that provided the impetus to revise the IPPC and substantially increase standard setting activities. There are now 170 contracting parties to the IPPC. Under the IPPC, internationally recognised standards covering a wide range of plant protection issues particularly relating to preventing or controlling pests moving internationally have been approved.

Biosecurity (regulatory regimes for food safety, animal and plant health, and genetically modified organisms) results in barriers to trade. Plant pest introductions cause substantial losses; growth of trade and international travel increases the risk of new introductions. Demand for agricultural produce follows world population growth and consumption pattern changes; international agricultural trade is vital to access to food. National and international biosecurity is fragmented, although nationally consolidation occurs. International biosecurity has changed over the last 15 years: the International Plant Protection Convention, the FAO/WHO Codex Alimentarius (Food Safety) and the World Organization for Animal Health, set Standards for national Sanitary and Phytosanitary measures, the WTO Sanitary and Phytosanitary (SPS) Agreement seeks to avoid arbitrary or unjustifiable discrimination among countries; the Convention on Biological Diversity, its Cartagena Protocol and the Biological Weapons Convention also address aspects of biosecurity: internationally, cooperation appears, at present, to be the most promising action. Developed and a limited number of mid-income developing countries raise phytosanitary trade concerns in the WTO SPS Committee. Certain food safety measures appear to become stricter, private standards more elaborate and small farmers have difficulties in adapting. Instead many phytosanitary standards aim to reduce unjustified national measures. National plant health is often under-funded, in particular non-export components including phytosanitary protection of biodiversity and natural ecosystems. Technical assistance to developing countries remains very limited. Plant health needs expertise in identification and epidemiology. Regional cooperation is vital. Opportunities exist for international action on pest free areas and certification of laboratories. It is unlikely that effects of climate change can be considered in Pest Risk Analysis.

Invasive alien plants may be introduced intentionally with trade (80% of current invasive alien plants in Europe were introduced as ornamental or agricultural plants) or unintentionally (as contaminants of grain, seeds, soil, machinery etc., or with travellers). Preventing the introduction of invasive alien plants is considered more cost-effective, from both environmental and economic points of view, than managing them after introduction. Pest risk analysis (PRA) standards have been developed by the International Plant Protection Convention (IPPC) and the European and Mediterranean Plant Protection Organization (EPPO) to allow assessment of the phytosanitary risk presented by invasive alien plants, and the development of appropriate measures to prevent their introduction and spread. These measures may in turn have an impact on international trade, and the obligations arising from trade agreements have also to be taken into account when phytosanitary measures are established. PRA basically consists in a framework for organizing biological and other scientific and economic information to assess risk. This leads to the identification of management options to reduce the risk to an acceptable level. Within the EPPO context, the results of these PRAs are translating into recommendations for countries to implement their national regulations. This article gives an overview of the international framework for regulation of invasive alien plants under the IPPC. It then presents the approach followed by EPPO for the evaluation and management of risks presented by such plants, as well as its application.

Currently Australia and New Zealand are free of Bursaphelenchus xylophilus, the pinewood nematode (PWN), which causes the pine wilt disease, and its primary vectors Monochamus spp. Both countries have increasing interest in this pest due to the significant area of exotic pine plantations, predominantly Pinus radiata, and native conifer species that could be susceptible to PWN. Although the current phytosanitary measures and surveillance programmes in New Zealand and Australia minimise the risk of entry of PWN, both countries have favourable climatic conditions for its establishment. In addition, there are significant uncertainties in the susceptibility of Pinus species to PWN and the potential vector status of native cerambycid beetles. A screening study was conducted in 2003 using 12-month old seedlings from Queensland of P. elliottii, P. caribaea. var hondurensis and $\mathrm{F}1$ and $\mathrm{F}2$ hybrid clones of these parents to determine their potential susceptibility to PWN and Bursaphelenchus mucronatus. Results illustrated the limitations of screening young plants in determining the susceptibility of older trees. The pest risk assessment conducted in 2004 and the screening study confirmed that maintenance of strong quarantine and surveillance programs and development of molecular or physiological markers for identification of PWN resistance in planting stocks of Pinus species and native species is indispensable to prevent the introduction of pine wilt disease in New Zealand and Australia.

The development of export strategies for horticultural products considered to be hosts of quarantine pests follows a logical series of steps. The foundation for the process is the completion of a commodity plant pest risk assessment that satisfies the needs of the national plant protection organization of the importing country. After identifying the significant pests, a set of proposed mitigation measures is designed. Regulatory officials are increasingly using systems approaches to supplement or substitute for direct postharvest treatments in developing export strategies. The components of systems approaches can be divided into a series of five categories of measures: field and production measures, preharvest measures, postharvest measures, inspection and certification measures, and shipping and distribution measures. The United States Department of Agriculture (USDA), working with the national plant protection organizations of other countries, has used the systems approach concept to develop quarantine strategies for both the domestic movement and the importation of fruit fly host commodities. The Mexican fruit fly Anastrepha ludens (Loew) programme in the Texas citrus production area of the lower Rio Grande River Valley is an example of a domestic systems approach. USDA allows the importation of papayas from Central America when produced in accordance with a systems approach that primarily targets the Mediterranean fruit fly Ceratitis capitata (Wiedemann). Systems approaches are groups of integrated pest risk management measures designed to provide importing countries with adequate phytosanitary security while facilitating trade in situations where direct postharvest commodity treatment is undesirable, not feasible or non-existent, or imported products are marginal hosts of the quarantine pest produced in a low-pest prevalence area.

The study of toxigenic fungi started at the end of 19th — at the beginning of 20th century after the appearance of Fusarium head blight epidemics in the Far East and after the registration of man and animal poisoning caused by infected grain (Voronin, 1890, Palchevsky, 1891, Jaczewski 1904, Naumov, 1916). More active researches of diseases caused by toxigenic fungi was developed in 30–40 years after the identification of stachybotryotoxicosis in horses and alimentary toxic aleukia in man (Sarkisov, 1947). In these years Sarkisov and his colleagues initiated the new direction of mycological researches, named mycotoxicology (Sarkisov, 1954). Presently in Russia mycotoxicological researches are carried out in different medical and agricultural laboratories. In this short review, information is provided about the phytosanitary state in Russia for the last 10–15 years in connection with the widespread of toxigenic fungi on cereals and contamination of food by mycotoxins.

The International Seed Health Initiative - Herbage Crops (ISHI-H) has initiated ringtesting to establish international standards for the detection of Clavibacter michiganensis subsp. insidiosus, the causal pathogen of bacterial wilt in lucerne-alfalfa [Medicago sativa) seed. Bacterial wilt of lucerne is a seedborne disease causing wilting, stunting and yellowing of plants. In the field disease symptoms are not severe until the second or third year when significant yield loss can occur. The bacterium can survive in the field for 8–10 years. Because infected seed is a possible source of international spread, phytosanitary regulations are in place in many importing countries. Currently there is no internationally accepted seed test for bacterial wilt.

Insects are no doubt the highest economically important phytosanitary problem for many cultivated plants; since they attacks different organs from the early development stages to the steps next to harvesting, thus causing severe losses in production as well as the product quality. Integrated pest management strategy (IPM), which aims at reducing the damage and insect population level is a useful strategy for this (Bustillo 2000). The control measures that have been considered are the chemical control, use of parasitoids, entomopathogens, cultural control and the development of plants resistant to plague.

The fourth International EFPP-Symposium focuses on status, recent developments and future trends in the field of diagnostics and identification of plant pathogens and parasites. It attains even more weight if it is placed in a broader spectrum of interests, more in particular phytosanitary subjects, quarantine problems and political objectives. In this paper seven subjects are being dealt with, ranging from innocent to major economic events: 1. new findings 2. Phoma spp 3. Meloidogyne chitwoodi and M. fallax 4. Thrips palmi 5. Mediterranean fruitfly 6. Brown rot of potatoes 7. Mad cow’s disease(BSE)

Our research objective was to develop a method for eradicating viable cells of Erwinia amylovora from symptomless scion budsticks of apple by immersion in hot water. The effect of duration of immersion in water at different temperatures on viability of dormant buds of apple cvs.‘Idared’ and‘Empire’ and on survival of E. amylovora were determined. Approximately 50% of buds of both cvs. survived 60 min at 48° C, but all buds were killed by 30 min at 50° C. Shoot tips of greenhouse-grown potted‘Idared’ and‘Empire’ trees were inoculated with E. amylovora. After lesions had ceased expanding 1 cm segments of the stem, directly below the lesion margin and including a node, were immersed in water at 48° C for 60 min. No viable E. amylovora was detected by direct planting of phosphat-buffer-ground tissue on CCT medium, or by plating of Kado 523 broth that had been inoculated with grindate and incubated for 18 hours. Unimmersed tissues yielded viable E. amylovora. Experiments to determine the time course of killing of E. amylovora cells infected apple tissue at 48° C indicated that after 60 min there was < 1 × 10-6 chance of a single viable cell remaining. A phytosanitary protocol for E. amylovora- free apple scions will be discussed.

Microtubers are of interest as propagules in seed potato production. With the increasing importance of their storability, phytosanitary advantages and ease of transportation, intensive attention is now being given to the production of microtubers. Culture environment and media including the use of growth regulators are the main factors exerting pronounced effects on tuberization. Varying concentrations of cytokinins, auxins and other regulatory substances have been used in culture media [4, 5, 11], while in the method developed at the International Potato Centre (CIP), the antigibberellin, chlorocholine chloride (CCC) together with BAP have been added [10]. A simple method for producing microtubers without growth regulators has been developed at Wye College [3] where all the twelve varieties tested produced microtubers in culture without difficulty [2]. The microtubers produced by the Wye College System have provided satisfactory results on field planting each year since 1984 [1]. The present research was undertaken to investigate the effects of growth regulators and day length and to compare the Wye College and CIP systems.

ZnSO4 5% plus phytosanitary pesticides at the dormancy stage, ZnSO4 1.5% plus break dormancy chemicals at the silver tip stage, and ZnSO4 0.2% with and without Ca(OH)2 0.2% or ZnO 0.1% or Zn chelate (7% Zn), 3L/ha, as 3 forthnightly sprays during the spring were sprayed in a Zn-deficient Golden Delicious apple orchard to control Zn deficiency and to evaluate possible phytotoxicity. All treatments significantly reduced Zn deficiency symtoms and none caused any visible phytotoxicity. Only spring sprays reduced necrotic leaf spot symptoms and increased Zn in the leaves at high levels suggesting leaf contamination. Leaf analysis was found not to be efficient to evaluate the Zn nutritional status of apple orchards. Sprays at the dormancy stage increased Zn in the top 20 cm soil layer suggesting rainfall leaching from the trees.