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The sanitary quality of seed is essential in agriculture. This is because pathogenic fungi compromise seed physiological quality and prevent the formation of plants in the field, which causes losses to farmers. Multispectral images technologies coupled with machine learning algorithms can optimize the identification of healthy peanut seeds, greatly improving the sanitary quality. The objective was to verify whether multispectral images technologies and artificial intelligence tools are effective for discriminating pathogenic fungi in tropical peanut seeds. For this purpose, dry peanut seeds infected by fungi (A. flavus, A. niger, Penicillium sp., and Rhizopus sp.) were used to acquire images at different wavelengths (365 to 970 nm). Multispectral markers of peanut seed health quality were found. The incubation period of 216 h was the one that most contributed to discriminating healthy seeds from those containing fungi through multispectral images. Texture (Percent Run), color (CIELab L*) and reflectance (490 nm) were highly effective in discriminating the sanitary quality of peanut seeds. Machine learning algorithms (LDA, MLP, RF, and SVM) demonstrated high accuracy in autonomous detection of seed health status (90 to 100%). Thus, multispectral images coupled with machine learning algorithms are effective for screening peanut seeds with superior sanitary quality.

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Anthracnose is the primary disease in Phaseolus lunatus cultures, causing severe losses. Inoculation techniques are vital for assessing genotype resistance and control methods at the early stages of seedling development. This study aimed to compare inoculation methods and exposure times of a lima bean seed variety to Colletotrichum truncatum using a completely randomized 4x5 factorial design with five replications. Seed inoculation methods by direct contact with mycelium, mannitol water restrictor, and sucrose water restrictor and immersion in conidia suspension were compared and submitted to substrates containing the developed pathogen or not at exposure times of 0, 36, 60, 84, and 108 hours. Evaluations were made by analyzing the severity, incidence, and disease index for anthracnose and seedling physiological quality under greenhouse conditions. The inoculation method by direct contact with sucrose solute for 36 hours was the most suitable for C. truncatum inoculation in lima bean seeds, providing a higher transmission rate but slightly affecting their physiological parameters. C. truncatum damage to lima bean seed performance increased with longer exposure times, regardless of the inoculation method.

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Seeds can harbor a wide range of microorganisms, especially fungi, which can cause different sanitary problems. Seed quality and seed longevity may be drastically reduced by fungi that invade seeds before or after harvest. Seed movement can be a pathway for the spread of diseases into new areas. Some seed-associated fungi can also produce mycotoxins that may cause serious negative effects on humans, animals and the seeds themselves. Seed storage is the most efficient and widely used method for conserving plant genetic resources. The seed storage conditions used in gene banks, low temperature and low seed moisture content, increase seed longevity and are usually favorable for the survival of seed-borne mycoflora. Early detection and identification of seed fungi are essential activities to conserve high-quality seeds and to prevent pathogen dissemination. This article provides an overview of the characteristics and detection methods of seed-borne fungi, with a special focus on their potential effects on gene bank seed conservation. The review includes the following aspects: types of seed-borne fungi, paths of infection and transmission, seed health methods, fungi longevity, risk of pathogen dissemination, the effect of fungi on seed longevity and procedures to reduce the harmful effects of fungi in gene banks.

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Plant diseases may survive and be spread by infected seeds. In this study we monitored the longevity of 14 seed-borne pathogens in 9 crop species commonly grown in the Nordic countries, in addition to a sample of sclerotia of Sclerotinia sclerotiorum. The data from the first 30 years of a 100-year seed storage experiment located in a natural -3.5 °C environment (permafrost) in Svalbard, Norway, are presented. To date, the pathogens, tested by traditional seed health testing methods (freezing blotter, agar plates, growing on tests), have survived. Linear regression analyses showed that the seed infection percentages of Drechslera dictyoides in meadow fescue, Drechslera phlei in timothy, and Septoria nodorum in wheat were significantly reduced compared to the percentages at the start of the experiment (from 63% to 34%, from 70% to 65%, and from 15% to 1%, respectively), and that Phoma betae in beet had increased significantly (from 43% to 56%). No trends in the infection percentage were observed over the years in Drechslera spp. in barley (fluctuating between 30% and 64%) or in Alternaria brassicicola in cabbage (fluctuating between 82% and 99%), nor in pathogens with low seed infection percentages at the start of the experiment. A major part of the stored sclerotia was viable after 30 years. To avoid the spread of seed-borne diseases, it is recommended that gene banks implement routines that avoid the use of infected seeds.

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Tomato (Solanum lycopersicum L.) is one of the most important vegetables in the world. However, tomato is also susceptible to many viral diseases. Several tobamoviruses, including tomato mosaic virus (ToMV), tomato mottle mosaic virus (ToMMV), and tomato brown rugose fruit virus (ToBRFV), are highly contagious pathogens that could result in significant economic losses if not controlled effectively. Tobamoviruses have been managed relatively well with broad adaptation of tomato cultivars with resistance genes. However, emergence of ToBRFV was shown to break down resistance conferred by the common resistance genes, resulting in serious outbreaks in many countries in Asia, Europe, and North America. The objective of this study was to conduct a comparative analysis of biological properties, including host range and disease resistance of ToMV, ToMMV, and ToBRFV. Results showed that despite many similarities in the host range, there were some unique host plant responses for each of the three viruses. In a comparative evaluation of disease resistance using the same tomato cultivars with or without Tm-22 gene, there was a striking difference in responses from tomato plants with Tm-22 gene inoculated with ToBRFV, ToMV, or ToMMV. Whereas these test plants were resistant to ToMV or ToMMV infection, all test plants were susceptible to ToBRFV. Further, for ToBRFV detection, a sensitive and reliable multiplex real-time reverse transcription (RT)-PCR assay using TaqMan probe with an internal 18S rRNA control was also developed. With simple modifications to RNA extraction and seed soaking, real-time RT-PCR could consistently detect the virus in single infested seed in varied levels of contamination, suggesting its usefulness for seed health assay.

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Seed systems research is central to achieving the United Nations Sustainable Development Goals. Improved varieties with promise for ending hunger, improving nutrition, and increasing livelihood security may be released, but how do they reach and benefit different types of farmers? Without widespread adoption the genetic gains achieved with improved crop varieties can never be actualized. Progress has been made toward demand responsive breeding, however the draft CGIAR 2030 Research and Innovation Strategy fails to recognize the complexity of seed systems and thus presents a narrow vision for the future of seed systems research. This points to the lack of evidence-based dialogue between seed systems researchers and breeders. This perspective paper presents findings from an interdisciplinary group of more than 50 CGIAR scientists who used a suite of seed systems tools to identify four knowledge gaps and associated insights from work on the seed systems for vegetatively propagated crops (VPCs), focusing on bananas (especially cooking bananas and plantains), cassava, potato, sweetpotato, and yam. We discuss the implications for thinking about and intervening in seed systems using a combined biophysical and socioeconomic perspective and how this can contribute to increased varietal adoption and benefits to farmers. The tools merit wider use, not only for the seed systems of VPCs, but for the seed of crops facing similar adoption challenges. We argue for deeper collaboration between seed systems researchers, breeders and national seed system stakeholders to address these and other knowledge gaps and generate the evidence and innovations needed to break through the 40% adoption ceiling for modern varieties, and ensure good quality seed once the new varieties have been adopted. Without this, the achievements of breeders may remain stuck in the seed delivery pipeline.

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Brazil is the largest producer, consumer, and exporter of forage seeds in the world. However, the high incidence of pathogens threatens the seed production system, resulting in decreases in production and quality. Currently, the emphasis has been placed on the use of alternative products as a way of maintaining productivity and environmental sustainability. Thus, the objective of this study was to evaluate the mycoflora associated with the commercial seeds of Brachiaria brizantha and Panicum maximum treated with chemical and alternative fungicide. To test with chemical and alternative treatments, seeds were subjected to fluazinam + thiophanate-methyl, fludioxonil + metalaxyl-M, noni essential oil, rosemary pepper essential oil, sulfuric acid. There was also an untreated control. The most efficient products in the control of incident fungi were, in decreasing order, fluazinam + thiophanate-methyl, fludioxonil + metalaxyl-M, noni essential oil, rosemary pepper essential oil, and sulfuric acid. Fungicides mainly controlled the genera Alternaria, Aspergillus, Bipolaris, Chaetomium, Cladosporium, Colletotrichum, Curvularia, Fusarium, Penicillium, Phoma and Rhizopus. Essential oils showed positive effects in controlling Bipolaris, Curvularia, and Fusarium. Sulfuric acid reduced the incidence of Bipolaris, Colletotrichum, Curvularia, Fusarium, Phoma, and Rhizopus. Seed treatment is one of the promising strategies for obtaining seeds of a healthy quality. As a result, the fungicides and botanical components listed above were promising in reducing fungi associated with forage seeds and can be used for seed treatments and in the development of new products with a natural base through essential oils.

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The inherent ability of seeds (orthodox, intermediate, and recalcitrant seeds and vegetative propagules) to serve as carriers of pests and pathogens (hereafter referred to as pests) and the risk of transboundary spread along with the seed movement present a high-risk factor for international germplasm distribution activities. Quarantine and phytosanitary procedures have been established by many countries around the world to minimize seed-borne pest spread by screening export and import consignments of germplasm. The effectiveness of these time-consuming and cost-intensive procedures depends on the knowledge of pest distribution, availability of diagnostic tools for seed health testing, qualified operators, procedures for inspection, and seed phytosanitation. This review describes a unique multidisciplinary approach used by the CGIAR Germplasm Health Units (GHUs) in ensuring phytosanitary protection for the safe conservation and global movement of germplasm from the 11 CGIAR genebanks and breeding programs that acquire and distribute germplasm to and from all parts of the world for agricultural research and food security. We also present the challenges, lessons learned, and recommendations stemming from the experience of GHUs, which collaborate with the national quarantine systems to export and distribute about 100,000 germplasm samples annually to partners located in about 90 to 100 countries. Furthermore, we describe how GHUs adjust their procedures to stay in alignment with evolving phytosanitary regulations and pest risk scenarios. In conclusion, we state the benefits of globally coordinated phytosanitary networks for the prevention of the intercontinental spread of pests that are transmissible through plant propagation materials.

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Maize chlorotic mottle virus (MCMV) has driven the emergence of maize lethal necrosis worldwide, where it threatens maize production in areas of East Africa, South America, and Asia. It is thought that MCMV transmission through seed may be important for introduction of the virus in new regions. Identification of infested seed lots is critical for preventing the spread of MCMV through seed. Although methods for detecting MCMV in leaf tissue are available, diagnostic methods for its detection in seed lots are lacking. In this study, ELISA, RT-PCR, and RT-qPCR were adapted for detection of MCMV in maize seed. Purified virions of MCMV isolates from Kansas, Mexico, and Kenya were then used to determine the virus detection thresholds for each diagnostic assay. No substantial differences in response were detected among the isolates in any of the three assays. The RT-PCR and a SYBR Green-based RT-qPCR assays were >3,000 times more sensitive than commercial ELISA for MCMV detection. For ELISA using seed extracts, selection of positive and negative controls was critical, most likely because of relatively high backgrounds. Use of seed soak solutions in ELISA detected MCMV with similar sensitivity to seed extracts, produced minimal background, and required substantially less labor. ELISA and RT-PCR were both effective for detecting MCMV in seed lots from Hawaii and Kenya, with ELISA providing a reliable and inexpensive diagnostic assay that could be implemented routinely in seed testing facilities.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

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Purple seed stain disease, caused by (Cercospora kukuchii), is a major concern in soybean (Glycine max (L.)) in Mississippi, USA, due to its effects on seed quality, reducing soybean seed grade and potential market price at elevators. Therefore, investigating the effects of purple seed stain (PSS) on seed quality (germination and vigor) and seed composition (nutrition) is critical. The objective of this study was to investigate the effects of PSS on seed harvest index, seed germination, seed vigor, and seed composition components (protein, oil, fatty acids, and sugars). A field experiment was initiated in 2019 in Stoneville, MS, at the Delta Research and Extension Center (DREC) on a Commerce silt loam soil (fine-silty, mixed, superactive, nonacid, thermic Fluventic Epiaquepts). Soybean variety Credenz 4748 LL was used. The results showed that infected (symptomatic) seed had a 5.5% greater Seed Index (based on 100 seed weight) when compared to non-infected (non-symptomatic, as control) seed. Non-infected seed had greater percent germination and seedling vigor when compared to infected seed. Germination was 30.9% greater and vigor was 58.3% greater in non-infected seed. Also, the results showed that infected seed with PSS had higher protein content and some amino acids. No changes in total oil and fatty acids. Sucrose and stachyose were lower in infected seed than in non-infected seed. The research showed that PSS impacted seed health and seed quality (germination and vigor) and seed composition (protein, sugars, and some amino acids). Purple stained seed should be avoided when planting and should be managed properly as low germination is a potential risk. Planting population should be adjusted accordingly due to lack of germination and vigor if PSS is present. This research help growers for purple seed management, and scientists to further understand the potential negative impact on seed quality and nutrition. Further research is needed before conclusive recommendations are made.

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Squash is one of the most important crops of tropical and temperate regions, and it can be affected by several fungal pathogens. Most of these pathogens infect the seeds, which become an efficient vehicle to disperse seedborne pathogens over long distances, with consequent severe crop losses. The main objective of this study was the identification of the principal seedborne fungi in seeds extracted from 66 samples of asymptomatic and symptomatic squash fruit (Cucurbita maxima, Cucurbita moschata) collected in two countries, Tunisia and Italy. The symptoms of fruit decay were identified and classified according to lesion size. Following the blotter test, 14 fungal species were detected from the seeds. Seedborne fungi were identified in all fruit samples tested, including asymptomatic fruit. The most frequent fungi from Tunisian seeds were Alternaria alternata (25.1%), followed by Stagonosporopsis cucurbitacearum (24.6%), Fusarium solani (16.6%), Rhizopus stolonifer (13.3%), F. fujikuroi (7.8%), Albifimbria verrucaria (3.3%), and Stemphylium vesicarium (2.3%). For the fruits from Italy, the most frequently identified fungal species in seed samples were Alternaria alternata (40.0%), followed by F. fujikuroi (20.8%), Stemphylium vesicarium (3.0%), and Curvularia spicifera (2.1%). Morphological identification was confirmed by molecular diagnosis using the available species-specific primers. Furthermore, specific primers were designed to identify Albifimbria verrucaria, Paramyrothecium roridum, and Stemphylium vesicarium. Application of seed-health testing methods, including such conventional and molecular diagnostic tools, will help to improve seed quality and crop yields.

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ABSTRACT: The aim of this research was to identify and quantify fungi infecting irrigated rice seeds produced in the 2015/16, 2016/17 and 2017/18 harvests in the pre-germinated system in the Alto Vale do Itajaí Region, Santa Catarina State, Brazil. A total of 479 lots of eight cultivars were analyzed. Seeds were disinfected and sown in potato-sucrose-agar culture medium with incubation for seven days at 25ºC ± 2ºC and photoperiod of 12 hours. The main fungus detected in the three crops season was Microdochium albescens with 54.9% of average incidence, followed by Alternaria padwikii (7.7%) and Bipolaris oryzae (3.3%).

RESUMO: O objetivo deste trabalho foi identificar e quantificar fungos infectando sementes de arroz irrigado, produzidas nas safras 2015/16, 2016/17 e 2017/18, no sistema pré-germinado na região do Alto Vale do Itajaí, estado de Santa Catarina. Foram analisados 479 lotes de oito cultivares, sendo as sementes desinfestadas e semeadas em meio de cultura de batata-sacarose-ágar com incubação por sete dias a 25ºC ± 2 ºC e fotoperíodo de 12 horas. O principal fungo detectado nas três safras foi Microdochium albescens com 54,9% de incidência média, seguido de Alternaria padwikii (7,7%) e Bipolaris oryzae (3,3%).

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The physiological performance of seeds is related to their physiological quality and seed vigor, while their health quality may interfere with germination and early seedling establishment in the field due to the interaction of microorganisms associated with seeds. The study aimed to evaluate the physiological performance and health quality of cucumber seeds and to verify the relationship between these attributes. The physiological quality of cucumber seeds was evaluated by standard germination tests, first count of germination, controlled deterioration test, electrical conductivity test, seedling emergence, emergence speed index, and traditional accelerated aging and aging modified with saline for 48, 72, and 96 h. The health quality of cucumber seed lots was evaluated by blotter test. Stratification of cucumber seed lots by seedling emergence was similar to seedling emergence by controlled deterioration test, first count of germination, and electrical conductivity results. The cucumber seed lots evaluated showed high germination rates; however, lots 1 and 3 had a better performance in vigor tests than lots 2 and 4. Fungi detected in the blotter test were Alternaria sp., Aspergillus sp., Cladosporium sp., and Penicillium sp. Seed lots 2 and 4 had low vigor evaluated by seedling emergence and controlled deterioration, and showed a higher incidence of Penicillium sp. in the evaluation of health quality of seeds. The incidence of Penicillium sp. may negatively affect the vigor of cucumber seeds evaluated by seedling emergence and by controlled deterioration test.(AU)

O desempenho fisiológico de sementes diz respeito à sua qualidade fisiológica e vigor, enquanto a qualidade sanitária pode interferir na germinação e estabelecimento inicial de plântulas no campo devido à interação dos micro-organismos associados às sementes. O trabalho teve por objetivo avaliar o desempenho fisiológico e a qualidade sanitária de sementes de pepino e verificar a relação entre esses atributos. A qualidade fisiológica de sementes de pepino foi avaliada pelos testes de germinação, primeira contagem de germinação, deterioração controlada, condutividade elétrica, emergência de plântulas, índice de velocidade de emergência e envelhecimento acelerado tradicional e modificado com solução salina por 48, 72 e 96 horas. A qualidade sanitária das sementes foi avaliada pelo método do papel-filtro. A estratificação de lotes de sementes de pepino ocorreu de maneira similar à emergência de plântulas pelos testes de deterioração controlada, primeira contagem da germinação e condutividade elétrica. Os lotes de sementes de pepino avaliados apresentaram alto percentual de germinação; no entanto, os lotes 1 e 3 apresentaram melhor desempenho nos testes de vigor que os lotes 2 e 4. Os fungos detectados no teste do papel-filtro foram Alternaria sp., Aspergillus sp., Cladosporium sp. e Penicillium sp. Os lotes 2 e 4 de menor vigor avaliados pelos testes de emergência de plântulas e de deterioração controlada foram os mesmos que apresentaram maior incidência de Penicillium sp. na avaliação da qualidade sanitária das sementes. A incidência de Penicillium sp. pode influenciar negativamente o vigor de sementes de pepino avaliado pela emergência de plântulas pelo teste de deterioração controlada.(AU)

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Pathogen buildup in vegetative planting material, termed seed degeneration, is a major problem in many low-income countries. When smallholder farmers use seed produced on-farm or acquired outside certified programs, it is often infected. We introduce a risk assessment framework for seed degeneration, evaluating the relative performance of individual and combined components of an integrated seed health strategy. The frequency distribution of management performance outcomes was evaluated for models incorporating biological and environmental heterogeneity, with the following results. (1) On-farm seed selection can perform as well as certified seed, if the rate of success in selecting healthy plants for seed production is high; (2) when choosing among within-season management strategies, external inoculum can determine the relative usefulness of 'incidence-altering management' (affecting the proportion of diseased plants/seeds) and 'rate-altering management' (affecting the rate of disease transmission in the field); (3) under severe disease scenarios, where it is difficult to implement management components at high levels of effectiveness, combining management components can be synergistic and keep seed degeneration below a threshold; (4) combining management components can also close the yield gap between average and worst-case scenarios. We also illustrate the potential for expert elicitation to provide parameter estimates when empirical data are unavailable. [Formula: see text] Copyright © 2017 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

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The seed-borne mycoflora of sorghum and foxtail millet collected from different growing areas in South Korea were isolated and taxonomically identified using dry inspection, standard blotter and the agar plate method. We investigated the in vitro and in vivo germination rates of disinfected and non-disinfected seeds of sorghum and foxtail millet using sterilized and unsterilized soil. The percent recovery of seed-borne mycoflora from the seed components of sorghum and foxtail millet seeds was determined and an infection experiment using the dominant species was evaluated for seedling emergence and mortality. A higher number of seed-borne fungi was observed in sorghum compared to that of foxtail millet. Eighteen fungal genera with 34 fungal species were identified from the seeds of sorghum and 13 genera with 22 species were identified from the seeds of foxtail millet. Five dominant species such as Alternaria alternata, Aspergillus flavus, Curvularia lunata, Fusarium moniliforme and Phoma sp. were recorded as seed-borne mycoflora in sorghum and 4 dominant species (Alternaria alternata, Aspergillus flavus, Curvularia lunata, Fusarium moniliforme) were observed in foxtail millet. The in vitro and in vivo germination rates were higher using disinfected seeds and sterilized soil. More seed-borne fungi were recovered from the pericarp compared to the endosperm and seed embryo. The percent recovery of seed-borne fungi ranged from 2.22% to 60.0%, and Alternaria alternata, Curvularia lunata and 4 species of Fusarium were isolated from the endosperm and embryo of sorghum and foxtail millet. Inoculation of the dominant seed-borne fungi showed considerable mortality of seedlings. All the transmitted seed-borne fungi might well be a primary source of infection of sorghum and foxtail millet crops.

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The seed-borne mycoflora of sorghum and foxtail millet collected from different growing areas in South Korea were isolated and taxonomically identified using dry inspection, standard blotter and the agar plate method. We investigated the in vitro and in vivo germination rates of disinfected and non-disinfected seeds of sorghum and foxtail millet using sterilized and unsterilized soil. The percent recovery of seed-borne mycoflora from the seed components of sorghum and foxtail millet seeds was determined and an infection experiment using the dominant species was evaluated for seedling emergence and mortality. A higher number of seed-borne fungi was observed in sorghum compared to that of foxtail millet. Eighteen fungal genera with 34 fungal species were identified from the seeds of sorghum and 13 genera with 22 species were identified from the seeds of foxtail millet. Five dominant species such as Alternaria alternata, Aspergillus flavus, Curvularia lunata, Fusarium moniliforme and Phoma sp. were recorded as seed-borne mycoflora in sorghum and 4 dominant species (Alternaria alternata, Aspergillus flavus, Curvularia lunata, Fusarium moniliforme) were observed in foxtail millet. The in vitro and in vivo germination rates were higher using disinfected seeds and sterilized soil. More seed-borne fungi were recovered from the pericarp compared to the endosperm and seed embryo. The percent recovery of seed-borne fungi ranged from 2.22% to 60.0%, and Alternaria alternata, Curvularia lunata and 4 species of Fusarium were isolated from the endosperm and embryo of sorghum and foxtail millet. Inoculation of the dominant seed-borne fungi showed considerable mortality of seedlings. All the transmitted seed-borne fungi might well be a primary source of infection of sorghum and foxtail millet crops.

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In just a few years, Pepino mosaic virus (PepMV) has become a major threat to greenhouse tomato production around the world. Although tomato seed is suspected to spread the disease, its importance as an initial virus inoculum for PepMV has not been established. To determine the potential for seed transmission, a tomato seed lot highly contaminated with PepMV was used for large-scale seedling grow-out tests. None of 10,000 grow-out seedlings was infected as determined by symptom expression, enzyme-linked immunosorbent assay (ELISA), or infectivity assay on Nicotiana benthamiana. Even though PepMV was not seed transmitted on tomato, the virus was effectively transmitted to tomato and N. benthamiana seedlings through mechanical transmission with seed extract. To examine the exact location where PepMV particles accumulated on the tomato seed, seed coats and embryos were carefully isolated and tested separately by ELISA, real-time RT-PCR, and bioassay on N. benthamiana. PepMV was detected in the seed coat fraction in both immature and mature tomato seeds, but not in the embryo. However, in N. benthamiana, the virus was neither seedborne nor seed-transmitted. Because PepMV is seedborne in tomato, efficient mechanical transmission of PepMV from the virus-contaminated tomato seed to seedlings could initiate a disease epidemic in a new tomato growing area. Thus, it is important to plant certified tomato seed that has been tested free of PepMV.