The use of cytoplasmic male-sterility in hybrid seed production

The practical utilization of heterosis in crop plants has been greatly facilitated during the past 15 years by the use of cytoplasmic male|sterility for low-cost, large-scale emasculation of the seed parents of hybrids. The method is now being used in commercial production of hybrid onions, sugar beets, field corn, grain sorghum, and petunias. Its use is contemplated for hybrid sweet corn, pop corn, red table beets, fodder beets, fodder sorghum, pearl millet, carrots, and garden peppers.     

Seeds of change: corn seed mixtures for resistance management and integrated pest management

The use of mixtures of transgenic insecticidal seed and nontransgenic seed to provide an in-field refuge for susceptible insects in insect-resistance-management (IRM) plans has been considered for at least two decades. However, the U.S. Environmental Protection Agency has only recently authorized the practice. This commentary explores issues that regulators, industry, and other stakeholders should consider as the use of biotechnology increases and seed mixtures are implemented as a major tactic for IRM. We discuss how block refuges and seed mixtures in transgenic insecticidal corn, Zea mays L., production will influence integrated pest management (IPM) and the evolution of pest resistance. We conclude that seed mixtures will make pest monitoring more difficult and that seed mixtures may make IRM riskier because of larval behavior and greater adoption of insecticidal corn. Conversely, block refuges present a different suite of risks because of adult pest behavior and the lower compliance with IRM rules expected from farmers. It is likely that secondary pests not targeted by the insecticidal corn as well as natural enemies will respond differently to block refuges and seed mixtures.     

Detection of Clavibacter michiganensis subsp. michiganensis in tomato seeds using immunomagnetic separation

The use of pathogen-free plant material is the main strategy for controlling bacterial canker of tomato caused by Clavibacter michiganensis subsp. michiganensis. However, detection and isolation of this pathogen from seeds before field or greenhouse cultivation is difficult when the bacterium is at low concentration and associated microbiota are present. Immunomagnetic separation (IMS), based on the use of immunomagnetic beads (IMBs) coated with specific antibodies, was used to capture C. michiganensis subsp. michiganensis cells, allowing removal of non-target bacteria from samples before plating on non-selective medium. Different concentrations of IMBs and of two antisera were tested, showing that IMS with 10(6)IMBs/ml coated with a polyclonal antiserum at 1/3200 dilution recovered more than 50% of target cells from initial inocula of 10(3) to 10(0)CFU/ml. Threshold detection was lower than 10CFU/ml even in seed extracts containing seed debris and high populations of non-target bacteria. The IMS permitted C. michiganensis subsp. michiganensis isolation from naturally infected seeds with higher sensitivity and faster than direct isolation on the semiselective medium currently used and could become a simple viable system for routinely testing tomato seed lots in phytosanitary diagnostic laboratories.     

Seed mixtures as a resistance management strategy for European corn borers (Lepidoptera: Crambidae) infesting transgenic corn expressing Cry1Ab protein

Dispersal of neonate European corn borers, Ostrinia nubilalis (Hübner), in seed mixtures of transgenic corn expressing Cry1Ab protein (Bt+) and nontransgenic corn (Bt-) was evaluated in a 2-yr field study. The main objective was to determine if larval dispersal limits the effectiveness of seed mixtures as a resistance management strategy. Mixtures evaluated included (1) all Bt+ plants, (2) every fifth plant Bt- with remaining plants Bt+, (3) every fifth plant Bt+ with remaining plants Bt-, and (4) all Bt- plants. The transformation events MON 802 (B73 BC1F2 x Mol7) and MON 810 (B73 BC1F1 x Mo17), which express the Cry1Ab endotoxin isolated from Bacillus thuringiensis subsp. kurstaki, were used as the sources of Bt+ seed in 1994 and 1995, respectively (YieldGard, Monsanto, St. Louis, MO). At corn growth stage V6-V8, subplots within each mixture (15-20 plants each) were infested so that every fifth plant in mixtures 1 and 4, every Bt- plant in mixture 2, and every Bt+ plant in mixture 3 received two egg masses. Larval sampling over a 21-d period indicated increased neonate dispersal off of Bt+ plants, reduced survival of larvae that dispersed from Bt+ plants to Bt- plants, and a low incidence of late-instar movement from Bt- plants to Bt+ plants. Computer simulations based on mortality and dispersal estimates from this study indicate that seed mixtures will delay the evolution of resistant European corn borer populations compared with uniform planting of transgenic corn. However, resistant European corn borer populations likely will develop faster in seed mixes compared with separate plantings of Bt and non-Bt corn.     

Application of Bacillus subtilis in the biological control of the phytopathogenic fungus Sporisorium reilianum

Sporisorium reilianum is the causal agent of head smut in corn, a worldwide disease that causes significant economic losses in the principal areas where maize is cultivated. To attack this problem, resistant commercial hybrids and synthetic fungicide seed treatment have been used, but with low effectiveness. In this study, a native strain of Bacillus subtilis isolated from the soil of corn crops in Mixquiahuala, Hidalgo, in central Mexico, was applied as a seed treatment in the field to test for the biological control of head smut. It significantly decreased the Smut Incidence Percentage (SIP) while increasing maize productivity. This work represents the first report in which B. subtilis has been used to control S. reilianum in the field.     

Western corn rootworm (Coleoptera: Chrysomelidae) dispersal and adaptation to single-toxin transgenic corn deployed with block or blended refuge

A simulation model of the temporal and spatial dynamics and population genetics of western corn rootworm, Diabrotica virgifera virgifera LeConte, was created to evaluate the use of block refuges and seed blends in the management of resistance to transgenic insecticidal corn (Zea mays L.). This Bt corn expresses one transgenic corn event, DAS-59122-7, that produces a binary insecticidal protein toxin (Cry34Ab1/Cry35Ab1) and provides host-plant resistance. The model incorporates the latest information about larval and adult behavior. Results of this modeling effort indicate that the seed-blend scenarios in many cases produced equal or greater durability than block refuges that were relocated each year. Resistance evolved in the most likely scenarios in 10-16 yr. Our standard analysis presumed complete adoption of 59122 corn by all farmers in our hypothetical region, no crop rotation, and 100% compliance with Insect Resistant Management (IRM) regulations. As compliance levels declined, resistance evolved faster when block refuges were deployed. Seed treatments that killed the pest when applied to all seeds in a seed blend or just to seeds in Bt corn blocks delayed evolution of resistance. Greater control of the pest population by the seed treatment facilitated longer durability of the transgenic trait. Therefore, data support the concept that pyramiding a transgenic insecticidal trait with a highly efficacious insecticidal seed treatment can delay evolution of resistance.     

Improved recovery of active recombinant laccase from maize seed

Lignolytic enzymes such as laccase have been difficult to over-express in an active form. This paper describes the expression, characterization, and application of a fungal laccase in maize seed. The transgenic seed contains immobilized and extractable laccase. Fifty ppm dry weight of aqueously extractable laccase was obtained, and the remaining solids contained a significant amount of immobilized laccase that was active. Although a portion of the extractable laccase was produced as inactive apoenzyme, laccase activity was recovered by treatment with copper and chloride. In addition to allowing the apoenzyme to regain activity, treatment with copper also provided a partial purification step by precipitating other endogenous corn proteins while leaving >90% of the laccase in solution. The data also demonstrate the application of maize-produced laccase as a polymerization agent. The apparent concentration of laccase in ground, defatted corn germ is approximately 0.20% of dry weight.     

Conventional and seed-based insect management strategies similarly influence nontarget coleopteran communities in maize

Seed-based pest management tools, such as transgenes and seed treatments, are emerging as viable alternatives to conventional insecticide applications in numerous crops, and often occur as coupled technologies. Seed-based technologies have been readily adopted in maize, for which ecological studies are needed to examine effects to farmland biodiversity. We compared the response of nontarget coleopteran communities in Cry1Ab/c sweet corn and Cry3Bb field corn to conventional pyrethroid applications and a control. Of particular interest was the Cry3Bb field corn, which was coupled with a neonicotinoid seed treatment and was not rotated across years. A functionally diverse subset of the coleopteran community, consisting of three families (Carabidae, Chrysomelidae, and Nitidulidae) and 9,525 specimens, was identified to species. We compared coleopteran diversity and dynamics using rarefaction and ordination techniques. There were no differences in species richness among treatments; however, higher activity densities were more common in the control. In the nonrotated field corn, principal response curves showed a consistent pattern of treatment communities deviating from the control communities over time, whereas crop rotation in the sweet corn negated treatment effects. Treatment effects could not be detected when beetles were grouped based on functional roles. Results indicate that neonicotinoid seed-based treatments may have effects on some nontarget coleopterans, but these effects are similar to conventional pyrethroid applications.     

木霉菌在玉米病害生物防治中的作用机制及应用

目前,世界上共有分属于10个属（Trichoderma,Gliocladium,Chaetomium,Bacillus,Burkhoderia,Streptomyces and Pseudomonas,Pantoea,Enterobacter,Macrobacterium）中的微生物被试验用于玉米病害的生物防治,其中细菌14种,真菌17种,放线菌1种。国际上由木霉菌开发的生物杀菌剂和生物肥料有50余种,其中以哈茨木霉T22菌株开发的产品最为著名。目前在我国也开发出了4种木霉菌剂型（可湿性粉剂、颗粒剂、水分散粒剂和种衣剂）,正式登记的木霉菌杀菌剂有7种,其中6种为可湿粉剂,1种为水分散粒剂,主要登记用于防治番茄、观赏百合、黄瓜的立枯病、猝倒病、根腐病、灰霉病、霜霉病以及小麦的纹枯病,但尚无木霉菌生物农药被登记用于防治玉米病害。以木霉菌为主要成分登记的菌肥产品有11种,其中在玉米上应用的有2种。由课题组研制的木霉菌颗粒剂和种衣剂通过土壤穴施和种子包衣可有效防治玉米茎腐病和纹枯病,其中木霉菌颗粒剂防效达65%-87%。近期研制的木霉菌可湿性粉剂对玉米小斑病的防效达50%-60%。国际上已鉴定出多种可诱导玉米获得系统抗性的木霉菌源激发子,其中包括Sm1、纤维素酶、疏水蛋白和Avr4 /Avr9等效应因子。本课题组近年鉴定出Thc6（锌指蛋白类转录因子）、PAF-AH和Thph1/Thph2的编码产物在系统诱导以JA/ET信号调控的玉米抗弯孢菌叶斑病中具有重要作用,符合植物免疫MAMPs模式,为全面认识木霉菌诱导免疫机理提供了重要理论依据。木霉菌诱导玉米从根至叶片的防御反应系统传导机制还需深入研究。     

Influence of plant severing on movement of Ostrinia nubilalis larvae in Zea mays hybrid seed production fields

Genetically engineered corn hybrids that contain a cry gene from the bacterium Bacillus thuringiensis Berliner (Bt) are gaining popularity for controlling the corn pest Ostrinia nubilalis (Hübner). Continuous use of Bt corn, however, could select for O. nubilalis that are resistant to this corn. Monitoring for insect resistance is important, because it could help maintain the Bt technology. A possible monitoring method is to collect larval insects in commercial drying bins after harvest from Bt seed production fields. A drawback to this method is that these collections may be contaminated by insects that moved as later instars from severed non-Bt male rows into the adjacent Bt female rows. These larvae have little to no exposure to Bt toxin, resulting in possible "false positives." The objectives of this study were to first find which combination of planting and severing dates produces the least number of larvae that move from non-Bt male plants to Bt female plants and to assess O. nubilalis larval movement from severed non-Bt male rows to Bt female rows. Field studies in 2002 and 2003 were designed to simulate a hybrid seed production field. Results suggest that movement of O. nubilalis larvae from male corn is minimized when corn is planted early and male plants are severed by 2 wk post-anthesis. This reduces the likelihood of false positives by reducing the number of susceptible larvae moving between Bt and non-Bt plants. Also, larvae moved to all four female rows that were adjacent to the severed rows, but there were significantly more larvae found in the closest row compared with the other three. These results could be used to develop a monitoring program to find O. nubilalis larvae with resistance to Bt corn in field populations of O. nubilalis.     

Agricultural Seed Biomass for Migrating and Wintering Waterfowl in the Southeastern United States

ABSTRACT Waterfowl frequently acquire high-energy agricultural seeds in harvested and unharvested croplands during migration and winter. Estimates of agricultural seed biomass in harvested and unharvested corn, soybean, and grain sorghum fields do not exist or are outdated for the southeastern United States. Therefore, we estimated seed biomass in 105 harvested and 59 unharvested corn, soybean, and grain sorghum fields across 4 climate regions in Tennessee, USA, from September through January 2006 and 2007. We also used estimates of seed biomass to calculate duck-energy days (DEDs) in December and January when migratory waterfowl abundance peaks in the southeastern United States. Mean biomass of corn, soybean, and grain sorghum seed in harvested fields declined 239 kg/ha to 39 kg/ha, 118 kg/ha to 26 kg/ha, and 392 kg/ha to 19 kg/ha, respectively, from postharvest to January. Continuous monthly rates of decline were 64% for corn, 84% for soybean, and 74% for grain sorghum. Agricultural seed biomass in harvested corn and grain sorghum fields dropped below the waterfowl giving-up density (i.e., 50 kg/ha) in 3 months; soybean dropped below this threshold 1 month postharvest. Mean DEDs/ha in harvested corn, soybean, and grain sorghum fields were low (274, 90, and 27, respectively) in January, and DEDs were zero in >85% of fields. In unharvested corn, soybean, and grain sorghum fields, mean DEDs/ha in January were high (69,000, 18,000, and 26,000, respectively), and continuous rates of decline (3%, 7%, and 18%, respectively) were much lower than for harvested crops. Waterfowl biologists in the Southeast should use our estimates of agricultural seed biomass in DED calculations. We also recommend that biologists provide unharvested grain fields and natural wetlands for migrating and wintering waterfowl because seed resources are low in harvested agricultural fields.     

Estimation of efficacy functions for products used to manage corn rootworm larval injury

Maize, Zea mays L., is an economically important crop grown throughout the world. Corn rootworm, Diabrotica spp. (Coleoptera: Chrysomelidae), larvae constitute a significant economic threat to maize production in the United States, where yield losses and management costs associated with corn rootworm species exceed $1 billion annually. Furthermore, the introduction of the western corn rootworm, D. virgifera virgifera LeConte, into maize‐producing regions of Europe has made managing corn rootworm larval injury an international concern. Larvae injure maize plants by feeding on root tissue and are the primary target of management activities. Products commonly used to protect root systems from injury include chemical insecticides (seed or soil applied) and genetically modified maize hybrids expressing toxins derived from Bacillus thuringiensis Berliner (Bt). The confirmation of field‐evolved resistance to various Bt toxins in populations of the western corn rootworm presents a significant management challenge. We performed a meta‐analysis to provide a broad understanding of the relative efficacy of the primary products currently being used to manage corn rootworm larval injury, including insecticidal seed treatments, soil insecticides and Bt hybrids (with and without the addition of soil insecticide). Our analysis is unique in the breadth of locations and years included – we analysed 135 individual trials conducted from 2003 through 2014 at multiple sites in both Illinois and Nebraska. Panel data were produced by pairing the mean node‐injury rating for each treatment of a given trial with the mean node‐injury rating for untreated maize. Linear regression models were developed to estimate the relationship between the potential for corn rootworm larval injury and product performance. For a given level of injury potential, the parameters estimated reveal differences in the degree of root protection offered by the various product categories analysed. Implications for developing long‐term, integrated, and sustainable practices for managing this important pest of maize are discussed.     

Components of reproductive isolation between the monkeyflowers Mimulus lewisii and M. cardinalis (Phrymaceae)

Evolutionists have long recognized the role of reproductive isolation in speciation, but the relative contributions of different reproductive barriers are poorly understood. We examined the nature of isolation between Mimulus lewisii and M. cardinalis, sister species of monkeyflowers. Studied reproductive barriers include: ecogeographic isolation; pollinator isolation (pollinator fidelity in a natural mixed population); pollen competition (seed set and hybrid production from experimental interspecific, intraspecific, and mixed pollinations in the greenhouse); and relative hybrid fitness (germination, survivorship, percent flowering, biomass, pollen viability, and seed mass in the greenhouse). Additionally, the rate of hybridization in nature was estimated from seed collections in a sympatric population. We found substantial reproductive barriers at multiple stages in the life history of M. lewisii and M. cardinalis. Using range maps constructed from herbarium collections, we estimated that the different ecogeographic distributions of the species result in 58.7% reproductive isolation. Mimulus lewisii and M. cardinalis are visited by different pollinators, and in a region of sympatry 97.6% of pollinator foraging bouts were specific to one species or the other. In the greenhouse, interspecific pollinations generated nearly 50% fewer seeds than intraspecific controls. Mixed pollinations of M. cardinalis flowers yielded >75% parentals even when only one-quarter of the pollen treatment consisted of M. cardinalis pollen. In contrast, both species had similar siring success on M. lewisii flowers. The observed 99.915% occurrence of parental M. lewisii and M. cardinalis in seeds collected from a sympatric population is nearly identical to that expected, based upon our field observations of pollinator behavior and our laboratory experiments of pollen competition. F1 hybrids exhibited reduced germination rates, high survivorship and reproduction, and low pollen and ovule fertility. In aggregate, the studied reproductive barriers prevent, on average, 99.87% of gene flow, with most reproductive isolation occurring prior to hybrid formation. Our results suggest that ecological factors resulting from adaptive divergence are the primary isolating barriers in this system. Additional studies of taxa at varying degrees of evolutionary divergence are needed to identify the relative importance of pre- and postzygotic isolating mechanisms in speciation.     

A Built-In Strategy to Mitigate Transgene Spreading from Genetically Modified Corn

Transgene spreading is a major concern in cultivating genetically modified (GM) corn. Cross-pollination may cause the spread of transgenes from GM cornfields to conventional fields. Occasionally, seed lot contamination, volunteers, mixing during sowing, harvest, and trade can also lead to transgene escape. Obviously, new biological confinement technologies are highly desired to mitigate transgene spreading in addition to physical separation and isolation methods. In this study, we report the development of a built-in containment method to mitigate transgene spreading in corn. In this method, an RNAi cassette for suppressing the expression of the nicosulfuron detoxifying enzyme CYP81A9 and an expression cassette for the glyphosate tolerant 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene G10 were constructed and transformed into corn via Agrobacterium-mediated transformation. The GM corn plants that were generated were found to be sensitive to nicosulfuron but resistant to glyphosate, which is exactly the opposite of conventional corn. Field tests demonstrated that GM corn plants with silenced CYP81A9 could be killed by applying nicosulfuron at 40 g/ha, which is the recommended dose for weed control in cornfields. This study suggests that this built-in containment method for controlling the spread of corn transgenes is effective and easy to implement.     

Selection for resistance to mCry3A-expressing transgenic corn in western corn rootworm

To investigate the development of resistance to mCry3A, a laboratory colony of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was established from field survivors of mCry3A-expressing (MIR604) corn, Zea mays L. Feral adults emerging from MIR604 (selected) and isoline (control) field plots were collected and returned to the laboratory. Progeny of each colony was reared one generation on isoline corn and then crossed reciprocally with a nondiapausing colony. The resulting nondiapausing progeny were then reared on greenhouse corn in accordance with the wild type parent's origin (on MIR604 or isoline corn). After four, seven, and 10 total generations of selection, the resistance ratio of the selected colony was 0.5, 4.3, and 15.4 in terms of lethal concentration (LC)50 values in toxicity assays, with the latter two LC50 values being significant. After seven generations of selection in total, selected and control colonies were screened on MIR604 and isoline corn under field conditions. There was a significant colony x corn pedigree interaction in terms of plant damage. There was no significant difference in damage between MIR604 and isoline corn, whereas this difference was significant for the control colony. After 14 generations of selection, a seedling bioassay was performed. Again, there was a significant colony x corn pedigree interaction, this time in terms of the number of larvae recovered. There was no significant difference in the number of larvae recovered from MIR604 and isoline corn for the selected colony, whereas this difference was significant for the control colony, although larval size was greater on isoline corn for both colonies. Resistance has developed in western corn rootworm laboratory colonies to all Bt proteins currently registered for corn rootworm management, which emphasizes the importance of adhering to resistance management plans for maintaining product efficacy.     

The mycobiota from seeds of Shrunken-2 (sh2) Sweet Corn

Sixteen species of fungi were isolated from the pericarp, endosperm, scutellum, and embryo of low (<75% germination), moderate (75–89%), and high vigor (>89%) seedlots of supersweet sweet corn (sh2) hybrids over two years. Most frequently isolated species werePenicillium oxalicum, Rhizopus arrhizus (14.5%), andRhizopus spp (17.4%).Fusarium moniliforme, a common inhabitant in field or dent corn, was isolated from only 2.4% of the samples. The low mean isolation values may be the result of poor conditions for infection or the data may reflect the methods used including sterilization techniques or random tissue selection.Aspergillus niger, F. moniliforme, andP. oxalicum, were isolated at a significantly greater level than other fungi from the high vigor hybrids at 0.89, 0.46 m and 4.46 respectively compared to 0.04, 0, and 1.82 for the low vigor hybrids. When Total Fungi were compared mean isolations were significantly greater from the high vigor hybrids at 11.96, the moderate 8.79, or low 4.86 vigor groups. When data from seed sources for all vigor groups were compared, significantly greater mean isolations were obtained from Illinois Foundation Seed hybrids forCladosporium sp,R. arrhizus, andRhizopus spp., but greater rates were obtained forFusarium oxysporum from the Asgrow hybrids. Isolation frequencies for the 16 species were not significantly different between the seed tissue types from any of the hybrids evaluated during this investigation. Results from this study showed that there is a diverse group of fungi present within thesh2 seed and seed treatments must be developed which will minimize seed rot and seedling blight from both internal seedborne and external pathogens.     

Dispersal behavior of neonate European corn borer (Lepidoptera: Crambidae) on Bt corn

European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), has historically been a significant economically important insect pest of corn (Zea mays L.) in the United States and Canada. The development in the 1990s of genetically modified corn expressing genes derived from Bacillus thuringiensis (Bt) that encodes insecticidal crystalline (Cry) proteins has proven to be effective in controlling this insect as well as other corn pests. The purpose of this study was to assess the movement and dispersal behavior of neonate European corn borer on Bt corn. We examined differences in neonate European corn borer dispersal behavior for the first 4 h after eclosion in the field among a stacked pyramid (Cry1F X Cry1Ab X Cry34/35Ab1) Bt corn, a Cry1F Bt corn, and a non-Bt sweet corn; and in the laboratory among a Bt corn hybrid containing Cry1F, a hybrid containing Cry1Ab, a pyramid combining these two hybrids (Cry1F X Cry1Ab), and a non-Bt near isoline corn. In field experiments, we found that dispersal was significantly higher on Bt corn compared with sweet corn. In laboratory experiments, dispersal was significantly higher on Cry1Ab Bt corn and Cry1F X Cry1Ab Bt corn than on non-Bt near isoline corn. Results indicated that neonate dispersal may be significantly greater in Bt cornfields compared with non-Bt cornfields. The findings on dispersal behavior in this study will be useful in evaluating the efficacy of a blended seed refuge system for managing European corn borer resistance in Bt corn.     

Development of an edible subunit vaccine in corn against enterotoxigenic strains of escherichia coli

Summary Advances in the development of subunit vaccines and in the production of foreign proteins in plants together offer the prospect of stable and inexpensive vaccine delivery systems. Various bacterial and viral proteins stably produced in plants have been shown to elicit immune responses in feeding trials. We have extended this approach by using Zea mays as the plant production system. Corn has several advantages as a vaccine delivery vehicle, most notably established technologies to generate transgenic plants, to optimize traits through breeding and to process the seed into a palatable form. Here we report on the production in corn seed of the GM₁ receptor binding (B) subunit of the heat-labile toxin (Lt) from enterotoxigenic strains of Escherichia coli. Versions of the Lt-B gene were synthesized to give optimum codon usage for corn and to target the protein to either the cell surface or the cytoplasm. These synthetic genes were fused to a strong promoter and transformed into corn. Lt-B was highly expressed in corn seed at up to 1.8% of the total soluble protein and this was further increased approximately five-fold through plant breeding. As in E. coli. Lt-B produced in corn forms a functional pentamer that can bind to the GM₁ receptor. Furthermore, Lt-B pentamer stored in corn seed is much more resistant to heat than is the pure protein, allowing the transgenic corn to be readily processed into an edible form. This work demonstrates the potential of using products derived from transgenic corn seed as delivery vehicles for subunit vaccines.     

Detection of Acidovorax valerianellae, the causing agent of bacterial leaf spots in corn salad [Valerianella locusta (L.) Laterr.], in corn salad seeds

Aim: The black leaf spot disease on corn salad caused by the bacterium Acidovorax valerianellae has been observed in Europe for several years and causes economic losses in corn salad cropping. Contaminated seeds or infested soil are considered as the major infection sources. The use of healthy seed material is the only way to prevent disease outbreaks. Therefore, a sensitive diagnostic method for seed testing should be developed. Methods and Results: Using a triple antibody sandwich ELISA with a high‐specific monoclonal antibody, a quick and reliable detection method for contamination of seed lots with the pathogen was developed. This method allowed to detect contaminated seed lots as well as contamination with A. valerianellae in single seeds. Furthermore, the occurrence and distribution of the pathogen could be shown in symptomatic corn salad leaves and in naturally infested seeds by transmission electron microscopy and immunogold labelling for the first time. Conclusion: Our results confirm the seed transmission of this corn salad disease. Pathogen load and distribution vary between positively tested seed lots. Significance and Impact of the study: With this method, not only routine testing of seed material to eliminate contaminated seed lots from production is possible but also the control of sanitation procedures to reduce contamination.     

Soil type does not affect seed ageing when soil water potential and temperature are controlled

To investigate the effects of soil type on seed persistence in a manner that controlled for location and climate variables, three weed species—Gomphocarpus physocarpus (swan plant), Avena sterilis ssp. ludoviciana (wild oat) and Ligustrum lucidum (broadleaf privet)—were buried for 21 months in three contrasting soils at a single location. Soil type had a significant effect on seed persistence and seedling vigour, but soil water content and temperature varied between soils due to differences in physical and chemical properties. Warmer, wetter conditions favoured shorter persistence. A laboratory-based test was developed to accelerate the rate of seed ageing within soils, using controlled superoptimal temperature and moisture conditions (the soil-specific accelerated ageing test, SSAAT). The SSAAT demonstrated that soil type per se did not influence seed longevity. Moreover, the order in which seeds aged was the same whether aged in the field or SSAAT, with L. lucidum being shortest-lived and A. sterilis being longest-lived of the three species.