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Background: While the relationship between pollen and respiratory allergies is well-documented, the role of short-term pollen exposure in food allergy and eczema flares has not previously been explored. We aimed to investigate these associations in a population-based sample of children., Methods: We investigated 1- (n = 1108) and 6-year-old (n = 675) children in the grass pollen season from the HealthNuts cohort. Grass pollen concentrations were considered on the day of testing (lag 0), up to three days before (lag 1-lag 3) and cumulatively (lag 0-3). Associations between grass pollen and food skin-prick test reactivity (SPT ≥ 2 mm at age 1 year and ≥ 3 mm at age 6 years), eczema flares, challenge-confirmed food allergy, reaction threshold to oral food challenges (OFC), and serum food-specific IgE levels were analyzed using either logistic or quantile regression models. Atopy and family history of allergic disease were considered as potent effect modifiers., Results: Grass pollen at lag 0-3 (every 20 grains/m 3 increase) was associated with an up to 1.2-fold increased odds of food SPT reactivity and eczema flares in 6-year-olds. In 1-year-olds, the associations were only observed for peanut in those with a family history of food allergy. Increasing grass pollen concentrations were associated with a lower reaction threshold to OFC and higher serum IgE levels in peanut-allergic 1-year-olds only., Conclusion: Increasing grass pollen concentration was associated with increased risk of food SPT reactivity and eczema flares in children. The associations in peanut-allergic infants may be related to immune activation and/or peanut and grass pollen cross-reactivity leading to a lower reaction threshold., (© 2022 The Authors. Pediatric Allergy and Immunology published by European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd.)

Background: Grass pollen exposure is a risk factor for childhood asthma hospital attendances. However, its short-term influence on lung function, especially among those with other allergic conditions, has been less well-studied., Objective: To investigate this association in a population-based sample of children., Methods: Within the HealthNuts cohort, 641 children performed spirometry during the grass pollen season. Grass pollen concentration was considered on the day of testing (lag 0), up to 3 days before (lag 1-lag 3), and cumulatively (lag 0-3). We used linear regression to assess the relevant associations and examined potential interactions with current asthma, hay fever or eczema, and food allergy., Results: Associations were observed only in children with allergic disease (P value for interaction ≤ 0.1). In children with food allergy, grass pollen concentration was associated with a lower ratio of forced expiratory volume in 1 second to forced vital capacity (FEV 1 /FVC) and lower mid-forced expiratory flows (FEF 25%-75% ) at all lags (eg, at lag 2, FEV 1 /FVC z-score = -0.50 [95% CI -0.80 to -0.20] and FEF 25%--75% z-score = -0.40 [-0.60 to -0.04] per 20 grains/m 3 pollen increase), and increased bronchodilator responsiveness (BDR) at lag 2 and lag 3 (eg, at lag 2, BDR = (31 [95% CI -0.005 to 62] mL). In children with current asthma, increasing grass pollen concentration was associated with lower FEF 25%-75% and increased BDR, whereas children with current hay fever or eczema had increased BDR only., Conclusions: A proactive approach needs to be enforced to manage susceptible children, especially those with food allergy, before high-grass pollen days., (Copyright © 2022 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)

Background: Asthma epidemics associated with thunderstorms have had catastrophic effects on individuals and emergency services. Seasonal allergic rhinitis (SAR) is present in the vast majority of people who develop thunderstorm asthma (TA), but there is little evidence regarding risk factors for TA among the SAR population., Objective: We sought to identify risk factors for a history of TA and hospital presentation in a cohort of individuals with SAR., Methods: This multicenter study recruited adults from Melbourne, Australia, with a past diagnosis of TA and/or self-reported SAR. Clinical information, spirometry results, white blood cell count, ryegrass pollen-specific (RGP-sp) IgE concentration, and fractional exhaled nitric oxide were measured to identify risk factors for a history of TA in individuals with SAR., Results: From a total of 228 individuals with SAR, 35% (80 of 228) reported SAR only (the I-SAR group), 37% (84 of 228) reported TA symptoms but had not attended hospital for treatment (the O-TA group), and 28% (64 of 228) had presented to the hospital for TA (the H-TA group). All patients in the H-TA group reported a previous asthma diagnosis. Logistic regression analysis of factors associated with O-TA and H-TA indicated that lower FEV 1 value and an Asthma Control Questionnaire score higher than 1.5 were associated with H-TA. Higher blood RGP-sp IgE concentration, eosinophil counts, and fractional exhaled nitric oxide level were significantly associated with both O-TA and H-TA. Receiver operating curve analysis showed an RGP-sp IgE concentration higher than 10.1 kU/L and a prebronchodilator FEV 1 value of 90% or lower to be biomarkers of increased H-TA risk., Conclusion: Clinical tests can identify risk of a history of TA in individuals with SAR and thereby inform patient-specific treatment recommendations., (Crown Copyright © 2021. Published by Elsevier Inc. All rights reserved.)

Background: Experimental challenge studies have shown that pollen can have early and delayed effects on the lungs and airways. Here, we qualitatively and quantitatively synthesize the evidence of outdoor pollen exposure on various lung function and airway inflammation markers in community-based studies., Methods: Four online databases were searched: Medline, Web of Science, CINAHL and Google Scholar. The search strategy included terms relating to both exposure and outcomes. Inclusion criteria were human-based studies published in English that were representative of the community. Additionally, we only considered cross-sectional or short-term longitudinal studies which investigated pollen exposure by levels or season. Study quality assessment was performed using the Newcastle-Ottawa scale. Meta-analysis was conducted using random-effects models., Results: We included 27 of 6551 studies identified from the search. Qualitative synthesis indicated associations between pollen exposure and predominantly type-2 inflammation in both the upper and lower airways, but little evidence for lung function changes. People with ever asthma and/or seasonal allergic rhinitis (SAR) were at higher risk of such airway inflammation. Meta-analysis confirmed a positive relationship between pollen season, eosinophilia and eosinophil cationic protein (ECP) in people with ever SAR but the results between studies were highly variable. Heterogeneity was reduced after further subgrouping by age, and the forest plots indicated that eosinophilic airway inflammation to outdoor pollen exposure increased with age., Conclusion: Among people with ever asthma and ever SAR, exposure to increased ambient pollen triggers type-2 upper and lower airway inflammation rather than a non-specific or innate inflammation. These findings can lead to the formulation of specific pollen immunotherapy for susceptible individuals. Future research should be directed towards investigating lagged associations and effect modifications using larger and more generalized populations., Systematic Review Registration: CRD42020146981 (PROSPERO)., (© 2021 John Wiley & Sons Ltd.)

The world's most severe thunderstorm asthma event occurred in Melbourne, Australia on 21 November 2016, coinciding with the peak of the grass pollen season. The aetiological role of thunderstorms in these events is thought to cause pollen to rupture in high humidity conditions, releasing large numbers of sub-pollen particles (SPPs) with sizes very easily inhaled deep into the lungs. The humidity hypothesis was implemented into a three-dimensional atmospheric model and driven by inputs from three meteorological models. However, the mechanism could not explain how the Melbourne event occurred as relative humidity was very low throughout the atmosphere, and most available grass pollen remained within 40 m of the surface. Our tests showed humidity induced rupturing occurred frequently at other times and would likely lead to recurrent false alarms if used in a predictive capacity. We used the model to investigate a range of other possible pollen rupturing mechanisms which could have produced high concentrations of SPPs in the atmosphere during the storm. The mechanisms studied involve mechanical friction from wind gusts, electrical build up and discharge incurred during conditions of low relative humidity, and lightning strikes. Our results suggest that these mechanisms likely operated in tandem with one another, but the lightning method was the only mechanism to generate a pattern in SPPs following the path of the storm. If humidity induced rupturing cannot explain the 2016 Melbourne event, then new targeted laboratory studies of alternative pollen rupture mechanisms would be of considerable value to help constrain the parameterisation of the pollen rupturing process., Competing Interests: The authors have declared that no competing interests exist.

Background: The association between grass pollen exposure and early markers of asthma exacerbations such as lung function changes and increase in airway inflammation is limited. We investigated the associations between short-term grass pollen exposure and lung function and airway inflammation in a community-based sample, and whether any such associations were modified by current asthma, current hay fever, pollen sensitization, age, and other environmental factors., Methods: Cross-sectional and short-term analyses of data from the Melbourne Atopy Cohort Study (MACS) participants (n = 936). Lung function was assessed using spirometry. Airway inflammation was assessed by fractional exhaled nitric oxide (FeNO) and exhaled breath condensate pH and nitrogen oxides (NOx). Daily pollen counts were collected using a volumetric spore trap. The associations were examined by linear regression., Results: Higher ambient levels of grass pollen 2 days before (lag 2) were associated with lower mid-forced expiratory flow (FEF 25%-75% ) and FEV 1 /FVC ratio (Coef. [95% CI] = -119 [-226, -11] mL/s and -1.0 [-3.0, -0.03] %, respectively) and also 3 days before (lag 3). Increased levels of grass pollen a day before (lag 1) were associated with increased FeNO (4.35 [-0.1, 8.7] ppb) and also at lag 2. Adverse associations between pollen and multiple outcomes were greater in adults with current asthma, hay fever, and pollen sensitization., Conclusion: Grass pollen exposure was associated with eosinophilic airway inflammation 1-2 days after exposure and airway obstruction 2-3 days after exposure. Adults and individuals with asthma, hay fever, and pollen sensitization may be at higher risk., (© 2020 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.)

Seasonal allergic rhinitis (AR), also known as hay fever, is a common respiratory condition brought on by a range of environmental triggers. Previous work has characterised the relationships between community-level AR symptoms collected using mobile apps in two Australian cities, Canberra and Melbourne, and various environmental covariates including pollen. Here, we build on these relationships by assessing the skill of models that provide a next-day forecast of an individual's risk of developing AR and that nowcast ambient grass pollen concentrations using crowd-sourced AR symptoms as a predictor. Categorical grass pollen forecasts (low/moderate/high) were made based on binning mean daily symptom scores by corresponding categories. Models for an individual's risk were constructed by forward variable selection, considering environmental, demographic, behaviour and health-related inputs, with non-linear responses permitted. Proportional-odds logistic regression was then applied with the variables selected, modelling the symptom scores on their original five-point scale. AR symptom-based estimates of today's average grass pollen concentration were more accurate than those provided by two benchmark forecasting methods using various metrics for assessing accuracy. Predictions of an individual's next-day AR symptoms rated on a five-point scale were correct in 36% of cases and within one point on this scale in 82% of cases. Both outcomes were significantly better than chance. This large-scale AR symptoms measurement program shows that crowd-sourced symptom scores can be used to predict the daily average grass pollen concentration, as well as provide a personalised AR forecast., Competing Interests: Declaration of competing interest The authors declare they have no actual or potential competing financial interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Allergic Rhinitis (AR) affects over half a billion people worldwide with an estimated prevalence of 1 in 5 individuals in developed countries. Although ambient pollen exposure is a causal factor in AR, the symptom-exposure relationship is typically not studied in the broader community but in small, well-characterised cohorts drawn from clinical populations. To identify relationships between AR symptoms in the community and a range of environmental factors, we used a database containing over 96,000 symptom score reports collected over a 3-year period (2014-2016) through freely available smartphone apps released in two Australian cities, Melbourne and Canberra. Ambient pollen levels and symptom scores were strongly related, with grass pollen explaining most of the symptom variation. Other factors correlated with higher symptom scores included temperature (R > 0.73) and wind speed (R > 0.75). In general, worse symptom scores were reported by younger participants, women, and those who had taken medication for AR in the preceding 24 h. The strength of this relationship varied between the two cities. Smartphone-based symptom surveys offer a cost-effective means of studying real-world risk factors for AR in a broader 'extra-clinical' population., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Thunderstorm asthma is defined as epidemics of asthma occurring shortly after a thunderstorm. While grass pollen has been implicated in thunderstorm asthma events, little is known about the role of fungi and studies have not been synthesised., Objective: This systematic review aims to evaluate whether grass pollen is necessary in thunderstorm asthma events and whether fungi also play a part in these associations., Methods: We conducted a systematic search using six electronic databases (i.e. CINAHL, Medline (Ovid), Web of Science, ProQuest Central, EMBASE and Google Scholar) and checked reference lists. The search terms used were pollen AND thunderstorm* AND asthma. The inclusion criteria were studies published in English with original human data relating to outdoor pollen and thunderstorm asthma., Results: Twenty of 2198 studies were eligible. Reported findings differed due to variation in methodological approaches and a meta-analysis was not possible. Nonetheless, of the 20 studies included, 15 demonstrated some relationship with nine demonstrating lagged effects up to four days for increasing grass pollen counts associated with increased risk of thunderstorm asthma. Of the 10 studies that examined fungi, nine demonstrated a positive relationship with thunderstorm asthma. The fungal taxa involved varied, depending on whether measurements were recorded before, during or after the thunderstorm. Nevertheless, none of the studies considered fungi as a potential effect modifier for the pollen-thunderstorm asthma association., Conclusion: We found evidence to suggest that grass pollen was a necessary factor for thunderstorm asthma but there are other as yet unrecognised environmental factors that may also be important. Further research is required to examine the role of fungi and other environmental factors such as air quality as potential effect modifiers of the association., Competing Interests: Declaration of competing interest Michael Abramson holds investigator-initiated grants from Pfizer and Boehringer-Ingelheim for unrelated research. He has also undertaken an unrelated consultancy for Sanofi., (Crown Copyright © 2019. Published by Elsevier Inc. All rights reserved.)

We examine the seasonality of asthma-related hospital admissions in Melbourne, Australia, in particular the contribution and predictability of episodic thunderstorm asthma. Using a time-series ecological approach based on asthma admissions to Melbourne metropolitan hospitals, we identified seasonal peaks in asthma admissions that were centred in late February, June and mid-November. These peaks were most likely due to the return to school, winter viral infections and seasonal allergies, respectively. We performed non-linear statistical regression to predict daily admission rates as functions of the seasonal cycle, weather conditions, reported thunderstorms, pollen counts and air quality. Important predictor variables were the seasonal cycle and mean relative humidity in the preceding two weeks, with higher humidity associated with higher asthma admissions. Although various attempts were made to model asthma admissions, none of the models explained substantially more variation above that associated with the annual cycle. We also identified a list of high asthma admissions days (HAADs). Most HAADs fell in the late-February return-to-school peak and the November allergy peak, with the latter containing the greatest number of daily admissions. Many HAADs in the spring allergy peak may represent episodes of thunderstorm asthma, as they were associated with rainfall, thunderstorms, high ambient grass pollen levels and high humidity, a finding that suggests thunderstorm asthma is a recurrent phenomenon in Melbourne that occurs roughly once per five years. The rarity of thunderstorm asthma events makes prediction challenging, underscoring the importance of maintaining high standards of asthma management, both for patients and health professionals, especially during late spring and early summer.

Grass pollen is a major trigger for allergic rhinitis and asthma, yet little is known about the timing and levels of human exposure to airborne grass pollen across Australasian urban environments. The relationships between environmental aeroallergen exposure and allergic respiratory disease bridge the fields of ecology, aerobiology, geospatial science and public health. The Australian Aerobiology Working Group comprised of experts in botany, palynology, biogeography, climate change science, plant genetics, biostatistics, ecology, pollen allergy, public and environmental health, and medicine, was established to systematically source, collate and analyse atmospheric pollen concentration data from 11 Australian and six New Zealand sites. Following two week-long workshops, post-workshop evaluations were conducted to reflect upon the utility of this analysis and synthesis approach to address complex multidisciplinary questions. This Working Group described i) a biogeographically dependent variation in airborne pollen diversity, ii) a latitudinal gradient in the timing, duration and number of peaks of the grass pollen season, and iii) the emergence of new methodologies based on trans-disciplinary synthesis of aerobiology and remote sensing data. Challenges included resolving methodological variations between pollen monitoring sites and temporal variations in pollen datasets. Other challenges included "marrying" ecosystem and health sciences and reconciling divergent expert opinion. The Australian Aerobiology Working Group facilitated knowledge transfer between diverse scientific disciplines, mentored students and early career scientists, and provided an uninterrupted collaborative opportunity to focus on a unifying problem globally. The Working Group provided a platform to optimise the value of large existing ecological datasets that have importance for human respiratory health and ecosystems research. Compilation of current knowledge of Australasian pollen aerobiology is a critical first step towards the management of exposure to pollen in patients with allergic disease and provides a basis from which the future impacts of climate change on pollen distribution can be assessed and monitored., (Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.)

Plants have been redesigned by humans since the advent of modern agriculture some 10000 years ago, to provide ever increasing benefits to society. The phenomenal success of the green revolution in converting biomass from vegetative tissues into grain yield has sustained a growing population. At the dawn of the 21st century the need to further optimise plant biomass (largely plant walls) for a sustainable future is increasingly evident as our supply of fossil fuels is finite and the quality of our crop-based foods (functional foods; also determined by the composition of walls) are critical to maintaining a healthy lifestyle. Our capacity to engineer 'designer walls' suited to particular purposes is challenging plant breeders and biotechnologists in unprecedented ways. In this review we provide an overview of the critical steps in the assembly and remodelling of walls, the success (or otherwise) of such approaches and highlight another complex network, the cell surface, as a cell wall integrity (CWI) sensor that exerts control over wall composition and will need to be considered in any future modification of walls for agro-industrial purposes., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The elongation (elo) mutants of barley (Hordeum vulgare cv 'Himalaya') are a class of dwarf plants with defects affecting cell expansion. The phenotypes of mutants in three of the elo loci (elo1, elo2 and elo3) are recessive to the wild-type allele, and the mutations at elo-4 and elo-5 are semi-dominant. Allelism tests showed that elo1, elo2 and elo3 were at separate loci, and mapping data indicated that elo-5 was possibly allelic to either elo1 or elo2. A phenotype common to all elo mutants was the presence of short, radially swollen cells on the leaf epidermis, indicating a defect in longitudinal cell expansion. In three of the mutants, elo1, elo3 and elo5, this was accompanied by a twisting growth habit. Two of the mutations, elo2 and elo-5, affected cell division, with aberrant periclinal cell division resulting in the formation of increased cell layers in the leaf epidermis of elo2 and elo-5 homozygotes and in the aleurone layer of elo2 grains. Misplaced anticlinal divisions also occurred in the elo-5 leaf epidermis. Leaf cell walls of all elo lines contained less cellulose than the wild- type, and the cortical microtubules in elongating root epidermal cells in some elo lines were more randomly oriented than in the wild-type, consistent with the presence of radially swollen cells. We discuss possible functions for the Elo genes in primary cell wall synthesis.

To identify integral and peripheral plasma membrane (PM) proteins from Oryza sativa (rice), highly enriched PM fractions from rice suspension cultured cells were analyzed using two complementary approaches. The PM was enriched using aqueous two-phase partitioning and high pH carbonate washing to remove soluble, contaminating proteins and characterized using enzymatic and immunological analyses. Proteins from the carbonate-washed PM (WPM) were analyzed by either one-dimensional gel electrophoresis (1D-SDS-PAGE) followed by tryptic proteolysis or proteolysis followed by strong cation exchange liquid chromatography (LC) with subsequent analysis of the tryptic peptides by LC-MS/MS (termed Gel-LC-MS/MS and 2D-LC-MS/MS, respectively). Combining the results of these two approaches, 438 proteins were identified on the basis of two or more matching peptides, and a further 367 proteins were identified on the basis of single peptide matches after data analysis with two independent search algorithms. Of these 805 proteins, 350 were predicted to be PM or PM-associated proteins. Four hundred and twenty-five proteins (53%) were predicted to be integrally associated with a membrane, via either one or many (up to 16) transmembrane domains, a GPI-anchor, or membrane-spanning beta-barrels. Approximately 80% of the 805 identified proteins were assigned a predicted function, based on similarity to proteins of known function or the presence of functional domains. Proteins involved in PM-related activities such as signaling (21% of the 805 proteins), transporters and ATPases (14%), and cellular trafficking (8%), such as via vesicles involved in endo- and exocytosis, were identified. Proteins that are involved in cell wall biosynthesis were also identified (5%) and included three cellulose synthase (CESA) proteins, a cellulose synthase-like D (CSLD) protein, cellulases, and several callose synthases. Approximately 20% of the proteins identified in this study remained functionally unclassified despite being predicted to be membrane proteins.

Barley endosperm begins development as a syncytium where numerous nuclei line the perimeter of a large vacuolated central cell. Between 3 and 6 days after pollination (DAP) the multinucleate syncytium is cellularized by the centripetal synthesis of cell walls at the interfaces of nuclear cytoplasmic domains between individual nuclei. Here we report the temporal and spatial appearance of key polysaccharides in the cell walls of early developing endosperm of barley, prior to aleurone differentiation. Flowering spikes of barley plants grown under controlled glasshouse conditions were hand-pollinated and the developing grains collected from 3 to 8 DAP. Barley endosperm development was followed at the light and electron microscope levels with monoclonal antibodies specific for (1-->3)-beta-D: -glucan (callose), (1-->3,1-->4)-beta-D: -glucan, hetero-(1-->4)-beta-D: -mannans, arabino-(1-->4)-beta-D: -xylans, arabinogalactan-proteins (AGPs) and with the enzyme, cellobiohydrolase II, to detect (1-->4)-beta-D: -glucan (cellulose). Callose and cellulose were present in the first formed cell walls between 3 and 4 DAP. However, the presence of callose in the endosperm walls was transient and at 6 DAP was only detected in collars surrounding plasmodesmata. (1-->3,1-->4)-beta-D: -Glucan was not deposited in the developing cell walls until approximately 5 DAP and hetero-(1-->4)-beta-D: -mannans followed at 6 DAP. Deposition of AGPs and arabinoxylan in the wall began at 7 and 8 DAP, respectively. For arabinoxylans, there is a possibility that they are deposited earlier in a highly substituted form that is inaccessible to the antibody. Arabinoxylan and heteromannan were also detected in Golgi and associated vesicles in the cytoplasm. In contrast, (1-->3,1-->4)-beta-D: -glucan was not detected in the cytoplasm in endosperm cells; similar results were obtained for coleoptile and suspension cultured cells.

A characteristic feature of grasses and commercially important cereals is the presence of (1,3;1,4)-beta-d-glucans in their cell walls. We have used comparative genomics to link a major quantitative trait locus for (1,3;1,4)-beta-d-glucan content in barley grain to a cluster of cellulose synthase-like CslF genes in rice. After insertion of rice CslF genes into Arabidopsis, we detected (1,3;1,4)-beta-d-glucan in walls of transgenic plants using specific monoclonal antibodies and enzymatic analysis. Because wild-type Arabidopsis does not contain CslF genes or have (1,3;1,4)-beta-d-glucans in its walls, these experiments provide direct, gain-of-function evidence for the participation of rice CslF genes in (1,3;1,4)-beta-d-glucan biosynthesis.

Convicilin, a trimeric globulin of pea (Pisum sativum L.) seeds, is closely related to vicilin and composed of polypeptides of 68.2 kilodaltons. A partial copy DNA (cDNA) clone encoding convicilin was isolated, sequenced, and used to select a convicilin gene from a pea genomic library. A part of the genomic clone was sequenced to obtain the coding sequences missing in the cDNA clone and a further 1 kilobase 5' to the start of transcription were also obtained. The entire sequence of convicilin was deduced from the combined genomic and cDNA sequences. The complete gene encoding convicilin was transferred to tobacco (Nicotiana tabacum L.) and the characteristics of its expression in the seeds of transgenic plants were studied. An unprocessed polypeptide, which was found only in the seeds of the transgenic plants, was identical in size to pea convicilin, and was recognized by vicilin antibodies. Convicilin, which does not undergo posttranslational cleavage in peas, was partially processed to polypeptides of a relative molecular mass (Mr) of approx. 50000 in transgenic tobacco seeds. There was a twofold variation in the level of convicilin accumulated by the mature seeds of a number of transgenic plants and this was well correlated with the number of gene copies incorporated in the different transformants. In the seeds of tobacco plants that contained a single copy of the transferred gene it was estimated that convicilin comprised up to 2% of the seed protein. Thus, using a combination of gene sequencing and expression in a heterologous host we believe we have characterized the gene corresponding to theCvc locus, whereas the gene described by D. Bown et al. (1988, Biochem J.,251, 717-726) probably encodes a minor convicilin-related protein.

A 5.5 kb Eco RI fragment containing a vicilin gene was selected from a Pisum sativum genomic library, and the protein-coding region and adjacent 5' and 3' regions were sequenced. A DNA construction comprising this 5.5 kb fragment together with a gene for neomycin phosphotransferase II was stably introduced into tobacco using an Agrobacterium tumefaciens binary vector, and the fidelity of expression of the pea vicilin gene in its new host was studied. The seeds of eight transgenic tobacco plants showed a sixteen-fold range in the level of accumulated pea vicilin. The level of accumulation of vicilin protein and mRNA correlated with the number of integrated copies of the vicilin gene. Pea vicilin was confined to the seeds of transgenic tobacco. Using immunogold labelling, vicilin was detected in protein bodies of eight out of ten embryos (axes plus cotyledons) and, at a much lower level, in two out of eleven endosperms. Pea vicilin was synthesized early in tobacco seed development; some molecules were cleaved as is the case in pea seeds, yielding a major parental component of M r∼50000 together with a range of smaller polypeptides.