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PubMed2007

Event-specific quantitative detection of nine genetically modified maizes using one novel standard reference molecule.

With the development of genetically modified organism (GMO) detection techniques, the Polymerase Chain Reaction (PCR) technique has been the mainstay for GMO detection, and real-time PCR is the most effective and important method for GMO quantification. An event-specific detection strategy based on the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene is being developed for its high specificity. This study establishes the event-specific detection methods for TC1507 and CBH351 maizes. In addition, the event-specific TaqMan real-time PCR detection methods for another seven GM maize events (Bt11, Bt176, GA21, MON810, MON863, NK603, and T25) were systematically optimized and developed. In these PCR assays, the fluorescent quencher, TAMRA, was dyed on the T-base of the probe at the internal position to improve the intensity of the fluorescent signal. To overcome the difficulties in obtaining the certified reference materials of these GM maizes, one novel standard reference molecule containing all nine specific integration junction sequences of these GM maizes and the maize endogenous reference gene, zSSIIb, was constructed and used for quantitative analysis. The limits of detection of these methods were 20 copies for these different GM maizes, the limits of quantitation were about 20 copies, and the dynamic ranges for quantification were from 0.05 to 100% in 100 ng of DNA template. Furthermore, nine groups of the mixed maize samples of these nine GM maize events were quantitatively analyzed to evaluate the accuracy and precision. The accuracy expressed as bias varied from 0.67 to 28.00% for the nine tested groups of GM maize samples, and the precision expressed as relative standard deviations was from 0.83 to 26.20%. All of these indicated that the established event-specific real-time PCR detection systems and the reference molecule in this study are suitable for the identification and quantification of these GM maizes.

Base Sequence↗

PubMedDate not supplied

Enumerating lepidopteran species associated with maize as a first step in risk assessment in the USA.

Pest management can have substantial impacts on non-target species both within and outside the units being managed. Assessment of these impacts is hampered by the lack of even the most basic checklist of the species present in most systems. The maize agroecosytem is of particular interest because of the large area covered and the intensity of widely varying forms of pest management. In this study a database of lepidopteran species that occur within the maize agroecosystem in the United States was compiled. The process was initiated by developing a list of plants present in maize using published sources and the first-hand knowledge of "weed" experts. This list of plant species associated with maize was then cross-listed with lepidopteran host feeding records using published sources. Finally, phenological profiles and conservation rankings were added. Although our list is not exhaustive, we found 132 plant species in 33 families associated with maize, and 229 lepidopteran species in 21 families that feed on these plants. The database of plants and lepidopteran species can be a starting point for assessment of risk to non-target Lepidoptera in maize from chemical control, biological control, and the use of transgenic Bt maize. The lepidopteran species associated with maize were found to be significantly less imperiled, as measured by their conservation rankings, than lepidopteran species as a whole in all habitats. This finding suggests that rare or endangered lepidopteran species are unlikely to be impacted by pest management in maize. Based on the likely lack of impact of pest management in maize on individual species, future studies should focus on potential impacts on the ecological services that lepidopteran species provide.

Agriculture↗

PubMedDate not supplied

Chemical composition and starch digestibility of tortillas prepared with non-conventional commercial nixtamalized maize flours.

Non-conventional nixtamalized maize flours elaborated by a factory in Mexico were used for tortilla preparation. Tortillas were stored at 4 degrees C for up to 72 h and the total starch, available starch, resistant starch and retrograded resistant starch were assessed. The traditional white tortilla, used as a control, showed higher protein and fat contents than blue maize tortilla, whereas a maize-bean mixed tortilla had the highest protein, ash and fat contents. Lower total starch was obtained in the maize-bean tortilla than in white and blue maize tortillas. The available starch content in all tortillas decreased with the cold-storage, although the change was more marked for blue-maize tortillas. The maize-bean mixed tortillas exhibited the lowest in vitro digestibility, which is consistent with the relatively high resistant starch levels in the bean. Differences in resistant starch content were found between the two maize tortillas, which might be related to the softer texture of blue-maize tortilla. The starch digestibility features of these new types of nixtamalized maize flours open up the possibility of producing tortillas with variable nutritional properties.

Bread↗

PubMed2002

Zinc absorption from a low-phytic acid maize.

BACKGROUND: Phytic acid reduction in cereal grains has been accomplished with plant genetic techniques. These low-phytic acid grains provide a strategy for improving the mineral (eg, zinc) status in populations that are dependent on grains, including maize (Zea mays L.), as major dietary staples. OBJECTIVE: The objective was to compare the fractional absorption of zinc from polenta prepared from maize low in phytic acid with that prepared from a wild-type isohybrid maize (control) after short-term consumption by adults whose habitual diet is low in phytic acid. DESIGN: Healthy adults served as their own control subjects in a crossover design. All meals on 1 d consisted of polenta prepared from a low-phytic acid maize homozygous for the recessive low phytic acid 1-1 (lpa1-1). On the preceding or following day, all meals consisted of polenta prepared from a sibling isohybrid homozygous wild-type maize with a "normal" phytic acid content. The low-phytic acid maize contained approximately 60% less phytic acid than did the wild-type maize. All test meals were extrinsically labeled with zinc stable-isotope tracers. The fractional absorption of zinc was determined on the basis of fecal enrichment. RESULTS: The molar ratios of phytic acid to zinc in the polenta prepared from lpa1-1 maize and the wild-type maize were 17:1 and 36:1, respectively. The corresponding fractional absorptions of zinc were 0.30 +/- 0.13 and 0.17 +/- 0.11, respectively (P < 0.005). CONCLUSION: Substitution of a low-phytic acid grain in a maize-based diet is associated with a substantial increase in zinc absorption.

Absorption↗

PubMed1997

Biodiversity of yeasts from Illinois maize.

Microflora in wound sites of preharvest maize (including bacteria, yeasts, and filamentous fungi) may play a role in attracting insects to maize plants and may also interact with growth and mycotoxin production by filamentous fungi. As little data are available about the yeasts occurring on maize from the U.S. corn belt, samples of milled maize from experimental plantings at the University of Illinois River Valley Sand Field were analyzed. Yeast counts showed slight yearly fluctuation and varied between 3.60 and 5.88 (log cfu/g maize). The majority of the yeasts were Candida guilliermondii (approximately 55%), Candida zeylanoides (24%), Candida shehatae (11%), and Debaryomyces hansenii (3%). Also present were Trichosporon cutaneum, Cryptococcus albidus var. aerius, and Pichia membranifaciens. The occurrence of killer yeasts was also evaluated. Killer yeasts were detected in maize for the first time and were identified as Trichosporon cutaneum and Candid zeylanoides. These were able to kill some representative yeasts isolated from maize, including Candida guilliermondii, Candida shehatae, and Cryptococcus albidus var. aerius. Other maize yeasts (Candida zeylanoides, Debaryomyces hansenii, Pichia membranifaciens) were not affected. The majority of yeasts found on maize were unable to ferment its major sugars, i.e., sucrose and maltose. Some (e.g., Candida zeylanoides) were not even able to assimilate these sugars. The importance of these properties in relation to insect attraction to preharvest ears of maize is discussed.

Yeasts↗

PubMed2001

Prospects for reducing fumonisin contamination of maize through genetic modification.

Fumonisins (FB) are mycotoxins found in (italic)Fusarium verticillioides-infected maize grain worldwide. Attention has focused on FBs because of their widespread occurrence, acute toxicity to certain livestock, and their potential carcinogenicity. FBs are present at low levels in most field-grown maize but may spike to high levels depending on both the environment and genetics of the host plant. Among the strategies for reducing risk of FB contamination in maize supplied to the market, development and deployment of Fusarium ear mold-resistant maize germplasm is a high priority. Breeding for increased ear mold tolerance and reduced mycotoxin levels is being practiced today in both commercial and public programs, but the amount of resistance achievable may be limited due to complicated genetics and/or linkage to undesirable agronomic traits. Molecular markers can be employed to speed up the incorporation of chromosomal regions that have a quantitative effect on resistance (quantitative trait loci). Transgenic approaches to ear mold/mycotoxin resistance are now feasible as well. These potentially include genetically enhanced resistance to insect feeding, increased fungal resistance, and detoxification/prevention of mycotoxins in the grain. An example of the first of these approaches is already on the market, namely transgenic maize expressing Bacillus thuringiensis (Bt) toxin, targeted to the European corn borer. Some Bt maize hybrids have the potential to reduce FB levels in field-harvested grain, presumably through reduced feeding of Bt-susceptible insects in ear tissues. However, improved ear mold resistance per se is still an important goal, as the plant will still be vulnerable to noninsect routes of entry to (italic)Fusarium. A second approach, transgene-mediated control of the ability of Fusarium to infect and colonize the ear, could potentially be achieved through overexpression of specific antifungal proteins and metabolites, or enhancement of the plant's own defense systems in kernel tissues. This has not yet been accomplished in maize, although promising results have been obtained recently in other monocots versus other fungal and bacterial pathogens. Achieving reproducible and stable enhanced ear mold resistance under field conditions will be immensely challenging for biotechnologists. A third approach, transgene strategies aimed at preventing mycotoxin biosynthesis, or detoxifying mycotoxins in planta, could provide further protection for the grower in environments where FBs present a risk to the crop even when the maize is relatively resistant to Fusarium mold. In one example of such a strategy, enzymes that degrade FBs have been identified in a filamentous saprophytic fungus isolated from maize, and corresponding genes have been cloned and are currently being tested in transgenic maize.

Animals↗

PubMed2006

Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) larval feeding behavior on transgenic maize (MON 863) and its isoline.

Diabrotica species (Coleoptera: Chrysomelidae) larval behavior studies have posed a challenge to researchers because of the subterranean life cycle of this pest. To fully understand how the western corn rootworm, Diabrotica virgifera virgifera LeConte, injures the maize, Zea mays L., root system, its behavior must be studied. For example, larvae that can detect an area of the root that has a lower amount of toxin, whether from an insecticide or a transgenic maize plant, have an increased chance of survival. This study assessed D. v. virgifera larval feeding behavior on rootworm-susceptible maize and maize containing a biotechnology-derived trait (MON 863) with resistance to D. v. virgifera first instar feeding. Maize plants were grown in a medium that allowed for direct observation and measurements during feeding of larval stadia. Neonates were placed on maize seedlings, and data were taken at 3, 6, 9, and 12 d postinfestation on resistant and susceptible maize. On rootworm-susceptible maize, neonate larvae aggregated at the root tips and began actively feeding, and then they moved to older root tissue. Conversely, some larvae that ingested Cry 3Bb1 from the resistant maize exhibited no movement. Other larvae on the resistant maize moved continuously, sampling root hairs or root tissue but not actively feeding. The continuously moving larvae had visibly empty guts, suggesting possible nonpreference for the resistant root. This study contributes to our understanding of D. v. virgifera larval behavior and provides insight into questions surrounding the potential evolution of behavioral and biochemical resistance to Cry3Bb1.

Animals↗

PubMed2002

Grass silage versus maize silage effects on retail packaged beef quality.

The effects of three preslaughter diets on heifer beef quality were investigated. Heifers (n = 45) were divided into three groups and fed for ad libitum consumption either maize silage, grass silage, or a 50:50 mixture of maize silage and grass silage. Meat quality was determined by measuring color, lipid oxidation, alpha-tocopherol levels, and fatty acid composition. Beef from the maize silage group had poorest color stability (P < 0.05), whereas beef from the grass silage diet had best (P < 0.05) color stability. The visual panel least preferred the maize silage group after 2 or more days of display, and lipid oxidation was significantly (P < 0.001) higher in this group compared to the 50:50 maize:grass silage and grass silage groups. There was a significant (P < 0.001) difference in the alpha-tocopherol levels detected in the meat from the three dietary groups. Alpha-tocopherol levels increased in the order: maize silage < 50:50 maize:grass silage < grass silage, at levels of 2.08, 2.95, and 3.84 microg/g meat, respectively. Fatty acid analysis indicated 18:3 was significantly (P < 0.001) lower in the maize silage-fed group than in the maize:grass silage and grass silage groups. However, 18:3 was significantly (P < 0.001) higher in the grass silage group than in the other two groups. There were no significant differences in all other fatty acids among the three dietary groups. It was concluded that beef from grass silage-fed animals had better overall quality in terms of color, lipid oxidation, and alpha-tocopherol levels than beef from maize silage fed animals.

Animals↗

PubMed2001

A multiplex PCR method of detecting recombinant DNAs from five lines of genetically modified maize.

Seven lines of genetically modified (GM) maize have been authorized in Japan as foods and feeds imported from the USA. We improved a multiplex PCR method described in the previous report in order to distinguish the five lines of GM maize. Genomic DNA was extracted from GM maize with a silica spin column kit, which could reduce experimental time and improve safety in the laboratory and potentially in the environment. We sequenced recombinant DNA (r-DNA) introduced into GM maize, and re-designed new primer pairs to increase the specificity of PCR to distinguish five lines of GM maize by multiplex PCR. A primer pair for the maize intrinsic zein gene (Ze1) was also designed to confirm the presence of amplifiable maize DNA. The lengths of PCR products using these six primer pairs were different. The Ze1 and the r-DNAs from the five lines of GM maize were qualitatively detected in one tube. The specific PCR bands were distinguishable from each other on the basis of the expected length. The r-DNA could be detected from maize samples containing 0.5% of each of the five lines of GM maize. The sensitivity would be acceptable to secure the verification of non-GMO materials and to monitor the reliability of the labeling system.

DNA Primers↗

PubMed2004

[Comparative analyses of QTL for important agronomic traits between maize and rice].

The objective of this study was to assess syntenic relationships of quantitative trait loci (QTL) for important agronomic traits between maize and rice based on the comparative genomic map of maize and rice using two F(2:3) populations. Through the comparisons, it was observed that there were extensive conserved relationships of maize QTL affected plant height, row number, and kernels per row with rice QTL affected plant height, tillers per plant, and grains per panicle respectively. Sixteen of 45 QTL affecting five different maize traits were conserved compared with 12 of 38 QTL affecting five different rice traits, which provided some useful information for locating, isolating and cloning maize QTL by using the rice genomic data. In this study, one QTL in rice usually had two conserved QTL in maize, further supporting the hypothesis that there is a polyploidization event during maize evolution. It was interested in observing that there were QTL rich regions on chromosomes in maize and rice, where QTL affecting different traits were usually clustered. These results revealed that the QTL affected the same or similar traits in maize and rice may have the common origin. These results will be helpful to map, isolate and clone QTL in large genome crops, such as maize, by using rice genome information, as well as to understand the evolutionary forces that structured the organization of the grass genomes.

Oryza↗

PubMed2005

Seed-specific expression of the lysine-rich protein gene sb401 significantly increases both lysine and total protein content in maize seeds.

The sb401 gene from potato (Solanum berthaultii) encoding a pollen-specific protein with high lysine content was successfully integrated into the genome of maize plants, and its expression was correlated with increased levels of lysine and total protein content in maize seeds. A plasmid vector containing the sb401 gene under the control of a maize seed-specific expression storage protein promoter (P19z) was constructed and introduced into maize calli by microprojectile bombardment. The integration of the sb401 gene into the maize genome was confirmed by Southern blot analysis, and its expression was confirmed by Western blot analysis. Quantification of the lysine and protein contents in R1 maize seeds showed that, compared with the nontransgenic maize control, the lysine content increased by 16.1% to 54.8% and the total protein content increased by 11.6% to 39.0%. There were no visible morphological changes in the vegetative parts and seeds of the transgenic maize plants. Lysine and protein analysis of the transgenic maize grains showed that the levels of lysine and total protein remained high for six continuous generations, indicating that the elevated lysine and total protein levels were heritable. These results indicate that the sb401 gene could be successfully employed in breeding programs aimed at improving the nutritional value of maize.

Consumer Product Safety↗

PubMed1992

Fungal transmission of a potyvirus: uredospores of Puccinia sorghi transmit maize dwarf mosaic virus.

Maize dwarf mosaic virus (MDMV) and maize rust, Puccinia sorghi Schw., occur as natural infections on cultivated maize in South Africa. P. sorghi often occurs as a secondary late infection on maize plants which have already been infected with MDMV earlier in the season, either seed or aphid transmitted. When MDMV isolates from maize plants naturally infected by both virus and fungus were propagated by sap inoculation in plant growth rooms, residual uredospores in the sap gave rise to the development of uredia under conditions of high humidity. When uredospores developing on MDMV-B-infected plants were germinated on virus free maize seedlings, these plants became infected with MDMV-B. Similarly, when uredospores, originating from maize plants infected with MDMV-A, were scattered onto virus free maize seedlings, these plants became infected with MDMV-A. The presence of virus on uredospores in infected plant tissue was visualized by indirect immunofluorescence. Identification of virus infection was by DAS-ELISA and immunoelectro-blotting utilizing strain-specific antisera. Virus transmission occurred between closely situated plants which had no actual contact (unaided transmission). MDMV-B transmission by uredospores, to new maize seedlings, has been maintained for three successive years (1988-1991) in a plant growth room. The MDMV-B isolate remained sap and non-persistently aphid transmissible.

Animals↗

PubMed2004

Rhizobium etli maize populations and their competitiveness for root colonization.

Rhizobium etli, which normally forms nitrogen-fixing nodules on Phaseolus vulgaris (common bean), is a natural maize endophyte. The genetic diversity of R. etli strains from bulk soil, bean nodules, the maize rhizosphere, the maize root, and inside stem tissue in traditional fields where maize is intercropped with P. vulgaris-beans was analyzed. Based on plasmid profiles and alloenzymes, it was determined that several R. etli types were preferentially encountered as putative maize endophytes. Some of these strains from maize were more competitive maize-root colonizers than other R. etli strains from the rhizosphere or from bean nodules. The dominant and highly competitive strain Ch24-10 was the most tolerant to 6-methoxy-2-benzoxazolinone (MBOA), a maize antimicrobial compound that is inhibitory to some bacteria and fungi. The R. tropici strain CIAT899, successfully used as inoculant of P. vulgaris, was also found to be a competitive maize endophyte in inoculation experiments.

DNA, Bacterial↗

PubMed2006

Assessing the effects of Bt Maize on the predatory mite Neoseiulus cucumeris.

The investigation of Neoseiulus cucumeris in the context of the ecological risk assessment of insect resistant transgenic plants is of particular interest as this omnivorous predatory mite species is commercially available and considered important for biological control. In a multitrophic feeding experiment we assessed the impact of Bt maize on the performance of N. cucumeris when offered spider mites (Tetranychus urticae) reared on Bt (Bt11, Syngenta) or non-Bt maize (near isogenic line) and Bt or non-Bt maize pollen as a food source. Various parameters including mortality, development time, oviposition rate were measured. Spider mites were used as a prey for N. cucumeris, since these herbivores are known to contain similar levels of Cry1Ab toxin, when reared on Bt maize, as those found in the transgenic leaf material. In contrast, toxin levels in pollen of this transgenic cultivar are very low. No differences in any of the parameters were found when N. cucumeris was fed with spider mites reared on Bt and non-Bt maize. Pollen was shown to be a less suitable food source for this predator as compared to spider mites. Moreover, subtle effects on female N. cucumeris (9% longer development time and 17% reduced fecundity) were measured when fed with pollen originating from Bt maize as compared to non-Bt maize pollen. Our findings indicate that the predatory mite N. cucumeris is not sensitive to the Cry1Ab toxin as no effects could be detected when offered Bt-containing spider mites, and that the effects found when fed with Bt maize pollen can be assigned to differences in nutritional quality of Bt and non-Bt maize pollen. The significance of these findings is discussed with regard to the ecological relevance for risk assessment of transgenic plants.

Animals↗

PubMed2005

Natural occurrence of Fusarium and subsequent fumonisin contamination in preharvest and stored maize in Benin, West Africa.

The natural occurrence of Fusarium and fumonisin contamination was evaluated from 1999 to 2003 in both preharvest and stored maize produced by small-scale farmers in four agroecological zones of Benin. Mycological analyses revealed a predominance of both Fusarium and Aspergillus in maize samples compared to other genera. The two Fusarium species most commonly isolated from maize were Fusarium verticillioides (68%) and Fusarium proliferatum (31%). Atypical isolates of F. verticillioides with some characteristics of Fusarium andiyazi but apparently closer to F. verticillioides, because the isolates were all high fumonisin producers, were also found only on preharvest maize. Study of F. verticillioides strains showed the presence of extremely high fumonisin producers in Benin with total fumonisin levels ranging from 8240 to 16,690 mg/kg. Apart from 2002-2003, Fusarium occurrence was not significantly different from one zone to another, although a slight decrease was observed from south, humid, to north, drier. Fusarium occurrence varied somewhat from one season to another. It significantly decreased over the 6 months of storage. Widespread fumonisin occurrence in maize was observed. Most of the maize samples collected were found positive for fumonisin with levels ranging from not detected to 12 mg/kg in 1999-2000, 6.7 mg/kg in 2000-2001 and 6.1 mg/kg in 2002-2003. Fumonisin levels in maize were found to be significantly higher in the two southern zones during all the surveys. The highest mean total fumonisin level was detected in 1999-2000 in maize samples from the southern Guinea Savannah (SGS) (12 mg/kg), whereas in both 2000-2001 and 2002-2003, it was in samples from the forest mosaic savannah (FMS) (6.7 and 6.1 mg/kg, respectively). Fumonisin levels varied from one season to another and, throughout the storage time, showing a decreasing trend in each zone. However, this decrease was not significant every season. An increasing trend was observed during some seasons in the SGS and northern Guinea Savannah (NGS) zones. The results of this study emphasise that farmers and consumers, not only in Benin but also in other West African countries, should be alerted to the danger of fumonisin contamination in maize.

Animals↗

PubMed2006

Maize meal predisposes to endemic squamous cancer of the oesophagus in Africa: breakdown of esterified linoleic acid to the free form in stored meal leads to increased intragastric PGE2 production and a low-acid reflux.

Endemic cancer of the oesophagus in Africa is associated with the use of maize as the staple. In one African community endemic cancer of the oesophagus has been shown to have a strong statistical association with the consumption of foods based on maize in the meal form. A strong association with consumption of maize meal has also been shown in a region of Italy. It has been argued in the past that the association with maize consumption is due to a fortuitous association; or due to the nutritional deficiencies of maize meal; or due to fungal contamination. We argue that maize meal is a distinctly different food from whole maize, and that nutritional content as much as deficiency is responsible for predisposition of the oesophagus to carcinogenesis. An important factor is the breakdown of esterified linoleic acid to the free form in stored maize meal. This leads to excess production of prostaglandin E2 in the stomach. The excess Prostaglandin E2 causes a low-acid duodenogastro-oesophageal reflux, which predisposes to carcinogenesis. Supporting evidence is available that the steps of this mechanism occur in endemic areas, and that they are associated with the carcinogenic process. Health measures including poverty alleviation, health education, and monitoring and control of maize meal storage and content may be required to reduce the incidence of this disease in Africa.

Africa↗

PubMed2006

Maize (Zea mays L.) genetic factors for preventing fumonisin contamination.

Fusarium moniliforme and Fusarium proliferatum are the most frequently isolated fungi from maize (Zea mays L.) in Spain. Both Fusarium species produce toxins potentially dangerous for animals and humans, the fumonisins being the most significant of those toxins. White maize is preferred for human consumption, and extra care should be taken to avoid kernel mycotoxin contamination. The objectives of this study were to identify and quantify kernel infection by Fusarium spp. and contamination by fumonisin on white maize hybrids, to search for white maize sources of resistance to infection by Fusarium spp. and mycotoxin contamination, and to preliminarily study the genetics involved in such resistances. Ten F(1) single crosses derived from a diallel mating design among five white maize inbreds were evaluated in a randomized complete block design with three replications in 2002 at two locations. Fusarium verticilloides and F. proliferatum were detected on kernels of white maize hybrids cultivated in northwestern Spain. No differences in fungal infection were found among maize genotypes, but differences in fumonisin contamination were significant and could be related, in part, to differences in husk tightness. Among the genotypes studied, general combining ability (GCA) effects were the most important for resistance to fumonisin contamination. Inbreds EP10 and EC22 showed the most favorable GCA effects for husk tightness and fumonisin content, and the cross between them, EP10 x EC22, had the most favorable specific combining ability (SCA) effect for husk tightness. Inbreds EP10 and EC22 showed favorable GCA effects for fumonisin contamination and husk tightness, and the cross EP10 x EC22 was the only one with an average fumonisin level below 1 mug/g. Although this should be confirmed with more extensive studies, white maize inbreds developed from white maize landraces could be sources of resistance to fumonisin contamination.

Breeding↗

PubMed2002

A functional antigen in a practical crop: LT-B producing maize protects mice against Escherichia coli heat labile enterotoxin (LT) and cholera toxin (CT).

We have produced a functional heat labile enterotoxin (LT-) B subunit of Escherichia coli in maize. LT-B is a multimeric protein that presents an ideal model for an edible vaccine, displaying stability in the gut and inducing mucosal and systemic immune responses. Transgenic maize was engineered to synthesize the LT-B polypeptides, which assembled into oligomeric structures with affinity for G(M1) gangliosides. We orally immunized BALB/c mice by feeding transgenic maize meal expressing LT-B or non-transgenic maize meal spiked with bacterial LT-B. Both treatments stimulated elevated IgA and IgG antibodies against LT-B and the closely related cholera toxin B subunit (CT-B) in serum, and elevated IgA in fecal pellets. The transgenic maize induced a higher anti-LT-B and anti-CT-B mucosal and serum IgA response compared to the equivalent amount of bacterial LT-B spiked into maize. Following challenge by oral administration of the diarrhea inducing toxins LT and CT, transgenic maize-fed mice displayed reduced fluid accumulation in the gut compared to non-immunized mice. Moreover, the gut to carcass ratio of immunized mice was not significantly different from the PBS (non-toxin) challenged control group. We concluded that maize-synthesized LT-B had features of the native bacterial LT-B such as molecular weight, G(M1) binding ability, and induction of serum and mucosal immunity. We have demonstrated that maize, a major food and feed ingredient, can be efficiently transformed to produce, accumulate, and store a fully assembled and functional candidate vaccine antigen.

Administration, Oral↗