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PubMedDate not supplied

The co-existence between transgenic and non-transgenic maize in the European Union: a focus on pollen flow and cross-fertilization.

The ongoing discussion on the co-existence between genetically modified (GM) and non-GM crops becomes more important in the European Union (EU). With the recent inscription of 17 GM maize varieties in the common EU catalogue of varieties of agricultural plant species, the acreage of transgenic maize for market purposes is expected to increase in some European countries. In the EU, specific tolerance thresholds have been established for the adventitious and technically unavoidable presence of GM material in non-GM produce, and member states are elaborating legal frames to cope with co-existence. As maize is a cross-pollinated crop relying on wind for the dispersal of its pollen, technical management measures will be imposed to reduce cross-fertilization between transgenic and non-transgenic maize. Various biological, physical and analytical parameters have been identified to play a role in the study of cross-fertilization in maize. This variability may hamper the comparison between research results and may complicate the definition of appropriate isolation distances and/or pollen barriers in order to limit out-crossing. The present review addresses these parameters and proposes containment measures in order to not exceed the legal labeling thresholds in maize.

Agriculture↗

PubMed2004

Contamination of refuges by Bacillus thuringiensis toxin genes from transgenic maize.

Transgenic crops producing insecticidal toxins from Bacillus thuringiensis (Bt) are widely used to control pests, but their benefits will be lost if pests evolve resistance. The mandated high-dose/refuge strategy for delaying pest resistance requires planting refuges of toxin-free crops near Bt crops to promote survival of susceptible pests. We report that pollen-mediated gene flow up to 31 m from Bt maize caused low to moderate Bt toxin levels in kernels of non-Bt maize refuge plants. Immunoassays of non-Bt maize sampled from the field showed that the mean concentration of Bt toxin Cry1Ab in kernels and the percentage of kernels with Cry1Ab decreased with distance from Bt maize. The highest Bt toxin concentration in pooled kernels of non-Bt maize plants was 45% of the mean concentration in kernels from adjacent Bt maize plants. Most previous work on gene flow from transgenic crops has emphasized potential effects of transgene movement on wild relatives of crops, landraces, and organic plantings, whereas implications for pest resistance have been largely ignored. Variable Bt toxin production in seeds of refuge plants undermines the high-dose/refuge strategy and could accelerate pest resistance to Bt crops. Thus, guidelines should be revised to reduce gene flow between Bt crops and refuge plants.

Animals↗

PubMed2006

Early maize (Zea mays L.) cultivation in Mexico: dating sedimentary pollen records and its implications.

A sedimentary pollen sequence from the coastal plain of Veracruz, Mexico, demonstrates maize cultivation by 5,000 years ago, refining understanding of the geography of early maize cultivation. Methodological issues related to bioturbation involved in dating that record combine with its similarity to a pollen sequence from the coastal plain of Tabasco, Mexico, to suggest that the inception of maize cultivation in that record occurred as much as 1,000-2,000 years more recently than the previously accepted 7,000 years ago. Our analysis thereby has substantive, theoretical, and methodological implications for understanding the complex process of maize domestication. Substantively, it demonstrates that the earliest securely dated evidence of maize comes from macrofossils excavated near Oaxaca and Tehuacán, Mexico, and not from the coastal plain along the southern Gulf of Mexico. Theoretically, that evidence best supports the hypothesis that people in the Southern Highlands domesticated this important crop plant. Methodologically, sedimentary pollen and other microfossil sequences can make valuable contributions to reconstructing the geography of early maize cultivation, but we must acknowledge the limits to precision that bioturbation in coastal lagoons imposes on the dating of such records.

Agriculture↗

PubMed1981

Overlapping divergent genes in the maize chloroplast chromosome and in vitro transcription of the gene for tRNA.

In the presence of the S polypeptide, maize chloroplast DNA-dependent RNA polymerase preferentially transcribes sequences within the 2200-nucleotide-pair-long maize chloroplast chromosome fragment Eco [unk] from a supercoiled chimeric plasmid cloned in Escherichia coli [Jolly, S. O. & Bogorad, L. (1980) Proc. Natl. Acad. Sci. USA 77, 822-826]. Eco [unk] contains one gene for tRNA(His) and one for a 1.6-kilobase RNA that includes an open reading frame. These two genes overlap by at least a few nucleotides and are transcribed divergently from complementary DNA strands. This indicates possible transcriptional regulation of chloroplast DNA at the nucleotide level. The 5' end of tRNA(His) (G-U-G) isolated from maize chloroplasts is indistinguishable from that of the transcript produced from Eco [unk] in vitro by maize chloroplast DNA-dependent RNA polymerase. This purified system initiates RNA synthesis faithfully and exhibits preference for some chloroplast genes. Maize chloroplast DNA for tRNA(His) lacks the sequence C-C-A at its 3' terminus; it is presumably added post-transcriptionally. Maize tRNA(His) has both prokaryotic and eukaryotic features.

Journal Article↗

PubMed1986

Differential expression of six light-harvesting chlorophyll a/b binding protein genes in maize leaf cell types.

Bundle sheath chloroplasts of maize leaves contain about one-fourth as much light-harvesting chlorophyll a/b binding protein of photosystem II (LHCP-II) as do mesophyll chloroplasts. We have determined that this difference is, in part, the result of differential expression of different LHCP-II genes. We have prepared and partially characterized cDNA clones specific for six LHCP-II genes of maize. Transcripts of these six LHCP-II genes are present at vastly different levels and account for about 95% of total LHCP-II mRNAs in bundle sheath and mesophyll cells of illuminated dark-grown maize leaves. Three genes are preferentially expressed in mesophyll cells, and their mRNAs constitute about 54% of the total LHCP-II transcripts in greening (24 hr) maize leaves. Two genes are expressed equally in bundle sheath and mesophyll cells. Most interestingly, the RNA of one gene that contributes about 8% of the total LHCP-II transcripts in leaves greening for 24 hr is present at a much higher level in bundle sheath than in mesophyll cells. Moreover, immunoblot analysis of maize thylakoids reveals at least five sizes of LHCP-II; these also differ from one another in their relative abundance in bundle sheath and mesophyll cells of developing maize leaves.

Chlorophyll↗

PubMed1995

Maize branching enzyme catalyzes synthesis of glycogen-like polysaccharide in glgB-deficient Escherichia coli.

The structure of alpha-glucan, isolated from wild-type Escherichia coli B, a glycogen branching enzyme (BE)-deficient E. coli AC71 (glgB-), or from AC71 transformed with genes coding for maize BEI and BEII individually as well as with both genes, was analyzed by high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection. Transformation of the maize BE gene(s) in AC71 (glgB-) showed complementation in branching activity. Analysis by HPAEC revealed different structures between glycogen of E. coli B and alpha-glucan of AC71 transformed with a different maize BE gene(s). The individual chains of the alpha-glucan debranched with isoamylase were distributed between chain length (CL) 3 and > 30 and the chain with CL 6 was the most abundant. In comparison with the glycogen of E. coli B, the alpha-glucan of AC71 transformed with the maize BE gene(s) consisted of a lesser amount of chains with CL 7-9 and a larger amount of chains with CL > 14. It also showed a broad peak with chains of CL 9-12 as in maize amylopectin. This study provides in vivo evidence that glycogen BE and maize BE isozymes may have different specificities in the length of chain transferred. Furthermore, this study suggests that the specificity of glycogen synthase and starch synthase and their concerted action with BE play an important role in determining the structure of the polysaccharide synthesized.

1,4-alpha-Glucan Branching Enzyme↗

PubMed1997

DNA sequence evidence for the segmental allotetraploid origin of maize.

It has long been suspected that maize is the product of an historical tetraploid event. Several observations support this possibility, including the fact that the maize genome contains duplicated chromosomal segments with colinear gene arrangements. Some of the genes from these duplicated segments have been sequenced. In this study, we examine the pattern of sequence divergence among 14 pairs of duplicated genes. We compare the pattern of divergence to patterns predicted by four models of the evolution of the maize genome-autotetraploidy, genomic allotetraploidy, segmental allotetraploidy, and multiple segmental duplications. Our analyses indicate that coalescent times for duplicated sequences fall into two distinct groups, corresponding to roughly 20.5 and 11.4 million years. This observation strongly discounts the possibility that the maize genome is the product of a genomic allotetraploid event, and it is also difficult to reconcile with either autotetraploidy or multiple independent segmental duplications. However, the presence of two (and only two) coalescent times is predicted by the segmental allotetraploid model. If the maize genome is the product of a segmental allotetraploid event, as these data suggest, then its two diploid progenitors diverged roughly 20.5 million years ago (Mya), and the allotetraploid event probably occurred approximately 11.4 Mya. Comparison of maize and sorghum sequences suggests that one of the two ancestral diploids shares a more recent common ancestor with sorghum than it does with the other ancestral diploid.

Journal Article↗

PubMed1997

A deletion in an indole synthase gene is responsible for the DIMBOA-deficient phenotype of bxbx maize.

The biosynthesis of DIMBOA, a pesticidal secondary metabolite of maize, branches off the tryptophan pathway. We have previously demonstrated that indole is the last intermediate common to both the tryptophan and hydroxamic acid pathways. The earliest discovered mutant in the DIMBOA pathway, bxbx (benzoxazineless), is deficient in the production of DIMBOA and related compounds. This paper presents evidence that a gene identified by Kramer and Koziel [Kramer, V. C. & Koziel, M. G. (1995) Plant Mol. Biol. 27, 1183-1188] as maize tryptophan synthase alpha (TSA) is the site of the genetic lesion in the DIMBOA-deficient mutant maize line bxbx. We demonstrate that the TSA gene has sustained a 924-bp deletion in bxbx compared with its counterpart in wild-type maize. We report that the TSA gene maps to the same location as the bxbx mutation, on the short arm of chromosome 4. We present evidence that the very early and very high level of expression of TSA corresponds to the timing and level of DIMBOA biosynthesis but is strikingly different from the expression of the maize tryptophan synthase beta (TSB) genes. We show that feeding indole to bxbx seedlings restores their ability to synthesize DIMBOA. We conclude that the maize enzyme initially named tryptophan synthase alpha in fact is a DIMBOA biosynthetic enzyme, and we propose that it be renamed indole synthase. This work confirms and enlarges upon the findings of Frey et al. [Frey, M., Chomet, P., Glawischniq, E., Stettner, C., Grün, S., Winklmair, A., Eisenreich, W., Bacher, A., Meeley, R. B., Briggs, S. P., Simcox, K. & Gierl, A. (1997) Science 277, 696-699], which appeared while the present paper was in review.

Benzoxazines↗

PubMed1997

Inhibition of the iron-induced ZmFer1 maize ferritin gene expression by antioxidants and serine/threonine phosphatase inhibitors.

Two pathways have been implicated in the regulation of maize ferritin synthesis in response to iron. One of them involves the plant hormone abscisic acid (ABA) and controls the expression of ZmFer2 gene(s). Another pathway, ABA-independent, has been characterized in a de-rooted maize plantlet system and involves an oxidative step. The ZmFer1 maize ferritin gene is not regulated by ABA, and it is shown in this paper that the corresponding mRNA accumulates in de-rooted maize plantlets and BMS (Black Mexican Sweet) maize cell suspension cultures in response to iron via the oxidative pathway described previously. To investigate ZmFer1 gene regulation further, the BMS cell system has been used to develop a transient expression assay using a ZmFer1-beta-glucuronidase fusion. Both iron induction and antioxidant inhibition of ZmFer1 gene expression were observed in this system. Using Northern blot analysis and transient expression experiments, it was shown that both okadaic acid and calyculin A, two serine/ threonine phosphatase inhibitors, specifically inhibit ZmFer1 gene expression. These data indicate that an okadaic acid-sensitive protein phosphatase activity is involved in the regulation of the ZmFer1 ferritin gene in maize cells, and this activity is required for iron-induced expression of this gene.

Abscisic Acid↗

PubMed2005

Abundance, dispersion and parasitism of the stem borer Busseola fusca (Lepidoptera: Noctuidae) in maize in the humid forest zone of southern Cameroon.

This study was conducted in the humid forest zone of Cameroon, in 2002 and 2003. The main objective was to investigate the effects of intercropping on infestation levels and parasitism of the noctuid maize stem borer Busseola fusca Fuller. Two trials were planted per year, one during the long and one during the short rainy season. Maize monocrops were compared with maize/legume or maize/cassava intercrops in two spatial arrangements: maize on alternate hills or in alternate rows. Spatial analyses showed that the stemborer egg batches were regularly dispersed in the maize monocrop and aggregated in the intercrops, as indicated by b, the index of dispersion of Taylor's power law. Depending on the crop association and planting pattern, intercrops reduced the percentage of plants with stem borer eggs by 47.4-58.4% and egg densities by 41.2-54.5% compared to monocropped maize. Consequently, larval densities were 44.4-61.5% lower in intercrops compared to monocrops. Intercropping maize with non-host plants did not affect larval parasitism. Up to two-fold higher levels of egg parasitism by scelionid Telenomus spp. were recorded in inter- compared to monocrops during the short rainy seasons of 2002 and 2003. No differences were found among the mixed cropping treatments and parasitism was lower during the long compared to the short rainy seasons. It was proposed that differences in levels of parasitism were due to density dependence effects rather than the effect of the presence of non-host plants in the system.

Agriculture↗

PubMed1981

Degradation of maize protein in rumen contents. Influence of ammonia concentration.

1. The influence of ammonia concentration on the distribution of nitrogen derived from opaque-2 maize uniformly-labelled with 15N has been investigated during short-term in vitro incubation of bovine rumen contents. 2. Less 15N derived from maize was found in the non-protein-N (NPN) fraction during incubation without added NH3 than with added NH3, due entirely to differences in the amount of N derived from maize in the NH3 fraction. 3. From calculations based on the transfer of N derived from maize to the NPN pool and to a bacterial fraction, it was concluded that degradation of maize protein was not influenced by NH3 concentration within the examined limits. 4. The decrease in relative amount of N derived from maize in the NH3 fraction at low concentrations of NH3, together with evidence for an increased fractional turnover rate of NH3-N suggests that a deficient supply of NH3 is compensated for by increased catabolism of nitrogenous compounds derived from the rumen micro-organisms.

Ammonia↗

PubMed1985

Insulin action and glucose metabolism in sheep fed on dried-grass or ground, maize-based diets.

The effect of an exogenous supply of glucose, provided by the digestion of maize starch in the small intestine, on endogenous glucose metabolism and insulin action was studied in sheep using the euglycaemic insulin clamp procedure. Insulin was infused intravenously at rates of 0.2, 0.5, 1.0 and 6.0 mU/min per kg live weight for four consecutive periods in each of four sheep fed on dried-grass and maize-based diets. Glucose was also infused intravenously at a variable rate, sufficient to maintain the plasma glucose concentration at basal levels. Whole-body rates of glucose metabolism were determined using a continuous infusion of [6-3H]glucose. From the resulting insulin dose-response curves, it was observed that, when the sheep were fed on the dried-grass diet, the responsiveness of glucose metabolism to insulin was less than that reported for non-ruminants. When fed the maize-based diet, the glucose metabolic clearance rates (MCR) observed during insulin infusions were significantly greater (P less than 0.05) than those observed for the dried-grass diet. However, after correcting for the non-insulin-mediated glucose disposal, differences between diets were not significant. The sensitivity of glucose utilization to insulin was not affected by diet. The plasma insulin concentrations causing half-maximal insulin-mediated glucose MCR were 103 (SE 21) and 85 (SE 11) mU/l for the dried-grass and maize-based diets respectively. The sensitivity of endogenous glucose production to insulin was also unaffected by diet. The plasma insulin concentrations resulting in the suppression of endogenous glucose production to half the basal level were 80 (SE 26) and 89 (SE 29) mU/l for the dried-grass and maize-based diets respectively. It is concluded that the observed increase in glucose utilization on the maize-based diet was due partly to a slight change in responsiveness to insulin and also partly to a change in the rate of non-insulin-mediated glucose disposal.

Animal Feed↗

PubMed1987

Influence of rumen ammonia concentration on the rumen degradation rates of barley and maize.

Four rumen-cannulated steers were given barley and maize diets supplemented with graded levels of an ammonium acetate solution. Animals were fed hourly from automatic feeders and water consumption was controlled to achieve steady-state conditions in the rumen. Dacron bags containing rolled barley or ground barley were incubated in the rumen of barley-fed steers, while ground maize and autoclaved maize were incubated in the rumen of maize-fed steers. Fractional degradation rates of dry matter were estimated for each cereal substrate incubated using a single-pol exponential decay model. No differences in degradation rate due to the method of feed processing were detected; however, barley was degraded at a faster rate than maize. Furthermore, the minimum rumen ammonia-nitrogen concentration required to maximize the degradation rate of barley (125 mg/l) was greater than that required to maximize the degradation rate of maize (61 mg/l). These results indicate that the optimal NH3-N concentration required to maximize the rate of grain digestion in the rumen is influenced by the chemical or structural characteristics of the grain.

Acetates↗

PubMed1994

Evaluation of fermentability of acid-treated maize husk by rat caecal bacteria in vivo and in vitro.

Fermentable energy in insoluble dietary fibre (DF) sources was evaluated by in vivo and in vitro methods using rats. Test diets contained 50 and 100 g maize husk or organic-acid-treated maize husk/kg diet. Soluble fractions were removed from both the DF sources by washing. The acid treatment increased digestibility by a microbial hemicellulase from 12.7% to 32.6%. The fermentability of DF was evaluated by measurement of the production rate of short-chain fatty acids (SCFA) in a short-term in vitro incubation of the caecal contents of rats fed on test diets for 22 d. The production rates of the major SCFA, acetic, propionic and butyric acids, were increased by feeding both DF sources, and these production rates in the acid-treated DF group were significantly higher than those in the untreated DF group. The production rate of a minor SCFA, isovaleric acid, was decreased by feeding both diets. The production rate of total SCFA in rats given the acid-treated maize husk was 32.6% higher than that in rats given the untreated maize husk. The fermentable energy in DF was estimated in vivo by subtracting the faecal excretion of DF energy from ingested DF energy. The fermentable energy in DF was increased by the acid treatment (32.5% in maize husk and 63.4% in acid-treated maize husk), which agreed with the SCFA production rate predicted in the caecum. These results indicate that a short-term incubation of caecal contents is a useful method for evaluation of the fermentability of DF sources, and that acid treatment can increase the fermentability of an insoluble DF source.

Acids↗

PubMed2003

Fate of genetically modified maize DNA in the oral cavity and rumen of sheep.

The polymerase chain reaction (PCR) technique was used to investigate the fate of a transgene in the rumen of sheep fed silage and maize grains from an insect-resistant maize line. A 1914-bp DNA fragment containing the entire coding region of the synthetic cryIA(b) gene was still amplifiable from rumen fluid sampled 5 h after feeding maize grains. The same target sequence, however, could not be amplified from rumen fluid sampled from sheep fed silage prepared from the genetically modified maize line. PCR amplification of a shorter (211-bp), yet still highly specific, target sequence was possible with rumen fluid sampled up to 3 and 24 h after feeding silage and maize grains, respectively. These findings indicate that intact transgenes from silage are unlikely to survive significantly in the rumen since a DNA sequence 211-bp long is very unlikely to transmit genetic information. By contrast, DNA in maize grains persists for a significant time and may, therefore, provide a source of transforming DNA in the rumen. In addition, we have examined the biological activity of plasmid DNA that had previously been exposed to the ovine oral cavity. Plasmid extracted from saliva sampled after incubation for 8 min was still capable of transforming competent Escherichia coli to kanamycin resistance, implying that DNA released from the diet within the mouth may retain sufficient biological activity for the transformation of competent oral bacteria.

Animal Feed↗

PubMed2003

Effects of graded levels of Fusarium toxin contaminated maize in diets for female weaned piglets.

A dose response study was carried out with piglets to examine the effects of increasing amounts of Fusarium toxins in the diet on performance, clinical serum characteristics, organ weights and residues of zearalenone (ZON) and deoxynivalenol (DON) and their metabolites in body fluids and tissues. For this purpose, Fusarium toxin contaminated maize (1.2 mg ZON and 8.6 mg DON per kg maize) was incorporated into a maize based diet for piglets at 0, 6, 12.5, 25 and 50% at the expense of control maize. The experimental diets were tested on 100 female piglets allotted to 20 boxes (five animals per box) covering a body weight range of 12.4 +/- 2.2 kg to 32.5 +/- 5.6 kg. Voluntary feed intake and, consequently, body weight gain of the animals receiving the highest proportion of Fusarium toxin contaminated maize were significantly decreased while the feed conversion ratio was not affected by the treatment. The mean weight of the uterus related to the body weight of the animals of the same group was increased by almost 100% as compared to the control. For this group, significantly decreased values of total serum protein were determined, while the serum activity of the liver enzyme glutamate dehydrogenase and the serum concentration of the follicle stimulating hormone were decreased for all treatment groups receiving 6% contaminated maize or more in the diet. Serum concentrations of immuneglobulins were not consistently altered by the treatment. Corresponding to the dietary exposure, increasing concentrations of ZON and alpha-zearalenol were detected in the bile fluid, liver and in pooled urine samples. The metabolite beta-zearalenol was detected only in bile fluid. The total concentration of ZON plus its metabolites in bile fluid correlated well with the diet contamination (r = 0.844). DON was found in serum, bile fluid and pooled urine samples while de-epoxy-DON was detected only in urine. The serum concentration of DON correlated well with the respective toxin intake 3-4 h prior to slaughtering (r = 0.957). For all mentioned analyses of residues it has to be noted that toxin residues were detectable even if negligible concentrations were present in the diet.

Animal Feed↗

PubMed2001

Nutrient digestibility and intestinal viscosities in broiler chickens fed on wheat diets, as compared to maize diets with added guar gum.

1. Three maize diets containing 0, 1 or 3 g of guar gum per kg and 2 wheat diets were given to male broiler chickens for measurements of growth performance and nutrient digestibility. 2. The intestinal viscosities found with wheat diets were within the range of those observed with the 3 maize diets. The correlations between in vivo and in vitro viscosities were higher with real than with potential applied viscosity. 3. The gain:food ratio decreased (P < 0.05) with the addition of 3 g guar gum/kg. The mean gain:food ratio observed for wheat diets was lower (P = 0.007) than the mean value found for the 3 maize diets. 4. The negative effects of increased intestinal viscosity due to guar gum addition on nutrient digestibility were highest for lipids and lowest for starch. 5. Wheat diets resulted in much lower (P < 0.001) starch apparent digestibility compared to maize diets with added guar gum despite similar mean intestinal viscosities in the 2 groups of diets. Apparent lipid digestibility with wheat diets was lower (P < 0.01) than the value predicted from intestinal viscosity, the predictive model having been calculated with the guar gum added maize diets. Apparent protein digestibility did not differ between wheat diets and guar gum added maize diets. 6. It was concluded that the low apparent digestibilities of starch and lipid observed with wheats could not be attributed only to intestinal viscosity and that other factors appear to be implicated in the low digestibilities observed with the wheat samples tested in the present experiment.

Animal Feed↗

PubMed2003

Supplements of transgenic malt or grain containing (1,3-1,4)-beta-glucanase increase the nutritive value of barley-based broiler diets to that of maize.

1. A diet with addition to normal barley of malt from transgenic barley expressing a protein engineered, thermotolerant Bacillus (1,3-1,4)-beta-glucanase during germination has previously been demonstrated to provide a broiler chicken weight gain comparable to maize diets. It also reduced dramatically the number of birds with adhering sticky droppings, but did not entirely eliminate sticky droppings. One of the objectives of the broiler chicken trials reported here was to determine if higher concentrations of transgenic malt could alleviate the sticky droppings. 2. Another aim was to investigate the feasibility of using mature transgenic grain containing the thermotolerant (1,3-1,4)-beta-glucanase as feed addition and to compare diets containing transgenic grain to a diet with the recommended amount of a commercial beta-glucanase-based product. 3. Inclusion of 75 or 151 g/kg transgenic malt containing 4.7 or 98 mg/kg thermotolerant (1,3-1,4)-beta-glucanase with 545 or 469 g/kg non-transgenic barley instead of maize yielded a weight gain in Cornish Cross broiler chickens indistinguishable from presently used maize diets. The gene encoding the enzyme is expressed in the aleurone with a barley alpha-amylase gene promoter and the enzyme is synthesised with a signal peptide for secretion into the endosperm of the malting grain. 4. Equal weight gain was achieved, when the feed included 39 g/kg transgenic barley grain [containing 66 mg/kg thermotolerant (1,3-1,4)-beta-glucanase] and 581 g/kg non-transgenic barley instead of maize. In this case, the gene encoding the enzyme has been expressed with the D-hordein gene (Hor3-1) promoter during grain maturation. The enzyme is synthesised as a precursor with a signal peptide for transport through the endoplasmic reticulum and targeted into the storage vacuoles. Deposition of the enzyme in the prolamin storage protein bodies of the endosperm protects it from degradation during the programmed cell death of the endosperm in the final stages of grain maturation and provides extraordinary heat stability. The large amount of highly active (1,3-1,4)-beta-glucanase in the mature grain allowed the reduction of the transgenic grain ingredient to 0.2 g/kg diet, thus making the ingredient comparable to that of the trace minerals added to standard diets. 5. A direct comparison using transgenic grain supplement at the level of 1 g/kg of feed with the standard recommended addition of the commercial enzyme preparation Avizyme 1100 at 1 g/kg yielded equal weight gain, feed consumption and feed efficiency in birds fed a barley-based diet. 6. The production of sticky droppings characteristic of broilers fed on barley diets was avoided with all 9 experimental diets and reduced to the level observed with a standard maize diet by supplementation with transgenic barley. 7. The excellent growth and normal survival of the 400 broilers tested on barley diets supplemented with transgenic grain or malt showed the grain and malt not to be toxic. 8. The barley feed with added transgenic grain or malt containing thermotolerant (1,3-1,4)-beta-glucanase provides an environmentally friendly alternative to enzyme additives, as it uses photosynthetic energy for production of the enzyme in the grain and thus avoids use of non-renewable energy for fermentation. The deposition of the enzyme in the protein bodies of the grain in the field makes coating procedures for stabilisation of enzyme activity superfluous. 9. Barley feed with the small amount of transgenic grain as additive to normal barley provides an alternative for broiler feed in areas where grain maize cannot be grown for climatic reasons or because of unsuitable soil and thus has to be imported.

Animal Feed↗