Isogenic Maize Research Articles

Different maize varieties have greater impact on Tetranychus urticae (Acari: Tetranychidae) than GE maize expressing Cry3Bb1 insecticidal protein

Genetically engineered (GE) maize expressing Bacillus thuringiensis Cry3Bb1 protein was developed to control Diabrotica spp. (Coleoptera: Chrysomelidae). However, Cry proteins have been reported to have effects on non-target arthropods. We therefore investigated whether the non-target pest Tetranychus urticae (Acari: Tetranychidae) was negatively affected by GE maize expressing the insecticidal Cry3Bb1 protein. Five treatments were used in the laboratory tests to determine the life history parameters of T. urticae on leaves of field-grown maize varieties: (1) GE maize MON 88017, (2) isogenic maize, (3) isogenic maize protected with the soil applied insecticide chlorpyrifos (Dursban 10G), and two unrelated varieties (4) Kipous and (5) PR38N86. Newly emerged T. urticae larvae were individually released on the upper surface of leaf discs placed on water saturated cotton wool. Immatures and adults survival, duration of developmental stages and female fecundity were recorded daily until T. urticae died. Age–stage, two-sex life table method and test for trends, revealed no significant differences in 13 of 18 studied parameters. The unrelated varieties Kipous and PR38N86 on one side and maize with the same genetic background, namely GE maize and isogenic maize with or without insecticide protection, on the other side, showed significant variations in male longevity, larval survival rate, preoviposition period, and fecundity. In addition to the differences between varieties, GE maize and insecticide-protected isogenic maize showed a substantial difference in age-specific fecundity, but not in the mean number of eggs laid by females.The obtained results do not indicate that consumption of Cry3Bb1 has negative effect on T. urticae and suggest that GE maize does not pose a risk with respect to the non-target mite pest T. urticae. The results may have implications for the approval and renewal of import and cultivation for GE crop in the European Union.

Phenotypic identification of near isogenic maize teosinte lines (Zea spp.) involved for resistance to gray leaf spot

AbstractWild germplasm of Zea is a source of variation that could be used in plant breeding to improve maize resistance to diseases such as gray leaf spot (GLS). The objectives were to develop a set of near‐isogenic lines (NILs) carrying introgressions from almost all known wild teosinte taxa in a common maize inbred line background (CML311) and to identify NILs expressing superior resistance to GLS. We developed 2,208 BC3‐derived NILs representing 79 different teosinte populations of 10 taxa as donor parents. We compared the means of NILs grouped by donor to the recurrent and identified donor population El Saucito, (Zea mays ssp. parviglumis race Balsas) as the donor source of NILs with highest mean resistance to GLS. Among the 2,208 NILs, 116 representing diverse introgressions of Zea donors were significantly more resistant than CML311. The most resistant individual line was T1662, derived from San Antonio Tlaltecahuacan donor (Zea mays ssp. mexicana, race Chalco). NILs were characterized for flowering time and yield, and we identified correlations between resistance to GLS and later flowering and lower grain yield.

PHYSIOLOGICAL RESPONSE OF CONVENTIONAL AND TRANSGENIC MAIZE TO WATER STRESS

Genetic modifications of plants can affect their ability to use environmental resources. It is known that the anthropic action has caused global climate changes, such as increases in temperature and drought. In view of this, the objective of this study was to evaluate if the addition of the Bt gene influences the physiological performance of isogenic maize hybrids grown under water stress and optimal conditions. Three Bt and non-Bt maize hybrids (Agromen 30A95, Dow 2B707 and BG 7049Y) were cultivated in soil with moisture content at 60% (water stress) and 100% of the field capacity (FC). Among the hybrids, the 30A95 presented greater variation between Bt and non-Bt hybrids in regard to physiological variables. The addition of the Bt gene practically had no impact on the photosynthetic performance of the 2B707 and BG7049 hybrids. The presence of the Bt gene increased the sensitivity to water stress in the hybrids, with reduction of maximal fluorescence and chlorophyll content in the plants. Regardless of the genetic factor (Bt or non-Bt), water stress reduced transpiration, stomatal conductance, photosynthetic rate, and maximum quantum yield of the PSII in maize plants. It was concluded that the presence of the Bt gene in maize indicates a reduction in the photosynthetic potential of the plant grown under water stress.

Arbuscular mycorrhizal colonization outcompetes root hairs in maize under low phosphorus availability.

An increase in root hair length and density and the development of arbuscular mycorrhiza symbiosis are two alternative strategies of most plants to increase the root-soil surface area under phosphorus (P) deficiency. Across many plant species, root hair length and mycorrhization density are inversely correlated. Root architecture, rooting density and physiology also differ between species. This study aims to understand the relationship among root hairs, arbuscular mycorrhizal fungi (AMF) colonization, plant growth, P acquisition and mycorrhizal-specific Pi transporter gene expression in maize. Using nearly isogenic maize lines, the B73 wild type and the rth3 root hairless mutant, we quantified the effect of root hairs and AMF infection in a calcareous soil under P deficiency through a combined analysis of morphological, physiological and molecular factors. Wild-type root hairs extended the rhizosphere for acid phosphatase activity by 0.5 mm compared with the rth3 hairless mutant, as measured by in situ zymography. Total root length of the wild type was longer than that of rth3 under P deficiency. Higher AMF colonization and mycorrhiza-induced phosphate transporter gene expression were identified in the mutant under P deficiency, but plant growth and P acquisition were similar between mutant and the wild type. The mycorrhizal dependency of maize was 33 % higher than the root hair dependency. The results identified larger mycorrhizal dependency than root hair dependency under P deficiency in maize. Root hairs and AMF inoculation are two alternative ways to increase Pi acquisition under P deficiency, but these two strategies compete with each other.

Expression profiling of key pathways in rat liver after a one-year feeding trial with transgenic maize MON810

In a recent one-year feeding study, we observed no adverse effects on tissue level in organs of rats fed with the genetically-modified maize MON810. Here, we assessed RNA expression levels of 86 key genes of the apoptosis-, NF-кB-, DNA-damage response (DDR)-, and unfolded-protein response (UPR) pathways by RT-qPCR in the rat liver. Male and female rats were fed either with 33% MON810 (GMO), isogenic- (ISO), or conventional maize (CONV) and RNAs were quantified from eight rats from each of the six feeding groups. Only Birc2 transcript showed a significant (p ≤ 0.05) consistent difference of ≥1.5-fold between the GMO and ISO groups in both sexes. Unsupervised cluster analysis showed a strong separation of male and female rats, but no clustering of the feeding groups. Individual analysis of the pathways did not show any clustering of the male or female feeding groups either, though transcript levels of UPR pathway-associated genes caused some clustering of the male GMO and CONV feeding group samples. These differences were not seen between the GMO and ISO control or within the female cohort. Our data therefore does not support an adverse effect on rat liver RNA expression through the long-term feeding of MON810 compared to isogenic control maize.

Integrating transcriptomic and proteomic analyses of photoperiod-sensitive in near isogenic maize line under long-day conditions

As a short-day plant species, maize requires an optimal photoperiod for inducing reproductive growth. However, there is a lack of information regarding photoperiod-induced changes in maize mRNA and protein levels. In this study, a photoperiod-insensitive maize inbred line and its near isogenic photoperiod-sensitive line were used. By integrating RNA-based transcriptomic and iTRAQ LC-MS/MS-based proteomic approaches, we generated a comprehensive inventory of the transcripts and proteins with altered abundances in response to a long photoperiod (LP) during growth stage transitions. We detected 22 000 transcripts in RNA-sequence runs and 5 259 proteins from an iTRAQ-based analysis. A weak correlation between mRNA- and protein-level changes was observed, suggesting the LP-induced transition between maize growth stages is largely regulated post-transcriptionally. Differentially expressed genes influenced by LP conditions were associated with several regulatory processes in both maize inbred lines, especially phosphate ion transport and the circadian rhythm. Additionally, 31 transcripts and six proteins related to photoperiodic flowering in maize were identified by comparing transcriptomic and proteomic data. This transcriptomic and proteomic analysis represents the first comprehensive and comparative study of gene/protein-level changes occurring in photoperiod-sensitive and -insensitive maize inbred lines during growth stage transitions under LP conditions.

Identification of Teosinte Alleles for Resistance to Southern Leaf Blight in Near Isogenic Maize Lines

Southern leaf blight ([SLB], causal agent Cochliobolus heterostrophus) is an important fungal disease of maize (Zea mays L.). Teosinte (Z. mays ssp. parviglumis), the wild progenitor of maize, offers a novel source of resistance alleles that may have been lost during domestication. The aims of this study were to identify teosinte alleles that, when present in a temperate maize background, confer a significant level of resistance to SLB. Ten populations of BC4S2 near isogenic lines (NILs), developed by crossing 10 different teosinte accessions to the maize inbred B73, comprising 774 lines in total, were screened for SLB resistance. Quantitative trait locus (QTL) analysis identified four significant QTL associated with SLB in bins 2.04, 3.04, 3.05, and 8.05. Sixteen individual NILs which were significantly different to the susceptible recurrent parent, B73 and which were carrying at least one of the teosinte‐derived resistance alleles were used to develop F2:3 populations by crossing each to B73 followed by two rounds of self‐pollination. These F2:3 populations were evaluated for SLB resistance and genotyped at the loci of interest. In 13 of 19 cases single marker analysis validated allelic substitution effects predicted from the original NIL population analysis, while in five cases we were not able to validate the effects and in one case a significant effect was detected in the opposite to the predicted direction. An allele at the QTL in bin 2.04 was shown to confer resistance to both SLB and a second maize foliar disease, gray leaf spot (GLS).

Body size inequality in ground beetle (Coleoptera: Carabidae) assemblages as a potential method to monitor environmental impacts of transgenic crops

Background and Purpose: Despite the obligatory post-market environmental monitoring of genetically modified (GM) crops in Europe, there are no available methodological guidelines or standards. Our aim was to examine the suitability of carabid body size inequality as a possible method for environmental monitoring. Material and Methods: We used carabids collected by pitfall traps in both insect-resistant GM (producing a Bacillus thuringiensis toxin) and isogenic maize plots at Flakkebjerg (Denmark), within the framework of the AMIGA Project. The body size distribution was calculated using various measures of size inequality: the Lorenz curve, the Gini and the Lorenz asymmetry coefficients every month during the summer 2014. Results: A total of 6339 carabids belonging to 38 species were captured and identified. The analysis detected a significant shift in size inequality between months, indicating the larger number of individuals of smaller-sized species later in the season, but no significant difference in inequality or mean body size was found between the assemblages in GM vs. isogenic maize plots. Conclusions: We concluded that the evaluation of body size inequality was sensitive to subtle changes in the structure of the carabid assemblages, and this method had the potential to be used during monitoring of the unanticipated environmental effects of GM plants.

Flea beetles (Coleoptera: Chrysomelidae, Alticinae) in Bt- (MON810) and near isogenic maize stands: Species composition and activity densities in Hungarian fields

Flea beetles (Chrysomelidae, Alticinae) were collected with Pherocon AM yellow sticky traps in maize plots to compare the assemblages from transgenic Bt- (genetic event MON810, producing Cry1Ab protein effective against lepidopteran pests) and near isogenic maize in Hungary. Altogether, 51,348 flea beetle individuals from 26 species were collected. The dominant species were Phyllotreta atra (F.) and Phyllotreta vittula (Redtenbacher). Their abundances along with other (non-P. atra and non-P. vittula) flea beetle species showed no significant differences between Bt- and isogenic maize plots. Similarly, no difference was found between Bt maize and isogenic maize plots in the species richness of the flea beetle assemblages.

Comparative data concerning aflatoxin contents in Bt maize and non-Bt isogenic maize in relation to human and animal health – a review

Transgenic Bt maize is a potentially important tool against insect pest in the EU and other countries. Bt maize (e.g. MON 810, Bt 11) which carries the Bt gene is highly resistant to larval feeding of European corn borer, stalk borer, and Southwestern corn borer, depending on Bt toxin (δ toxin) production. Effective measures used to fight pests may often have positive side-effects in that they may also contribute to reducing mycotoxin concentrations. A systematic review has been used for the purposes of evaluating the studies on the reduction of aflatoxins in Bt maize. According to five studies, Bt maize has significantly lower concentrations of aflatoxins than non-Bt maize hybrids, only one study has shown no significant effect of Bt maize. Other studies have shown mixed results (four studies). The results of these studies were influenced by the year of sampling or by using maize breeding lines selected for resistance to aflatoxin accumulation.

Effect of stacking insecticidal cry and herbicide tolerance epsps transgenes on transgenic maize proteome.

BackgroundThe safe use of stacked transgenic crops in agriculture requires their environmental and health risk assessment, through which unintended adverse effects are examined prior to their release in the environment. Molecular profiling techniques can be considered useful tools to address emerging biosafety gaps. Here we report the first results of a proteomic profiling coupled to transgene transcript expression analysis of a stacked commercial maize hybrid containing insecticidal and herbicide tolerant traits in comparison to the single event hybrids in the same genetic background.ResultsOur results show that stacked genetically modified (GM) genotypes were clustered together and distant from other genotypes analyzed by PCA. Twenty-two proteins were shown to be differentially modulated in stacked and single GM events versus non-GM isogenic maize and a landrace variety with Brazilian genetic background. Enrichment analysis of these proteins provided insight into two major metabolic pathway alterations: energy/carbohydrate and detoxification metabolism. Furthermore, stacked transgene transcript levels had a significant reduction of about 34% when compared to single event hybrid varieties.ConclusionsStacking two transgenic inserts into the genome of one GM maize hybrid variety may impact the overall expression of endogenous genes. Observed protein changes differ significantly from those of single event lines and a conventional counterpart. Some of the protein modulation did not fall within the range of the natural variability for the landrace used in this study. Higher expression levels of proteins related to the energy/carbohydrate metabolism suggest that the energetic homeostasis in stacked versus single event hybrid varieties also differ. Upcoming global databases on outputs from “omics” analyses could provide a highly desirable benchmark for the safety assessment of stacked transgenic crop events. Accordingly, further studies should be conducted in order to address the biological relevance and implications of such changes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-014-0346-8) contains supplementary material, which is available to authorized users.

Arbuscular mycorrhizal fungi colonisation of Cry3 toxin-producing Bt maize and near isogenic maize

Despite the fact that, on average, approximately 5–6 metric tons/ha of Bt maize stubble enter the soil on more than 170 million of hectares worldwide, the environmental impact of genetically modified maize plants on the arbuscular mycorrhizal fungi (AMF) is poorly known. In this study, the mycorrhizal colonisation on the roots of Bt maize (DAS-59122-7) and its near isogenic line was examined during the whole vegetation period. Cry3 toxin-producing Bt maize and its near isogenic line were grown in an experimental field in Julianna-major, Nagykovácsi, Hungary. DAS-59122-7 maize produces Cry34Ab1, Cry35Ab1 toxins and pat proteins for herbicide tolerance. The study assessed whether similar arbuscular mycorrhizal colonisation can be observed on the root of the Bt and near isogenic maize line and whether there are any differences in the temporal dynamics of AMF development. The arbuscular, hyphal and the arbuscular mycorrhizal fungi colonisation were higher in the near isogenic line as compared to its Bt counterpart, but no significant effect of the maize line was found as regards vesicle colonisation. The intensity of the arbuscular infection increased over time during plant maturation. DAS-59122-7 Bt maize had a negative effect on the initial development of AMF under field conditions, but no difference was seen in the case of the last two sampling dates (day 82 and 135). The reason of the latter is still not known.

Genetic Improvement of Root Growth Contributes to Efficient Phosphorus Acquisition in maize (Zea mays L.)

Maize plants adapt to low phosphorus (P) stress by increasing root growth. It is of importance to know the extent to which genetic improvement of root growth can enhance P acquisiton. In the present study, the contribution of root growth improvement to efficient P acquisition was evaluated in two soils using T149 and T222, a pair of near isogenic maize testcrosses which were derived from a backcross BC4F3 population. T149 and T222 showed no difference in shoot biomass and leaf area under normal growth conditions, but differed greatly in root growth. T149 had longer lateral roots and a larger root surface area compared to T222. In calcareous soil, when P was insufficient, i.e., when P was either supplied as KH2PO4 at a concentration of 50 mg P kg−1 soil, or in the form of Phy-P, Ca3-P or Ca10-P, a 43% increase in root length in T149 compared to T222 resulted in an increase in P uptake by 53%, and shoot biomass by 48%. In acid soil, however, when P supply was insufficient, i.e., when P was supplied as KH2PO4 at a concentration of 100 mg P kg−1 soil, or in the form of Phy-P, Fe-P or Al-P, a 32% increase in root length in T149 compared to T222 resulted in an increase in P uptake by only 12%, and shoot biomass by 7%. No significant differences in the exudation of organic acids and APase activity were found between the two genotypes. It is concluded that genetic improvement of root growth can efficiently increase P acquisition in calcareous soils. In acid soils, however, improvements in the physiological traits of roots, in addition to their size, seem to be required for efficient P acquisition.

Porównanie wybranych elementów bionomii Ostrinia nubilalis Hbn. (Lep., Crambidae) na kukurydzy Bt i non-Bt w południowo-wschodniej Polsce

A study on the dynamics of the oviposition and hatching of Ostrinia nubilalis Hbn. larvae on transgenic maize line MON 810 (DKC 3421YG cultivar) and isogenic maize (DKC 3420 cultivar) was carried out in 2008-2011 in Głuchów (south-eastern Poland). No significant differences were found between the number of egg clusters deposited by O. nubilalis on Bt and non-Bt maize based on the carried out observations. A small, statistically significant difference in the number of egg clusters between the studied cultivars was found only in 2011 and was most likely associated with weather changes. Female O. nubilalis did not significantly prefer any of the cultivars when depositing eggs. In addition, we found no differences in the dynamics of oviposition and larvae hatching between Bt and non-Bt maize. The first egg clusters, their maximum number, and the last egg clusters were found on similar dates for both studied cultivars. Empty egg clusters proving larval hatching were also found on similar dates for Bt and non-Bt maize.

Comparison of some aspects of the bionomy of Ostrinia nubilalis Hbn. (Lep., Crambidae) on Bt and non-Bt maize in south-eastern Poland

A study on the dynamics of the oviposition and hatching of Ostrinia nubilalis Hbn. larvae on transgenic maize line MON 810 (DKC 3421YG cultivar) and isogenic maize (DKC 3420 cultivar) was carried out in 2008-2011 in Gluchow (south-eastern Poland). No significant differences were found between the number of egg clusters deposited by O. nubilalis on Bt and non-Bt maize based on the carried out observations. A small, statistically significant difference in the number of egg clusters between the studied cultivars was found only in 2011 and was most likely associated with weather changes. Female O. nubilalis did not significantly prefer any of the cultivars when depositing eggs. In addition, we found no differences in the dynamics of oviposition and larvae hatching between Bt and non-Bt maize. The first egg clusters, their maximum number, and the last egg clusters were found on similar dates for both studied cultivars. Empty egg clusters proving larval hatching were also found on similar dates for Bt and non-Bt maize.

WPLYW GENETYCZNIE MODYFIKOWANEJ KISZONKI Z KUKURYDZY NA ZAWARTOSC KWAS�W TLUSZCZOWYCH W TKANKACH JAGNIAT

The aim of this work was the evaluation of fatty acids contents in meat and selected offal in lambs fed a diet containing silage of whole plants of genetically modified maize (Bt) MON 810 line. The material consisted of 14 Polish Merino lambs of mean start body weight 24 kg. There were two feeding groups selected of 7 lambs each. In the control group (K) the lambs were fed isogenic maize silage, which in the second group (GMO) was substituted with the modified maize silage (Bt) MON 810 line. After 70 days of feeding (feed portions were standardised according to the DLG system) the lambs were slaughtered and dissected. The results were evaluated statistically and the significance of differences was calculated with the two factor variation analysis (nutrition, tissue). Feeding genetically modified maize silage did not change, in a statistically significant way, the contents of any main fatty acids in the pool of all acids nor the contents of the totals and their proportions, while the factor causing clear differences was the tissue. Differences for the majority of the results were statistically significant. Statistically significant interactions noted (nutrition x tissue) are probably due to different values of these traits in the analysed tissues.

The role of maize root size in phosphorus uptake and productivity of maize/faba bean and maize/wheat intercropping systems

Interspecific root/rhizosphere interactions affect phosphorus (P) uptake and the productivity of maize/faba bean and maize/wheat intercropping systems. The aim of these experiments was to determine whether manipulation of maize root growth could improve the productivity of the two intercropping systems. Two near isogenic maize hybrids (the larger-rooted T149 and smaller-rooted T222) were intercropped with faba bean and wheat, under conditions of high- and low-P availability. The larger-rooted T149 showed greater competitive ability than the smaller-rooted T222 in both maize/faba bean and maize/wheat intercropping systems. The higher competitive ability of T149 improved the productivity of the maize/faba bean intercropping system in P-sufficient conditions. In maize/wheat intercropping systems, root growth, shoot biomass, and P uptake of maize were inhibited by wheat, regardless of the P-supply. Compared with T222, the larger-rooted T149 suffered less in the intercropping systems. The total biomass of the maize/wheat intercropping system was higher for wheat/T149 than for wheat/T222 under low-P conditions. These data suggested that genetic improvement of maize root size could enhance maize growth and its ability to compete for P resources in maize/faba bean and maize/wheat intercropping systems. In addition, depending on the P availability, larger maize roots could increase the productivity of intercropping systems.

Transgenerational effects of feeding genetically modified maize to nulliparous sows and offspring on offspring growth and health1

This study assessed the effect of feeding genetically modified maize expressing a truncated form of the Cry1Ab protein from Bacillus thuringiensis (Bt MON810 maize) to sows during gestation and lactation and their offspring from weaning to 115 d postweaning on offspring growth and health. After weaning at approximately 28 d of age (d 0), individually penned, mixed sex pigs (approximately 8 kg BW) from sows fed isogenic or Bt maize diets were blocked by sow treatment, sex, and BW and randomly assigned to Bt or isogenic maize diets as follows: i) isogenic maize-fed sow/isogenic maize-fed offspring (iso/iso); ii) isogenic maize-fed sow/Bt maize-fed offspring (iso/Bt); iii) Bt maize-fed sow/isogenic maize-fed offspring (Bt/iso); and iv) Bt maize-fed sow/Bt maize-fed offspring (Bt/Bt). Growth performance was recorded at intervals to harvest at approximately 105 kg BW (n=15/treatment) and blood samples were taken for biochemical analysis on d 0, 30, 70, 100, and 115 postweaning (n=10/treatment). Pigs were harvested on d 115 postweaning (n=10/treatment), and carcass weight, backfat depth, and organ weights (heart, kidney, spleen, and liver) were recorded. Kidney, liver, lymph nodes, and small intestine were collected for histological analysis. Offspring from Bt maize-fed sows were heavier than offspring from isogenic maize-fed sows on d 30 (P<0.05), 100 (P<0.05), and 115 postweaning (P<0.05) and had greater overall ADG (P<0.05). Overall ADFI was greater for offspring from sows fed Bt maize (P<0.05) and for Bt maize-fed pigs (P<0.05). Offspring from Bt maize-fed sows had greater carcass (P<0.05) and lighter spleen (P<0.05) weights. Dressing percentage was greater for Bt maize-fed pigs than isogenic maize-fed pigs (P<0.05), and livers were lighter for pigs in the Bt/Bt group than pigs in the iso/Bt or Bt/iso group (P<0.05). Offspring from Bt maize-fed sows also had greater duodenal crypt depths (P<0.05) and lower villus height/crypt depth ratios (P<0.05). No pathology was observed in the organs, and serum biochemistry values generally remained within normal limits and no overall differences were observed, with the exception of overall γ glutamyltransferase, which was less for pigs on the Bt/Bt treatment than pigs on the iso/Bt and Bt/iso treatments. These results indicate that transgenerational consumption of Bt maize diets is not detrimental to pig growth and health.

Evidence for a Natural Allelic Series at the Maize Domestication Locus teosinte branched1

Despite numerous quantitative trait loci and association mapping studies, our understanding of the extent to which natural allelic series contribute to the variation for complex traits is limited. In this study, we investigate the occurrence of a natural allelic series for complex traits at the teosinte branched1 (tb1) gene in natural populations of teosinte (Zea mays ssp. parviglumis, Z. mays ssp. mexicana, and Z. diploperennis). Previously, tb1 was shown to confer large effects on both plant architecture and ear morphology between domesticated maize and teosinte; however, the effect of tb1 on trait variation in natural populations of teosinte has not been investigated. We compare the effects of nine teosinte alleles of tb1 that were introgressed into an isogenic maize inbred background. Our results provide evidence for a natural allelic series at tb1 for several complex morphological traits. The teosinte introgressions separate into three distinct phenotypic classes, which correspond to the taxonomic origin of the alleles. The effects of the three allelic classes also correspond to known morphological differences between the teosinte taxa. Our results suggest that tb1 contributed to the morphological diversification of teosinte taxa as well as to the domestication of maize.

Electrochemical detection of 0.6% genetically modified maize MON810 in real flour samples

We demonstrate an approach for the electrochemical detection of genetically modified maize in real maize flour samples by means of square-wave voltammetry (SWV). After labeling the asymmetric PCR-amplified targets with osmium tetroxide bipyridine [OsO4(bipy)], they were hybridized with immobilized oligonucleotide probes on gold electrodes. We could detect the maize genes ivrp and SSIIb in near isogenic maize. The transgene cryIa/b and the MON810 specific fragment were detected in all transgenic maize samples, down to a content of 0.6% of MON810 in mixed samples. While it was possible to detect all sequences in the samples containing 100% near isogenic or respectively transgenic maize after a hybridization time of less than 10min, a hybridization time of 30min was necessary for the detection of the genetic modifications in samples containing only 0.9 to 0.6% of transgenic maize. No significant detection of the transgene cryIa/b or MON810 was possible when only 0.5% of transgenic maize was present in the sample, most likely due to insufficient amplification of the template DNA.