Fungal growth and fusarium mycotoxin content in isogenic traditional maize and genetically modified maize grown in France and Spain

Fungi of the genus Fusarium are common fungal contaminants of maize and are also known to produce mycotoxins. Maize that has been genetically modified to express a Bt endotoxin has been used to study the effect of insect resistance on fungal infection of maize grains by Fusarium species and their related mycotoxins. Maize grain from Bt hybrids and near-isogenic traditional hybrids was collected in France and Spain from the 1999 crop, which was grown under natural conditions. According to the ergosterol level, the fungal biomass formed on Bt maize grain was 4-18 times lower than that on isogenic maize. Fumonisin B(1) grain concentrations ranged from 0.05 to 0.3 ppm for Bt maize and from 0.4 to 9 ppm for isogenic maize. Moderate to low concentrations of trichothecenes and zearalenone were measured on transgenic as well as on non-transgenic maize. Nevertheless, significant differences were obtained in certain regions. The protection of maize plants against insect damage (European corn borer and pink stem borer) through the use of Bt technology seems to be a way to reduce the contamination of maize by Fusarium species and the resultant fumonisins in maize grain grown in France and Spain.     

Comparison of the nutritional profile of glyphosate-tolerant corn event NK603 with that of conventional corn (Zea mays L.)

The composition of glyphosate-tolerant (Roundup Ready) corn event NK603 was compared with that of conventional corn grown in the United States in 1998 and in the European Union in 1999 to assess compositional equivalence. Grain and forage samples were collected from both replicated and nonreplicated field trials, and compositional analyses were performed to measure proximates, fiber, amino acids, fatty acids, vitamin E, nine minerals, phytic acid, trypsin inhibitor, and secondary metabolites in grain as well as proximates and fiber in forage. Statistical analysis of the data was conducted to assess statistical significance at the p < 0.05 level. The values for all of the biochemical components assessed for corn event NK603 were similar to those of the nontransgenic control or were within the published range observed for nontransgenic commercial corn hybrids. In addition, the compositional profile of Roundup Ready corn event NK603 was compared with that of traditional corn hybrids grown in Europe by calculating a 99% tolerance interval to describe compositional variability in the population of traditional corn varieties in the marketplace. These comparisons, together with the history of the safe use of corn as a common component of animal feed and human food, support the conclusion that Roundup Ready corn event NK603 is compositionally equivalent to, and as safe and nutritious as, conventional corn hybrids grown commercially today.     

Composition of grain and forage from corn rootworm-protected corn event MON 863 is equivalent to that of conventional corn (Zea mays l.)

Insect-protected corn hybrids containing event MON 863 protect corn plants against feeding damage from corn rootworm (Diabrotica), a major North American insect pest. Corn event MON 863 contains a gene that expresses an amino acid sequence variant of the wild-type Cry3Bb1 insecticidal protein from Bacillus thuringiensis. The purpose of this study was to compare the composition of corn containing event MON 863 with that of conventional nontransgenic corn. Compositional analyses were conducted to measure proximates, fiber, amino acids, fatty acids, minerals, folic acid, thiamin, riboflavin, vitamin E, antinutrients, and certain secondary metabolites in grain and proximates and fiber content in forage collected from a total of eight field sites in the U.S. and Argentina. Compositional analyses demonstrated that the grain and forage of event MON 863 are comparable in their nutritional content to the control corn hybrid and conventional corn. These comparisons, together with the history of the safe use of corn as a common component of animal feed and human food, support the conclusion that corn event MON 863 is compositionally equivalent to, and as safe and nutritious as, conventional corn hybrids grown commercially today.     

Fate of Bt Protein and Influence of Corn Hybrid on Ethanol Production

Corn hybrids were compared to determine the fate of recombinant Bt protein (CRY1Ab from Bacillus thuringiensis) in coproducts from dry grind and wet‐milled corn during production of fuel ethanol. Two pairs of Bt and non‐Bt hybrids were wet milled, and each fraction was examined for the presence of the Bt protein. Bt protein was found in the germ, gluten, and fiber fractions of Bt hybrids. In addition, one set of Bt and non‐Bt hybrids were treated by the dry‐grind ethanol process and Bt protein was monitored during each step of the process. The Bt protein was not detected after liquefaction. Subsequent experiments determined that the Bt protein is rapidly denatured at liquefaction temperatures. Finally, five hybrids were compared for ethanol yield after dry grinding. Analysis of fermentation data with an F‐test revealed the percent of total starch available for conversion into ethanol varied significantly among the hybrids (P < 0.002), indicating ethanol yield is not exclusively dependent on starch content. No difference, however, was observed between Bt and non‐Bt corn hybrids for either ethanol productivity or yield.     

Soil microbial properties in Bt (Bacillus thuringiensis) corn cropping systems

Growing Bt crops reduces the use of insecticides applied to them, but these crops could affect soil microorganisms and their activities. We evaluated the effects of Bt (Cry1Ab) corn (Zea mays L.) and deltamethrin ([S]-α-cyno-3-phenoxybenzyl [1R, 3R]-3-[2,2-dibromovinyl]-2,2-dimethylcyclopropane-1-carboxylate) insecticide application on soil microbial biomass C (MBC), β-glucosidase enzyme activity (final season only), bacterial functional diversity, and bacterial community-level physiological profiles (CLPPs) in corn monoculture in five seasons. We also determined if growing Bt corn in crop rotation would alter these effects. Statistical analysis of pooled data across seasons did not show any effects of Bt technology, insecticide application or crop rotation on soil microbial biomass or diversity even though differences between seasons and between the rhizosphere and bulk soil were observed. Annual analyses of results also showed that neither the Bt technology nor insecticide application affected soil MBC, enzyme activity, or functional diversity of bacteria in corn rhizosphere, but shifts in bacterial CLPPs due to Bt trait were observed in one year. Crop rotation effects on soil microbial properties were not observed in most cases. Where effects were observed, Bt corn grown in rotation resulted in greater MBC, enzyme activity and functional diversity than Bt corn grown in monoculture or conventional corn grown in rotation, and these effects were observed only in bulk soil. Therefore, the Bt technology is safe with respect to the non-target effects measured in this study. However, the effects of repeated use of Bt crops over many years on the soil environment should continue to be monitored.     

Comparison of the forage and grain composition from insect-protected and glyphosate-tolerant MON 88017 corn to conventional corn (Zea mays L.)

The next generation of biotechnology-derived products with the combined benefit of herbicide tolerance and insect protection (MON 88017) was developed to withstand feeding damage caused by the coleopteran pest corn rootworm and over-the-top applications of glyphosate, the active ingredient in Roundup herbicides. As a part of a larger safety and characterization assessment, MON 88017 was grown under field conditions at geographically diverse locations within the United States and Argentina during the 2002 and 2003-2004 field seasons, respectively, along with a near-isogenic control and other conventional corn hybrids for compositional assessment. Field trials were conducted using a randomized complete block design with three replication blocks at each site. Corn forage samples were harvested at the late dough/early dent stage, ground, and analyzed for the concentration of proximate constituents, fibers, and minerals. Samples of mature grain were harvested, ground, and analyzed for the concentration of proximate constituents, fiber, minerals, amino acids, fatty acids, vitamins, antinutrients, and secondary metabolites. The results showed that the forage and grain from MON 88017 are compositionally equivalent to forage and grain from control and conventional corn hybrids.     

Dynamics of mycotoxin and Aspergillus flavus levels in aging Bt and non-Bt corn residues under Mississippi no-till conditions

Mycotoxin and Aspergillus flavus levels in soil-surface corn debris left by no-till agriculture methods (stover, cobs, and cobs with grain) were determined during the December-March fallow period for near-isogenic Bt and non-Bt hybrid corn. By December, average mycotoxin levels in non-Bt corn were many times higher in cobs with grain than in grain harvested in September (total aflatoxins, 774 vs 211 ng/g; total fumonisins, 216 vs 3.5 microg/g; cyclopiazonic acid, 4102 vs 72.2 microg/g; zearalenone, 0.2 vs < 0.1 microg/g). No trichothecenes were detected. Levels of mycotoxins and A. flavus propagules were approximately 10- to 50-fold lower in cobs without grain and stover, respectively, for all mycotoxins except zearalenone. Mycotoxin levels in corn debris fractions decreased during winter but began to rise in March. Levels of all mycotoxins and A. flavus propagules were lower in harvested grain and debris from Bt than non-Bt corn, but differences were significant (p < 0.05) only for aflatoxins.     

Reduction of Aflatoxin and Fumonisin Contamination in Yellow Corn by High‐Speed Dual‐Wavelength Sorting

A high‐speed dual‐wavelength sorter was tested for removing corn contaminated in the field with aflatoxin and fumonisin. To achieve accurate sorting, single kernel reflectance spectra (500–1,700 nm) were analyzed to select the optimal pair of optical filters to detect mycotoxin‐contaminated corn during high‐speed sorting. A routine, based on discriminant analysis, was developed to select the two absorbance bands in the spectra that would give the greatest classification accuracy. In a laboratory setting, and with the kernels stationary, absorbances at 750 and 1,200 nm could correctly identify >99% of the kernels as aflatoxin‐contaminated (>100 ppb) or uncontaminated. A high‐speed sorter was tested using the selected filter pair for corn samples inoculated with Aspergillus flavus; naturally infested corn grown in central Illinois; and naturally infested, commercially grown and harvested corn from eastern Kansas (2002 harvest). For the Kansas corn, the sorter was able to reduce aflatoxin levels by 81% from an initial average of 53 ppb, while fumonisin levels in the same grain samples were reduced an average of 85% from an initial level of 17 ppm. Similar reductions in mycotoxin levels were observed after high‐speed sorting of A. flavus inoculated and naturally mold‐infested corn grown in Illinois.     

Evaluation of compositional equivalence for multitrait biotechnology crops

Compositional analysis is an important tool in the evaluation of the safety and nutritional status of biotechnology-derived crops. As part of the comparative assessment of a biotechnology-derived crop, its composition is evaluated by quantitative measurement of the levels of key nutrients, antinutrients, and secondary metabolites and compared to that of conventional crops. To evaluate the effect of combining multiple biotech traits through conventional breeding, the forage and grain compositions of the double combinations MON 810 × NK603, MON 863 × MON 810, and MON 863 × NK603 and the triple combination MON 863 × NK603 × MON 810 were compared to their respective near-isogenic, conventional control hybrids. Overall, a total of 241 statistical comparisons between the multitrait biotechnology crop and its corresponding conventional controls were conducted. Of these comparisons 192 (79.7%) were not statistically significantly different (p > 0.05), and all 49 of the differences were within the 99% tolerance interval for commercial hybrids grown in the same field or related field trials. These data on combined trait biotechnology-derived products demonstrated that the forage and grain were compositionally equivalent to their conventional comparators, indicating the absence of any influence of combining insect protection and herbicide tolerance traits by conventional breeding on compositional variation.     

Assessment of the arbuscular mycorrhizal fungal community in roots and rhizosphere soils of Bt corn and their non-Bt isolines

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) in combination with sequencing of amplified partial 18S ribosomal DNA was used to assess the effects of transgenic Bt corn 5422Bt1 (event Bt 11) and 5422CBCL (event MON810) on the community structure of a non-target microorganism, namely the arbuscular mycorrhizal fungi (AMF) Glomus in corn roots and rhizosphere soils, relative to their non-Bt isolines 5422 (conventional parent) and 5422wx (conventional hybrid). AMF colonization in roots of different corn genotypes was also assessed using microscopic visualization. No adverse effect was detected on the indigenous AMF colonization of the roots of Bt hybrids 5422Bt1 and 5422CBCL. Two-way indicator species analysis (TWINSPAN^®) and detrended correspondence analysis (DCA) of the DGGE data from corn roots presented differences between Bt and non-Bt corn isolines (5422Bt1 vs. 5422wx, and 5422CBCL vs. 5422wx). However, differences were also recognized between the two non-Bt corn cultivars (5422 vs. 5422wx), and between the two Bt corn lines (5422Bt1 vs. 5422CBCL) in roots. Our results suggest that corn genotypes may have a greater influence on the AMF community structure of plant roots and rhizosphere soils than other factors, such as the age of the growing plants.     

Assessment of the arbuscular mycorrhizal fungal community in roots and rhizosphere soils of Bt corn and their non-Bt isolines

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) in combination with sequencing of amplified partial 18S ribosomal DNA was used to assess the effects of transgenic Bt corn 5422Bt1 (event Bt 11) and 5422CBCL (event MON810) on the community structure of a non-target microorganism, namely the arbuscular mycorrhizal fungi (AMF) Glomus in corn roots and rhizosphere soils, relative to their non-Bt isolines 5422 (conventional parent) and 5422wx (conventional hybrid). AMF colonization in roots of different corn genotypes was also assessed using microscopic visualization. No adverse effect was detected on the indigenous AMF colonization of the roots of Bt hybrids 5422Bt1 and 5422CBCL. Two-way indicator species analysis (TWINSPAN^®) and detrended correspondence analysis (DCA) of the DGGE data from corn roots presented differences between Bt and non-Bt corn isolines (5422Bt1 vs. 5422wx, and 5422CBCL vs. 5422wx). However, differences were also recognized between the two non-Bt corn cultivars (5422 vs. 5422wx), and between the two Bt corn lines (5422Bt1 vs. 5422CBCL) in roots. Our results suggest that corn genotypes may have a greater influence on the AMF community structure of plant roots and rhizosphere soils than other factors, such as the age of the growing plants.     

转Bt基因玉米自交系及其杂交种的抗螟性鉴定

采用基因枪法将Bt基因(GFM CrylA)导入东北春玉米自交系铁7922的幼胚中,获得转基因植株。经过连续3年的田间接卵、室内饲喂玉米螟鉴定,获得稳定抗虫转基因系。以此为亲本与常规自交系配制了四密25Bt、通单24Bt、吉单209Bt3个杂交种。转基因自交系和杂交种的抗螟性鉴定结果表明：转基因系间抗虫性差异极显著,同一转基因系单株间和配制的3个杂交种间抗虫性也存在差异;与对照相比,转基因系配制的杂交种抗虫性明显提高,差异显著。农艺性状鉴定结果显示,转基因系配制的杂交种与对照在株高、穗长、穗行数、行粒数、百粒重上无显著差异。本研究认为,GFM CrylA(Bt)基因可用于玉米的抗螟性改良,也为转Bt基因玉米自交系能够直接用于育种提供了依据。     

Iron efficiency of different corn hybrids grown in nutrient solution culture

Growing Fe-efficient genotype(s) could be considered as a preferred genetic approach to tackle the widespread constraint of Fe-deficiency-/lime-induced chlorosis in crop grown on alkaline soil. This study aimed to investigate morphological and physiological traits linked to expression of Fe deficiency among four corn (Zea mays) including sweet (Z. mays sacchrata cvs. H403 and H404) and grain (Z. mays indentata cvs. H500 and H700) hybrids grown in nutrient solution using two Fe concentrations (5 and 50 µM Fe-ethylenediaminetetraacetic acid (Fe-EDTA)). Significant variation was found among studied hybrids in their tolerance to Fe-deficiency stress. Sweet corn hybrids were more sensitive to Fe deficiency as compared with grain corn hybrids and greater reduction was observed in their shoot dry matter at the 5 µM Fe-EDTA treatment. The greatest decrease in plant height, leaf area, and root and shoot dry matter weight under Fe-deficiency condition was found for H403 hybrid. No significant correlation was found between shoot and root Fe concentration with crop tolerance to Fe deficiency. Furthermore, different response of corn hybrids to Fe deficiency is an important factor, which has to be considered in Fe fertilizer recommendation as well as breeding programs.     

Elemental and/or cation ratio efficiency of corn hybrids grown on an infertile soil inadequate in magnesium

Abstract

Magnesium deficiency in corn (Zea mays L.) is often attributed to the low levels of Mg in soils. This study was conducted to determine elemental and/or cation balance efficiency of corn hybrids grown on a soil low in available Mg, Among the 15 hybrids tested, no differential efficiency in ear leaf concentration of P was found at two planting dates. Leaf concentrations of Zn and Fe were influenced by planting date for all hybrids but interactions between hybrids and planting date were found only for K, Ca, Mg concentrations and the sum of the Meq Ca + Mg/100 g, the Meq K + Ca + Mg/100 g, and the K/Ca and K/Ca + Mg ratios. Planting dates did not Influence the K/Mg ratio among hybrids. Large differences in efficiency of K, Ca, and Mg were found but these cations were found to have large Interactions. Data show that Ca and Mg efficient hybrids are less efficient in K than Ca and Mg inefficient hybrids. It should be possible to breed corn hybrids for better cation balance efficiency for use on infertile soils low in avallable Mg.     

Effects of nitrogen,phosphorus, and potassium nutrition on yield,rates of kernel growth and grain filling periods of two corn hybrids

Abstract

Depression of corn grain yields from nutrient stress has been studied extensively, but effects of nutrient stress on rates of corn development and yield determinants are less well understood. Nutritional effects on the number of kernels/unit area, growth rate/kernel, and duration of growth have implications concerning fertilization practices and yield potentials of crops. Two corn hybrids with equivalent silking dates but having different grain filling periods were grown in a field experiment. Fertility treatments consisted of a N series receiving 0, 112, or 336 kg of N/ha and a P‐K series receiving factorial combinations of 0, 22, or 112 kg of P/ha and 0, 56, or 224 kg of K/ha. Dates for grain initiation and maturity were determined for each plot along with tissue analyses of ear leaves, grain yields, and kernel weights. Concentrations of N and K in ear leaves generally corresponded to treatment levels of these nutrients, although Pioneer Hi‐bred 3390 appeared to be less efficient than Pioneer Hi‐bred 3334 in K uptake. Effects of nutrient stress on yield determinants depended on the determinant and nutrient under consideration. Severe N stress did not change length of grain filling periods, but decreased kernel numbers 30 to 70%. Stress for K, on the other hand, shortened grain filling periods about 13% and had only a slight effect on kernel number. Negligible P stress occurred in the experiment. The two hybrids produced equal quantities of grain/ha/day but the hybrid with a longer filling period (Pioneer 3334) filled many more kernels at a slightly slower rate and for a longer period of time to give a significantly greater grain yield compared to Pioneer 3390.     

Antifungal proteins in commercial hybrids and elite sorghums

The antifungal proteins (AFP) in the caryopsis of commercial and elite sorghums grown in several environments were related to grain mold resistance. Previous studies revealed that improved grain mold resistance was correlated with higher levels of AFP at combine harvest maturity [50 days after anthesis (DAA)] or with better retention of AFP after physiological maturity (30 DAA). Commercial hybrids and public parental lines and hybrids were grown in College Station, TX, during 2000, 2001, and 2002. Samples of caryopses were collected at 30 and 50 DAA, and caryopsis proteins were extracted, blotted, identified with immunoassays, and quantified for two AFP (chitinase and sormatin). Sorghums varied in amounts of AFP and their ability to retain AFP after physiological maturity. The environment in 2002 was conducive for fungal deterioration of grain, and a wide range of AFP and grain molding was observed at harvest maturity. High levels of AFP and low levels of grain molding were observed in non-mold-conducive environments in 2000 and 2001. Commercial and elite sorghums with higher levels and increased retention of AFP had less grain molding in the mold-conducive environment.     

Chemical composition of glyphosate-tolerant soybean 40-3-2 grown in Europe remains equivalent with that of conventional soybean (Glycine max L.)

The composition of glyphosate-tolerant (Roundup Ready) soybean 40-3-2 was compared with that of conventional soybean grown in Romania in 2005 as part of a comparative safety assessment program. Samples were collected from replicated field trials, and compositional analyses were performed to measure proximates (moisture, fat, ash, protein, and carbohydrates by calculation), fiber, amino acids, fatty acids, isoflavones, raffinose, stachyose, phytic acid, trypsin inhibitor, and lectin in grain as well as proximates and fiber in forage. The mean values for all biochemical components assessed for Roundup Ready soybean 40-30-2 were similar to those of the conventional control and were within the published range observed for commercial soybean. The compositional profile of Roundup Ready soybean 40-3-2 was also compared to that of conventional soybean varieties grown in Romania by calculating a 99% tolerance interval to describe compositional variability in the population of traditional soybean varieties already on the marketplace. These comparisons, together with the history of the safe use of soybean as a common component of animal feed and human food, lead to the conclusion that Roundup Ready soybean 40-3-2 is compositionally equivalent to and as safe and nutritious as conventional soybean varieties grown commercially.     

Composition of forage and grain from second-generation insect-protected corn MON 89034 is equivalent to that of conventional corn (Zea mays L.)

Insect-protected corn hybrids containing Cry insecticidal proteins derived from Bacillus thuringiensis have protection from target pests and provide effective management of insect resistance. MON 89034 hybrids have been developed that produce both the Cry1A.105 and Cry2Ab2 proteins, which provide two independent modes of insecticidal action against the European corn borer ( Ostrinia nubilalis ) and other lepidopteran insect pests of corn. The composition of MON 89034 corn was compared to conventional corn by measuring proximates, fiber, and minerals in forage and by measuring proximates, fiber, amino acids, fatty acids, vitamins, minerals, antinutrients, and secondary metabolites in grain collected from 10 replicated field sites across the United States and Argentina during the 2004-2005 growing seasons. Analyses established that the forage and grain from MON 89034 are compositionally comparable to the control corn hybrid and conventional corn reference hybrids. These findings support the conclusion that MON 89034 is compositionally equivalent to conventional corn hybrids.     

A comparison of the chemical composition of the corn leaf and the grain sorghum leaf

Abstract

Leaf samples were taken from corn and grain sorghum being grown in adjacent fields with the same soil type and with the same cultural practices insofar as possible. The samples were analyzed for N, P, K, Ca, Mg, Fe, Zn and Mn. Results show that the nutrient composition of the leaves of corn and grain sorghum were consistently different for most nutrients. The differences were sufficiently great and consistent to strongly suggest that critical nutrient levels need to be established for grain sorghum instead of using diagnostic criteria established for corn.     

Root growth and mineral nutrition of corn hybrids as affected by phosphorus and lime

Abstract

The effects of triple superphosphate (TS) and liming on macronutrient accumulation and root growth of Pioneer 3072 and Cargill 505 corn hybrids were studied. Corn plants were grown up to 30 days in pots with 7 L of a dark red Latosol sandy loam (Haplortox). Lime was applied to raise base saturation to 30, 50, and 70%, in two levels of phosphorus (P) fertilization with TS (0 and 200 ppm P). There was an increase in root surface due to lime only in pots without TS, with no effects on plant growth or nutrition. Both corn hybrids responded to P fertilization, but Pioneer yielded more dry matter than Cargill. The roots of Cargill were thicker and, when in TS presence, were longer and had a larger surface than Pioneer. There was an increase in macronutrient uptake in the P fertilized pots. Pioneer required more nutrients and showed a higher efficiency in acquiring and utilizing the nutrients from the soil. A higher response of Pioneer in dry matter and nutrient acquisition was more related to the physiological efficiency than to root morphology.