Metabolite analyses of grain from maize hybrids grown in the United States under drought and watered conditions during the 2002 field season

Understanding natural variation in the composition of conventional crop germplasms is critical in establishing a baseline for comparison of biotechnology-derived crops. This is particularly relevant to such traits as tolerance to drought stress. Thus, there is both a need to understand the contribution of stress conditions to natural variation in plant nutritional components and to determine whether levels of small molecule metabolites such as osmoprotectants and stress metabolites are also affected. As a first step in developing such information for maize, seven conventional hybrids were grown under different moisture regimens and the impact of moisture on composition was assessed. The regimens included well-watered conditions, water restriction during the vegetative phase, and water restriction during grain fill. Compositional analyses of the harvested grain included assessments of the levels of proximates (moisture, protein, oil, starch) and small molecule metabolites such as fatty acids, free amino acids, organic acids, sugars, total glycerol, glycine betaine, and abscisic acid. Ranges for these analytes were determined across all moisture regimens, and the effect of the different water regimens on these analytes was also evaluated. The number and type of grain analytes that showed statistically significant differences in levels between different water regimens differed quite markedly by maize hybrid. However, the magnitude of mean differences between well-watered and water-restricted samples was typically small, and statistically significant differences for any given analyte were typically observed in only one to three of the seven maize hybrids. Only two analytes, free glutamine and free proline, showed a significant drought-induced difference in at least four maize hybrids.     

Impact of genetics and environment on nutritional and metabolite components of maize grain

The Organization for Economic Co-operation and Development (OECD) recommends the measurement of specific plant components for compositional assessments of new biotechnology-derived crops. These components include proximates, nutrients, antinutrients, and certain crop-specific secondary metabolites. A considerable literature on the natural variability of these components in conventional and biotechnology-derived crops now exists. Yet the OECD consensus also suggests measurements of any metabolites that may be directly associated with a newly introduced trait. Therefore, steps have been initiated to assess natural variation in metabolites not typically included in the OECD consensus but which might reasonably be expected to be affected by new traits addressing, for example, nutritional enhancement or improved stress tolerance. The compositional study reported here extended across a diverse genetic range of maize hybrids derived from 48 inbreds crossed against two different testers. These were grown at three different, but geographically similar, locations in the United States. In addition to OECD analytes such as proximates, total amino acids and free fatty acids, the levels of free amino acids, sugars, organic acids, and selected stress metabolites in harvested grain were assessed. The major free amino acids identified were asparagine, aspartate, glutamate, and proline. The major sugars were sucrose, glucose, and fructose. The most predominant organic acid was citric acid, with only minor amounts of other organic acids detected. The impact of genetic background and location was assessed for all components. Overall, natural variation in free amino acids, sugars, and organic acids appeared to be markedly higher than that observed for the OECD analytes.     

Compositional equivalency of Cry1F corn event TC6275 and conventional corn (Zea mays L.)

Maize (Zea mays L.) plants have been transformed to express a Cry1F insecticidal crystal protein originally isolated from Bacillus thuringiensis Berliner. This protein controls lepidopteran pests of maize, including the European corn borer, Ostrinia nubilalis (Hübner). As part of the safety assessment for crops containing transgenes, a compositional analysis of the food and feed is conducted. This analysis is designed to detect unintended changes in the nutrient and antinutrient content of the raw commodities produced by the crop due to the insertion of the genes into the genomic DNA of the plant (pleotropic effects). Samples of transgenic and nontransgenic maize forage and grain were collected from six field sites located in the U.S. and Canada. Forage samples were analyzed for proximates and minerals, and grain was further analyzed for fatty acids, amino acids, vitamins, secondary metabolites, and antinutrients. Results demonstrated that maize expressing the Cry1F protein was equivalent to nontransgenic maize with respect to these important components. Comparison of the variability within the nontransgenic and transgenic hybrid, as compared to composition values reported in the literature, suggest that factors other than transgenes may contribute more substantially to the composition of crops.     

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.     

Plant development,agronomic performance and nutritive value of forage maize depending on hybrid and marginal site conditions at high latitudes

Abstract

The objectives of this investigation were to study the effects of marginal site conditions and hybrid on plant development, agronomic performance and nutritive characteristics of forage maize (Zea mays L.) at high latitudes. Field experiments were conducted in 2008 and 2009 at three experimental sites, Kristianstad, Skara and Västerås, at increasing latitudes from 55°–60° N. Experimental design used two replicated randomized complete blocks at each site with three maize hybrids, Avenir (FAO 180), Isberi (FAO 190) and Burli (FAO 210), which were continuously assessed for plant development and harvested at various levels of maturity. The chemical composition and nutritional characteristics of harvested plant materials were analysed and hybrid responses to advancing maturity in terms of yield and nutritional qualities were evaluated. Results showed that maize hybrids required different numbers of accumulated thermal units at sites on varying latitudes to achieve developmental stages. Lowest thermal unit requirements among hybrids were observed for hybrid Avenir, and for sites it was highest for plants grown in the most northern site, Västerås. The most southern site, Kristianstad, was the only site at which all hybrids reached the dent stage (c. 450 g kg^?1 kernel DM), a recommended maturity for ensiling. The DM yields of early maturing hybrid Avenir were consistently lower than those for Isberi and Burli at all the sites. Results also revealed nutritional differences among maize hybrids at a given maturity (DM, g kg^?1), indicating that the effects of maturation should be factored into design of hybrid performance trials. This study highlights the effects of marginal site conditions and hybrids on plant development, agronomic performance and nutritional characteristics of maize hybrids at high latitudes. Further studies on marginal sites are recommended to enlighten the understanding of interaction between environmental and genetic factors on the performance of forage maize.     

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.     

Variability in European Maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) Landraces under High and Low Nitrogen Inputs

An European maize (Zea mays L.) landrace core collection (EMLCC) was formed with samples from several countries. Evaluation of the EMLCC may contribute to broad the genetic base of maize breeding programs. The objective of this study was to assess the variability of EMLCC under low nitrogen (N) in relation to high N input. Eighty-five landraces of the EMLCC, grouped in four maturity groups, and three check hybrids were evaluated for response to low (0 kg ha⁻¹) and high (150 kg ha⁻¹) N in Spain and Greece. Five plant size traits (plant height, ear height, leaf length, leaf width and leaf area index), two grain traits (1000-kernel weight and grain yield), and two agronomic traits [growing degree units (GDU) and lodging] were studied. Overall means of plant size and grain traits increased when genotypes were grown at 150-N relative to 0-N input. The relative increase for grain traits was smaller in landraces than in hybrids. This suggests that landraces had lower grain yield response to N supply compared to hybrids. Linear regressions of plant size traits on GDU indicated that vegetative development was primarily associated with flowering lateness. The maturity group was the main source of variation for all traits. Landrace variability within maturity groups was significant for all traits across environments, despite significant landrace × environment interactions. Estimates of genetic and genotype × environment variances, and heritabilities at both high and low N inputs were not significantly different from each other. However estimates were generally larger at high N. Genetic and phenotypic correlation coefficients between the two N levels were very high for all traits.     

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.     

Role of maize hybrids and their chemical composition in Fusarium infection and fumonisin production

This study was designed to investigate the role of hybrids in maize Fusarium section Liseola interaction and fumonisin production, with particular emphasis on the occurrence and accumulation of hidden fumonisins in maize (masking phenomenon). In this 2 year study, naturally infected field crops were chosen with 10 maize hybrids, six of them grown in both years. Maize samples collected in 2010 showed a higher incidence of fungal infection as well as higher fumonisin contamination than those obtained in 2009 but a very similar incidence of F. section Liseola. Fumonisin masking was confirmed in raw maize, with a lower amount of hidden forms as compared to free fumonisins detected in the year with higher contamination. The chemical composition of the different hybrids was determined and correlated with the contamination data: the results obtained highlight the main role of fatty acids, with a higher fumonisin contamination in hybrids showing a higher linoleic acid content and a higher masking action in hybrids with higher oleic to linoleic ratio. These results represent a good basis to explain maize hybrid susceptibility to F. section Liseola infection, fumonisin contamination, and masking not related to a specific commercial hybrid but extendable to all hybrids.     

Classification of Dry-Milled Maize Grit Yield Groups Using Quadratic Discriminant Analysis and Decision Tree Algorithm

A genetically and environmentally diverse collection of maize (Zea maize L.) samples was evaluated for physical properties and grit yield to help develop a standard set of criteria to identify grain best suited for dry-milling. Application of principal component analysis (PCA) reduced a set of approximately 500 samples collected from six states to 154 maize hybrids. Selected maize hybrids were placed into seven groups according to their dry-milled grit yields. Regression analysis explained only 50% of the variability in dry-milling grit yield. Patterns of differences in the physical properties for the seven grit yield groups implied that the seven yield groups could be placed into two or three groups. Using two pattern recognition techniques for improving classification accuracy, quadratic discriminant analysis and the classification and regression tree (CART) model, dry-milled grit yield groups were predicted. The estimated correct classification rates were 69–80% when the samples were divided into three yield groups and 81–90% when samples were divided into two yield groups. The results indicated the comparable success of both techniques and the superiority of the decision tree algorithm to quadratic discriminant analysis by offering higher accuracy and clearer classification rules in differentiating among dry-milled grit yield groups.     

EFFECT OF POTASSIUM NUTRITION ON SOLUTE ACCUMULATION,ION COMPOSITION AND YIELD OF MAIZE HYBRIDS GROWN UNDER SALINE CONDITIONS

An experiment was conducted to study the response of two maize hybrids to external potassium (K) application under saline conditions. The data showed that there was an increase in the organic solute contents and sodium ion under salinity stress, though potassium, calcium, nitrogen and phosphorus were decreased. There was a non-significant effect of K application on glycinebetaine and total soluble sugar, however; the proline, protein and total free amino acids were increased with the application of external K. The enzymatic activity like nitrate reductase and nitrite reductase activity were severely reduced under salinity stress and improved by K application. The maize hybrids differed significantly for all the parameters discussed in the study except sugar, phosphorus and number of grain rows per cob. The increase in yield parameters was more pronounced under control than under saline conditions. The enhanced yield and yield components of these maize hybrids might be due to the quick response to external K application, resulting in high contents of leaf potassium, calcium, nitrogen and phosphorus. The results indicated that the maize hybrid ‘Pioneer32B33’ might perform better than ‘Dekalb979’ under saline conditions when sufficient potassium is applied in the rooting medium.     

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.     

Agronomic evaluation of testcrosses of drought-tolerant maize (Zea mays L.) inbred lines using different selection index methods

The present study aims at identifying potentially high-yielding testcross maize hybrids using different selection indices that consider agronomic traits other than grain yield alone. Forty-one (41) testcross hybrids developed from 20 elite drought-tolerant maize inbred lines and two inbred testers plus three hybrid checks were evaluated in the rainy seasons of 2013 and 2014 in Ogbomoso in the derived savanna agroecological zone of Nigeria. Hybrids were planted in each year in 11 × 4 lattices with two replications. Testcross hybrids and checks exhibited significant differences for all measured traits except ear height, husk cover and number of ears per plant. Grain yield of hybrids and checks averaged over the 2 years varied from 3117 to 8393 kg ha^?1 with a mean of 5935 kg ha^?1. Top 11 hybrids produced vigorous plants, each yielding >7000 kg ha^?1. Base index and multivariate index methods identified EXL11 × 9071 whereas base index and Smith–Hazel selection index methods identified EXL18 × 9071 as superior hybrids at 5% selection intensity. These hybrids will be considered for advanced breeding program and release to farmers as high-yielding drought-tolerant hybrid maize varieties in the increasingly drought-prone derived savanna agroecology of Nigeria.     

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.     

Physical,Compositional, and Wet Milling Characteristics of Grain from Crosses of Corn Inbreds with Exotic and Nonexotic Background

Corn breeders have developed hybrids with enhanced compositional characteristics, but exotic germplasm represents little of the germplasm base used to produce these hybrids. Effects of the exotic germplasm on physical, compositional, and wet‐milling properties as well as the proximate composition of recovered fractions need to be determined before these materials are of value to the corn processing industry. Ten lines from the Germplasm Enhancement of Maize (GEM) project with exotic germplasm introgressed from Argentina, Chile, Uruguay, Cuba, and Florida were crossed to three adapted inbred lines (testers) and grain from the resulting 30 hybrids were evaluated for physical, compositional, and wet‐milling characteristics and the expression of heterosis in these variables. The B73xMo17 adapted public hybrid was used as control. Grain obtained by self‐pollination of the hybrid plants was analyzed using near‐infrared transmittance (NIT) technology and a 100‐g wet‐milling procedure. There was great variation among physical, compositional, and wet‐milling characteristics, and some of the experimental hybrids with exotic origin had better starch yield and starch recovery than B73xMo17, which suggests that wet‐milling characteristics of U.S. hybrids can be improved by breeding with exotic germplasm. In particular, GEM breeding crosses AR16035:S19, CH05015:N15, CUBA117:S1520, and FS8B(T):N1802 could be valuable germplasm sources to produce inbreds with good milling properties. Testers varied in ability to produce hybrids with good milling properties, indicating that choice of tester is an important factor when evaluating this end use. Although general trait trends for mid‐ and high‐parent heterosis were revealed, individual variation among hybrids and testers was large for most traits. This demonstrates the importance of analyzing individual hybrids that are intended for the wet‐milling industry when breeding with exotic introgressed lines.     

Polyphenols distribution in juices from Citrus allotetraploid somatic hybrids and their sexual hybrids

The polyphenol profile of an interspecific allotetraploid somatic hybrid, achieved by protoplast fusion, the 'Valencia' sweet orange ( Citrus sinensis L. Osbeck) + 'Femminello' lemon ( Citrus limon L. Burm), and three sexual hybrids obtained by backcrosses between Femminello lemon and the allotetraploid somatic hybrid ('Valencia' + 'Femminello') was studied by liquid chromatography-ultraviolet-diode array detector-mass spectrometry (LC-UV-DAD-MS). The aim of the work was to evaluate whether superior traits and improved performance can be observed in these new genotypes. Ten flavonoids (TF), comprising seven flavanones and three flavones, and four hydroxycinnamic acids (HCA) have been characterized and quantified in all hybrids and compared with those of the respective parents. The 'Valencia' + 'Femminello' somatic hybrid shows an intermediate polyphenol composition with respect to those of the parents, with a slight prevalence of lemon influence. The three sexual hybrids show, instead, different and more complex chromatographic profiles.     

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.     

Metabolite profiling of maize kernels--genetic modification versus environmental influence

A metabolite profiling approach based on gas chromatography-mass spectrometry (GC-MS) was applied to investigate the metabolite profiles of genetically modified (GM) Bt-maize (DKC78-15B, TXP 138F) and Roundup Ready-maize (DKC78-35R). For the comparative investigation of the impact of genetic modification versus environmental influence on the metabolite profiles, GM maize was grown together with the non-GM near-isogenic comparators under different environmental conditions, including several growing locations and seasons in Germany and South Africa. Analyses of variance (ANOVA) revealed significant differences between GM and non-GM maize grown in Germany and South Africa. For the factor genotype, 4 and 3%, respectively, of the total number of peaks detected by GC-MS showed statistically significant differences (p < 0.01) in peak heights as compared to the respective isogenic lines. However, ANOVA for the factor environment (growing location, season) revealed higher numbers of significant differences (p < 0.01) between the GM and the non-GM maize grown in Germany (42%) and South Africa (10%), respectively. This indicates that the majority of differences observed are related to natural variability rather than to the genetic modifications. In addition, multivariate data assessment by means of principal component analysis revealed that environmental factors, that is, growing locations and seasons, were dominant parameters driving the variability of the maize metabolite profiles.     

Variability in Starch Acetylation Efficiency from Commercial Waxy Corn Hybrids

Raw material variability is common for starch processors and is responsible for increased processing costs. In this study, variability of starch acetylation due to hybrid influence was quantified. Six waxy corn (maize) hybrids from 1998 and five waxy corn hybrids from 1999 were wet‐milled in the laboratory. Starch obtained from each hybrid was modified according to a laboratory‐scale acetylation procedure. To evaluate reaction efficiency, reaction rate, acetyl content, pH, and amount of NaOH used were recorded for each reaction. After modification, a Rapid Visco Analyser (RVA) was used to characterize modified starches and determine differences in modified starches from different hybrids. Using the same acetylation protocol, reaction efficiencies were observed at 47–73%. Reaction efficiencies were significantly lower for 1998 hybrids (50.0%) compared with the efficiency observed for the same hybrids grown in 1999 (62.7%). Acetylated starch from 1999 had increased peak, trough and final viscosities and increased reaction efficiency as compared with acetylated starch from 1998. Differences in setback were observed among 1998 hybrids for acetylated samples. Differences in trough and final viscosity were observed among 1999 hybrids for acetylated and native (unmodified) samples. Differences in breakdown among 1999 hybrids also were observed for native samples.     

INTER-RELATIONSHIPS OF APPLIED NITROGEN,SPAD, AND YIELD OF LEAFY AND NON-LEAFY MAIZE GENOTYPES

The SPAD chlorophyll meter was found to be a reliable, quick, and non-destructive tool used for directly measuring leaf chlorophyll and indirectly assessing the proportional parameter of leaf, and by extension, plant nitrogen (N) status. The meter has been used successfully to assess leaf N in conventional maize crops, but it has not been used with new maize (Zea mays L.) genotypes containing leafy (L) and reduced stature (RS) traits. SPAD meter readings were collected on the uppermost fully developed leaves (before silking) and on the ear leaf (after silking) of field grown maize genotypes with and without the L and RS traits. The experiment was conducted during 1996 and 1997 at two sites in Eastern Canada (Ottawa and Montreal). At each site in each year, a split plot arrangement of two treatment factors was used in a randomized complete block design with four blocks. The main plot treatments were levels of N (0, 85, 170, and 255 kg ha^?1), with six maize genotypes as subplot treatments. The hybrids included: (i) leafy reduced-stature, LRS, (ii) non-leafy normal stature, NLNS, (iii) leafy normal stature, (LNS), (iv) non-leafy reduced-stature, NLRS, (v) conventional commercial hybrids, Pioneer 3905 as the hybrid check for late maturity, and (vi) Pioneer 3979, a check for early maturity. The hybrids were chosen on the basis of their contrasting canopies and root architecture. The SPAD meter readings were collected on the same five plant genotypes over time (six times per site per year, except four times for the Ottawa site in 1997). All genotypes showed increasing meter reading values as plants aged until silking. In general, SPAD meter readings increased as N fertilization level increased at each measurement date for both sites and years. In general, LNS and P3905 hybrids showed greater SPAD meter readings than other hybrids at all sampling dates for both sites and growing seasons. Applied N rates were significantly correlated with the SPAD meter readings. More highly significant relationships were found for N fertilizer levels and SPAD meter readings for the hybrids in 1997 than for the hybrids in 1996. For the Montreal site in 1997, LRS, LNS and P3905 hybrids were among those showing the highest r values between N level and SPAD readings. The correlation coefficients between SPAD readings and grain yield were generally lower. However, the NLNS hybrid had a high SPAD-yield correlation at the Macdonald site in 1997.