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International shipping of fumonisins from maize extracts on C18 sorbent.

Fumonisins are mycotoxins found in maize. In developing countries, the resources required for analysis are often lacking, and the shipping of maize between countries can be difficult since the importation of plant materials requires permits/inspection to prevent the entry of pests that frequently infest maize. A simple, safe and legal method for shipping maize extracts to the USA was needed to conduct a survey of fumonisins in Central America. The objective was to develop a method for isolating and shipping maize extracts for fumonisin analysis so as to facilitate a survey of fumonisin exposure. The results indicate that fumonisins in acetonitrile:water extracts of maize can be isolated on C18 cartridges, held for at least 3 days at 22 degrees C and then an additional 4 days at 4 degrees C before elution and analysis with no losses. This method allows the importation and analysis of maize samples from foreign locations without complications from international safety concerns.

Carbon↗

Weed interference on maize (Zea mays) under no tillage system.

A field trial was carried out in Brazil in March 2002 with the aim to evaluate the effects of different timing and extension of weedy period on maize productivity. The hybrid Pioneer 30K75 was sowed under 7 t ha(-1) mulching promoted by glyphosate spraying. The treatments were divided in two groups: In the first group, weeds were maintained since the maize sowing until different periods in the crop cycle: 0, 14, 28, 42, 56, 70, and 150 days (harvesting time). In the second group, the maize crop was kept weed free for the same periods of the first group. Weed control was done through hand hoeing. A complete randomized blocks experimental design with five replications was used for plots distribution in the field. Nonlinear regression model was used to study the effects of weedy or weedfree periods on maize productivity. Weed community included 13 families and 31 species. Asteraceae, Poaceae, and Euphorbiaceae were the most abundant families. Results showed that under no tillage condition with 7 t ha(-1) mulching at sowing time, the maize crop could cohabit with weed community for 54 days without any yield lost. On the other hand, if the crop was kept weed free for 27 days, the weed interference was not enable to reduce maize production. According to these results one weed control measure between 27 and 54 days after crop emergence could be enough to avoid any reduction in maize productivity.

Agriculture↗

Zinc absorption from low-phytate hybrids of maize and their wild-type isohybrids.

BACKGROUND: Identification of allelic variants in a single gene that determine the phytate content of maize kernels and the subsequent breeding of low-phytate maize have facilitated studies designed to determine quantitatively the effects of maize phytate on the bioavailability of minerals in maize. OBJECTIVE: The objective was to determine the relation between the fractional absorption of zinc (FAZ) and the phytate content and phytate:zinc molar ratios of maize tortillas prepared from hybrids with different phytate contents. DESIGN: Six healthy adults were fed, as the only food for 2 d, maize tortillas prepared from 1 of 2 low-phytate mutants: lpa1-1 (lpa1-1-LP) or Nutridense Low Phytate (ND-LP), which have phytate reductions of approximately 60% and approximately 80%, respectively, compared with their respective wild-type isohybrids. Four additional subjects were fed tortillas prepared from the corresponding wild-type isohybrids (lpa1-1-WT and ND-WT) according to the same study design. Meals were extrinsically labeled with zinc stable isotopes, and FAZ was determined with a dual-isotope-tracer ratio technique. Overall FAZ values were examined in relation to dietary phytate and phytate:zinc molar ratios by using a mixed nonlinear regression model. RESULTS: The mean (+/-SD) FAZ values from tortillas prepared from ND-LP, lpa1-1-LP, lpa1-1-WT, and ND-WT were 0.38 +/- 0.07, 0.28 +/- 0.04, 0.15 +/- 0.07, and 0.13 +/- 0.05, respectively. A negative relation (P < 0.001) was found between FAZ and both dietary phytate and the phytate:zinc molar ratio. The effect of dietary zinc (8-14 mg Zn/d) under these experimental conditions was not significant. CONCLUSIONS: FAZ from maize tortillas is positively related to the extent of phytate reduction achieved with low-phytate hybrids.

Adult↗

Complex structure of knob DNA on maize chromosome 9. Retrotransposon invasion into heterochromatin.

The recovery of maize (Zea mays L.) chromosome addition lines of oat (Avena sativa L.) from oat x maize crosses enables us to analyze the structure and composition of specific regions, such as knobs, of individual maize chromosomes. A DNA hybridization blot panel of eight individual maize chromosome addition lines revealed that 180-bp repeats found in knobs are present in each of these maize chromosomes, but the copy number varies from approximately 100 to 25, 000. Cosmid clones with knob DNA segments were isolated from a genomic library of an oat-maize chromosome 9 addition line with the help of the 180-bp knob-associated repeated DNA sequence used as a probe. Cloned knob DNA segments revealed a complex organization in which blocks of tandemly arranged 180-bp repeating units are interrupted by insertions of other repeated DNA sequences, mostly represented by individual full size copies of retrotransposable elements. There is an obvious preference for the integration of retrotransposable elements into certain sites (hot spots) of the 180-bp repeat. Sequence microheterogeneity including point mutations and duplications was found in copies of 180-bp repeats. The 180-bp repeats within an array all had the same polarity. Restriction maps constructed for 23 cloned knob DNA fragments revealed the positions of polymorphic sites and sites of integration of insertion elements. Discovery of the interspersion of retrotransposable elements among blocks of tandem repeats in maize and some other organisms suggests that this pattern may be basic to heterochromatin organization for eukaryotes.

Avena↗

Expression patterns and mutant phenotype of teosinte branched1 correlate with growth suppression in maize and teosinte.

The evolution of domesticated maize from its wild ancestor teosinte is a dramatic example of the effect of human selection on agricultural crops. Maize has one dominant axis of growth, whereas teosinte is highly branched. The axillary branches in maize are short and feminized whereas the axillary branches of teosinte are long and end in a male inflorescence under normal growth conditions. Previous QTL and molecular analysis suggested that the teosinte branched1 (tb1) gene of maize contributed to the architectural difference between maize and teosinte. tb1 mutants of maize resemble teosinte in their overall architecture. We analyzed the tb1 mutant phenotype in more detail and showed that the highly branched phenotype was due to the presence of secondary and tertiary axillary branching, as well as to an increase in the length of each node, rather than to an increase in the number of nodes. Double-mutant analysis with anther ear1 and tassel seed2 revealed that the sex of the axillary inflorescence was not correlated with its length. RNA in situ hybridization showed that tb1 was expressed in maize axillary meristems and in stamens of ear primordia, consistent with a function of suppressing growth of these tissues. Expression in teosinte inflorescence development suggests a role in pedicellate spikelet suppression. Our results provide support for a role for tb1 in growth suppression and reveal the specific tissues where suppression may occur.

Biological Evolution↗

Influence of maize/lablab intercropping on lepidopterous stem borer infestation in maize.

Lepidopterous stem borers seriously affect production of maize, Zea mays L., in sub-Saharan Africa. Intercropping maize with legumes such as lablab, Lablab purpurens (L.), is one of the effective systems to control stem borers. Sole culture maize and maize/lablab intercrop system of different lablab densities were planted at two locations to investigate the effects of intercrop system on incidence and severity of stem borers with particular reference to Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae). Stem borer infestation was found to be more severe in sole culture maize than maize in maize/lablab intercrop. There was a significantly negative relationship between lablab densities and maize grain yields, suggesting a possible competition for resources between the two crops. It was concluded that density of lablab and date of planting of lablab in maize/lablab intercropping have significant affects on stem borer populations and maize grain yields.

Agriculture↗

MaizeGDB, the community database for maize genetics and genomics.

The Maize Genetics and Genomics Database (MaizeGDB) is a central repository for maize sequence, stock, phenotype, genotypic and karyotypic variation, and chromosomal mapping data. In addition, MaizeGDB provides contact information for over 2400 maize cooperative researchers, facilitating interactions between members of the rapidly expanding maize community. MaizeGDB represents the synthesis of all data available previously from ZmDB and from MaizeDB-databases that have been superseded by MaizeGDB. MaizeGDB provides web-based tools for ordering maize stocks from several organizations including the Maize Genetics Cooperation Stock Center and the North Central Regional Plant Introduction Station (NCRPIS). Sequence searches yield records displayed with embedded links to facilitate ordering cloned sequences from various groups including the Maize Gene Discovery Project and the Clemson University Genomics Institute. An intuitive web interface is implemented to facilitate navigation between related data, and analytical tools are embedded within data displays. Web-based curation tools for both designated experts and general researchers are currently under development. MaizeGDB can be accessed at http://www.maizegdb.org/.

Computational Biology↗

Characterization of a highly conserved sequence related to mutator transposable elements in maize.

Mutator stocks of maize exhibit a high mutation rate correlated with the activity of a family of transposable elements. Mu1 and, to a lesser extent, the closely related Mu1.7 elements are responsible for most mutator-induced mutations that have been characterized. These elements are found in 10-60 copies in mutator stocks, and zero to a few intact elements exist in nonmutator maize stocks. Additionally, the component parts of Mu elements exist separately in the maize genome. The Mu terminal inverted repeats are found in multiple copies in all maize lines and related Zea species tested, and Mu internal sequences exist unassociated with Mu termini. In the present paper, we describe the structure and genomic distribution of one Mu-homologous sequence termed MRS-A (for Mu-related sequence). DNA sequencing shows that MRS-A is closely related to the internal region of Mu1 and Mu1.7 elements. However, it has no Mu termini and does not have the structure of a transposable element. This sequence is present in one or two copies in all maize lines and is highly conserved in the genus Zea. A similar sequence exists in a species within the genus most closely related to Zea, Tripsacum dactyloides, although the T. dactyloides genome does not contain any Mu termini or intact Mu elements. Furthermore, an RNA transcript homologous to MRS-A and its flanking DNA is found in both mutator and nonmutator maize plants. These results suggest that MRS-A represents a stable, functional region of the maize genome, and we speculate that a similar sequence was encompassed by Mu termini to generate a Mu transposable element.

Base Sequence↗

FUM1--a gene required for fumonisin biosynthesis but not for maize ear rot and ear infection by Gibberella moniliformis in field tests.

We have analyzed the role of fumonisins in infection of maize (Zea mays) by Gibberella moniliformis (anamorph Fusarium verticillioides) in field tests in Illinois and Iowa, United States. Fumonisin-nonproducing mutants were obtained by disrupting FUM1 (previously FUM5), the gene encoding a polyketide synthase required for fumonisin biosynthesis. Maize ear rot, ear infection, and fumonisin contamination were assessed by silk-channel injection in 1999 and 2000 and also by spray application onto maize silks, injection into maize stalks, and application with maize seeds at planting in 1999. Ear rot was evaluated by visual assessment of whole ears and by calculating percentage of symptomatic kernels by weight. Fumonisin levels in kernels were determined by high-performance liquid chromatography. The presence of applied strains in kernels was determined by analysis of recovered isolates for genetic markers and fumonisin production. Two independent fumonisin-nonproducing (fum1-3 and fum1-4) mutants were similar to their respective fumonisin-producing (FUM1-1) progenitor strains in ability to cause ear rot following silk-channel injection and also were similar in ability to infect maize ears following application by all four methods tested. This evidence confirms that fumonisins are not required for G. moniliformis to cause maize ear rot and ear infection.

Fumonisins↗

Patterns of chromosomal duplication in maize and their implications for comparative maps of the grasses.

The maize genome contains extensive chromosomal duplications that probably were produced by an ancient tetraploid event. Comparative cereal maps have identified at least 10 duplicated, or homologous, chromosomal regions within maize. However, the methods used to document chromosomal homologies from comparative maps are not statistical, and their criteria are often unclear. This paper describes the development of a simulation method to test for the statistical significance of marker colinearity between chromosomes, and the application of the method to a molecular map of maize. The method documents colinearity among 24 pairs of maize chromosomes, suggesting homology in maize is more complex than represented by comparative cereal maps. The results also reveal that 60%-82% of the genome has been retained in colinear regions and that as much as a third of the genome could be present in multiple copies. Altogether, the complex pattern of colinearity among maize chromosomes suggests that current comparative cereal maps do not adequately represent the evolution and organization of the maize genome.

Chromosome Mapping↗

Uneven chromosome contraction and expansion in the maize genome.

Maize (Zea mays or corn), both a major food source and an important cytogenetic model, evolved from a tetraploid that arose about 4.8 million years ago (Mya). As a result, maize has extensive duplicated regions within its genome. We have sequenced the two copies of one such region, generating 7.8 Mb of sequence spanning 17.4 cM of the short arm of chromosome 1 and 6.6 Mb (25.6 cM) from the long arm of chromosome 9. Rice, which did not undergo a similar whole genome duplication event, has only one orthologous region (4.9 Mb) on the short arm of chromosome 3, and can be used as reference for the maize homoeologous regions. Alignment of the three regions allowed identification of syntenic blocks, and indicated that the maize regions have undergone differential contraction in genic and intergenic regions and expansion by the insertion of retrotransposable elements. Approximately 9% of the predicted genes in each duplicated region are completely missing in the rice genome, and almost 20% have moved to other genomic locations. Predicted genes within these regions tend to be larger in maize than in rice, primarily because of the presence of predicted genes in maize with larger introns. Interestingly, the general gene methylation patterns in the maize homoeologous regions do not appear to have changed with contraction or expansion of their chromosomes. In addition, no differences in methylation of single genes and tandemly repeated gene copies have been detected. These results, therefore, provide new insights into the diploidization of polyploid species.

Base Sequence↗

The polar auxin transport inhibitor N-1-naphthylphthalamic acid disrupts leaf initiation, KNOX protein regulation, and formation of leaf margins in maize.

Maize (Zea mays) leaves develop basipetally (tip to base); the upper blade emerges from the shoot apical meristem (SAM) before the expansion of the lower sheath. Founder cells, leaf initials located in the periphery of the SAM, are distinguished from the SAM proper by the differential accumulation of KNOX proteins. KNOX proteins accumulate in the SAM, but are excluded from maize leaf primordia and leaf founder cells. As in Arabidopsis and tomato (Lycopersicon esculentum), maize shoots failed to initiate new leaves when cultured in the polar auxin transport inhibitor N-1-naphthylphthalamic acid (NPA). We demonstrate that NPA-induced arrest of leaf initiation in maize is correlated with the failure to down-regulate KNOX accumulation in the SAM. In addition, NPA-cultured shoots formed abnormal tubular leaf bases in which the margins failed to separate in the lower leaf zone. The tubular leaf bases always formed in the fourth leaf from the arrested meristem. Moreover, the unseparated margin domains of these tubular leaf bases accumulated ectopic KNOX protein(s). Transfer of NPA-cultured apices to NPA-free media resulted in the resumption of leaf initiation from the SAM and the restoration of normal patterns of KNOX down-regulation, accordingly. These data suggest that the lower sheath margins emerge from the leaf base late in maize leaf development and that the separation of these leaf margin domains is correlated with auxin transport and down-regulation of KNOX proteins. In addition, these results suggest that the down-regulation of KNOX accumulation in maize apices is not upstream of polar auxin transport, although a more complicated feedback network may exist. A model for L1-derived margin development in maize leaves is presented.

Biological Transport↗

The maize root transcriptome by serial analysis of gene expression.

Serial Analysis of Gene Expression was used to define number and relative abundance of transcripts in the root tip of well-watered maize seedlings (Zea mays cv FR697). In total, 161,320 tags represented a minimum of 14,850 genes, based on at least two tags detected per transcript. The root transcriptome has been sampled to an estimated copy number of approximately five transcripts per cell. An extrapolation from the data and testing of single-tag identifiers by reverse transcription-PCR indicated that the maize root transcriptome should amount to at least 22,000 expressed genes. Frequency ranged from low copy number (2-5, 68.8%) to highly abundant transcripts (100-->1,200; 1%). Quantitative reverse transcription-PCR for selected transcripts indicated high correlation with tag frequency. Computational analysis compared this set with known maize transcripts and other root transcriptome models. Among the 14,850 tags, 7,010 (47%) were found for which no maize cDNA or gene model existed. Comparing the maize root transcriptome with that in other plants indicated that highly expressed transcripts differed substantially; less than 5% of the most abundant transcripts were shared between maize and Arabidopsis (Arabidopsis thaliana). Transcript categories highlight functions of the maize root tip. Significant variation in abundance characterizes transcripts derived from isoforms of individual enzymes in biochemical pathways.

Base Sequence↗

Purification and cDNA cloning of maize Poly(ADP)-ribose polymerase.

Poly(ADP)-ribose polymerase (PADPRP) has been purified to apparent homogeneity from suspension cultures of the maize (Zea mays) callus line. The purified enzyme is a single polypeptide of approximately 115 kD, which appears to dimerize through an S-S linkage. The catalytic properties of the maize enzyme are very similar to those of its animal counterpart. The amino acid sequences of three tryptic peptides were obtained by microsequencing. Antibodies raised against peptides from maize PADPRP cross-reacted specifically with the maize enzyme but not with the enzyme from human cells, and vice versa. We have also characterized a 3.45-kb expressed-sequence-tag clone that contains a full-length cDNA for maize PADPRP. An open reading frame of 2943 bp within this clone encodes a protein of 980 amino acids. The deduced amino acid sequence of the maize PADPRP shows 40% to 42% identity and about 50% similarity to the known vertebrate PADPRP sequences. All important features of the modular structure of the PADPRP molecule, such as two zinc fingers, a putative nuclear localization signal, the automodification domain, and the NAD+-binding domain, are conserved in the maize enzyme. Northern-blot analysis indicated that the cDNA probe hybridizes to a message of about 4 kb.

Amino Acid Sequence↗

Stomatal Behavior and Water Status of Maize, Sorghum, and Tobacco under Field Conditions: I. At High Soil Water Potential.

Diurnal changes in the vertical profiles of irradiance incident upon the adaxial leaf surface (I), stomatal resistance (r(s)), leaf water potential (psi), osmotic potential (pi), and turgor potential (P) were followed concurrently in crops of maize (Zea mays L. var. Pa 602A), sorghum (Sorghum bicolor [L.] Moench var. RS610), and tobacco (Nicotiana tabacum L. var. Havanna Seed 211) on several days in 1968 to 1970 when soil water potentials were high. In all three crops the r(s), measured with a ventilated diffusion porometer, the psi, measured with the pressure chamber, the pi, measured with a vapor pressure osmometer, and the calculated P, decreased from sunrise to reach minimum values near midday and then increased again in the afternoon. The diurnal range of all the variables was greater for leaves in the upper canopy than for those in the lower canopy. P was observed to decrease with decreasing psi, but never became zero. Sorghum had a higher P at a psi of, say -10 bars, than did maize, and maize had a higher P than tobacco at the same psi. Moreover, at the same psi the upper leaves in all canopies had a higher P than the lower leaves. When compared at high irradiances, r(s) did not increase as psi declined to -13, -15, and -10 bars or as P declined to 0.3, 3.5, and 1.2 bars in maize, sorghum, and tobacco, respectively. The relation between r(s) and I in the upper, nonsenescent leaves of all three crops fits a hyperbolic curve, but the response varied with species and leaf senescence. The adaxial and abaxial epidermises had the same response of r(s) to I in maize and sorghum, whereas in tobacco the adaxial epidermis had a higher r(s) than the abaxial epidermis at all values of I. At equal values of I, tobacco had the lowest leaf resistance (r(l)) and maize had the highest r(l). Senescent maize leaves had nonfunctional stomata, whereas the lowermost sorghum leaves had higher stomatal resistances on average than the other leaves.

Journal Article↗

Maize centromeres: organization and functional adaptation in the genetic background of oat.

Centromeric DNA sequences in multicellular eukaryotes are often highly repetitive and are not unique to a specific centromere or to centromeres at all. Thus, it is a major challenge to study the fine structure of individual plant centromeres. We used a DNA fiber-fluorescence in situ hybridization approach to study individual maize (Zea mays) centromeres using oat (Avena sativa)-maize chromosome addition lines. The maize centromere-specific satellite repeat CentC in the addition lines allowed us to delineate the size and organization of centromeric DNA of individual maize chromosomes. We demonstrate that the cores of maize centromeres contain mainly CentC arrays and clusters of a centromere-specific retrotransposon, CRM. CentC and CRM sequences are highly intermingled. The amount of CentC/CRM sequence varies from approximately 300 to >2800 kb among different centromeres. The association of CentC and CRM with centromeric histone H3 (CENH3) was visualized by a sequential detection procedure on stretched centromeres. The analysis revealed that CENH3 is always associated with CentC and CRM but that not all CentC or CRM sequences are associated with CENH3. We further demonstrate that in the chromosomal addition lines in which two CenH3 genes were present, one from oat and one from maize, the oat CENH3 was consistently incorporated by the maize centromeres.

Adaptation, Physiological↗

Evolution of DNA sequence nonhomologies among maize inbreds.

Allelic chromosomal regions totaling more than 2.8 Mb and located on maize (Zea mays) chromosomes 1L, 2S, 7L, and 9S have been sequenced and compared over distances of 100 to 350 kb between the two maize inbred lines Mo17 and B73. The alleles contain extended regions of nonhomology. On average, more than 50% of the compared sequence is noncolinear, mainly because of the insertion of large numbers of long terminal repeat (LTR)-retrotransposons. Only 27 LTR-retroelements are shared between alleles, whereas 62 are allele specific. The insertion of LTR-retrotransposons into the maize genome is statistically more recent for nonshared than shared ones. Most surprisingly, more than one-third of the genes (27/72) are absent in one of the inbreds at the loci examined. Such nonshared genes usually appear to be truncated and form clusters in which they are oriented in the same direction. However, the nonshared genome segments are gene-poor, relative to regions shared by both inbreds, with up to 12-fold difference in gene density. By contrast, miniature inverted terminal repeats (MITEs) occur at a similar frequency in the shared and nonshared fractions. Many times, MITES are present in an identical position in both LTRs of a retroelement, indicating that their insertion occurred before the replication of the retroelement in question. Maize ESTs and/or maize massively parallel signature sequencing tags were identified for the majority of the nonshared genes or homologs of them. In contrast with shared genes, which are usually conserved in gene order and location relative to rice (Oryza sativa), nonshared genes violate the maize colinearity with rice. Based on this, insertion by a yet unknown mechanism, rather than deletion events, seems to be the origin of the nonshared genes. The intergenic space between conserved genes is enlarged up to sixfold in maize compared with rice. Frequently, retroelement insertions create a different sequence environment adjacent to conserved genes.

Alleles↗

Screening the allergenic repertoires of wheat and maize with sera from double-blind, placebo-controlled food challenge positive patients.

BACKGROUND: Food allergy to wheat and maize is an increasing factor of deterioration of life quality, especially childhood and can, in rare cases, even induce anaphylaxis. Although omega-5 gliadin from wheat and maize lipid transfer protein have been characterized as major cereal allergens on the molecular level, the list of food allergens is far to be complete. METHODS: To identify the IgE-binding repertoires of wheat and maize we screened respective cDNA libraries displayed on phage surface with sera from patients with a confirmed food allergy. The study included six patients with a positive double-blind, placebo-controlled food challenge (DBPCFC) to wheat, nine patients with a positive DBPCFC to maize, and six patients with anaphylactic reactions after ingestion of wheat. RESULTS: The enriched sequences encoding IgE-binding proteins showed heterogeneous repertoires for both, wheat and maize. The selected wheat repertoire yielded 12, the maize repertoire 11 open reading frames. Among these we identified allergens belonging to already characterized allergens families, such as gliadin, profilin and beta-expansin. Besides, we found novel proteins with high cross-reactive potential, such as thioredoxins, as well as sequences that had so far not been related to cereal allergy at all. The IgE-binding capacity of some selected proteins was evaluated in vitro and cross-reactivity was demonstrated by competition ELISA. CONCLUSION: With regard to the heterogeneity of the characterized sequences as well as to the biochemical nature of the new allergens detected we conclude that wheat and maize-related food allergy is more complex than so far anticipated.

Adolescent↗