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PubMedDate not supplied

Development and mapping of SSR markers for maize.

Microsatellite or simple sequence repeat (SSR) markers have wide applicability for genetic analysis in crop plant improvement strategies. The objectives of this project were to isolate, characterize, and map a comprehensive set of SSR markers for maize (Zea mays L.). We developed 1051 novel SSR markers for maize from microsatellite-enriched libraries and by identification of microsatellite-containing sequences in public and private databases. Three mapping populations were used to derive map positions for 978 of these markers. The main mapping population was the intermated B73 x Mo17 (IBM) population. In mapping this intermated recombinant inbred line population, we have contributed to development of a new high-resolution map resource for maize. The primer sequences, original sequence sources, data on polymorphisms across 11 inbred lines, and map positions have been integrated with information on other public SSR markers and released through MaizeDB at URL:www.agron.missouri.edu. The maize research community now has the most detailed and comprehensive SSR marker set of any plant species.

Chromosome Mapping↗

PubMed2002

A chimeric ecdysone receptor facilitates methoxyfenozide-dependent restoration of male fertility in ms45 maize.

A mutation in the maize Ms45 gene results in abortion of microspore development and a male-sterile phenotype. MS45 protein has been localized to the tapetum and maximally expressed in anthers at the early vacuolate stage of microspore development. Molecular complementation analysis determined that a transformed copy of the gene fully restored fertility to ms45 maize. In this report, using phenotypic complementation as an assay, chimeric transcriptional activators were expressed to regulate a gal:MS45 gene and test the ability of a multi-component system to restore male fertility. A high frequency of phenotypic complementation was observed when either C1-GAL4 or VP16-GAL4 activators were transcribed by promoters that expressed at a stage of anther development that precedes the early vacuolate stage of microsporogenesis. For the conditional regulation of male fertility, these transcriptional activators were modified by the addition of regions that include the ligand-binding domain from the European corn borer ecdysone receptor to generate the nuclear receptors C1-GAL4-EcR (CGEcR) and VP16-GAL4-EcR (VGEcR). These chimeric receptors were introduced with the gal:MS45 gene into ms45 maize, and in the absence of ligand, these plants were male sterile. In contrast, application of the ecdysone agonist, methoxyfenozide, to plants containing either a constitutive (Ubiquitin1) or anther-specific (maize 5126) VGEcR resulted in the restoration of fertility to ms45 plants grown in either the greenhouse or in the field.

Chimera↗

PubMed2003

Fungus- and wound-induced accumulation of mRNA containing a class II chitinase of the pathogenesis-related protein 4 (PR-4) family of maize.

Pathogenesis-related (PR) proteins are plant proteins that are induced in response to pathogen attack. PR proteins are grouped into independent families based on their sequences and properties. The PR-4 family comprises class I and class II chitinases. We have isolated a full-length cDNA encoding a chitinase from maize which shares a high degree of nucleotide and amino acid sequence homology with the class II chitinases of the PR-4 family of PR proteins. Our results indicate that fungal infection, and treatment either with fungal elicitors or with moniliformin, a mycotoxin produced by the fungus Fusarium moniliforme, increase the level of ZmPR4 mRNA. In situ mRNA hybridization analysis in sections obtained from fungus-infected germinating embryos revealed that ZmPR4 mRNA accumulation occurs in those cell types that first establish contact with the pathogen. ZmPR4 mRNA accumulation is also stimulated by treatment with silver nitrate whereas the application of the hormones gibberellic acid or acetylsalicylic acid has no effect. Wounding, or treatment with abscisic acid or methyl jasmonate, results in accumulation of ZmPR4 mRNA in maize leaves. Furthermore, the ZmPR4 protein was expressed in Escherichia coli, purified and used to obtain polyclonal antibodies that specifically recognized ZmPR4 in protein extracts from fungus-infected embryos. Accumulation of ZmPR4 mRNA in fungus-infected maize tissues was accompanied by a significant accumulation of the corresponding protein. The possible implications of these findings as part of the general defence response of maize plants against pathogens are discussed.

Amino Acid Sequence↗

PubMedDate not supplied

An assessment of the effects of crude oil pollution on soil properties, germination and growth of maize (Zea mays) using two crude types--Forcados light and Escravos light.

The effect of crude oil pollution on soil properties, germination and height of maize (Zea mays) was investigated under natural environment in three separate pot experiments. Two crude oil types--Forcados light and Escravos light were used. In Experiment 1, the effect of crude oil application on germination using high pollution levels of 5, 15, 25 and 40 mL kg(-1) of soil was assessed. In Experiment 2, the effect of crude oil application on maize plant height using the same pollution levels was assessed. The last experiment (Experiment 3) was used to test for the effects of crude oil application on maize plant height using lower levels of pollution (1.0, 1.5, 2.0 and 2.5 mL kg(-1) of soil). Soil samples were collected before, during and after the experiment and analyzed for some physical and chemical properties. Results obtained showed variation in chemical properties of soil. % organic matter increased from 1.34 to 2.62% in polluted soils. Available P decreased from 15.00 ppm in control to between 7.34 and 5.42 in soils polluted with 'high' levels of crude oil. Statistical analysis of height data showed that crude oil inhibited the growth of maize at high pollution levels. High levels of pollution also inhibited germination and for Escravos light there was no germination at 40 mL kg(-1) soil level of crude oil pollution.

Dose-Response Relationship, Drug↗

PubMed2004

Mycoflora of Iranian maize harvested in the main production areas in 2000.

Maize is one of the most important cereals produced in, and imported into, Iran. The incidences of seed-borne fungi were determined in Iranian maize harvested in 2000 from four major production areas with different climatic conditions, namely Fars, Khuzestan, Kermanshah and Mazandaran provinces. This is the first study to compare the mycoflora of maize in the aforementioned areas. Mycological analyses showed a predominance of Fusarium species (38.5%), followed by Aspergillus species (8.7%), Rhizopus species (4.8%), Penicillium species (4.5%), Mucor species (1.1%), and four other fungal genera. Fusarium verticillioides was the most prevalent species (83% of Fusarium isolates and 52% of the total isolations), with the highest incidence in Mazandaran (59%), a region of Iran with the highest rainfall and relative humidity, high rate of esophageal cancer (EC) and high levels of fumonisins in maize. Aspergillus flavus was the most widely recovered Aspergillus species and 38% of samples were contaminated with this potentially aflatoxigenic fungus. The incidence of A. flavus was highest in Kermanshah, the province with lowest mean minimum temperature. Penicillium species were seen in all the samples and Fars had the highest incidence, with highly significant differences when compared to the other three provinces. Diplodia species were not isolated from any of the samples examined.

Climate↗

PubMed2003

Dual positional specificity and expression of non-traditional lipoxygenase induced by wounding and methyl jasmonate in maize seedlings.

Lipoxygenases (LOXs) catalyze the formation of fatty acid hydroperoxides involved in responses to stresses. This study examines the expression of a non-traditional dual positional specific maize LOX in response to wounding or methyl jasmonate (MeJA). Full-length maize LOX cDNA was expressed in Escherichia coli, and recombinant LOX was purified and characterized enzymatically. RP-HPLC and GC-MS analysis showed that the purified LOX converts alpha-linolenic acid into 13-hydroperoxylinolenic acid and 9-hydroperoxylinolenic acid in a 6:4 ratio. LOX mRNA accumulated rapidly and transiently in response to wounding reaching a peak of expression about 3 h after wounding. This increase followed an initial increase in endogenous jasmonic acid (JA) 1 h after wounding (JA burst). However, the expression of LOX induced by MeJA lasted longer than the expression induced by wounding, and the MeJA-induced expression seemed to be biphasic pattern composed of early and late phases. The expression of LOX in the presence of inhibitors of JA biosynthesis was not completely inhibited, but delayed in wound response and the expression period was shortened in MeJA response. These results suggest that wound-responsive JA burst may trigger the early phase of LOX expression which facilitates biosynthesis of endogenous JA through its 13-LOX activity, and subsequently leads to the activation of the late phase LOX expression in MeJA-treated maize seedlings. Implications of dual positional specificity of maize LOX in the observed expression kinetics are discussed.

Acetates↗

PubMed2004

Serine:glyoxylate aminotransferases from maize and wheat leaves: purification and properties.

Photorespiratory enzyme serine:glyoxylate aminotransferase (SGAT, EC 2.6.1.45) was purified from green parts of seedlings of two Gramminae species with different photosynthetic pathways, maize (Zea mays L., C(4) species) and wheat (Triticum aestivum L., C(3) species). The preparation from wheat was homogeneous as judged by SDS-PAGE with silver staining for proteins; however, the same method revealed approximately 9% contamination in a highly purified maize preparation. Molecular masses of SGAT from maize and wheat were estimated by SDS-PAGE to be 44.1 and 44.6 kDa, respectively. C(4) enzyme exhibited a specific activity in homogenates that was seven times lower than wheat, and this was associated with lower K (m) values for all substrates examined as well as a more than two times lower turnover number k (cat) with serine and glyoxylate as a pair of substrates. In contrast, the ratio of the turnover number to K (m)(Ser)(k (cat)/K (m)(Ser)) for C(4) aminotransferase proved to be about two times higher than for C(3) aminotransferase. The sensitivity of two enzymes to some inhibitors, especially aminooxyacetate, was different and they also differed with respect to thermal stability and pH optimum - the maize enzyme required 0.6 unit higher pH (8.6) for maximal activity and was more heat-resistant.

Journal Article↗

PubMed2000

Identification of immunogenic maize proteins in a casein hydrolysate formula.

Cow's milk-based formulas used for infants with cow's milk allergy are based on hydrolyzed proteins. The formulas that are successful in preventing allergic responses are extensively hydrolyzed. Nevertheless, reactions to such formulas are occasionally reported, and protein material of higher molecular weight than expected has been detected by binding immunoglobulin E (IgE) from patients' sera. This paper presents the identification of high-molecular-weight material in the extensively hydrolyzed casein formula, Nutramigen. The material was concentrated by simple centrifugation. The proteins in the pellet were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein-containing bands were analyzed by protein sequencing after electroblotting. The proteins were identified as maize zeins, which are water-insoluble proteins of apparent M(r) 20,000 and 23,000, presumably originating from the maize starch in Nutramigen. Rabbits immunized with this formula developed antibodies against zeins but not against milk proteins. The maize zeins are probably identical to the recently reported components in Nutramigen (1), detected by binding of IgE from milk allergic patients, but not correlated to clinical allergic reactivity. The clinical relevance of maize proteins in Nutramigen remains to be established.

Animals↗

PubMed2002

Allelochemical stress produced by the aqueous leachate of Callicarpa acuminata: effects on roots of bean, maize, and tomato.

The in vitro effects of an aqueous leachate (1%) of Callicarpa acuminata Kunth. (Verbenaceae) on radicle growth, protein expression, catalase activity, free radical production and membrane lipid peroxidation in roots of bean, maize, and tomato were examined. Aqueous extract of C. acuminata inhibited the radicle growth of tomato by 47%, but had no effect on root growth of maize and beans. 2D-PAGE and densitometry analysis showed that C. acuminata aqueous leachate modified the expression of various proteins in the roots of all treated plants. In treated bean roots, microsequencing analysis of an 11.3-kDa protein, whose expression was enhanced by leachate treatment, revealed a 99% similarity with subunits of alpha-amylase inhibitor of other beans. A 27.5-kDa protein induced in treated tomato showed 69-95% similarity to glutathione-S-transferases (GST) of other Solanaceae. Spectrophotometric analysis and native gels revealed that catalase activity was increased by 2.2-fold in tomato roots and 1.4-fold in bean roots. No significant changes were observed in treated maize roots. Luminol chemiluminescence levels, a measure of free radicals, increased 3.8-fold in treated tomato roots and 2.1-fold in treated bean roots. Oxidative membrane damage in treated roots was measured by lipid peroxidation rates. In tomato we observed a 2.4-fold increase in peroxidation, however, no effect was observed in maize or beans.

Journal Article↗

PubMed1995

A glutathione S-transferase involved in vacuolar transfer encoded by the maize gene Bronze-2.

Glutathione S-transferases (GSTs) are enzymes that detoxify heterocyclic compounds (xenobiotics) by covalently linking glutathione to the substrate, forming a glutathione S-conjugate. A glutathione pump in the vacuolar membrane of barley actively sequesters herbicide-glutathione S-conjugates; glutathionation allows recognition and entry of the conjugates into vacuoles. The protein encoded by the Bronze-2 gene in maize performs the last genetically defined step in anthocyanin biosynthesis, resulting in the deposition of red and purple pigments in the vacuoles of maize tissues. We show here that Bz2 encodes a GST with activity in maize, transformed Arabidopsis thaliana plants and Escherichia coli. We demonstrate that anthocyanins extracted from maize protoplasts expressing BZ2 are conjugated with glutathione, and that vanadate, a known inhibitor of the glutathione pump in plant vacuolar membranes, inhibits the accumulation of anthocyanins in the vacuole. These results provide a biochemical function for BZ2, and suggest a common mechanism for the ability of plants to sequester structurally similar but functionally diverse molecules in the vacuole.

Amino Acid Sequence↗

PubMed1997

The evolution of apical dominance in maize.

The domestication of crop plants has often involved an increase in apical dominance (the concentration of resources in the main stem of the plant and a corresponding suppression of axillary branches). A striking example of this phenomenon is seen in maize (Zea mays spp. mays), which exhibits a profound increase in apical dominance compared with its probable wild ancestor, teosinte (Zea mays ssp. parviglumis). Previous research has identified the teosinte branched1 (tb1) gene as a major contributor to this evolutionary change in maize. We have cloned tb1 by transposon tagging and show here that it encodes a protein with homology to the cycloidea gene of snapdragon. The pattern of tb1 expression and the morphology of tb1 mutant plants suggest that tb1 acts both to repress the growth of axillary organs and to enable the formation of female inflorescences. The maize allele of tb1 is expressed at twice the level of the teosinte allele, suggesting that gene regulatory changes underlie the evolutionary divergence of maize from teosinte.

Alleles↗

PubMed2004

The role of barren stalk1 in the architecture of maize.

The architecture of higher plants is established through the activity of lateral meristems--small groups of stem cells formed during vegetative and reproductive development. Lateral meristems generate branches and inflorescence structures, which define the overall form of a plant, and are largely responsible for the evolution of different plant architectures. Here, we report the isolation of the barren stalk1 gene, which encodes a non-canonical basic helix-loop-helix protein required for the initiation of all aerial lateral meristems in maize. barren stalk1 represents one of the earliest genes involved in the patterning of maize inflorescences, and, together with the teosinte branched1 gene, it regulates vegetative lateral meristem development. The architecture of maize has been a major target of selection for early agriculturalists and modern farmers, because it influences harvesting, breeding strategies and mechanization. By sampling nucleotide diversity in the barren stalk1 region, we show that two haplotypes entered the maize gene pool from its wild progenitor, teosinte, and that only one was incorporated throughout modern inbreds, suggesting that barren stalk1 was selected for agronomic purposes.

Amino Acid Sequence↗

PubMed1993

Purification and characterization of a phytase (myo-inositol-hexakisphosphate phosphohydrolase) accumulated in maize (Zea mays) seedlings during germination.

Phytase (myo-inositol-hexakisphosphate phosphohydrolase, EC 3.1.3.8) has been purified from 5-7-day-old maize (Zea mays) seedlings, using a four-step purification procedure. The native protein has a molecular mass of about 76 kDa and is built up from two 38 kDa subunits. The pH and temperature optima of the purified enzyme were respectively 4.8 and 55 degrees C. The apparent Km for phytate was estimated to be 117 microM. Like other acidic phytases, the maize seedling enzyme exhibited a broad affinity for various phosphorylated substrates and especially for penta- and tri-phosphate esters of myo-inositol. The amino acid composition of the h.p.l.c.-purified protein indicated a high hydrophobicity (44% non-polar amino acids). Rabbit antibodies were produced in response to maize seedling phytase. Western-blot analyses clearly demonstrate that the increase of phytase activity observed during the first 7 days of germination corresponded to an accumulation of the protein in maize seedlings. Phytase accumulated essentially in the shoots (mesocotyl plus coleoptiles.

6-Phytase↗

PubMed2001

ZmES genes encode peptides with structural homology to defensins and are specifically expressed in the female gametophyte of maize.

All four members of a gene family, which are highly expressed in the cells of the female gametophyte (ZmES1--4: Zea mays embryo sac), were isolated from a cDNA library of maize egg cells. High expression of ZmES genes in the synergids around the micropylar region was detected in thin sections of maize ovaries. Single-cell RT--PCR analyses with the various cells of the female gametophyte confirmed the expression in synergids and also showed expression in the egg cell and central cell, and low expression in the antipodals. The expression of the whole gene family is suppressed after fertilization of the embryo sac, and expression in two-cell or later embryo stages or other tissues of maize could not be detected. In order to investigate ZmES mRNA gradients in the highly polarized and vacuolized cells of the maize embryo sac, a whole-mount in situ protocol with isolated single cells was developed: as for total RNA, ZmES transcripts are uniformly distributed in the cytoplasm of egg cell, synergids and central cell. ZmES genes encode small, cysteine-rich proteins with an N-terminal signal peptide, probably for translocation into the embryo sac cell wall. The four ZmES proteins display high sequence identity with each other, and the proposed tertiary structure of the mature peptides is similar to that of plant and animal defensins. The function of ZmES1-4 during the fertilization process is discussed.

Amino Acid Sequence↗

PubMed2001

Is maize B chromosome preferential fertilization controlled by a single gene?

In previous work, genotypes for high and low B chromosome transmission rate were selected from a native race of maize. It was demonstrated that the B transmission is genetically controlled. The present work reports the fourth and fifth generations of selection and the F1 hybrids between the lines. The native B is characterized by a constant behaviour, with normal meiosis and nondisjunction in 100% of postmeiotic mitosis. It is concluded that genetic variation for B transmission between the selected lines is due to the preferential fertilization process. The F1 hybrids show intermediate B transmission rate between the lines. They are uniform, the variance of the selected character being one order of magnitude lower than that of the native population. In addition, 0B x 2B and 2B x 2B crosses were made to study the effect of the presence of B chromosomes in the female parent, resulting in non-significant differences. Several crosses were made both in Buenos Aires and in Madrid to compare the possible environmental effect, but significant differences were not found. Our results are consistent with the hypothesis of a single major gene controlling B transmission rate in maize, which acts in the egg cell at the haploid level during fertilization. It is also hypothesized that maize Bs use the normal maize fertilization process to promote their own transmission.

Argentina↗

PubMed1998

Brown-midrib maize (bm1)--a mutation affecting the cinnamyl alcohol dehydrogenase gene.

Brown-midrib (bm) mutants of maize have modified lignin of reddish-brown colour. Although four independent bm loci are known, only one of the mutant genes has been previously identified. We report here that maize bm1, one of the less characterised mutants, shows severely reduced CAD activity in lignified tissues, resulting in the production of a modified lignin. Both the total lignin content and the structure of the polymer are altered by the mutation. We further describe the isolation and characterisation of the maize CAD cDNA and mapping of the CAD gene. CAD maps very closely to the known location of bm1 and co-segregates with the bm1 locus in two independent recombinant inbred populations. These data strongly support the premise that maize bm1 directly affects expression of the CAD gene.

Alcohol Oxidoreductases↗

PubMed1998

Suppression of endogenous alpha and beta tubulin synthesis in transgenic maize calli overexpressing alpha and beta tubulins.

Maize Black Mexican Sweetcorn cells have been transformed with constructs containing alpha and beta tubulin coding sequences either singly or together. It is shown that recovery of stable maize transformants is dependent on the co-expression of transfected alpha and beta tubulin in the same lines, indicating that plant cells cannot tolerate an imbalance in the ratio of alpha tubulin to beta tubulin within the cytoplasm. The co-expression of transfected alpha and beta tubulin in maize cells results in an increase in the overall tubulin content (approximately threefold). The transfected alpha and beta tubulins are incorporated into cortical, spindle and phragmoplast microtubule arrays indicating that they are functional. Furthermore, the co-expression of the transfected alpha and beta tubulins results in the suppression of endogenous alpha and beta tubulin synthesis. This suppression increases both with the strength of the promoter in the constructs and with the number of copies of the transgenes inserted into the maize genome. The implications for the post-transcriptional and post-translational regulation of tubulin synthesis in plant cells are discussed.

Acetyltransferases↗

PubMed2000

Immunological analysis of the phosphorylation state of maize C4-form phosphoenolpyruvate carboxylase with specific antibodies raised against a synthetic phosphorylated peptide.

The phosphoenolpyruvate carboxylase (PEPC) isozyme involved in C4 photosynthesis is known to undergo reversible regulatory phosphorylation under illuminated conditions, thereby decreasing the enzyme's sensitivity to its feedback inhibitor, L-malate. For the direct assay of this phosphorylation in intact maize leaves, phosphorylation state-specific antibodies to the C4-form PEPC were prepared. The antibodies were raised in rabbits against a synthetic phosphorylated 15-mer peptide with a sequence corresponding to that flanking the specific site of regulatory phosphorylation (Ser15) and subsequently purified by affinity-chromatography. Specificity of the resulting antibodies to the C4-form PEPC phosphorylated at Ser15 was established on the basis of several criteria. The antibodies did not react with the recombinant root-form of maize PEPC phosphorylated in vitro. By the use of these antibodies, the changes in PEPC phosphorylation state were semi-quantitatively monitored under several physiological conditions. When the changes in PEPC phosphorylation were monitored during the entire day with mature (13-week-old) maize plants grown in the field, phosphorylation started before dawn, reached a maximum by mid-morning, and then decreased before sunset. At midnight dephosphorylation was almost complete. The results suggest that the regulatory phosphorylation of C4-form PEPC in mature maize plants is controlled not only by a light signal but also by some other metabolic signal(s). Under nitrogen-limited conditions the phosphorylation was enhanced even though the level of PEPC protein was decreased. Thus there seems to be some compensatory regulatory mechanism for the phosphorylation.

Amino Acid Sequence↗