Shoot and root growth of hydroponic maize (Zea mays L.) as influenced by K deficiency

Potassium (K) has major biophysical and biochemical functions in plant physiology. However, plant responses to K deficiency at the whole plant level are not always clearly related to these well-known functions of K at the cellular level. The objective of this study was to investigate the morphological response of maize to increasing K deficiency and test to what extent this morphological response can be interpreted in the light of the simple model proposed by Leigh and Wyn Jones, suggesting that biophysical functions are affected first. Maize was grown in a greenhouse under hydroponic conditions. For half of the plants, K was removed from the nutrient solution from the 4th visible leaf stage. The K content in the starved plants dropped from 100 to 30 mM, and was not fully compensated by an increase in other cations. Leaf elongation rates were reduced on K-deprived plants, whereas axile root elongation rates were slightly increased between 45°C days and 75°C days after starvation, and reduced thereafter. During the first part of the starvation period, i.e. under moderate K deficiency (K concentration above 40 mM), all measured variables suggest that the whole plant response may be interpreted as the consequence of the reduced leaf growth, probably due to insufficient turgor pressure or cell-wall extensibility. This general pattern of response is in agreement with the model of Leigh and Wyn Jones. However, during the second part of the starvation period, i.e. under more severe K deficiency (K concentration below 40 mM), malfunction of additional physiological processes (mostly related to biochemical functions like photosynthetic processes) must be considered to explain the plant morphological response.     

Relationship between gene expression and hybrid vigor in primary root tips of young maize (Zea mays L.) plantlets

Summary To provide an insight into the molecular basis of heterosis, we investigated gene expression in primary root tips of a heterotic maize hybrid (B73 × Mo17) and its parental lines (B73 and Mo17). This analysis was carried out (i) by differential plaque hybridization of a recombinant cDNA library made to poly(A) RNA isolated from B73 × Mo17 primary root tips, and (ii) by comparing with two-dimensional gel electrophoresis proteins synthesized in vitro in the rabbit reticulocyte system by poly(A) RNA isolated, at different stages of development, from the three genotypes. The results showed that there are sets of proteins and mRNAs that are differentially synthesized and expressed in the F₁ primary root tips in comparison to the parental lines. Moreover, results from the survey of 21 major in-vitrosynthesized polypeptide variants, from mRNAs of primary root tips of the parental lines and their F₁ hybrid, indicated that in seven instances hybrid proteins translated in vitro were more abundant or possibly new. In most of the remaining cases, hybrid spots were similar in intensity to the same protein produced by one of the two parental lines.     

Wide hybridization: pollination of Zea mays L. by Sorghum bicolor (L.) Moench

Summary Foreign pollen tubes in the stigma of Zea mays can be prevented from reaching the ovary cavity by the unusual length of the pollen tube pathway. A simple and rapid procedure is described for overcoming this difficulty by pollinating the basal parts of the stigmas without removing the ensheathing bracts (husks). The method maintains high humidity in the vicinity of the ovaries, and by conserving photosynthetic tissues probably also ensures a more normal O₂ /CO₂ balance in the neighbourhood of the stigmas than do bagging procedures. It is shown that Sorghum pollen tubes readily reach the ovary after pollination by the method. Their presence induces some of the characteristic post-pollination effects caused by Zea pollen tubes, but they frequently also stimulate premature enlargement of the nucellus and lysis of nucellar cells. Although Sorghum tubes have been traced across the inner ovary wall, they have not been seen to enter the micropyle, and hybrid embryos have not yet been obtained.     

Studies on nitrate uptake by solution grown corn (Zea mays) L. genotypes

Nitrate depletion from the media by intact corn seedlings grown under solution culture was measured over a period of time. Two methods were assessed to differentiate corn genotypes for nitrate uptake ability: (1) Nitrate uptake per day per plant basis. (2) Nitrate uptake per day per gram root dry matter basis. The former method was found to be superior as it gave significant and positive correlations with dry matter, nitrate, and reduced nitrogen accumulation in stem, leaf and roots. Nitrate uptake was found to vary with plant age. Root mass and efficiency of roots appear to contribute to the total nitrate uptake ability of the genotype. Corn genotypes studied exhibited marked differences in nitrate uptake ability.     

Purification and characterization of a novel antimicrobial peptide from maize (Zea mays L.) kernels.

Several small, acid-soluble, basic peptides with anti-microbial properties have been isolated from maize (inbred B73) kernels. One of these peptides (MBP-1) has been purified to homogeneity and characterized. The peptide has a molecular weight of 4127.08 as determined by plasma desorption mass spectroscopy, has no free cysteines, and is predominantly alpha-helical as determined by circular dichroism. The primary sequence of the peptide (33 residues) has been determined by Edman degradation and shows no homology to the thionins, a group of cysteine-rich peptides found in some cereals including wheat, barley, and sorghum, as well as several dicot species. Like the thionins, however, MBP-1 has been found to have antimicrobial properties in vitro. MBP-1 inhibits spore germination or hyphal elongation of several plant pathogenic fungi, including two seed pathogens of maize (Fusarium moniliforme Sheld. and Fusarium graminearum (Gibberella zeae (Schw.) Petsch)), and several bacteria, including a bacterial pathogen of maize (Clavibacter michiganense ssp. nebraskense). A synthetic MBP-1 peptide, air-oxidized and purified by reverse phase chromatography, was equally antifungal as compared with the naturally occurring peptide.     

Developmental disaster1: A novel mutation causing defects during vegetative and inflorescence development in maize (Zea mays, Poaceae)

Axillary meristems, which give rise to branches and flowers, play a critical role in plant architecture and reproduction. To understand how axillary meristems initiate, we have screened for mutants with defects in axillary meristem initiation to uncover the genes controlling this process. These mutants, called the barren class of mutants in maize (Zea mays), have defects in axillary meristem initiation during both vegetative and reproductive development. Here, we identify and characterize a new member of the barren class of mutants named Developmental disaster1 (Dvd1), due to the pleiotropic effects of the mutation. Similar to the barren mutants, Dvd1 mutants have fewer branches, spikelets, florets, and floral organs in the inflorescence due to defects in the initiation of axillary meristems. Furthermore, double mutant analysis with teosinte branched1 shows that dvd1 also functions in axillary meristems during vegetative development. However, unlike the barren mutants, Dvd1 mutants are semidwarf due to the production of shorter internodes, and they produce leaves in the inflorescence due to the outgrowth of bract leaf primordia. The suite of defects seen in Dvd1 mutants, together with the genetic interaction of Dvd1 with barren inflorescence2, suggests that dvd1 is a novel regulator of axillary meristem and internode development.     

Identification and characterization of a novel adenine phosphoribosyltransferase gene (ZmAPT2) from maize (Zea mays L.).

Adenine phosphoribosyltransferase (APRT) is the key enzyme that converts adenine to adenosine monophosphate (AMP) in the purine salvage pathway. It was found that several different forms of APRT gene exist in plants, but no APRT gene in maize has been reported up to now. In this study, a novel maize APRT gene was cloned and characterized through a combination of bioinformatic, RT-PCR and RACE strategies. The full length of APRT cDNA sequence is 1202 nucleotides, with an ORF encoding 214 amino acid residues. Alignment of the deduced protein with that of other plant APRT genes indicates that the new gene is the form 2 of maize APRT, thus it was named ZmAPT2. Through basic local alignment search tool, search in the genomic survey sequence database of MaizeGDB, the putative genomic sequence of ZmAPT2 was obtained. Comparison of the cDNA and genomic sequence of the ZmAPT2 gene revealed that it contained seven exons and six introns. The locations of the introns within the maize ZmAPT2 coding region were consistent with those in the previously isolated APRTs of arabidopsis and rice. RT-PCR analysis showed that ZmAPRT was constitutively expressing in different organs under high temperature and salt stresses. Southern blot analysis indicated that at least three APRT genes existed in maize genome. These results confirmed that the novel maize ZmAPT2 gene was truly identified, and its potential role in maize growth and development was discussed.     

Comparison of K+-channel activation and deactivation in guard cells from a dicotyledon (Vicia faba L.) and a graminaceous monocotyledon (Zea mays)

We describe and compare inward and outward whole-cell K⁺ currents across the plasma membrane surrounding guard-cell protoplasts from the dicotyledon, Vicia faba, and the graminaceous monocotyledon, Zea mays. Macrosopic whole-cell current is considered in terms of microscopic single-channel activity, which involves discrete steps between conducting (open) and nonconducting (closed) states of the channel protein. Kinetic equations are used to model the number of open and closed states for channels conducting K⁺ influx (K(in)) and K⁺ efflux (K(out)) in the two species, and to calculate the rate at which open-closed transitions occur. The opening and closure of K(in) channels in both Vicia and Zea follow single-exponential timecourses, indicating that K(in)-channel proteins in each species simply fluctuate between one open and one closed state. In both species, opening of K(in) channels is voltage-independent, but closure of K(in) channels is faster at more positive membrane potentials. In response to identical voltage stimuli, K(in) channels in Zea open and close approximately three times as fast as in Vicia. In contrast to K(in), K(out) channels in Zea open and close more slowly than in Vicia. The closure of K(out) channels follows a single-exponential timecourse in each species, indicating one open state. The kinetics of K(out)-channel opening are more complicated and indicate the presence of at least two (Vicia) or three (Zea) closed states. The authors thank Professor N.A. Walker and Dr. D.R. Laver for the use of laboratory equipment, for helpful discussion and for provision of the program, GETHH. Thanks also to Dr. R.J. Ritchie for assistance with statistical analyses and to Ms. Janet Sherwood for maintenance of Vicia and Zea plants. This work was supported by grants from the National Science Foundation (DCB-89-04041) and the McKnight Foundation (S.M.A) and by a Charles Gilbert Heydon Travelling Fellowship (K.F-G).     

Analysis of nonadditive protein accumulation in young primary roots of a maize (Zea mays L.) F(1)-hybrid compared to its parental inbred lines

Heterosis describes the superior performance of heterozygous F(1)-hybrids compared to their homozygous parental inbred lines. Heterosis is already manifested during early maize (Zea mays L.) primary root development. In this study, the most abundant soluble proteins have been investigated before the phenotypic manifestation of heterosis in 3.5-day-old primary roots in the flint inbred line UH002, the dent inbred line UH301 and the corresponding hybrid UH301 x UH002. In CBB-stained 2-DE gels, 150 of 304 detected proteins (49%) were accumulated in a nonadditive fashion in the hybrid compared to the average of their parental inbred lines (Student's t-test: p < 0.05). Remarkably, expression of 51% (76/150) of the nonadditively accumulated proteins exceeded the high parent or was below the low parent. ESI-MS/MS identified 75 of the 76 proteins that belonged to these expression classes. The most abundant functional classes among the 75 proteins that were encoded by 60 different genes were metabolism (58%) and disease and defense (19%). Nonadditive protein accumulation in primary roots of maize hybrids might be associated with heterosis manifestation. Identification of these proteins could therefore contribute to the better understanding of the molecular basis of heterosis.     

Depolymerization of cortical microtubules is not a primary cause of chilling injury in corn (Zea mays L. cv Black Mexican Sweet) suspension culture cells

Cold-induced depolymerization of cortical microtubules were examined in suspension culture cells of corn (Zea mays L. cv Black Mexican Sweet) at various stages of chilling. In an attempt to determine whether microtubule depolymerization contributes to chilling injury, experiments were carried out with and without abscisic acid (ABA) pretreatment, since ABA reduces the severity of chilling injury in these cells. Microtubule depolymerization was detectable after 1 h at 4°C and became more extensive as the chilling was prolonged. There was little chilling injury after 1 d at 4°C in either ABA-treated or non-ABA-treated cells. After 3 d at 4°C, there was about 26% injury for ABA-treated and 40% injury for non-ABA-treated cells, as evaluated by 2,3,5-triphenyl-tetrazolium chloride reduction and by regrowth. After 1d at 4°C, less than 10% of cells retained full arrays of microtubules in both ABA-treated and non-ABA-treated cells, the remainder having either partial arrays or no microtubules. After 3d at 4°C, about 90% of cells showed complete or almost complete depolymerization of microtubules in both ABA-treated and non-ABA-treated cells. ABA did not stabilize the cortical microtubules against cold-induced depolymerization. In about 66% of ABA-treated cells and 57% of non-ABA-treated cells that had been held at 4°C for 3d, repolymerization of cortical microtubules occurred after 3h at 28°C. These results argue against the hypothesis that depolymerization of cortical microtubules is a primary cause of chilling injury.     

Early responses to gibberellic acid in a dwarf maize mutant (Zea mays L. d 1)

Early effects of gibberellic acid (GA₃) (1–4 h treatment) on the ion ratios in a dwarf maize mutant (Zea mays L. d ₁) showing normal growth after hormone treatment, have been investigated by electron microprobe analysis. GA₃ exerts a different effect on the ion ratios in plastids, cytoplasm and vacuoles in short term experiments. The Cl content of chloroplasts and cytoplasm increases without a lag phase after GA₃ treatment. The K content of plastids increases after a lag phase of 2 h, whereas in the cytoplasm an increase can be observed immediately after GA₃ addition. The hormone has only little influence on the Ca content of the cell compartments investigated. Control experiments with water and the physiologically inactive GA₃ methylester confirm the specifity of the short-term actions of GA₃ on the ion ratios. The primary action of GA₃ at the membrane level is discussed.     

A novel morphological response of maize (Zea mays) adult roots to heterogeneous nitrate supply revealed by a split‐root experiment

Approximately 35–55% of total nitrogen (N) in maize plants is taken up by the root at the reproductive stage. Little is known about how the root of an adult plant responds to heterogeneous nutrient supply. In this study, root morphological and physiological adaptations to nitrate‐rich and nitrate‐poor patches and corresponding gene expression of ZmNrt2.1 and ZmNrt2.2 of maize seedlings and adult plants were characterized. Local high nitrate (LoHN) supply increased both lateral root length (LRL) and density of the treated nodal roots of adult maize plants, but only increased LRL of the treated primary roots of seedlings. LoHN also increased plant total N acquisition but not N influx rate of the treated roots, when expressed as per unit of root length. Furthermore, LoHN markedly increased specific root length (m g^?1) of the treated roots but significantly inhibited the growth of the lateral roots outside of the nitrate‐rich patches, suggesting a systemic carbon saving strategy within a whole root system. Surprisingly, local low nitrate (LoLN) supply stimulated nodal root growth of adult plants although LoLN inhibited growth of primary roots of seedlings. LoLN inhibited the N influx rate of the treated roots and did not change plant total N content. The gene expression of ZmNrt2.1 and ZmNrt2.2 of the treated roots of seedlings and adult plants was inhibited by LoHN but enhanced by LoLN. In conclusion, maize adult roots responded to nitrate‐rich and nitrate‐poor patches by adaptive morphological alterations and displayed carbon saving strategies in response to heterogeneous nitrate supply.     

The effect of auxins (IAA and 4-Cl-IAA) on the redox activity and medium pH of Zea mays L. root segments

Indole-3-acetic acid (IAA) and 4-chloroindole-3-acetic acid (4-Cl-IAA) were tested at different concentrations and times for their capacity to change the redox activity and medium pH of maize root segments. The dose-response surfaces (dose-response curves as a function of time) plotted for redox activity and changes in medium pH (expressed as ΔpH) had a similar shape for both auxins, but differed significantly at the optimal concentrations. With 4-Cl-IAA, the maximal values of redox activity and medium pH changes were observed at 10⁻¹⁰ M, which was a 100-fold lower concentration than with IAA. Correlations were observed between redox activity and medium pH changes at the optimal concentrations of both IAA and 4-Cl-IAA. The results are discussed herein, taking into account both the concentration of the auxins and the effects produced by them.     

Cell-cell recognition of host surfaces by pathogens. The adsorption of maize (Zea mays) root mucilage by surfaces of pathogenic fungi.

The adsorption of radioactive mucilage by pathogenic fungi was shown to be dependent upon time, the composition of mucilage, the type of fungal surface (conidia, hyphae, hyphal apices), fungal species, pH and bivalent cations. All fungal adhesins were inactivated by either proteinase or polysaccharase treatments. Adsorption was not inhibited by the numberous mono-, di- and oligo-saccharides that were tested individually, but it was inhibited absolutely by several polysaccharides. This suggested that adsorption of mucilage by pathogens involved conformational and ionic interactions between plant and fungal polymers but not fungal lectins bound to sugar residues of mucilage. Several fractionation schemes showed that pathogens bound only the most acidic of the variety of polymers that comprise mucilage. There was not any absolute distinction between ability to bind radioactive mucilage and type of pathogen or non-pathogen. However, there were notable differences in characteristics of adsorption between two types of pathogen. Differences were revealed by comparison of the adsorption capacities of conidia and germinant conidia and chromatography of radioactive mucilage on germinant conidia. An ectotrophic root-infecting fungus (a highly specialized pathogen) bound a greater proportion of mucilage than did a vascular-wilt fungus (of catholic host and tissue range) with more than one class of site for adsorption. In contrast with the vascular-wilt fungus, sites for adsorption on the specialized pathogen were present solely on surfaces formed by germination.     

Mechanistic studies of perfluorooctane sulfonate, perfluorooctanoic acid uptake by maize (Zea mays L. cv. TY2)

Aims

There is a need to predict trace metal concentration in plant organs at given development stages. The aim of this work was to describe the Cd hyperaccumulation kinetics in the different plant organs, throughout the complete cultivation cycle, independently of a possible soil effect.

Methods

Plants of Noccaea caerulescens were exposed in aeroponics to three constantly low Cd concentrations and harvested at 6 to 11 dates, until siliquae formation.

Results

Dry matter allocation between roots and shoots was constant over time and exposure concentrations, as well as Cd allocation. However 86 % of the Cd taken up was allocated to the shoots. Senescent rosette leaves showed similar Cd concentrations to the living ones, suggesting no redistribution from old to young organs. The Cd root influx was proportional to the exposure concentration and constant over time, indicating that plant development had no effect on this. The bio-concentration factor (BCF), i.e. [Cd]_plant/[Cd²⁺]_solution for the whole plant, roots or shoots was independent of the exposure concentration and of the plant stage.

Conclusions

Cadmium uptake in a given plant part could therefore be predicted at any plant stage by multiplying the plant part dry matter by the corresponding BCF and the Cd²⁺ concentration in the exposure solution.