Development of Endopeptidase Activities in Maize (Zea mays L.) Endosperms

An activity stain was used after native polyacrylamide gel electrophoresis, and at least 17 different endopeptidase activities were detected in maize (Zea mays L.) endosperm extracts prepared during the first 6 d after imbibition. The enzymes detected were classified into four groups based on their time of appearance and on their mobility in polyacrylamide gels. The first group, which included two enzymes present in dry endosperms, disappeared soon after imbibition. The second group, comprising five activity bands, appeared during the first 2 to 3 d after imbibition and then disappeared. The third set of enzymes increased continuously throughout the experimental period. The fourth group appeared after d 3 and remained at a constant level after that time. The endopeptidase activities were characterized by the effect of specific inhibitors on their activities. The two enzymes of the first group are metalloendopeptidases based on their sensitivity to ethylenediaminetetracetate (EDTA). Enzymes of the second, third, and fourth groups are sulfhydryl-endopeptidases as judged by their sensitivity to antipain, chymostatin, leupeptin, and E-64 and by their requirement for 2-mercaptoethanol. Pepstatin, phenylmethylsulfonyl fluoride, or EDTA had no effect on these enzymes. Many of the second, third, and fourth group enzymes cleaved [alpha]-zein-rich proteins as well as such easily obtained proteins as gelatin (used in our standard assay) and hemoglobin. The second group had a high affinity for [gamma]-zein, whereas none of the bands in the fourth group of enzymes cleaved this type of zein. The two metalloenzymes of the first group cleaved neither [alpha]- nor [gamma]-zeins.     

Biochemical Characteristics of Membrane Fractions Isolated from Maize (Zea mays L.) Roots

Membrane fractions have been isolated from maize (Zea mays L.)roots by discontinuous density gradient centrifugation and phaseseparation methods. A number of approaches were tried with theaim of identifying specific membrane types, especially the plasmamembrane. These included the use of enzymic markers, determinationof glucose and leucine incorporation, separation of membraneproteins by SDS-PAGE, and attempts to identify the plasma membranefraction by cell surface labelling. The results are discussedin relation to the usefulness of membrane markers and the difficultiesof isolating surface membranes from higher plant tissues.     

Low Water Potential Disrupts Carbohydrate Metabolism in Maize (Zea mays L.) Ovaries

Water deficit during pollination increases the frequency of kernel abortion in maize (Zea mays L.). Much of the kernel loss is attributable to lack of current photosynthate, but a large number of kernels fail to develop on water-deficient plants even when assimilate supply is increased. We examined the possibility that assimilate utilization by developing ovaries might be impaired at low water potential ([Psi]w). Plants were grown in the greenhouse in 20-L pots containing 22 kg of amended soil. Water was withheld on the first day silks emerged, and plants were hand-pollinated 4 d later when leaf [Psi]w decreased to approximately - 1.8 MPa and silk [Psi]w was approximately -1.0 MPa. Plants were rehydrated 2 d after pollination. The brief water deficit inhibited ovary growth (dry matter accumulation) and decreased kernel number per ear by 60%, compared to controls. Inhibition of ovary growth was associated with a decrease in the level of reducing sugars, depletion of starch, a 75-fold increase in sucrose concentration (dry weight basis), and inhibition of acid invertase (EC 3.2.1.26) activity. These results indicate that water deficits during pollination disrupt carbohydrate metabolism in maize ovaries. They suggest that acid invertase activity is important for establishing and maintaining reproductive sink strength during pollination and early kernel development.     

The Growth and Development of Maize (Zea mays L.) at Five Temperatures

The objectives of this work were to measure growth and developmentrates over a range of temperatures and to identify processeswhich may limit vegetative yield of maize (Zea mays L.). Twosingle cross Corn Belt Dent maize hybrids were grown from sowingin a diurnal temperature regime of 16/6 °C day/night andin constant temperature environments of 16, 20, 24 and 28 °C.The 16/6 °C environment was close to the minimum for sustainedgrowth and 28 °C was near the optimum. Entire plants wereharvested at stages with 4, 6, 7 and 8 mature leaves in alltemperature treatments except 20 °C in which the final twoharvests were carried out at 9 and 10 mature leaves. Mean totalleaf number varied between 19.5 and 16.0 with the maximum occurringat 16/6 °C. Although harvests were carried out at comparableleaf numbers, and hence at similar developmental stages, thetime interval between sowing and harvest decreased considerablyas temperatures increased. The relative rates of dry weight and leaf area accumulationwith time increased with a Q₁₀ of 2.4 between 16 and 28 °C,while leaf appearance rate increased with a Q₁₀ of 2.9 overthe same range; both rates were highest at 28 °C. Althoughdry matter partitioning to the shoots increased with temperature,the area of individual leaves varied in a systematic patternwhich resulted in maximum leaf area, leaf area duration andconsequently dry weight being realized at 20 °C for anygiven stage of development. Zea mays, corn, low temperature stress, temperature response, growth, development     

Differentiation of the Properties of the Branching Isozymes from Maize (Zea mays)

The multiple forms of branching enzyme (BE) from developing maize (Zea mays) endosperm were purified by modification of previous procedures such that amylase activity could be eliminated completely from the BE preparation. Three distinct assays for BE activity (phosphorylase a stimulation assay, BE linkage assay, and iodine stain assay) were used to characterize and differentiate the properties of the BE isoforms. This study presents the first evidence that the BE isoforms differ in their action on amylopectin. BEI had the highest activity in branching amylose, but its rate of branching amylopectin was less than 5% of that of branching amylose. Conversely, BEII isoforms had lower rates in branching amylose (about 9-12% of that of BEI) and had higher rates of branching amylopectin (about 6-fold) than BEI. The implication of these findings to the mechanism of amylopectin synthesis in vivo are discussed.     

Fertility of Deep-frozen Maize (Zea mays L.) Pollen

On average, 50 per cent of maize pollen grains can be kept viableand almost 30 per cent remain fertile for up to a year whenthe water content on a fresh weight basis is reduced to about30 per cent of the original and the pollen is stored at –76or –196 °C. Over this period no significant differencewas found between storage at either temperature. Zea mays L., maize, pollen, fertility, viability     

Effect of the Osmotic Environment on Assimilate Transport in Isolated Developing Embryos of Maize (Zea mays)

Embryos obtained from developing kernels of maize (Zea maysL.) were incubated in a bathing medium, to measure the effectof the osmotic environment on the balance between uptake andrelease of assimilates by the embryo. Net efflux of sucroseand amino acids from the embryo decreased with increasing mannitolconcentration in the bathing medium and net uptake of [¹⁴C]valine increased with increasing mannitol concentration. Therole of a high osmolality of the seed apoplast in seed developmentis discussed Zea mays, maize, embryo, seed development, assimilate transport, turgor-sensitive transport     

Endopolyploidy in Diploid and Tetraploid Maize (Zea mays L.)

Nuclei from different tissues such as stem, mesocotyl, nodalroot and root tip of diploid and tetraploid maize were isolated,stained with propidium iodide and passed through an EPICS-751flow-cytometer cell sorter. Variations in flow histograms wereobserved in different tissues. Stem tissues of both the diploidand tetraploid had two peaks representing G1 and G2 somaticnuclei. The remaining tissues in both the diploids and tetraploidsexhibited three peaks. The first peak observed in these tissuesrepresents G1 somatic nuclei of the lowest ploidy level. Thesecond peak represent G2 somatic nuclei of the lowest ploidylevel+G1 somatic nuclei of the next ploidy level. The thirdpeak represents G2 of the higher ploidy level+G1 somatic nucleiof the next higher ploidy level. Statistically significant differenceswere observed between the diploid and tetraploid maize tissueswith respect to nuclei distribution in the higher ploidy levelpeaks implying variation in the degree of endopolyploidy inthe diploid and tetraploid maize. The results of this studysuggest that the amount of endopolyploid observed in maize genotypeshas an effect on their overall agronomic performance under thefield conditions.Copyright 1993, 1999 Academic Press Zea mays L., maize, endopolyploidy, diploid, tetraploid, flow cytometry     

Solubilization of a Proline Dehydrogenase from Maize (Zea mays L.) Mitochondria

L-Proline is oxidized to pyrroline-5-carboxylic acid in intact plant mitochondria by a proline dehydrogenase (EC 1.4.3) that is bound to the matrix side of the inner mitochondrial membrane (TE Elthon, CR Stewart [1981] Plant Physiol 67: 780-784). This investigation reports the first solubilization of the L-proline dehydrogenase (PDH) from plant mitochondria. The supernatant from NP-40-treated etiolated shoot mitochondria of maize, Zea mays L., reduced iodonitrotetrazolium violet in a proline dependent manner. The pH optimum for this activity was 8. The apparent K_m for proline was 6.6 millimolar. When supplied with proline, this solubilized PDH activity also synthesized pyrroline-5-carboxylic acid. The PDH activity was inhibited in vitro by 300 millimolar potassium chloride but not by 300 millimolar potassium acetate. The PDH activity had a molecular mass that was greater than 150 kilodaltons. Mitochondria were prepared from etiolated shoots grown in 100% water-saturated vermiculite (control) and 16% water-saturated vermiculite (stress). The specific activity of solubilized PDH from the stress treatment was 11% of the same activity from the control treatment. Oxygen uptake in the presence of proline and ADP (state 3 proline oxidation) by mitochondria from the stress treatment was 25% of the same rate by mitochondria from the control treatment. Mitochondria were also prepared 16 hours after rewatering the seedlings growing in the stress treatment. Both the solubilized PDH specific activity and state 3 proline oxidation returned to the control levels. The specific activities of the NAD⁺-dependent pyrroline-5-carboxylic acid dehydrogenase and cytochrome c oxidase in the solubilized preparations were unaffected by these stress and recovery treatments. Oxygen uptake rates by intact mitochondria in the presence of ADP and NADH, succinate or malate-pyruvate were also unaffected by these treatments.     

Effects of Moniliformin on Mitosis in Maize (Zea mays L.)

Maize (Zea mays L. ‘Norfolk White’) roots were treatedwith solutions of moniliformin (a metabolite of Fusarium moniliformeSheldon) at 0.0001 M and 0.001 M for 8, 24, and 48 h. Only aslight inhibition of division was noted in root tips treatedwith the lower concentration. The higher concentration causeda disruption of the spindle apparatus and consequent C-mitosis,and metaphase accumulation. (Received February 14, 1984; Accepted May 18, 1984)     

Abscisic Acid Biosynthesis in Isolated Embryos of Zea mays L

Previous labeling experiments with ¹⁸O₂ have supported the hypothesis that stress-induced abscisic acid (ABA) is synthesized through an indirect pathway involving an oxygenated carotenoid (xanthophyll) as a precursor. To investigate ABA formation under nonstress conditions, an ¹⁸O₂ labeling experiment was conducted with isolated embryos from in vitro grown maize (Zea mays L.) kernels. Of the ABA produced during the incubation in ¹⁸O₂, three-fourths contained a single ¹⁸O atom located in the carboxyl group. Approximately one-fourth of the ABA synthesized during the experiment contained two ¹⁸O atoms. These results suggest that ABA synthesized in maize embryos under nonstress conditions also proceeds via the indirect pathway, requiring a xanthophyll precursor. It was also found that the newly synthesized ABA was preferentially released into the surrounding medium.     

Acetolactate Synthase Activity in Developing Maize (Zea mays L.) Kernels

Acetolactate synthase (EC 4.1.3.18) activity was examined in maize (Zea mays L.) endosperm and embryos as a function of kernel development. When assayed using unpurified homogenates, embryo acetolactate synthase activity appeared less sensitive to inhibition by leucine + valine and by the imidazolinone herbicide imazapyr than endosperm acetolactate synthase activity. Evidence is presented to show that pyruvate decarboxylase contributes to apparent acetolactate synthase activity in crude embryo extracts and a modification of the acetolactate synthase assay is proposed to correct for the presence of pyruvate decarboxylase in unpurified plant homogenates. Endosperm acetolactate synthase activity increased rapidly during early kernel development, reaching a maximum of 3 micromoles acetoin per hour per endosperm at 25 days after pollination. In contrast, embryo activity was low in young kernels and steadily increased throughout development to a maximum activity of 0.24 micromole per hour per embryo by 45 days after pollination. The sensitivity of both endosperm and embryo acetolactate synthase activities to feedback inhibition by leucine + valine did not change during kernel development. The results are compared to those found for other enzymes of nitrogen metabolism and discussed with respect to the potential roles of the embryo and endosperm in providing amino acids for storage protein synthesis.     

Alterations of Free Amide and Amino Acid Contents During the Development of Maize Plants, Zea mays L.

Concentrations of free amino acids and amides were measuredin organs of maize plants, Zea mays L. in the period from 14d before pollen liberation until complete seed maturation. Inthis time anthesis took place and only the ovaries of ear 9(numbered from below) developed into seeds. In mature leaf bladesNH₄ ion assimilation had ceased and asparagine and glutaminewere not found there. N redistribution induced the occurrenceof large amounts of aspartate, glutamate and alanine. The amountsof amides in leaf sheaths and stem parts depended on the neighbourhoodof generative parts. The generative plant parts can be distinguishedfrom adjacent vegetative plant parts in concentrations of freeproline or asparagine. Proline occurred in pollen but not inthe empty anthers. Ears had a small, early peak amount of prolinemostly before pollination. Only the ninth ear had a first maximumproline amount after the fifth day of pollen liberation. A secondproline peak in the ears coincided with the period of maximumincrease in d. wt. The occurrence of proline in the generativeorgans relative to metabolic processes inducing fertility orseed maturation is discussed. Zea mays L., amides, amino acids, amino-transferring components, asparagine, glutamine, proline     

Purification and Partial Characterization of Maize (Zea mays L.) beta-Glucosidase

Maize (Zea mays L.) β-glucosidase (β-d-glucoside glucohydrolase, EC 3.2.1.21) was extracted from the coleoptiles of 5- to 6-day-old maize seedlings with 50 millimolar sodium acetate, pH 5.0. The pH of the extract was adjusted to 4.6, and most of the contaminating proteins were cryoprecipitated at 0°C for 24 hours. The pH 4.6 supernatant from cryoprecipitation was further fractionated by chromatography on an Accell CM column using a 4.8 to 6.8 pH gradient of 50 millimolar sodium acetate, which yielded the enzyme in two homogeneous, chromatographically different fractions. Purified enzyme was characterized with respect to subunit molecular weight, isoelectric point, amino acid composition, NH₂-terminal amino acid sequence, pH and temperature optima, thermostability, and activity and stability in the presence of selected reducing agents, metal ions, and alkylating agents. The purified enzyme has an estimated subunit molecular mass of 60 kilodaltons, isoelectric point at pH 5.2, and pH and temperature optima at 5.8 and 50°C, respectively. The amino acid composition data indicate that the enzyme is rich in Glx and Asx, the sum of which approaches 25%. The sequence of the first 20 amino acids in the N-terminal region was H₂N-Ser-Ala-Arg-Val-Gly-Ser-Gln-Asn-Gly-Val-Gln-Met-Leu-Ser-Pro-(Ser?) -Glu-Ile-Pro-Gln, and it shows no significant similarity to other proteins with known sequence. The enzyme is extremely stable at 0 to 4°C up to 1 year but loses activity completely at and above 55°C in 10 minutes. Likewise, the enzyme is stable in the presence of or after treatment with 500 millimolar 2-mercaptoethanol, and it is totally inactivated at 2000 millimolar 2-mercaptoethanol. Such metal ions as Hg²⁺ and Ag⁺ reversibly inhibit the enzyme at micromolar concentrations, and inhibition could be completely overcome by adding 2-mercaptoethanol at molar excess of the inhibitory metal ion. The alkylating agents iodoacetic acid and iodoacetamide irreversibly inactivate the enzyme and such inactivation is accelerated in the presence of urea.     

不同基因型玉米间作的群体质量

采用大田试验,研究了不同基因型玉米间作的群体质量特征.结果表明,HF9‖XD20间作,植株中部叶片平均叶龄延长,而对下部和上部叶片影响不大;ZD958‖LD981间作,ZD958植株下部叶片平均叶龄延长,而中、上部叶片则缩短,LD981植株下、中、上部叶片平均叶龄均有所延长.吐丝前,群体叶面积指数(LAI)单间作无明显差异,吐丝后,HF9和LD981的LAI分别大于和显著大于单作群体,而ZD958和XD20则分别小于和显著小于单作群体.紧凑型品种和半紧凑型品种间作增加了群体透光率,吐丝后10d,4个品种棒三叶叶色值(SPADR)均有所增加,并且除ZD958外,其余3个品种棒三叶净光合速率均有所增加,其中LD981增加显著.间作对吐丝以前的群体干物质积累量影响不大,吐丝后,半紧凑型品种(HF9和LD981)的干物质积累量增加,其中LD981增加显著,而紧凑型品种(XD20和ZD958)的干物质积累量减少,其中ZD958显著减少;间作还提高了收获指数,并且两种间作群体的土地当量比(LER)均大于1.结果提示,紧凑型与半紧凑型玉米品种的间作可以提高群体质量,延长叶片功能期,提高光合效率,增加籽粒产量.     

Dynamics of the Elongation of Internodes in Maize (Zea mays L.): Analysis of Phases of Elongation and their Relationships to Phytomer Development

The kinetics of elongation of individual internodes of maizestems were studied under field conditions. Thermal time coursesof internode length were recorded using non-destructive methods,based on direct measurement of X-ray photographs or on indirectestimates using heights of leaf collars. These data were complementedby serial dissections of maize stems, and by precise observationof the process of sheath emergence, to specify its role in thekinetics of internode elongation. The kinetics of elongationwere found to be composed of four phases. The rate of elongationrises exponentially during phase I, and then increases sharplyduring a short period (phase II), which is followed by a majorperiod of constant growth rate (phase III) and a shorter periodin which the rate declines (phase IV). During phase I, elongationappears to be integrated at the level of the whole apical cone.From phase II onwards, elongation becomes determined at thelevel of the phytomer. The emergence of the sheath attachedto the internode appears to be a possible trigger for the transitionbetween phase I and phase II, and it may also be involved invariation in final length among phytomers. Copyright 2000 Annalsof Botany Company Zea mays L., internode, elongation, modelling, dynamics, X-rays, collar, phytomer, stem, thermal time, phasic development     

Further Studies on Mosaic Disease of Maize (Zea mays L.)

Schizaphis graminum (Rondani) is proved to be an additional vector of maize mosaic virus (MMV). The pH range for the infectivity of the virus in extracted juice is found to be from 4.4 to 9.0, the optimum being 5.6 to 7.2. Effect of certain chemicals on the virusin vitro has also been studied. Cross protection between MMV and Sugar-cane mosaic virus (SMV) indicated positive results. It has been concluded on the basis of similar physical properties, tolerance towards certain chemicals, host range, symptomatology, aphid vectors and positive immunological tests, that MMV and SMV are related viruses.     

Bacterial Streak Disease of Maize (Zea mays L.) in South Africa

Bacterial streak disease of maize is currently causing some concern among breeders in South Africa. The causal organism of this previously undescribed disease was successfully isolated and its pathogenicity established using KoCH's postulates. Standard physiological and biochemical tests used to identify phytopathogenic bacteria indicated that the bacterium is a Xanthomonas campestris pathovar. Comparisons between this organism and other recognized X. campestris pathovars of the Poaceae indicated that apart from some minor differences the maize streak pathogen is physiologically similar to X. campestris pv. holcicola. However, in repeated reciprocal inoculation experiments all attempts to induce disease symptoms in sorghum with the maize streak pathogen were unsuccessful. Conversely, X. campestris pv. holcicola did produce symptoms in maize leaves. In all the maize cultivars tested the symptoms produced by the maize streak pathogen were, however, always considerably more severe than those caused by X. campestris pv. holcicola. Notwithstanding its physiological similarity to X. campestris pv. holicola it would appear that on the grounds of host specificity the maize streak pathogen warrants new pathovar status. The name X. campestris pv. zeae is proposed.     

玉米雌穗发育期基因差异表达与杂种优势的研究

杂种优势在提高粮食作物特别是玉米的产量方面具有重要的作用。然而,杂种优势的原理却仍然是一个世界性的难题。用12个玉米自交系及其按不完全双列杂交组配的33个杂交种为材料,分4个不同发育时期取杂交种及其亲本的雌穗组织,利用差异显示技术,分析杂种与亲本的基因差异表达类型及其与7个主要农艺性状的杂种表现和杂种优势的相关关系。发现1):在5种表达类型中单态表达(基因在杂交种和双亲中同时表达的类型)的数量最大,这说明杂种优势的形成不仅与基因的表达与否相关,还与大量基因的上调或下调表达相关;2):在玉米雌幼穗的发育初期杂交种与双亲的基因表达差异最大,这可能与雌穗发育初期器官的形成和发育相关,因此这一时期差异表达(在质的方面)的基因对产量性状和杂种优势的形成具有密切关系;3):综合各种基因表达类型与产量性状和杂种优势的关系,发现某些基因在杂种中的沉默表达可以促进籽粒的发育和抑制幼穗中小花的发育。