Showing papers on "Endosperm published in 1990"

A water relations analysis of seed germination rates.

Abstract: Seed germination culminates in the initiation of embryo growth and the resumption of water uptake after imbibition. Previous applications of cell growth models to describe seed germination have focused on the inhibition of radicle growth rates at reduced water potential (Psi). An alternative approach is presented, based upon the timing of radicle emergence, to characterize the relationship of seed germination rates to Psi. Using only three parameters, a ;hydrotime constant' and the mean and standard deviation in minimum or base Psi among seeds in the population, germination time courses can be predicted at any Psi, or normalized to a common time scale equal to that of seeds germinating in water. The rate of germination of lettuce (Lactuca sativa L. cv Empire) seeds, either intact or with the endosperm envelope cut, increased linearly with embryo turgor. The endosperm presented little physical resistance to radicle growth at the time of radicle emergence, but its presence markedly delayed germination. The length of the lag period after imbibition before radicle emergence is related to the time required for weakening of the endosperm, and not to the generation of additional turgor in the embryo. The rate of endosperm weakening is sensitive to Psi or turgor.

Identification and molecular characterization of shrunken-2 cDNA clones of maize.

Abstract: Mutation at the shrunken-2 (Sh2) locus of maize, a gene described more than 40 years ago, greatly reduces starch levels in the endosperm through its effect on the starch synthetic enzyme ADP-glucose pyrophosphorylase, an enzyme thought to be regulatory in this biosynthetic pathway. Although our previous work has suggested that Sh2 is a structural gene for this enzyme, we have also reported data compatible with Sh2 acting post-transcriptionally. In this study, we took advantage of a transposable element-induced Sh2 allele, its progenitor, and revertants to identify a clone for this locus. Although the cloning and identification were done independently of any knowledge concerning the product of this gene, examination of the deduced amino acid sequence revealed much similarity to known ADP-glucose pyrophosphorylase subunits of plants and bacteria, including regions involved in substrate binding and activator binding. Little sequence similarity, however, was found at the DNA level. These observations provide direct evidence that Sh2 encodes a subunit for endosperm ADP-glucose pyrophosphorylase. Analysis of several phenotypically wild-type alleles arising from a mutable sh2-Ds allele revealed one unexpected case in which DNA sequences of Sh2 were rearranged in comparison with the progenitor Sh2. In contrast to wild type, the Ds-induced sh2 allele conditions at least two transcripts in the endosperm.

Primary structure and inhibition of protein synthesis in eukaryotic cell-free system of a novel thionin, γ-hordothionin, from barley endosperm.

Abstract: A new sulfur-rich and basic polypeptide, designated as gamma-hordothionin, has been isolated from barley endosperm by a semi-preparative purification consisting of extraction with a volatile salt solution followed by high-performance liquid chromatography using a reversed-phase C4 column. The isolated polypeptide was found to be homogeneous by micro-two-dimensional gel electrophoresis in the presence of sodium dodecyl sulfate. The complete primary structure of gamma-hordothionin was determined by automatic degradation of the intact, S-carboxymethylated and S-pyridylethylated gamma-hordothionin and fragments obtained by proteolytic cleavage. gamma-Hordothionin consists of a single polypeptide chain of 47 amino acids with a calculated molecular mass of 5250 Da and contains four disulfide bridges. gamma-Hordothionin inhibits translation in cell-free systems derived from mammalian (rabbit reticulocyte, mouse liver) as well as non-mammalian (Artemia embryo) cells, at several levels. At low concentrations (1-10 microM) the protein seems to affect mainly the polypeptide-chain-initiation process, although it might also act at the elongation level. At higher concentrations (20-80 microM) this inhibitor induces activation of an eukaryotic polypeptide-chain initiation factor 2 alpha-subunit (eIF-2 alpha) kinase in hemin-supplemented reticulocyte lysates, as does hemin deficiency. The presence of the disulfide bridges in gamma-hordothionin appears to be essential for the eIF-2 alpha kinase activation. Based on its similarity at both the structural and functional level with the different genetic variants of thionins (alpha and beta-thionins, from wheat and barley), gamma-hordothionin is a putative member of the thionin family.

Scavenging of Hydrogen Peroxide in the Endosperm of Ricinus communis by Ascorbate Peroxidase

Abstract: In this report we provide evidence, that in the endosperm of Ricinus also the H 2 O 2 scavenging pathway involving ascorbate and glutathione plays a significant role in H 2 O 2 degradation

Size-distribution of wheat starch granules during endosperm development.

Abstract: Starch granule development in hard red winter wheat was followed by light and electron microscopy and quantitative image analysis. Proplastids in the cytoplasm of the coenocytic endosperm two days after flowering (DAF) differentiated into amyloplasts by 4 DAF. Quantitative image analysis showed that the starch granules initiated during the first 4 DAF increased in size to mean diameter of 5.6 μm by 7 DAF (...)

Expression of CaMV35S-GUS gene in transgenic rice plants

Abstract: To understand the properties of the cauliflower mosaic virus (CaMV) 35S promoter in a monocotyledonous plant, rice (Oryza sativa L.), a transgenic plant and its progeny expressing the CaMV35S-GUS gene were examined by histochemical and fluorometric assays. The histochemical study showed that β-glucuronidase (GUS) activity was primarily localized at or around the vascular tissue in leaf, root and flower organs. The activity was also detected in the embryo and endosperm of dormant and germinating seeds. The fluorometric assay of various organs showed that GUS activity in transgenic rice plants was comparable to the reported GUS activity in transgenic tobacco plants expressing the CaMV35S-GUS gene. The results indicate that the level of expression of the CaMV 35S promoter in rice is similar to that in tobacco, a dicotyledonous plant, suggesting that it is useful for expression of a variety of foreign genes in rice plants.

Effects of Priming and Endosperm Integrity on Seed Germination Rates of Tomato Genotypes II. GERMINATION AT REDUCED WATER POTENTIAL

Abstract: Seed germination rates (GR = inverse of time to germination) are sensitive to genetic, environmental, and physiological factors. We have compared the GR of tomato (Lycopersicon esculentum Mill.) seeds of cultivar T5 to those of rapidly germinating L. esculentum genotypes PI 341988 and PI 120256 over a range of water potential ( were also assessed. Germination time-courses at different 0's were analysed according to a model that identified a base, or minimum, \j> allowing germination of a specific percentage (g) of the seed population (ifib(g)), and a 'hydrotime constant' (9H) indicating the rate of progress toward germination per MPa.h. The distribution of if>b(g) determined by probit analysis was characterized by a mean base b) and the standard deviation in b among seeds ( . A normalized time-scale for comparing germination responses to reduced ij> is introduced. The time to germination at any tp (t g(tfi)) can be normalized to be equivalent to that observed in water (/g(0)) according to the equation fg(0) = (l -^(ip/Ms)))^^)- PI 341988 seeds were more tolerant of reduced > b(g), but did not lower the minimum b between intact and cut seeds incubated at high 0 was much less («0-l MPa), indicating that at the time of radicle protrusion, the endosperm had weakened to the point where it constituted only a small mechanical barrier. In the intact seed, endosperm weakening and the downward adjustment in embryo ifib ceased at

Identification of an Enhancer Element for the Endosperm-Specific Expression of High Molecular Weight Glutenin

Abstract: Genes encoding high molecular weight (HMW) glutenin, a wheat seed storage protein, are expressed only in the developing endosperm. It was previously demonstrated that sequences essential for endosperm-specific transcription reside within 436 base pairs upstream of the initiation codon for HMW glutenin translation. We have further analyzed this region by testing the ability of a series of truncated HMW glutenin promoter fragments to enhance transcription from an adjacent heterologous promoter. The activity of these hybrid promoters was determined by measuring the expression of a linked beta-glucuronidase (GUS) reporter gene in transgenic tobacco plants. An HMW glutenin promoter fragment spanning nucleotides -375 to -45 relative to the transcription start site was found to stimulate GUS expression in tobacco seeds when inserted in either orientation upstream of the heterologous promoter. Furthermore, this fragment could also potentiate transcription when located 3' to the GUS reporter gene. Stimulation of GUS gene expression in transgenic tobacco seeds did not occur until 9 days to 12 days after anthesis, coincident with the onset of storage protein synthesis in the developing tobacco and wheat seed, and was confined to the endosperm tissue. By testing progressively shorter promoter fragments, the enhancer element responsible for this pattern of expression was localized to a 40-base pair region some 170 base pairs upstream of the start site for HMW glutenin transcription.

Effects of Priming and Endosperm Integrity on Seed Germination Rates of Tomato Genotypes

Abstract: The bases of differences in germination rates (GRg = inverse of time to germination [/g] of percentage g) among three cold/salt tolerant tomato (Lycopersicon esculentum Mill.) accessions (PI 341988, PI 120256, and PI 174263) and one cold/salt-sensitive tomato cultivar (T5) were investigated. The effects of seed priming (6 d imbibition in aerated - 1-2 MPa polyethylene glycol solution at 20 °C followed by redrying) and of removing the endosperm/testa cap covering the radicle on the temperature sensitivity of GRg, and the interaction of these treatments with genotypes, were also examined. GRg decreased linearly with decreasing temperature for all genotypes and seed treatments. The minimum or base temperatures for germination (Th) varied by Tb, but did not lower the minimum temperature at which germination could occur. Priming increased the GRg of T5 seeds to equal or exceed those of control (non-primed) seeds of the cold/salt-tolerant genotypes at any T> Tb, but the PI lines exhibited an even greater response to priming. Times to germination within each seed lot were normally distributed on a logarithmic scale. Priming increased the variance in tg within a seed lot when compared to control seeds. However, the variation in thermal time for germination between the 10th and 90th percentiles of the seed population (0r(lO-9O)) was relatively unaffected_by priming due to the reduction in 0r in primed seeds. Removing the endosperm cap and testa opposite the radicle tip decreased dT almost 6-fold and and reduced Tb by 5 °C in T5 and PI 341988, implicating processes in the endosperm/testa as the limiting factors in germination at suboptimal temperatures.

Maize sucrose synthase-1 promoter directs phloem cell-specific expression of Gus gene in transgenic tobacco plants.

Abstract: Tobacco plants were transformed by using a chimeric gene construction, in which a corn sucrose synthase-1 gene (Sh) promoter was used to direct expression of the beta-glucuronidase (Gus) reporter gene. Expression of Sh-Gus activity in these plants was found to be cell type specfifc. GUS activity was detected only in the phloem cells but not in any other cell types of vegetative tissues. In addition, Sh-Gus expression was found to be anaerobically inducible in tobacco roots. Sh-Gus was also expressed at high levels in the endosperm tissue of maturing tobacco seeds. We thus demonstrated that the corn Sh promoter can direct cell-type-specific and inducible expression in a heterologous dicotyledonous plant.

Effect of ruminal microbial colonization on cereal grain digestion.

Abstract: The objective of this study was to examine the microbial digestion and colonization of whole (W), halved (H) and quartered (Q) cereal grains within the rumen. Barley (Hordeum vulgare L.), maize (Zea mays L.), sorghum (Sorghum bicolor (L.) Moench), and wheat (Triticum aestivum L.) were placed in nylon bags in the rumen of a fistulated steer. In sacco DM disappearance (ISDMD) of W grain was lower (P < 0.001) than that of H or Q grain. Once sectioned, wheat was most susceptible to microbial digestion followed by barley, sorghum and maize, respectively. Scanning electron microscopy showed that microbial colonization of W grain was restricted to fractured areas of the pericarp. Fracturing of the pericarp is necessary to allow rumen bacteria to gain access to the rapidly digestible nutrients of the endosperm. Initial colonization (2 h) of the endosperm of H and Q grains by rumen bacteria tended to be between large starch granules. After 24 h of exposure in the rumen, the endosperm of barley, wheat and sorghum w...

Degradation of Native Starch Granules by Barley α-Glucosidases

Abstract: The initial hydrolysis of native (unboiled) starch granules in germinating cereal kernels is considered to be due to α-amylases. We report that barley (Hordeum vulgare L.) seed α-glucosidases (EC 3.2.1.20) can hydrolyze native starch granules isolated from barley kernels and can do so at rates comparable to those of the predominant α-amylase isozymes. Two α-glucosidase charge isoforms were used individually and in combination with purified barley α-amylases to study in vitro starch digestion. Dramatic synergism, as much as 10.7-fold, of native starch granule hydrolysis, as determined by reducing sugar production, occurred when high pl α-glucosidase was combined with either high or low pl α-amylase. Synergism was also found when low pl α-glucosidase was combined with α-amylases. Scanning electron micrographs revealed that starch granule degradation by α-amylases alone occurred specifically at the equatorial grooves of lenticular granules. Granules hydrolyzed by combinations of α-glucosidases and α-amylases exhibited larger and more numerous holes on granule surfaces than did those granules attacked by α-amylase alone. As the presence of α-glucosidases resulted in more areas being susceptible to hydrolysis, we propose that this synergism is due, in part, to the ability of the α-glucosidases to hydrolyze glucosidic bonds other than α-1,4- and α-1,6- that are present at the granule surface, thereby eliminating bonds which were barriers to hydrolysis by α-amylases. Since both α-glucosidase and α-amylase are synthesized in aleurone cells during germination and secreted to the endosperm, the synergism documented here may function in vivo as well as in vitro.

Abscisic acid and osmoticum prevent germination of developing alfalfa embryos, but only osmoticum maintains the synthesis of developmental proteins

Abstract: Developing seeds of alfalfa (Medicago sativa L.) acquire the ability to germinate during the latter stages of development, the maturation drying phase. Isolated embryos placed on Murashige and Skoog medium germinate well during early and late development, but poorly during mid-development; however, when placed on water they germinate well only during the latter stage of development. Germination of isolated embryos is very slow and poor when they are incubated in the presence of surrounding seed structures (the endosperm or seed coat) taken from the mid-development stages. This inhibitory effect is also achieved by incubating embryos in 10−5 M abscisic acid (ABA). Endogenous ABA attains a high level during mid-development, especially in the endosperm. Seeds developing in pods treated with fluridone (1-methyl-3-phenyl-5[3-(trifluoromethyl)-phenyl]-4(1H)-pyridinone) contain low levels of ABA during mid-development, and the endosperm and seed coat only weakly inhibit the germination of isolated embryos. However, intact seeds from fluridone-treated pods do not germinate viviparously, which is indicative that ABA alone is not responsible for maintaining seeds in a developing state. Application of osmoticum (e.g. 0.35 M sucrose) to isolated developing embryos prevents their germination. Also, in the developing seed in situ the osmotic potential is high. Thus internal levels of osmoticum may play a role in preventing germination of the embryo and maintaining development. Abscisic acid and osmoticum impart distinctly different metabolic responses on developing embryos, as demonstrated by their protein-synthetic capacity. Only in the presence of osmoticum do embryos synthesize proteins which are distinctly recognizable as those synthesized by developing embryos in situ, i.e. when inside the pod. Abscisic acid induces the synthesis of a few unique proteins, but these arise even in mature embryos treated with ABA. Thus while both osmoticum and ABA prevent precocious germination, their effects on the synthetic capacity of the developing embryo are quite distinct. Since seeds with low endogenous ABA do not germinate, osmotic regulation may be the more important of these two factors in controlling seed development.

Cloning of the y1 Locus of Maize, a Gene Involved in the Biosynthesis of Carotenoids.

Abstract: The y1 gene is one of the genes responsible for the production of [beta]-carotene in the endosperm and leaves of maize. We have cloned a Robertson9s Mutator-tagged allele of the y1 gene (y1-mum) by using a Mu3 element as a hybridization probe. We substantiate that the cloned sequence is a portion of the y1 gene by molecular analyses of a revertant of a putative Mutator-induced y1 allele and the incidence of insertions within the cloned y1 sequence from several independently derived Mutator-induced y1 mutant stocks. The y1-mum sequence was used to isolate the standard Y1 allele, which conditions the presence of [beta]-carotene in the endosperm of the maize kernel.

Gametic imprinting in maize in relation to the angiosperm life cycle

Abstract: Differences in the activity of maternally and paternally derived genomes in maize endosperm have been observed at three levels of genetic manipulation. When the balance of entire chromosome sets departs from the standard ratio of two of maternal origin to one of paternal origin, development is impaired, often leading to seed failure. At the level of individual chromosomes, absence of a paternal representative for 8 of the 19 chromosome arms tested causes a marked reduction in kernel size. Replacement of the missing arms by ones of maternal origin does not complement this defect. At the gene level, some alleles of R confer solid coloration on the aleurone layer when transmitted maternally but patchy coloration (mottled) when transmitted via pollen. In contrast with the endosperm, no effect of parentage on R phenotype has been detected in embryonic and seedling tissues. Furthermore, gynogenetic and androgenetic haploid plants are viable in maize and are similar in appearance. The detection of parental effects in the endosperm, but not the embryo, points to the few cell divisions of the gametophytes as a critical stage in imprinting. Chromosomally based epigenetic variation originating at this stage would be reflected as imprinting effects. A separate fertilization establishes a line of genetic descent in the embryo that appears to be relatively free of imprinted genes.

Effects of Priming and Endosperm Integrity on Seed Germination Rates of Tomato Genotypes I. GERMINATION AT SUBOPTIMAL TEMPERATURE

Abstract: The bases of differences in germination rates (GR g ) among three cold/salt-tolerant tomato accessions and one cold/salt-sensitive tomato cultivar were investigated. The effects of seed priming (6 d imbibition in aerated −1.2 MPa polyethylene glycol solution at 20°C followed by redrying) and of removing the endosperm/testa cap covering the radicle on the temperature sensitivity of GR g , and the interaction of these treatments with genotypes, were also examined

Endoreduplication of nuclear DNA in the developing maize endosperm

Abstract: Multiparametric flow cytometry was used to analyze the development of the endosperm in Zea mays L. during the period from 8 to 20 days after pollination (dap). Nuclear size, DNA content per nucleus, and frequencies of nuclei with varying properties were measured in preparations that included all of the endosperm nuclei of single kernels of the inbred strain Al88. Characteristics of nuclear populations from different kernels on the same ear showed minimal variation. The dynamic changes of non-mitotic cells involved in endosperm development consisted of alternating periods of DNA replication with non-replication. Seven rounds of DNA replication had occurred in some nuclei in the later developmental stages with the rate averaging approximately one round per 24-hour period. Analysis of the DNA levels in the nuclei showed an exact doubling pattern indicating an endoreduplication process, that is, replication of the entire genome during each round. The loosely organized polytenization of the chromatin occurred to varying extents among the nuclei within an endosperm. A weak positive correlation existed between DNA content and size of nuclei suggesting that DNA increases and nuclear growth may not be highly coordinated in this tissue. Increased proportions of the larger nuclei occurred in the later stages of endosperm development. Considering the entire endosperm, the average DNA content per nucleus at the 15-dap peak level was approximately 12.8 C constituting a 2.7-fold overall increase from 8 dap.

Molecular Characterization of the Brittle-2 Gene Effect on Maize Endosperm ADPglucose Pyrophosphorylase Subunits.

Abstract: Activity of the enzyme ADPglucose pyrophosphorylase is known to be reduced in maize (Zea mays L.) endosperm mutants at two independent loci, Shrunken-2 (Sh(2)) and Brittle-2 (Bt(2)). Spinach leaf ADPglucose pyrophosphorylase has previously been shown to comprise two subunits of 51 and 54 kilodaltons. Anti-bodies raised to each of the two subunits of spinach leaf ADPglucose pyrophosphorylase were found to cross-react to different bands on Western blots prepared from polyacrylamide gel electrophoresis separated wild-type maize endosperm proteins. The anti-spinach leaf 51 kilodalton subunit antibody cross-reacted with a 55 kilodalton maize endosperm protein and the anti-spinach leaf 54 kilodalton subunit antibody cross-reacted with a 60 kilodalton maize endosperm protein. These immunological reactions were observed in maize endosperm extracts and with a highly purified preparation of maize endosperm ADPglucose pyrophosphorylase. Mutant bt(2) endosperm lacked the 55 kilodalton subunit while mutant sh(2) endosperm lacked the 60 kilodalton subunit on Western blots. These results suggest that the maize endosperm ADPglucose pyrophosphorylase is made up of two immunologically dissimilar subunits and that the bt(2) and sh(2) mutations cause reduction in ADPglucose pyrophosphorylase activity through the lack of one of these two subunits. An ADPglucose pyrophosphorylase cDNA clone antigenically selected from a rice seed cDNA expression library was found to hybridize strongly with a cDNA corresponding to a maize endosperm transcript which is absent in a W64A bt(2) mutant. Thus, the bt(2) mutant causes the absence not only of the small subunit but of the corresponding transcript. Bt(2) is implicated as the structural gene for the small (54 kilodalton) subunit of maize endosperm ADPglucose pyrophosphorylase.

Lack of fructose-1,6-bisphosphatase in a range of higher plants that store starch

Abstract: The aim of this work was to discover whether fructose-1,6-bisphosphatase (FBPase) is present in higher-plant cells that synthesize storage starch. The following were examined: suspension cultures of soybean (Glycine max), tubers of potato (Solanum tuberosum), florets of cauliflower (Brassica oleracea), developing endosperm of maize and of sweet corn (Zea mays), roots of pea (Pisum sativum), and the developing embryos of round and wrinkled varieties of pea. Unfractionated extracts of each tissue readily converted fructose 1,6-bisphosphate to fructose 6-phosphate in assays for both plastidic and cytosolic FBPase. These conversions were not inhibited by 20 microM-fructose 2,6-bisphosphate. Except in extracts of pea embryos and sweet-corn endosperm, treatment with affinity-purified antibodies to pyrophosphate: fructose-6-phosphate 1-phosphotransferase reduced the above fructose 6-phosphate production to the rate found with boiled extracts. The antibody-resistant activity from sweet corn was slight. In immunoblot analyses, antibody to plastidic FBPase did not react positively with any protein in extracts of soybean cells, potato tuber, cauliflower florets, maize endosperm and pea roots. Positive reactions were found for extracts of embryos of both round and wrinkled varieties of peas and endosperm of sweet corn. For pea embryos, but not for sweet-corn endosperm, the Mr of the recognized protein corresponded to that of plastidic FBPase. It is argued that soybean cells, potato tuber, cauliflower florets, maize (var. White Horse Tooth) endosperm and pea roots lack significant activity of plastidic FBPase, but that this enzyme is present in developing embryos of pea. The data for sweet corn (var. Golden Bantam) are not decisive. It is also argued that, where FBPase is absent, carbon for starch synthesis does not enter the amyloplast as triose phosphate.

Digestion of barley, maize, and wheat by selected species of ruminal bacteria.

Abstract: Differences in the digestion of barley, maize, and wheat by three major ruminal starch-digesting bacterial species, Streptococcus bovis 26, Ruminobacter amylophilus 50, and Butyrivibrio fibrisolvens A38, were characterized. The rate of starch digestion in all cereal species was greater for S. bovis 26 than for R. amylophilus 50 or B. fibrisolvens A38. Starch digestion by S. bovis 26 was greater in wheat than in barley or maize, whereas starch digestion by R. amylophilus 50 was greater in barley than in maize or wheat. B. fibrisolvens A38 digested the starch in barley and maize to a similar extent but was virtually unable to digest the starch in wheat. The higher ammonia concentration in cultures of B. fibrisolvens A38 when grown on wheat than when grown on barley or maize suggests that B. fibrisolvens A38 utilized wheat protein rather than starch. Scanning electron microscopy revealed that B. fibrisolvens A38 initially colonized cell wall material, while S. bovis 26 randomly colonized the endosperm and R. amylophilus 50 preferentially colonized starch granules. There was subsequent colonization but only superficial digestion of wheat starch granules by B. fibrisolvens A38. Variation in the association between starch and protein within the endosperm of cereal grains contributes to the differential effectiveness with which amylolytic species can utilize cereal starch.

Induction of triploid Citrus plants from endosperm calli in vitro

Abstract: Triploid hybrid Citrus plants were regenerated by somatic embryogenesis in vitro from endosperm derived calli. A sequence of media formulations was used to induce and support proliferation of primary callus from endosperm, to induce embryogenesis from primary callus, and to allow embryo development leading to viable plantlets. Calli were induced from cellular endosperm of Citrus sinensis (sweet orange), C. Xparadisi (grapefruit), and C. grandis (pummelo) excised 12–14 weeks post-anthesis. Induction of embryogenesis from sweet orange and pummelo primary calli required gibberellic acid and double mineral nutrient concentrations. Embryogenesis was not induced from grapefruit calli in these experiments. Only sweet orange embryos developed sufficiently to allow plant regeneration. Triploid axillary buds were minigrafted onto etiolated diploid rootstock seedlings in vitro in order to transfer triploid regenerants to soil and the external environment. Triploidy (2n = 3x = 27) was observed consistently in all phases of regeneration and in recovered plants. These results demonstrate that triploid hybrid plant recovery from Citrus endosperm can overcome barriers to sexual hybridization resulting from apomixis.

Abscisic Acid Inhibition of Endosperm Cell Division in Cultured Maize Kernels

Abstract: The response of developing maize (Zea mays L.) endosperm to elevated levels of abscisic acid (ABA) was investigated. Maize kernels and subtending cob sections were excised at 5 days after pollination (DAP) and placed in culture with or without 90 micromolar (±)-ABA in the medium. A decreased number of cells per endosperm was observed at 10 DAP (and later sampling times) in kernels cultured in medium containing ABA from 5 DAP, and in kernels transferred at 8 DAP to medium containing ABA, but not in kernels transferred at 11 DAP to medium containing ABA. The number of starch granules per endosperm was decreased in some treatments, but the reduction, when apparent, was comparable to the decreased number of endosperm cells. The effect on endosperm fresh weight was slight, transient, and appeared to be secondary to the effect on cell number. Mature endosperm dry weight was reduced when kernels were cultured continuously in medium containing ABA. Endosperm (+)-ABA content of kernels cultured in 0, 3, 10, 30, 100, or 300 micromolar (±)-ABA was measured at 10 DAP by indirect ELISA using a monoclonal antibody. Content of (+)-ABA in endosperms correlated negatively (R = −0.92) with endosperm cell number. On the basis of these studies we propose that during early kernel development, elevated levels of ABA decrease the rate of cell division in maize endosperm which, in turn, could limit the storage capacity of the kernel.

Comparative studies of the development of endosperm-degrading enzymes in malting sorghum and barley.

Abstract: The endosperm cell walls of barley are degraded extensively during malting whilst those of sorghum are not. Malting barley produced endo-β-1,3:1,4-glucanase, endo-β-1,3-glucanase and pentosanase in large quantities. In contrast, malting sorghum developed mainly endo-β-1,3-glucanase and pentosanase. Although the limited break-down of the endosperm cell walls of sorghum may reflect sub-optimal activities of β-glucanases, such as endo-β-1,3:1,4-glucanases, it is possible that the highly intractable nature of the cell walls and their high protein content (approx. 60%) may contribute to the low susceptibility of sorghum endosperm cell walls to enzymic degradation during malting.

Changes in the Endosperm Cell Walls of Two Datura Species before Radicle Protrusion

Abstract: The possibility of an association between changes in cell walls of the micropylar portion of the endosperm and the induction of germination was explored in seeds of Datura ferox and Datura stramonium The structure of the inner surface of the endosperm was studied by scanning electron microscopy and the composition of cell wall polysaccharides analyzed by gas chromatography and gas chromatography-mass spectrometry Both scanning electron microscope images and chemical analysis showed changes in the micropylar portion of the endosperm in induced seeds before radicle protrusion The inner surface of the endosperm appeared eroded, and in some areas, wall material seemed to be missing The content of the main component of the cell wall polysaccharides, containing predominantly 4-linked mannose, decreased well before the emergence of the radicle through the endosperm We propose that the degradation of a mannan type polysaccharide is an important factor in the reduction in mechanical strength of the endosperm, thus facilitating germination

A plant serpin gene : structure, organization and expression of the gene encoding barley protein z4

Abstract: A 3133-bp nucleotide sequence of the gene Paz1 on chromosome 4 of barley, encoding endosperm protein Z4, has been determined. The sequence includes 1079 bp 5' upstream and 523 bp 3' downstream of the coding region. The 1079-bp 5' upstream region of the gene shows little similarity to 5' regions of other sequences genes expressed in the developing cereal endosperm. The coding sequence is interrupted by one 334-bp-long intron (bases 1497-1830). The deduced amino acid sequence, which was corroborated by peptide sequences, consists of 399 amino acids and has a molecular mass of 43,128 Da. This sequence confirms protein Z4 to be a member of the serpin superfamily of proteins. The similarity with other members of the family expressed as amino acids in identical positions is in the order of 25-30% and pronounced in the carboxy-terminal half of the molecule. Sequence residues assumed to form clusters stabilizing the tertiary structure are highly conserved. Protein Z4 is synthesized in the developing endosperm without a signal peptide and protein Z4 mRNA was evenly distributed among the free and membrane-bound polyribosomes of the endosperm cell. An internal hydrophobic region of 21 amino acids (residues 36-56) may serve as a signal for targeting the polypeptide into the lumen of the endoplasmic reticulum. The gene for protein Z4 could not be detected in the barley variety Maskin and some of its descendants. The 'high-lysine' allees, lys1 (Hiproly barley) and lys3a (Bomi mutant 1508) on chromosome 7, enhance and repress, respectively, the expression of the protein Z4 gene. Also, 1554 bp of another 8-kbp fragment of the barley genome Paz psi, similar to the protein-Z4-coding region, have been determined. Small insertions and deletions and the presence of an internal stop codon identify this fragment as part of a pseudogene related to the protein Z4 gene.

Purification and characterization of zein-degrading proteases from endosperm of germinating maize seeds.

Abstract: We have purified a group of four proteases (molecular mass 26-33 kilodalton) from germinating seeds of maize (Zea mays L. var W64A) using ammonium sulfate and isoelectric precipitations, anion exchange chromatography, and electroelution from preparative nondenaturing polyacrylamide gels. Their appearance in the endosperm of germinating seeds coincides with the onset of zein degradation. We have shown that these proteases degrade zeins dissolved in alcoholic solution as well as aggregated in protein bodies from developing maize kernels. The apparent molecular weights and net negative charges of each of these proteases are very similar. Additionally, they are inhibited by thiol-blocking agents and activated by reducing compounds. These characteristics suggest that they are a group of cysteine proteases involved in the first steps of storage protein degradation.

Gene expression in seed development and germination.

Abstract: Publisher Summary This chapter explains the regulatory aspects of the expression of genes for proteins synthesized during development and germination. It reviews the synthesis of some important classes of storage proteins during seed development, and discusses the cessation of protein synthesis during maturation drying and the revival of synthesis during germination, leading to seedling establishment. Gene expression during germination has been studied to a far lesser extent, for unlike in development, there are no proteins synthesized in large amount. Post-germinative physiology, involving the mobilization of stored reserves to provide for seeding growth, requires the expression of genes for hydrolytic enzymes. Studies on this subject, especially in relation to the synthesis of α-amylase in the aleurone layer of the cereal endosperm, remain a major part of this aspect of seed biology. During development, an endosperm is formed from a triploid fusion nucleus. The endosperm may or may not persist in the mature seed, but when present, it usually is the tissue in which the stored reserves are deposited; otherwise, the cotyledons are the major store. In seeds of monocotyledonous plants (e.g., cereals), the embryo contains an axial region, but the cotyledon is modified to form a relatively diminutive scutellum; the endosperm is a large structure at maturity, and is the important storage tissue.

Response of Enzymes and Storage Proteins of Maize Endosperm to Nitrogen Supply

Abstract: To examine the effects of N nutrition upon endosperm development, maize (Zea mays) kernels were grown in vitro with either 0, 3.6, 7.1, 14.3, or 35.7 millimolar N. Kernels were harvested at 20 days after pollination for determination of enzyme activities and again at maturity for quantification of storage products and electrophoretic separation of zeins. Endosperm dry weight, starch, zein-N, and nonzein-N all increased in mature kernels as N supply increased from zero to 14.3 millimolar. The activities of sucrose synthase, aldolase, phosphoglucomutase, glutamate-pyruvate transaminase, glutamate-oxaloacetate transaminase, and acetolactate synthase increased from 1- to 2.5-fold with increasing N supply. Adenosine diphosphate-glucose pyrophosphorylase and both ATP- and PPi-dependent phosphofructokinases increased to lesser extents, while no significant response was detected for hexose kinases and glutamine synthetase. Nitrogen-induced changes in enzyme activities were often highly correlated with changes in final starch and/or zein-N contents. Separation of zeins indicated that these peptides were proportionately enhanced by N supply, with the exception of C-zein, which was relatively insensitive to N. These data indicate that at least a portion of the yield increase in maize produced by N fertilization is induced by a modification of kernel metabolism in response to N supply.