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Early Events in Maize Seed Development : 1-Methyl-3-phenyl-5-(3-[trifluoromethyl]phenyl)-4-(1H)-Pyridinone Induction of Vivipary.

Preharvest sprouting or vivipary is induced in developing maize (Zea mays, inbred Tx 5855 and Va 35) seeds by fluridone, a pyridinone inhibitor of carotenoid biosynthesis. Fluridone has a maximal effect on vivipary at 11 days after pollination (DAP) and little effect at 13 DAP in the inbred maize line Tx 5855. Abscisic acid partially reversed the chemically induced vivipary. Though the precise mechanism of fluridone-induced vivipary is unknown, these results indicate that there are important developmental changes occurring at 11 DAP which reversibly commit the immature embryo to vivipary or dormancy.

Journal Article↗

Rice has two distinct classes of protein kinase genes related to SNF1 of Saccharomyces cerevisiae, which are differently regulated in early seed development.

We have isolated five cDNA clones (osk1-5) for protein kinases from rice which are related to SNF1 protein kinase of Saccharomyces cerevisiae. Based on the sequence homology, these cDNAs can be classified into two groups, group 1 (osk1) and group 2 (osk2-5). The products of these genes were demonstrated to be functional SNF1-related protein kinases by in vitro and in vivo experiments. Recombinant proteins expressed from both groups of genes were fully active as protein kinases and could phosphorylate SAMS peptide, a substrate specific for the SNF1/AMPK family, as well as themselves (autophosphorylation). Moreover, expression of osk3 cDNA in yeast snf1 mutants restored SNF1 function. Northern blot analyses showed differential expression of these two gene groups; group 1 is expressed uniformly in growing tissues (young roots, young shoots, flowers, and immature seeds), whereas group 2 is strongly expressed in immature seeds. SNF1-related protein kinases have been reported from different plant species, such as rye, barley, Arabidopsis, tobacco, and potato, while the type of gene strongly expressed in immature seeds is known only in cereals such as rye, barley, and, from our findings, in rice. Expression levels of the group 2 genes were further analyzed in seeds during seed maturation. Expression is transiently increased in the early stages of seed maturation and then decreases. The expression peak precedes those of the sbe1 and waxy genes, which are involved in starch synthesis in rice. Taken together, these findings suggest that group 2 OSK genes play important roles in the early stages of endosperm development in rice seeds.

Carbohydrate Metabolism↗

Gibberellins and seed development in maize. II. Gibberellin synthesis inhibition enhances abscisic acid signaling in cultured embryos.

Abscisic acid (ABA) is required for seed maturation in maize (Zea mays L.) and other plants. Gibberellins (GAs) are also present in developing maize embryos, and mutual antagonism of GAs and ABA appears to govern the choice between precocious germination or quiescence and maturation. Exogenous ABA can also induce quiescence and maturation in immature maize embryos in culture. To examine the role of GAs versus ABA in regulating maize embryo maturation, the effects of modulating GA levels were compared with those of ABA in embryos cultured at successive stages of development. The effects of GA synthesis inhibition or exogenous GA application differed markedly in embryos at different stages of development, indicating changes in both endogenous GA levels and in the capacity for GA synthesis as embryogenesis and maturation progress. In immature embryos, the inhibition of GA synthesis mimicked the effects of exogenous ABA, as shown by the suppression of germination, the acquisition of anthocyanin pigments, and the accumulation of a variety of maturation-phase mRNAs. We suggest that GA antagonizes ABA signaling in developing maize embryos, and that the changing hormone balance provides temporal control over the maturation phase.

Abscisic Acid↗

Oxygen-depleted zones inside reproductive structures of Brassicaceae: implications for oxygen control of seed development.

Growth of Arabidopsis thaliana (L.) Heynh. in decreasing oxygen partial pressures revealed a linear decrease in seed production below 15 kPa, with a complete absence of seed production at 2.5 kPa oxygen. This control of plant reproduction by oxygen had previously been attributed to an oxygen effect on the partitioning between vegetative and reproductive growth. However, plants grown in a series of decreasing oxygen concentrations produced progressively smaller embryos that had stopped developing at progressively younger stages, suggesting instead that their growth is limited by oxygen. Internal oxygen concentrations of buds, pistils, and developing siliques of Brassica rapa L. and siliques of Arabidopsis were measured using a small-diameter glass electrode that was moved into the structures using a micromanipulator. Oxygen partial pressures were found to be lowest in the developing perianth (11.1 kPa) and pistils (15.2 kPa) of the unopened buds. Pollination reduced oxygen concentration inside the pistils by 3 kPa after just 24 h. Inside Brassica silique locules, partial pressures of oxygen averaged 12.2 kPa in darkness, and increased linearly with increasing light levels to 16.2 kPa. Measurements inside Arabidopsis siliques averaged 6.1 kPa in the dark and rose to 12.2 kPa with light. Hypoxia in these microenvironments is postulated to be the point of control of plant reproduction by oxygen.

Arabidopsis↗

Dynamics of inositol phosphate pools (tris-, tetrakis- and pentakisphosphate) in relation to the rate of phytate synthesis during seed development in common bean (Phaseolus vulgaris).

Four cultivars of Phaseolus vulgaris were grown in a greenhouse and each flower was Labeled with date of anthesis. Seeds were collected at six different stages of development and inositol phosphates (InsPs) were analyzed by ion-pair reversed-phase HPLC. Phytate accumulation was similar in all cultivars, and the specific rate of phytate synthesis (Rs) peaked at about 22 days after flowering (DAF). Variations in the concentrations of the InsP3 and InsP4 pools matched changes in Rs in cultivars Una and Aruã. These results suggest mass-action effects. Thus, the rates of conversion of InsP3 to InsP5 appeared to be at least partly dependent on substrate concentration. Proportional increases in size of all InsP pools up to 21 DAF are also consistent with Little regulation in this part of the pathway. However, this did not appear to be the case in cv. Diamante Negro or with the conversion of InsP5 to InsP6 in all cultivars, where concentrations of the InsP precursor pools peaked earlier or even dropped as Rs peaked, suggesting activation of enzyme activity. Therefore, the evidence is consistent with a control point regulating this metabolic route upstream of InsP3 and possibly in the conversion of InsP5 to InsP6.

Flowers↗

Analysis of amylose accumulation during seed development in maize.

Starch, which includes amylose and amylopectin, is the most important component in maize (Zea mays L.) seeds. The accumulation of amylose in maize seeds was examined in this study. The percentage of amylose content gradually increased in seeds from day 10 to day 25 after pollination, which is consistent with the changes of GBSS activity. The transcripts of GBSSI were detected in both the endosperm and embryo of wild-type maize. However, its transcripts, GBSS activity, and amylose were not detected in either the endosperm or embryo of waxy maize. These results indicate that the accumulation of amylose is controlled by GBSSI expression in the seeds of maize.

Amylose↗

[Synthesis and degradation of the peanut storage proteins during seed development and germination].

Three polypeptides, 41 kD and 38.5 kD subunits of arachin and 60.5 kD subunit of conarachin in peanut (Arachis hypogaea L. Shanyou 523) seeds were purified by gel filtration and SDS-PAGE. Polyclonal antibodies against these subunits were raised in mice. Western blot showed that the subunits appeared in axes and cotyledons at the tissue differentiation stage. The 60.5 kD subunit was firstly synthesized and accumulated in considerable quantity in axes and cotyledons, and then the 41 kD and 38.5 kD subunits increased during the development of peanut embryos. The degradation patterns of these three subunits were different during the germination of peanut seeds. The 41 kD and 38.5 kD subunits in the axes and cotyledons were degraded earlier than the 60.5 kD subunit.

Allergens↗

Water Relations of Seed Development and Germination in Muskmelon (Cucumis melo L.) : V. Water Relations of Imbibition and Germination.

The initiation of radicle growth during seed germination may be driven by solute accumulation and increased turgor pressure, by cell wall relaxation, or by weakening of tissues surrounding the embryo. To investigate these possibilities, imbibition kinetics, water contents, and water (Psi) and solute (psi(s)) potentials of intact muskmelon (Cucumis melo L.) seeds, decoated seeds (testa removed, but a thin perisperm/endosperm envelope remains around the embryo), and isolated cotyledons and embryonic axes were measured. Cotyledons and embryonic axes excised and imbibed as isolated tissues attained water contents 25 and 50% greater, respectively, than the same tissues hydrated within intact seeds. The effect of the testa and perisperm on embryo water content was due to mechanical restriction of embryo swelling and not to impermeability to water. The Psi and psi(s) of embryo tissues were measured by psychrometry after excision from imbibed intact seeds. For intact or decoated seeds and excised cotyledons, Psi values were >-0.2 MPa just prior to radicle emergence. The Psi of excised embryonic axes, however, averaged only -0.6 MPa over the same period. The embryonic axis apparently is mechanically constrained within the testa/perisperm, increasing its total pressure potential until axis Psi is in equilibrium with cotyledon Psi, but reducing its water content and resulting in a low Psi when the constraint is removed. There was no evidence of decreasing psi(s) or increasing turgor pressure (Psi-psi(s)) prior to radicle growth for either intact seeds or excised tissues. Given the low relative water content of the axes within intact seeds, cell wall relaxation would be ineffective in creating a Psi gradient for water uptake. Rather, axis growth may be initiated by weakening of the perisperm, thus releasing the external pressure and creating a Psi gradient for water uptake into the axis. The perisperm envelope contains a cap of small, thin-walled endosperm cells adjacent to the radicle tip. We hypothesize that weakening or separation of cells in this region could initiate radicle expansion.

Journal Article↗

Protein Synthesis in Cotyledons of Pisum sativum L: I. Changes in Cell-Free Amino Acid Incorporation Capacity during Seed Development and Maturation.

The changes in protein content of pea cotyledons have been followed during the period from 9 to 33 days after flowering. Initially protein content increased gradually with a rapid period of deposition occurring between days 21 and 27 after flowering. After the 28th day the rate of accumulation of protein declined as the seed dehydrated and matured. At maturity the pea cotyledon contained approximately 25% protein which was divided into albumins and globulins in the ratio of 1:1.4.Analytical data and the incorporation of exogenously supplied (14)C-leucine indicated that albumins were synthesized early in cotyledon development whereas globulin synthesis predominated with increasing maturity.Ribosomal preparations extracted from seeds during the period of rapid protein synthesis contained a high percentage of polysomes. Preparations from older cotyledons with a declining capacity for protein synthesis had few polysomes and an abundance of monosomes. The amino acid-incorporating capacity of ribosomal preparations from cotyledons of varying age was related to the polysomic content. The phenylalanine-incorporating capacity of ribosomal preparations from mature pea seed could be stimulated by the addition of polyuridylic acid. The distribution of polysomes and the in vitro incorporation data suggested that protein synthesis could be partially restricted by the availability of messenger RNA at maturity.However, reciprocal mixing experiments of supernatant and ribosomal fractions from cotyledons of different developmental age indicated that the supernatant fractions have varying capacities to stimulate in vitro amino acid incorporation. Thus the possibility of the regulation of protein synthesis at the translational level was not precluded.

Journal Article↗

Petunia Ap2-like genes and their role in flower and seed development.

We have isolated three Apetala2 (Ap2)-like genes from petunia and studied their expression patterns by in situ hybridization. PhAp2A has a high sequence similarity to the A function gene Ap2 from Arabidopsis and a similar expression pattern during flower development, suggesting that they are cognate orthologs. PhAp2B and PhAp2C encode for AP2-like proteins that belong to a different subgroup of the AP2 family of transcription factors and exhibit divergent, nearly complementary expression patterns during flower development compared with PhAp2A. In contrast, all three PhAp2 genes are strongly expressed in endosperm. The phenotype of the petunia A-type mutant blind cannot be attributed to mutations in the petunia Ap2 homologs identified in this study, and reverse genetics strategies applied to identify phap2a mutants indicate that PhAp2A might not be essential for normal perianth development in petunia. Nevertheless, we show that PhAp2A is capable of restoring the homeotic transformations observed in flowers and seed of the ap2-1 mutant of Arabidopsis. Although the interspecific complementation proves that PhAp2A encodes a genuine Ap2 ortholog from petunia, additional factors may be involved in the control of perianth identity in this species.

Amino Acid Sequence↗

A sucrose-synthase gene of Vicia faba L.: expression pattern in developing seeds in relation to starch synthesis and metabolic regulation.

Copy-DNA clones encoding a single class of sucrose-synthase (SUCS; EC 2.4.1.13) subunit have been isolated and sequenced from a Vicia faba L. seed cotyledonary library. Southern analyses indicated the existence of only one gene. Transcript levels determined by Northern blot hybridisation steadily increased until the middle of development [25-35 days after flowering (DAF)] and declined thereafter. Sucrose levels approximately paralleled levels of SUCS mRNA. The activity of SUCS increased with decreasing fructose and glucose concentrations and peaked about 10 d later than mRNA levels. In-vitro culture experiments demonstrated that increasing the sucrose concentration leads to increased levels of SUCS mRNA. The SUCS mRNA was also synthesised in seed-coat tissue, but in lower amounts than in cotyledons and with a different developmental profile. The early peak level of SUCS mRNA (20 DAF) in seed coats coincided with the peak in the amount of sucrose and with a peak of transiently synthesised starch.

Amino Acid Sequence↗

Carbon supply for storage-product synthesis in developing seeds of oilseed rape.

The aim of this work was to find out how the sugars in the endosperm of oilseed rape contribute to the flux of oil synthesis. While the hexose content of the liquid endosperm decreased during development the sucrose content increased. It is important to understand the relative rates of use of the endosperm sugars for two reasons. Firstly we need to know which sugars are used, and at what stages in development, in order to understand the roles of enzymes involved in their metabolism. Secondly, changes in sugar concentration have been implicated in the regulation of expression of genes determining storage-product synthesis [see Weber, Borisjuk and Wobus (1997) Trends Plant Sci. 2, 169-174, for review]. The rate of consumption of sugar is one factor governing its concentration. We present data showing both the concentration-dependence of conversion of sugar to oil, and the in vivo concentrations of sugars; we relate these data sets to each other and discuss the effects of the intracellular pool of sucrose. Glucose, fructose and sucrose are all substrates for oil synthesis, but the rates of their use (particularly sucrose) are underestimated because of dilution by sucrose from the intracellular pool.

Brassica↗