Kernel Abortion in Maize : II. Distribution of C among Kernel Carbohydrates

This study was designed to compare the uptake and distribution of ¹⁴C among fructose, glucose, sucrose, and starch in the cob, pedicel, and endosperm tissues of maize (Zea mays L.) kernels induced to abort by high temperature with those that develop normally. Kernels cultured in vitro at 30 and 35°C were transferred to [¹⁴C]sucrose media 10 days after pollination. Kernels cultured at 35°C aborted prior to the onset of linear dry matter accumulation. Significant uptake into the cob, pedicel, and endosperm of radioactivity associated with the soluble and starch fractions of the tissues was detected after 24 hours in culture on labeled media. After 8 days in culture on [¹⁴C]sucrose media, 48 and 40% of the radioactivity associated with the cob carbohydrates was found in the reducing sugars at 30 and 35°C, respectively. This indicates that some of the sucrose taken up by the cob tissue was cleaved to fructose and glucose in the cob. Of the total carbohydrates, a higher percentage of label was associated with sucrose and a lower percentage with fructose and glucose in pedicel tissue of kernels cultured at 35°C compared to kernels cultured at 30°C. These results indicate that sucrose was not cleaved to fructose and glucose as rapidly during the unloading process in the pedicel of kernels induced to abort by high temperature. Kernels cultured at 35°C had a much lower proportion of label associated with endosperm starch (29%) than did kernels cultured at 30°C (89%). Kernels cultured at 35°C had a correspondingly higher proportion of ¹⁴C in endosperm fructose, glucose, and sucrose. These results indicate that starch synthesis in the endosperm is strongly inhibited in kernels induced to abort by high temperature even though there is an adequate supply of sugar.     

The free movement of14C-labelled organic compounds from intact apple seeds to growing fruitlets and shoots

Indole-3-acetic acid (IAA-1¹⁴C, IAA-2¹⁴C) and gamma-aminobutyric acid labelled with¹⁴C were applied in lanoline to the surface of intact seeds or inserted into seeds of growing apple fruitlets or to the cut surface of the pericarp. Their translocation in trees was checked by means of autoradiography or by a low background Geiger-Müller counter. Auxin applied to the top of intact seeds, or inserted into the seeds was translocated and distributed within the pericarp, spur tissues and shoots below. The translocation of gamma-aminobutyric acid from seeds suggests that the capacity of apple seeds for extensive interchange with the surrounding tissues of the fruit and the spur is more general than suspected. A much poorer or no translocation to the spur was found when radioactive compounds were applied to the cut surface of the pericarp, while the seeds were left untouched. The results suggest two mechanisms of metabolite translocation in the apple fruit: two-directional for seeds and one-directional for the pericarp.     

Translocation and Metabolism of Endosperm-Applied [2-C] Indoleacetic Acid in Etiolated Avena sativa L. Seedlings

The role of free indole-3-acetic acid (IAA) in the endosperm of Avena sativa L. seedlings was investigated to determine its contribution to free IAA in the shoot. [2-¹⁴C]IAA was injected into the endosperm of darkgrown seedlings and the transport and metabolism of the [¹⁴C]-labeled compounds determined. It was concluded that translocation of free IAA directly from the endosperm is probably not a significant source of free IAA in the shoot, mainly because even small amounts of [¹⁴C]IAA introduced into the endosperm were rapidly metabolized. This suggested that, in Avena, free IAA does not normally exist in the liquid endosperm.     

Light or high (30°C) temperature effects on l-[U-C]leucine incorporation and protein patterns in embryos and endosperm during the early phase of germination of the negatively photoblastic and thermosensitive seeds of Phacelia tanacetifolia

The activities of l-[U-¹⁴C]leucine uptake and incorporation into proteins of embryos and endosperm of seeds of Phacelia tanacetifolia Benth. cv Bleu-Clair were analysed during the first 24 h of incubation under conditions optimal for germination (16°C in darkness) and in two inhibitory conditions: 16°C in the light and 30°C in darkness. Blocking germination induced by light or 30°C was accompanied by the inhibition of l-[U-¹⁴C]leucine uptake and incorporation in embryos. In the endosperm, the activation of l-[U-¹⁴C]leucine uptake was of the same magnitude for the non-inhibited and the light-inhibited seeds and much higher for the 30°C-inhibited seeds; the activation of l-[U-¹⁴C]leucine incorporation was quantitatively similar in all three conditions, with the patterns of newly synthesised proteins qualitatively different in the endosperm from light- or 30°C-inhibited seeds. The results showed that germination of P. tanacetifolia seeds is controlled by light or super-optimal temperature through the inhibition of the activation of transport and protein synthetic activities in embryo without effect on the endosperm. We suggest, on the basis of the translational activity, the possibility that in the inhibitory conditions the blockade of the embryo to operate as a sink affects the transition of the endosperm to operation as a source.     

Myo-Inositol Esters of Indole-3-acetic Acid as Seed Auxin Precursors of Zea mays L

Indole-3-acetyl-myo-inositol esters constitute 30% of the low molecular weight derivatives of indole-3-acetic acid (IAA) in seeds of Zea mays. [¹⁴C]Indole-3-acetyl-myo-inositol was applied to a cut in the endosperm of the seed and found to be transported from endosperm to shoot at 400 times the rate of transport of free IAA. The rate of transport of indole-3-acetyl-myo-inositol from endosperm to shoot was 6.3 picomoles per shoot per hour and thus adequate to serve as the seed auxin precursor for the free IAA diffusing downward from the shoot tip. Indole-3-acetyl-myo-inositol is the first seed auxin precursor to be identified.     

Factors Affecting Dalapon Absorption and Translocation in Johnsongrass

The factors affecting the absorption and translocation of ¹⁴C-dalapon (2.2-dichloropropionic acid) in johnsongrass were studied. Johnsongrass [Sorghum halepense (L.) Pers.] was first pot-grown in a greenhouse and then treated and placed in controlled-environment chambers. Absorption of ¹⁴C-dalapon into johnsongrass leaves and subsequent translocation occurred continuously within the plant during a 48-h period after treatment. Gas chromatographic analysis of johnsongrass extracts showed that the dalapon molecule was translocated intact. Absorption and translocation of ¹⁴C-dalapon increased as the droplet volume of the diluent was increased from 0.2 to 5.0 μl per treated spot. At 21 and 32°C, translocation of ¹⁴C-dalapon from a 2-cm treated leaf section into the plant was greater at 100% than at 35% relative humidity. At 38°C, translocation was greater at 35% than at 100% relative humidity. The addition of 0.5% surfactant to the dalapon solution increased translocation under all environmental conditions studied. The addition of 0.1 M KH₂PO₄ to dalapon-surfactant solutions increased ⁴-dalapon translocation under high temperature (38°C), especially at 35% relative humidity.     

Metabolism of C-Maltose in Avena fatua Seeds During Germination

Non-dormant and dormant seeds of Avena fatua metabolize ¹⁴C-maltose in different ways: in non-dormant seeds, ¹⁴C-maltose administered to the endosperm is readily converted to sucrose in the scutellum and translocated to the embryo; in dormant seeds, little sucrose is synthesized from ¹⁴C-maltose, and maltose and glucose tend to accumulate in the endosperm. It is suggested that biosynthesis of sucrose is essential for effective transport of the endosperm reserve to the embryonic axis in germinating seeds.     

The amino acid permease AAP8 is important for early seed development in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The development of seeds depends on the import of carbohydrates and amino acids supplied by the maternal tissue via the phloem. Several amino acid transporters have been reported to be expressed during seed and silique development in Arabidopsis thaliana (L.) Heynh. Here we show that mutants lacking the high affinity amino acid permease 8 (At1g10010) display a severe seed phenotype. The overall number of seeds and the number of normally developed seed is reduced by ∼50% in siliques of the Ataap8 T-DNA insertion mutant. This result could be reproduced in plants where expression of AtAAP8 is targeted with an RNAi approach. The seed phenotype is correlated with a specifically altered amino acid composition of young siliques. Aspartic acid and glutamic acid are significantly reduced in young siliques of the mutants. In correlation with the fact that AAP8 is a high affinity transporter for acidic amino acids, translocation of ¹⁴C-labelled aspartate fed via the root system to seeds of the mutants is reduced. AAP8 plays a crucial role for the uptake of amino acids into the endosperm and supplying the developing embryo with amino acids during early embryogenesis.     

Sugar Uptake and Metabolism in the Developing Endosperm of Tassel-seed Tunicate (Ts-5 Tu) Maize

Factors regulating assimilate transport into developing maize (Zea mays L.) kernels have been difficult to determine because of the structural complexity of basal kernel tissues and the damage that results from tissue dissection. The sensitivity of maize kernels to experimental manipulation is such that substantial maternal tissue is required to support kernel growth in vitro. Consequently, sugar transport experiments with isolated seed tissues or detached kernels have not unequivocally demonstrated how sugar transport occurs. In the present study, Tassel-seed Tunicate (Ts-5 Tu) maize kernels were investigated as a model system for introducing test solutions into the pedicel apoplast with minimal wounding. Transpiration in leafy glumes drew ¹⁴C-sugar solutions up the 8- to 10-millimeter-long pedicel stalks into the basal endosperm transfer cell region. ¹⁴C from fructose was incorporated into starch for 8 days. Sugar uptake into endosperm and embryo tissue showed specificity and inhibitor sensitivity. In particular, p-chloromercuribenzene sulfonate partially inhibited fructose uptake into the endosperm but had no effect on the metabolic conversion of that fructose that entered the endosperm. These results are consistent with active, carrier-mediated sugar transport, but a definitive determination would require more detailed tissue analysis. We propose that further refinement of the incubation solution may allow long-term kernel growth without cob tissue and thus provide a more precise determination of which maternal factors influence seed development.     

Chlorophenoxyacetic acid and chloropyridylphenylurea accelerate translocation of photoassimilates to parthenocarpic and seeded fruits of muskmelon (Cucumis melo)

We compared the effect of p-chlorophenoxyacetic acid (p-CPA) and 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU) on parthenocarpic and seeded muskmelon (Cucumis melo) fruits in regards to fruit development and the transport of photoassimilates from leaves exposed to ¹⁴CO₂ to the developing fruits. Ten days after anthesis (DAA), the fresh weight, total ¹⁴C-radioactivity and contents of ¹⁴C-sucrose and ¹⁴C-fructose were higher in the CPPU-induced parthenocarpic fruits than in seeded fruits. However, at 35 DAA, fresh weight and sucrose content in mesocarp, placenta and empty seeds of the parthenocarpic fruits were lower than in seeded fruits. Also, total ¹⁴C-radioactivity and ¹⁴C-sugar content of the parthenocarpic fruits were lower as well as the translocation rate of ¹⁴C-photoassimilates into these fruits. Application of p-CPA to the parthenocarpic fruits at 10 and 25 DAA increased fresh weight and sugar content. Moreover, these treatments elevated the total ¹⁴C-radioactivity, ¹⁴C-sucrose content and the translocation rate of ¹⁴C-photoassimilates. The ¹⁴C-radioactivity along the translocation pathway from leaf to petiole, stem, lateral shoot and peduncle showed a declining pattern but dramatically increased again in the fruits. These results suggest that the fruit's sink strength was regulated by the seed and enhanced by the application of p-CPA.     

Soluble and particulate lysophospholipase in the aleurone and endosperm of germinating barley

Lysophospholipase was measured in extracts of germinating barley by determining the amount of free [¹⁴C]palmitate released from [1-¹⁴C] 1-palmitoyl-lysophosphatidylcholine (LPC). Soluble and particulate lysophospholipase activity was measured at 1-day intervals in extracts from the aleurone and endosperm of barley seeds germinated for 8 days. The soluble and particulate activities of the aleurone increase approximately in parallel with one another and after 8 days of germination have 20–30 times more activity than at day 1. The activity profiles and the distribution of the activity between the soluble and particulate forms of lysophospholipase in the endosperm are markedly different. With the exception of the first 2 days when the aleurone activity is low, the endosperm activity is less than that associated with the aleurone. The soluble activity increases during the first 3 days and is more active than that of the aleurone. Thereafter it diminishes and remains low. The particulate enzyme, however, increases dramatically between days 4 and 5 and remains moderately high. The fourth and fifth day represent that stage of germination when starch-bound LPC is released in concert with the increase in amylase activity. It is proposed that it is this particulate form of the endosperm activity which may be responsible for maintaining the level of free LPC low in the endosperm of the germinating seed.     

Subcellular site of carotenoid biosynthesis in Neurospora crassa

A cell-free system prepared from Neurospora crassa mycelia was capable of incorporating radioactivity from [¹⁴C]-mevalonic acid into phytoene and to a much lesser extent into more unsaturated carotenoids. Whereas carotenogenic activities were only minimal in extracts from dark-grown cultures, they were several-fold increased following a short in vivo illumination; this photo-induced increase was inhibited by cycloheximide. Subcellular localization of carotenogenic enzyme activities was investigated using incubations of particular isolated fractions, together with [¹⁴C]-mevalonic acid and a geranylgeranyl-pyrophosphate-synthesizing system provided by the endosperm of maturing pumpkin seeds. Maximum carotenogenic activity (ca 80%) was localized in two membrane fractions previously shown to contain plasma membranes and, in particular, membranes of the endoplasmic reticulum. The lipid layer, containing the bulk of carotenoid pigments, possesses only trace amounts of enzyme activities.     

Characterization of the activity of fatty-acid epoxide hydrolase in seeds of castor bean (Ricinus communis L.)

Epoxide hydrolase (EC 3.3.2.3) activity was measured with [1-¹⁴C]cis-9,10-epoxystearic acid as the substrate. Homogenates were prepared from the endosperm tissue of germinating seeds of castor bean (Ricinus communis L. zanzibariensis). The activity of fatty-acid epoxide hydrolase was characterized with respect to dependence on time, amount of protein, pH and temperature. Analyses of enzyme distribution in endosperm, cotyledons, root and hypocotyl showed the highest total activity in the endosperm, less in the cotyledons and low activity in the root and hypocotyl. The specific activity was similar for cotyledons and endosperm. Analysis of the temporal expression of the enzyme in the endosperm during germination revealed high activity already in the imbibed seed. Activity was maximal between days four to six and then decreased at the end of one week. Subcellular fractionation of endosperm revealed a dual distribution of activity between the glyoxysomal and the cytosolic fractions.     

Seasonal differences in mineral content, distribution and leakage of sweet pepper seeds

Pepper seed quality is sensitive to variations in climatic conditions during seed development, which might be associated with accumulation, distribution and leakage of mineral elements from the seeds. This was examined in hybrid seeds of sweet pepper (Capsicum annuum L. cv.‘Hazera’ 1195) in two experiments during two growing seasons. The mean daily temperature (day/night) and daily total radiation receipt during seed development were 27.9/23.2°C and 8.63 kW m^?2 in the summer and 18.3/ 14.9°C and 3.18 kW m^?2 in the winter, respectively. Seeds developed in the summer had lower percentage of seedling emergence and leaked a larger portion (45%) of their K content into the water medium than in winter‐developed seeds. Summer seeds accumulated more K and Cl, but less P, Mg, Ca and the weight ratio of linoleic acid to oleic acid was lower than in the winter seeds. The season did not significantly affect N, S and total fatty acids. The most abundant element on the seed coat surface was K in the summer and Ca in the winter seeds. The cotyledon and endosperm of the summer seeds contained relatively higher ratios of K and Ca and lower ratios of P and Mg than the winter seeds. Transportation of mineral nutrients appeared to be involved in the effect of heat and moisture stresses on emergence quality of the pepper seeds.     

Uptake and Translocation of Benthiocarb Herbicide by Plants

The uptake and translocation of ¹⁴C-benthiocarb labelled at benzyl methylene by rice plant, bamyardgrass, wild amaranth, smart weed and lambsquarters were investigated, ¹⁴C-Benthiocarb was absorbed through the roots and the radioactivity was translocated into whole plants. The rate of absorption and translocation varied by the kind of plants. The translocation was occurred not only from roots into leaves, but from a leaf into other leaves, and even into roots of some kinds of plant. The absorption and translocation was more easy in barnyard-grass than in rice plant. Benthiocarb was rapidly absorbed by seeds and accumulated mostly in the embryo. The uptake of benthiocarb by seedlings decreased with the order of mesocotyl (bamyardgrass only), coleoptyl, root and leaf. Benthiocarb was degraded rapidly in plants.     

Growth induced translocation effectively directs an amino acid analogue to developing zones in Agaricus bisporus

The vegetative mycelium of Agaricus bisporus supplies developing white button mushrooms with water and nutrients. However, it is not yet known which part of the mycelium contributes to the feeding of the mushrooms and how this depends on growth conditions. Here we used photon counting scintillation imaging to track translocation of the ¹⁴C-radiolabeled metabolically inert amino acid analogue α-aminoisobutyric acid (¹⁴C-AIB). Translocation to the periphery of the mycelium was observed in actively growing vegetative mycelium with a velocity of up to 6.6 mm h⁻¹, which was 30-fold higher than the growth rate. Furthermore, ¹⁴C-AIB translocated to neighboring colonies after fusion by anastomosis depending on the relative growth rate in these colonies. When mushrooms started to develop, translocation of ¹⁴C-AIB was redirected to the fruiting bodies via mycelium and hyphal cords. More abundant mycelial cord formation and a 5-fold higher rate of translocation was observed for cultures growing directionally from inoculum located at one side of the substrate, when compared to non-directional growth (inoculum mixed throughout the substrate). The maximum translocation distance was also greater (≥50 and 22 cm, respectively). In conclusion, ¹⁴C-AIB translocation switches between vegetative growth and towards developing mushrooms, especially via cords and when source–sink relationships change.     

Movement of C-labeled Assimilates into Kernels of Zea mays L: III. AN ANATOMICAL EXAMINATION AND MICROAUTORADIOGRAPHIC STUDY OF ASSIMILATE TRANSFER

Basal tissue of developing maize kernels was examined by light micros-copy and by scanning and transmission electron microscopy. Plasmodesmata occur in pedicel and endosperm parenchyma but were not seen between the placento-chalazal cells and basal endosperm transfer cells. A layer of noncellular material separates the transfer cells from the placentochalazal cells. Microautoradiography of ¹⁴C-labeled assimilates entering the kernels revealed that incoming sugars are not confined to the apoplast, but rather are present in the cytoplasm and vacuoles of pedicel and endosperm cells. No specific accumulation of radioactivity was seen in any particular tissue, although at later sampling times, a higher grain density in the pedicel than the endosperm indicated a general buildup of sugars in the pedicel. A possible model for sugar movement into developing kernels is discussed.     

The iron-chelate transporter OsYSL9 plays a role in iron distribution in developing rice grains

Key message

Rice OsYSL9 is a novel transporter for Fe(II)-nicotianamine and Fe(III)-deoxymugineic acid that is responsible for internal iron transport, especially from endosperm to embryo in developing seeds.

Abstract

Metal chelators are essential for safe and efficient metal translocation in plants. Graminaceous plants utilize specific ferric iron chelators, mugineic acid family phytosiderophores, to take up sparingly soluble iron from the soil. Yellow Stripe 1-Like (YSL) family transporters are responsible for transport of metal-phytosiderophores and structurally similar metal-nicotianamine complexes. Among the rice YSL family members (OsYSL) whose functions have not yet been clarified, OsYSL9 belongs to an uncharacterized subgroup containing highly conserved homologs in graminaceous species. In the present report, we showed that OsYSL9 localizes mainly to the plasma membrane and transports both iron(II)-nicotianamine and iron(III)-deoxymugineic acid into the cell. Expression of OsYSL9 was induced in the roots but repressed in the nonjuvenile leaves in response to iron deficiency. In iron-deficient roots, OsYSL9 was induced in the vascular cylinder but not in epidermal cells. Although OsYSL9-knockdown plants did not show a growth defect under iron-sufficient conditions, these plants were more sensitive to iron deficiency in the nonjuvenile stage compared with non-transgenic plants. At the grain-filling stage, OsYSL9 expression was strongly and transiently induced in the scutellum of the embryo and in endosperm cells surrounding the embryo. The iron concentration was decreased in embryos of OsYSL9-knockdown plants but was increased in residual parts of brown seeds. These results suggested that OsYSL9 is involved in iron translocation within plant parts and particularly iron translocation from endosperm to embryo in developing seeds.

     

Role of transamination in the mobilization of respiratory substrates in germinating seeds of castor oil plants

The metabolism of 1,4-¹⁴C-succinate and 2,3-¹⁴C-succinate and the activity of succinic semialdehyde dehydrogenase (EC 1.2.1.16) were studied in germinating seeds of castor oil plants (Ricinus communis L.). Succinate metabolism involved succinate dehydrogenase and was sensitive to metabolites of the γ-aminobutyric acid shunt. Considerable accumulation of the label in amino acids reflected the progression of transamination reactions. Succinic semialdehyde dehydrogenase was purified from the endosperm of castor oil plants. Kinetic characteristics of the enzyme were evaluated. Our study indicates that the mobilization of respiratory substrates during germination of castor oil plants is related to active transamination of ketoacids in the Krebs cycle and involves the γ-aminobutyric acid shunt.     

Transport and Metabolism of a Sucrose Analog (1'-Fluorosucrose) into Zea mays L. Endosperm without Invertase Hydrolysis

1′-Fluorosucrose (FS), a sucrose analog resistant to hydrolysis by invertase, was transported from husk leaves into maize (Zea mays L., Pioneer Hybrid 3320) kernels with the same magnitude and kinetics as sucrose. ¹⁴C-Label from [¹⁴C]FS and [¹⁴C]sucrose in separate experiments was distributed similarly between the pedicel, endosperm, and embryo with time. FS passed through maternal tissue and was absorbed intact into the endosperm where it was metabolized and used in synthesis of sucrose and methanol-chloroform-water insolubles. Accumulation of [¹⁴C] sucrose from supplied [¹⁴C]glucosyl-FS indicated that the glucose moiety from the breakdown of sucrose (here FS), which normally occurs in the process of starch synthesis in maize endosperm, was available to the pool of substrates for resynthesis of sucrose. Uptake of FS into maize endosperm without hydrolysis suggests that despite the presence of invertase in maternal tissues and the hydrolysis of a large percentage of sucrose unloaded from the phloem, hexoses are not specifically needed for uptake into maize endosperm.