Sterol and triterpene synthesis in the developing and germinating pea seed.
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ABSTRACT:
SUBMITTER: Baisted DJ
PROVIDER: S-EPMC1177152 | biostudies-other | 1971 Sep
REPOSITORIES: biostudies-other
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Project description:An important function of the seed coat is to deliver nutrients to the embryo. To relate this function to anatomical characteristics, the developing seed coat of pea (Pisum sativum L.) was examined by light- and cryo-scanning electron microscopy (cryo-SEM) from the late pre-storage phase until the end of seed filling. During this time the apparently undifferentiated seed coat tissues evolve into the epidermal macrosclereids, the hypodermal hourglass cells, chlorenchyma, ground parenchyma and branched parenchyma. Using the fluorescent symplast tracer 8-hydroxypyrene-1,3,6-trisulfonic acid, it could be demonstrated that solutes imported by the phloem move into the chlorenchyma and ground parenchyma, but not into the branched parenchyma. From a comparison with literature data of common bean (Phaseolus vulgaris L.) and broad bean (Vicia faba L.), it is concluded that in the three species different parenchyma layers, but not the branched parenchyma, may be involved in the post-phloem symplasmic transport of nutrients in the seed coat. In pea, the branched parenchyma dies during the storage phase, and its cell wall remnants then form the boundary layer between the living seed coat parenchyma cells and the cotyledons. Using cryo-SEM, clear images were obtained of this boundary layer which showed that many intracellular spaces in the seed coat parenchyma are filled with an aqueous solution. This is suggested to facilitate the diffusion of nutrients from the site of unloading towards the cotyledons.
| S-EPMC4242349 | biostudies-literature
Project description:The synthesis of lipids and acyl thioesters was studied in microsomal preparations from germinating pea (Pisum sativum cv. Feltham First) seeds. Under conditions of maximal synthesis (in the presence of exogenous acyl-carrier protein) acyl-acyl-carrier proteins accounted for about half the total incorporation from [14C]malonyl-CoA. Decreasing the concentrations of exogenous acyl-carrier protein lowered the overall synthesis of fatty acids by decreasing, almost exclusively, the radioactivity associated with acyl-acyl-carrier proteins. A time-course experiment showed that acyl-acyl-carrier proteins accumulated most of the radioactive label at the beginning of the incubation but, eventually, the amount of radioactivity in that fraction decreased, while a simultaneous increase in the acyl-CoA and lipid fractions was noticed. Addition of exogenous CoA (1 mM) produced a decrease of total incorporation, but an increase in the radioactivity incorporated into acyl-CoA. The microsomal preparations synthesized saturated fatty acids up to C20, including significant proportions of pentadecanoic acid and heptadecanoic acid. Synthesis of these 'odd-chain' fatty acids only took place in the microsomal fraction. In contrast, when the 18,000g supernatant (containing the microsomal and soluble fractions) was incubated with [14C]malonyl-CoA, the radioactive fatty acid and acyl classes closely resembled the patterns produced by germinating in the presence of [14C]acetate in vivo. The results are discussed in relation to the role of acyl thioesters in the biosynthesis of plant lipids.
| S-EPMC1163353 | biostudies-other
Project description:1. The effect of exogenous lipases on fatty acid synthesis from [14C]malonyl-CoA by the microsomal and soluble fractions from germinating peas was studied. 2. Addition of phospholipase A2 or the lipase from Rhizopus arrhizus had no effect on total fatty acid synthesis by the soluble fraction but caused severe inhibition of that by the microsomal fraction. 3. The addition of enzymes with phospholipase activity particularly inhibited the microsomal stearate elongase. 4. Control studies indicated that the phospholipase-induced inhibition of fatty acid synthesis was due to the location of fatty acid synthetase, palmitate elongase and stearate elongase on the outside of the microsomal vesicles. 5. Experiments with a trypsin-like proteinase showed that approximately half the microsomal fatty acid synthesis was resistant to proteolysis. 6. Although addition of exogenous phospholipases had no effect on total fatty acid synthesis by the soluble fraction, it did increase alpha-hydroxylation of newly-formed palmitate and stearate. 7. The results provide further evidence for differences between the soluble and particulate fatty acid synthetase and palmitate elongase activities of germinating pea.
| S-EPMC1158373 | biostudies-other
Project description:The tissue-specific syntheses of seed storage proteins in the cotyledons of developing pea (Pisum sativum L.) seeds have been demonstrated by estimates of their qualitative and quantitative accumulation by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and rocket immunoelectrophoresis respectively. Vicilin-fraction proteins initially accumulated faster than legumin, but whereas legumin was accumulated throughout development, different components of the vicilin fraction had their predominant periods of synthesis at different stages of development. The translation products in vitro of polysomes isolated from cotyledons at different stages of development reflected the synthesis in vivo of storage-protein polypeptides at corresponding times. The levels of storage-protein mRNA species during development were estimated by 'Northern' hybridization using cloned complementary-DNA probes. This technique showed that the levels of legumin and vicilin (47000-Mr precursors) mRNA species increased and decreased in agreement with estimated rates of synthesis of the respective polypeptides. The relative amounts of these messages, estimated by kinetic hybridization were also consistent. Legumin mRNA was present in leaf poly(A)+ RNA at less than one-thousandth of the level in cotyledon poly(A)+ (polyadenylated) RNA, demonstrating tissue-specific expression. Evidence is presented that storage-protein mRNA species are relatively long-lived, and it is suggested that storage-protein synthesis is regulated primarily at the transcriptional level.
| S-EPMC1153937 | biostudies-other
Project description:The synthesis of fatty acids from [14C]malonyl-CoA was studied with a high-speed particulate fraction from germinating pea (Pisum sativum). The variety used (Feltham First) produced mainly saturated fatty acids with palmitate (30--40%) and stearate (40--60%) predominating. Several palmitate-containing lipids stimulated overall synthesis and, in addition, increased the percentage of label in stearate. The production of stearate was severely inhibited by preincubation of the microsomal fraction with snake venom phospholipase A2 or by incubation with Rhizopus arrhizus lipase. Addition of a series of di-saturated phosphatidylcholines, with different acyl constituents, resulted in stimulation of overall fatty acid synthesis as well as an increase in the radiolabelling of the fatty acid two carbon atoms longer than the acyl chain added. This chain lengthening of fatty acids donated from phosphatidylcholine was due to the action of both fatty acid synthetase and palmitate elongase. The latter would utilize dipalmitoyl phosphatidylcholine and was sensitive to arsenite whereas fatty acid synthetase would use dilauroyl phosphatidylcholine and was sensitive to cerulenin. The results are discussed in relation to previous data obtained in vivo on plant fatty acid synthesis and current suggestions for the role of phosphatidylcholine in this process.
| S-EPMC1162279 | biostudies-other
Project description:1. Fatty acid synthesis was studied in microsomal preparations from germinating pea (Pisum sativum). 2. The preparations synthesized a mixture of saturated fatty acids up to a chain length of C(24) from [(14)C]malonyl-CoA. 3. Whereas hexadecanoic acid was made de novo, octadecanoic acid and icosanoic acid were synthesized by elongation. 4. The products formed during [(14)C]malonyl-CoA incubation were analysed, and unesterified fatty acids and polar lipids were found to be major products. [(14)C]Palmitic acid represented a high percentage of the acyl-carrier protein esters, whereas (14)C-labelled very-long-chain fatty acids were mainly present as unesterified fatty acids. CoA esters were minor products. 5. The addition of exogenous lipids to the incubation system usually resulted in stimulation of [(14)C]malonyl-CoA incorporation into fatty acids. The greatest stimulation was obtained with dipalmitoyl phosphatidylcholine. Both exogenous palmitic acid and dipalmitoyl phosphatidylcholine increased the amount of [(14)C]-stearic acid synthesized, relative to [(14)C]palmitic acid. Addition of stearic acid increased the amount of [(14)C]icosanoic acid formed. 6. [(14)C]Stearic acid was elongated more effectively to icosanoic acid than [(14)C]stearoyl-CoA, and its conversion was not decreased by addition of unlabelled stearoyl-CoA. 7. Incorporation of [(14)C]malonyl-CoA into fatty acids was markedly decreased by iodoacetamide and 5,5'-dithiobis-(2-nitrobenzoic acid). Palmitate elongation was sensitive to arsenite addition, and stearate elongation to the presence of Triton X-100 or fluoride. The action of fluoride was not, apparently, due to chelation. 8. The microsomal preparations differed from soluble fractions from germinating pea in (a) synthesizing very-long-chain fatty acids, (b) not utilizing exogenous palmitate-acyl-carrier protein as a substrate for palmitate elongation and (c) having fatty acid synthesis stimulated by the addition of certain complex lipids.
| S-EPMC1183759 | biostudies-other
Project description:12plex_pea_2013_02 - 12plex_pea_2013_02_g - What is the effect of a moderate water stress on seed filling (reserve accumulation) and nitrogen remobilisation in pea (Pisum sativum) - Pea plants (genotype Cameor) were subjected to a moderate water stress at the beggining of the seed filling period (12 Days After Pollination) of the second flowering node for a period of 8 days. Samples were collected from Well Watered (WW) plants at the beginning of the stress imposition (point A, T=0), and from Water-Stressed (WS) and WW control plants at the end of the drought period (point B, T=+8). Samples named SEED consisted of seeds from the pod of the second flowering node (seed-WW-A, seed-WW-B and Seed-WS-B). Samples named LEAF consisted of the leaves and stem sections from the two vegetative nodes below the first flowering node (leaf-WW-A, Leaf-WW-B and Leaf-WS-B). Each sample consited of a pool of 3 individual plants and 4 repetitions per condition were carried out.
2017-04-05 | GSE97237 | GEO
Project description:12plex_pea_2013_02 - 12plex_pea_2013_02_f - What is the effect of a moderate water stress on seed filling (reserve accumulation) and nitrogen remobilisation in pea (Pisum sativum) - Pea plants (genotype Cameor) were subjected to a moderate water stress at the beggining of the seed filling period (12 Days After Pollination) of the second flowering node for a period of 8 days. Samples were collected from Well Watered (WW) plants at the beginning of the stress imposition (point A, T=0), and from Water-Stressed (WS) and WW control plants at the end of the drought period (point B, T=+8). Samples named SEED consisted of seeds from the pod of the second flowering node (seed-WW-A, seed-WW-B and Seed-WS-B). Samples named LEAF consisted of the leaves and stem sections from the two vegetative nodes below the first flowering node (leaf-WW-A, Leaf-WW-B and Leaf-WS-B). Each sample consited of a pool of 3 individual plants and 4 repetitions per condition were carried out.
2017-02-01 | GSE93630 | GEO
Project description:The existing variation among pea protein isolates' functionality limits their application in food formulations. The source and extent of variations among yellow pea protein profiles was assessed in 10 single seeds of two varieties with different size and weight. A new approach was developed to analyze proteins of yellow pea combining three analytical methods of size exclusion chromatography (SEC), reverse phase high performance liquid chromatography (RP-HPLC), and microfluidic SDS-PAGE, to achieve the highest separation resolution. A high variation of protein concentration was observed not only between varieties, but also among seeds of the same variety. Vicilin to legumin ratio was between 2.72-4.19, and 1.70-2.22 among the individual seeds of AC Agassiz and CDC Saffron varieties, respectively. V/L ratio was significantly different among the individual seeds for both varieties. The amount of some protein fractions/subunits were correlated with seeds' size and weight for AC Agassiz, while such correlations were not observed for CDC Saffron.
| S-EPMC9352015 | biostudies-literature
Project description:Polycyclic triterpenes are members of the terpene family produced by the cyclization of squalene. The most representative polycyclic triterpenes are hopanoids and sterols, the former are mostly found in bacteria, whereas the latter are largely limited to eukaryotes, albeit with a growing number of bacterial exceptions. Given their important role and omnipresence in most eukaryotes, contrasting with their scant representation in bacteria, sterol biosynthesis was long thought to be a eukaryotic innovation. Thus, their presence in some bacteria was deemed to be the result of lateral gene transfer from eukaryotes. Elucidating the origin and evolution of the polycyclic triterpene synthetic pathways is important to understand the role of these compounds in eukaryogenesis and their geobiological value as biomarkers in fossil records. Here, we have revisited the phylogenies of the main enzymes involved in triterpene synthesis, performing gene neighborhood analysis and phylogenetic profiling. Squalene can be biosynthesized by two different pathways containing the HpnCDE or Sqs proteins. Our results suggest that the HpnCDE enzymes are derived from carotenoid biosynthesis ones and that they assembled in an ancestral squalene pathway in bacteria, while remaining metabolically versatile. Conversely, the Sqs enzyme is prone to be involved in lateral gene transfer, and its emergence is possibly related to the specialization of squalene biosynthesis. The biosynthesis of hopanoids seems to be ancestral in the Bacteria domain. Moreover, no triterpene cyclases are found in Archaea, invoking a potential scenario in which eukaryotic genes for sterol biosynthesis assembled from ancestral bacterial contributions in early eukaryotic lineages.
| S-EPMC7306690 | biostudies-literature