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Ethylene-promoted conversion of 1-aminocyclopropane-1-carboxylic Acid to ethylene in peel of apple at various stages of fruit development.

Internal ethylene concentration, ability to convert 1-amino-cyclopropane-1-carboxylic acid (ACC) to ethylene (ethylene-forming enzyme [EFE] activity) and ACC content in the peel of apples (Malus domestica Borkh., cv Golden Delicious) increased only slightly during fruit maturation on the tree. Treatment of immature apples with 100 microliters ethylene per liter for 24 hours increased EFE activity in the peel tissue, but did not induce an increase in ethylene production. This ability of apple peel tissue to respond to ethylene with elevated EFE activity increased exponentially during maturation on the tree. After harvest of mature preclimacteric apples previously treated with aminoethoxyvinyl-glycine, 0.05 microliter per liter ethylene did not immediately cause a rapid increase of development in EFE activity in peel tissue. However, 0.5 microliter per liter ethylene and higher concentrations did. The ethylene concentration for half-maximal promotion of EFE development was estimated to be approximately 0.9 microliter per liter. CO(2) partially inhibited the rapid increase of ethylene-promoted development of EFE activity. It is suggested that ethylene-promoted CO(2) production is involved in the regulation of autocatalytic ethylene production in apples.

Journal Article↗

Ascorbic acid concentration in Cv. conference pears during fruit development and postharvest storage.

L-ascorbic acid (L-AA) concentration changes during the development of cv. Conference pears and the influence of postharvest handlings (gas condition, cooling rate, cooling duration) on L-AA breakdown were studied. L-AA concentration fluctuates in young fruits, remains stable during fruit maturation, and starts to decline 1 week before commercial harvest. The most rapid decrease in L-AA concentration was found during immediate controlled atmosphere. During short-term storage, only the gas condition was found to influence L-AA breakdown; no significant difference between gradually or immediately cooled pears was determined. Under air conditions, both cooling strategies did not differ from the L-AA breakdown in pears allowed to ripen on the tree up until 3 weeks after the optimal harvest date. During long-term storage, the cooling duration (1-3 weeks) had no effect whereas both O2 and CO2 had a significant effect on L-AA retention. After 7 months of storage, no difference was found in dehydroascorbic acid concentration; the L-AA and total L-AA concentrations, in contrast, were significantly lower in the 5% CO2 conditions.

Ascorbic Acid↗

Flower and fruit development in Arabidopsis thaliana.

The study of flower development has experienced great advances over the last 15 years. The most important landmark was the proposal of the ABC model in which three different functions of overlapping activities account for the development of the four rings of organs of the eudicot flower. Most interestingly, during recent years this simple and elegant model has been broadly accepted and is applicable to a wide range of plant species. However, recent advances in the characterization of protein interactions and the discovery of the SEPALLATA genes that are required for proper floral organ development have led to a revision of the ABC model. The largely accepted floral quartet model, which includes the new SEPALLATA function, postulates that the development of a specific floral organ is achieved by the formation of a single complex of different MADS-box proteins. The ultimate fate of the flower is to become a fruit, ensuring dispersal of the seeds and therefore survival of the species. The Arabidopsis fruit is a silique or pod. Only in the last five years important advances have been made in establishing the differentiation of the tissues required for the opening of the fruit: the valve margins and dehiscence zone. Classical genetic analyses and molecular biology approaches have pointed to the involvement of the transcription factors SHP, ALC and IND in the formation of these tissues and of FUL and RPL in repressing this identity in the bordering tissues, valves and replum, respectively.

Arabidopsis↗

Temporally extended gene expression of the ADP-Glc pyrophosphorylase large subunit (AgpL1) leads to increased enzyme activity in developing tomato fruit.

Tomato plants (Solanum lycopersicum) harboring the allele for the AGPase large subunit (AgpL1) derived from the wild species Solanum habrochaites (AgpL1 ( H )) are characterized by higher AGPase activity and increased starch content in the immature fruit, as well as higher soluble solids in the mature fruit following the breakdown of the transient starch, as compared to fruits from plants harboring the cultivated tomato allele (AgpL1 ( E )). Comparisons of AGPase subunit gene expression and protein levels during fruit development indicate that the increase in AGPase activity correlates with a prolonged expression of the AgpL1 gene in the AgpL1 ( H ) high starch line, leading to an extended presence of the L1 protein. The S1 (small subunit) protein also remained for an extended period of fruit development in the AgpL1 ( H ) fruit, linked to the presence of the L1 protein. There were no discernible differences between the kinetic characteristics of the partially purified AGPase-L1(E) and AGPase-L1(H) enzymes. The results indicate that the increased activity of AGPase in the AgpL1 ( H ) tomatoes is due to the extended expression of the regulatory L1 and to the subsequent stability of the heterotetramer in the presence of the L1 protein, implying a role for the large subunit not only in the allosteric control of AGPase activity but also in the stability of the AGPase L1-S1 heterotetramer. The introgression line of S. lycopersicum containing the wild species AgpL1 ( H ) allele is a novel example of transgressive heterosis in which the hybrid multimeric enzyme shows higher activity due to a modulated temporal expression of one of the subunits.

Alleles↗

A mutant Arabidopsis heterotrimeric G-protein beta subunit affects leaf, flower, and fruit development.

A genetic screen was performed to find new mutants with an erecta (er) phenotype and to identify genes that may function with ER, a receptor-like kinase. These mutants were named elk (for erecta-like) and were placed into five complementation groups. We positionally cloned ELK4 and determined that it encodes AGB1, a putative heterotrimeric G-protein beta subunit. Therefore, elk4 was renamed agb1. agb1-1 plants express similar fruit phenotypes, as seen in er plants, but differ from er in that the stem is only slightly shorter than that in the wild type, the pedicel is slightly longer than that in the wild type, and the leaves are rounder than those in er mutants. Molecular analysis of agb1-1 indicates that it is likely a null allele. AGB1 mRNA is expressed in all tissues tested but is highest in the silique. Analysis of agb1-1 er double mutants suggests that AGB1 may function in an ER developmental pathway regulating silique width but that it functions in parallel pathways affecting silique length as well as leaf and stem development. The finding that AGB1 is involved in the control of organ shape suggests that heterotrimeric G-protein signaling is a developmental regulator in Arabidopsis.

Arabidopsis↗

Temporal and transient expression of olive enoyl-ACP reductase gene during flower and fruit development.

Enoyl-ACP reductase is a catalytic component of the fatty acid synthetase (FAS) type II system in plants that is involved in the de novo fatty acid biosynthesis in plastids. A cDNA encoding an enoyl-ACP reductase responsible for the removal of the trans-unsaturated double bonds to form saturated acyl-ACP has been isolated from a library made from ripening fruits of Olea europaea L. The predicted protein contains 393 amino acid residues including a consensus chloroplast specific transit peptide. A strong homology was observed when olive enoyl-ACP reductase aligned with other plant sequences. Southern hybridization analysis revealed that enoyl-ACP reductase is encoded by a single gene in olives. Northern hybridization showed a transient expression of the enoyl-ACP reductase (ENR) gene at early stages of drupe (5-7 weeks after flowering, WAF), embryo and endosperm (13-16 WAF) while in mesocarp (13-19 WAF) the expression remained at high levels. In situ hybridization showed particularly prominent expression in the palisade and vascular tissue of young leaves, the tapetum, developing pollen grains and vascular tissue of anthers and to less extent in the embryo sac and transmitting tissue of the carpel. The distinctive spatial and temporal regulation of the ENR gene is consistent with major roles, not only in thylakoid membrane formation and fatty acid deposition, but also in the provision of precursor molecules for the biosynthesis of oxilipins that are important in plant tissues involved in transportation and reproduction.

Amino Acid Sequence↗

Temporal and spatial gene expression of cytochrome B5 during flower and fruit development in olives.

We report the characterisation of two cytochrome b5 genes and their spatial and temporal patterns of expression during development in olive, Olea europaea. A PCR-generated probe, based on a tobacco cytochrome b5 sequence, was used to isolate two full-length cDNA clones (cytochrome b5-15 and cytochrome b5-38) from a library derived from 13 WAF olive fruits. The cDNAs encoded proteins of 17.0 and 17.7 kDa, which contained all the characteristic motifs of cytochromes b5 from other organisms and exhibited 63% identity and 85% similarity with each other. The olive cytochrome b5-15 cDNA was then used as a probe for more detailed analysis. Southern blotting revealed a gene family of at least 4-6 members while northern blotting and in situ hybridisation showed a highly specific pattern of gene expression. Very low levels of cytochrome b5 mRNA were detected in tissues characterised by high rates of lipid accumulation, such as young expanding leaves, maturing seeds and ripening mesocarp. The cytochrome b5 genes were not induced at 6 degrees C and their response to ABA was relatively slow compared with fatty acid desaturase genes. In contrast, high levels of cytochrome b5 gene expression were found in young fruits at the pattern formation (globular/heart) stage of embryogenesis and in vascular and transmitting tissues of male and female reproductive organs. The data are consistent with a major role for cytochrome b5 in developmental processes related to plant reproduction in addition to being an electron donor to microsomal desaturases.

Abscisic Acid↗

Morphology and anatomy of developing fruits and seeds of Vismia guianensis (Aubl.) Choisy (Clusiaceae).

Morphological, structural and developmental features of fruits and seeds of Vismia guianensis (Aubl.) Choisy. are here presented, with the purpose to elucidate their structure and to contribute to taxonomical and ecological studies of the family. The fruit is a berry and the "rind" is constituted by the exocarp and by the subepidermal layers which constitute the mesocarp. The reddish pulp, rich in starch, is constituted by the parenchymatic mesocarp, with branched secretory ducts and vascular bundles, joined with the endocarp, which arises from a ventral meristem. The bitegmic, anatropous ovules, provided by the endothelium, develop into anatropous, bitegmic and exalbuminous seeds. The reddish uniseriate testa shows phenolic contents. The tegmen becomes crushed resting only the uniseriate exotegmen with undulate, thick walled and lignified cells, which contain a number of calcium oxalate prismatic crystals. The embryo, rich in lipids, is straight and shows foliaceous cotyledons.

Ericales↗

Isolation and transcript analysis of gibberellin 20-oxidase genes in pea and bean in relation to fruit development.

PCR was used with degenerate primers based on conserved amino acid sequences in gibberellin (GA) 20-oxidases to isolate cDNA clones for these enzymes from young seeds of pea (Pisum sativum) and developing embryos of French bean (Phaseolus vulgaris). One GA 20-oxidase cDNA (Ps27-12) was obtained from pea and three (Pv 15-11, Pv73-1 and Pv85-26) from bean. Their identities were confirmed by demonstrating that fusion proteins expressed in Escherichia coli exhibited GA 20-oxidase activity, converting [14C]GA12 to [14C]GA9. The intermediates in this three-step reaction, GA15 and GA24, were also identified as products. The expression proteins from three of the clones (Ps27-12, Pv15-11 and Pv73-1) were also shown to convert GA53 to GA20, as effectively as they did GA12. On the basis of transcript levels measured by northern blot analysis, the pea GA 20-oxidase gene is most highly expressed in young leaves, fully expanded internodes, very young seeds (until 4 days after anthesis) and expanding pods (from 3 days after anthesis at least until day 6). Expression in pods from 3-day-old unpollinated ovaries is higher than in those from pollinated ovaries. Treatment of unpollinated ovaries with GA3 to induce parthenocarpic fruit-set severely reduced the amount of GA 20-oxidase mRNA, whereas treatment with 2,4-D, although inducing fruit-set, did not reduce the levels of these transcripts. Plant decapitation above an unpollinated ovary resulted in very high levels of GA 20-oxidase mRNA in the pod. The three GA 20-oxidase genes from French bean showed very different patterns of expression: Pv 15-1 was expressed in the roots, young leaves, and developing seeds, but most highly in immature cotyledons, while Pv73-1 has a similar expression pattern to Ps27-12, with transcripts found only in young seeds and young leaves, where it was particularly abundant. Transcripts corresponding to Pv85-26 were detected in developing seeds, and just traces in the young leaves. Southern blot analysis indicated that the bean GA 20-oxidases are each encoded by single-copy genes, whereas one more gene, homologous to Ps27-12, could also exist in pea.

Amino Acid Sequence↗

Anatomical, histological, and histochemical changes in grape seeds from Vitis vinifera L. cv Cabernet franc during fruit development.

Cabernet franc berries were sampled at five stages from berry set to harvest from an experimental vineyard in mid Loire Valley. Seeds were collected from representative berries in term of stage of development. The evolution of seed was followed both macro- and microscopically. For microscopy analysis, seeds were cut, put in a fixation solution, and cut into thin sections with a microtome. Five staining solutions were used for each seed sample: toluidine blue O, phloroglucinol, periodic acid-Schiff's reagent and naphtol blue black, vanillin, and p-dimethylaminocinnamaldehyde. Toluidine blue O staining revealed the evolution of tissue structures during grape seed development. We studied the changes in chemical compounds (lignin, polysaccharides, proteins, and tannins) with the other reagents. Seed lignification was achieved at veraison. Proanthocyanidins were localized in epidermis, inner cells of the soft seed coat, and inner cell layer of the inner integument. Finally, the localization of flavan-3-ols was linked with changes in cell walls of the outer integument.

France↗

Evidence for apoplasmic phloem unloading in developing apple fruit.

The phloem unloading pathway remains unclear in fleshy fruits accumulating a high level of soluble sugars. A structural investigation in apple fruit (Malus domestica Borkh. cv Golden Delicious) showed that the sieve element-companion cell complex of the sepal bundles feeding the fruit flesh is symplasmically isolated over fruit development. 14C-autoradiography indicated that the phloem of the sepal bundles was functional for unloading. Confocal laser scanning microscopy imaging of carboxyfluorescein unloading showed that the dye remained confined to the phloem strands of the sepal bundles from the basal to the apical region of the fruit. A 52-kD putative monosaccharide transporter was immunolocalized predominantly in the plasma membrane of both the sieve elements and parenchyma cells and its amount increased during fruit development. A 90-kD plasma membrane H(+)-ATPase was also localized in the plasma membrane of the sieve element-companion cell complex. Studies of [14C]sorbitol unloading suggested that an energy-driven monosaccharide transporter may be functional in phloem unloading. These data provide clear evidence for an apoplasmic phloem unloading pathway in apple fruit and give information on the structural and molecular features involved in this process.

Biological Transport↗

Energy conservation and dissipation in mitochondria isolated from developing tomato fruit of ethylene-defective mutants failing normal ripening: the effect of ethephon, a chemical precursor of ethylene.

Alternative oxidase (AOX) and uncoupling protein (UCP) are present simultaneously in tomato fruit mitochondria. In a previous work, it has been shown that protein expression and activity of these two energy-dissipating systems exhibit large variations during tomato fruit development and ripening on the vine. It has been suggested that AOX and UCP could be responsible for the respiration increase at the end of ripening and that the cytochrome pathway could be implicated in the climacteric respiratory burst before the onset of ripening. In this study, the use of tomato mutants that fail normal ripening because of deficiencies in ethylene perception or production as well as the treatment of one selected mutant with a chemical precursor of ethylene have revealed that the bioenergetics of tomato fruit development and ripening is under the control of this plant hormone. Indeed, the evolution pattern of bioenergetic features changes with the type of mutation and with the introduction of ethylene into an ethylene-synthesis-deficient tomato fruit mutant during its induced ripening.

Carrier Proteins↗

Triacylglycerol and phospholipid composition of hazelnut (Corylus avellana L.) lipid fraction during fruit development.

We analyzed the triacylglycerol and phospholipid contents of hazelnuts from early development to maturity. Both were analyzed by high-performance liquid chromatography coupled to a light scattering detector. Trioleylglycerol, linoleyl-dioleylglycerol, and palmitoyl-dioleylglycerol were the most predominant triacylglycerols throughout development. Triacylglycerols showed small variation during hazelnut development. Phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol were the most abundant phospholipids. Traces of phosphatidic acid were also detected. The statistical analysis showed that the positive correlation among the individual phospholipid contents was significant. Phospholipid contents showed a steep decrease during hazelnut development. Triacylgycerols and phosphatidylcholine were isolated by preparative thin-layer chromatography, and their fatty acid profile was determined by gas-liquid chromatography. Triacylglycerols showed a high percentage of monounsaturated fatty acid moieties, whereas phosphatidylcholine had the highest percentage of saturated and monounsaturated fatty acid moieties. The polyunsaturated fatty acid moiety showed low percentages in the triacylglycerol and phospholipid backbone.

Chromatography, High Pressure Liquid↗

Sucrose uptake, invertase localization and gene expression in developing fruit of Lycopersicon esculentum and the sucrose-accumulating Lycopersicon hirsutum.

By using immunolocalization and differential extraction methods we show that only apoplastic invertase, but not vacuolar invertase, was present in the mature, sucrose-accumulating L. hirsutum pericarp. In contrast, in the hexose-accumulating L. esculentum fruit, both the apoplastic and vacuolar invertase activities and protein content increase in the mature fruit. Quantitative expression studies of the soluble invertase gene (TIV1) and the apoplastic invertase genes (LINs) showed that only TIV1 gene expression could account for the species and developmental differences of both soluble and insoluble enzyme activity of the pericarp. The expression of the LIN genes encoding for apoplastic tomato invertases was unrelated to the differences in bound enzyme activity and could not account for the rise in bound invertase activity in the mature L. esculentum fruit. Evidence is presented that the bound invertase activity of tomato fruit is also the TIV1 gene product. The presence of apoplastic invertase in the mature sucrose-accumulating L. hirsutum fruit suggests a hydrolysis-resynthesis mechanism of sucrose uptake. In order to test this hypothesis, we studied short- and long-term uptakes of asymmetrically labelled 3H-fructosyl-sucrose accompanied by compartmental analysis of the sugars in attached whole fruits of L. hirsutum and L. esculentum. The results indicate that hydrolysis-resynthesis is slow in the sucrose-accumulating fruit but is not an integral part of an uptake and compartmentation mechanism.

Journal Article↗