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PubMed2002

Distinct physiological roles of fructokinase isozymes revealed by gene-specific suppression of Frk1 and Frk2 expression in tomato.

There are two divergent fructokinase isozymes, Frk1 and Frk2 in tomato (Lycopersicon esculentum Mill.) plants. To investigate the physiological functions of each isozyme, the expression of each fructokinase mRNA was independently suppressed in transgenic tomato plants, and the respective phenotypes were evaluated. Suppression of Frk1 expression resulted in delayed flowering at the first inflorescence. Suppression of Frk2 did not effect flowering time but resulted in growth inhibition of stems and roots, reduction of flower and fruit number, and reduction of seed number per fruit. Localization of Frk1 and Frk2 mRNA accumulation by in situ hybridization in wild-type tomato fruit tissue indicated that Frk2 is expressed specifically in early tomato seed development. Fruit hexose and starch content were not effected by the suppression of either Frk gene alone. The results collectively indicate that flowering time is specifically promoted by Frk1 and that Frk2 plays specific roles in contributing to stem and root growth and to seed development. Because Frk1 and Frk2 gene expression was suppressed individually in transgenic plants, other significant metabolic roles of fructokinases may not have been observed if Frk1 and Frk2 play, at least partially, redundant metabolic roles.

Enzyme Activation↗
PubMed2003

Expression of plastid-encoded photosynthetic genes during chloroplast or chromoplast differentiation in Cucurbitae pepo L. fruits.

The objective of the study was to determine the patterns of expression of two photosynthetic genes rbcL and psbA, during chloroplast and chromoplast differentiation in fruit tissues of three Cucurbitae pepo L. cultivars: Early Prolific, Foodhook Zucchini and Bicolor Gourds. In two Early Prolific isogenic lines, YYBB and YYB+B+, the steady-state amounts of rbcL and psbA transcripts increased with fruit development upto 14 days post-pollination. The YYB+B+ line in which chloroplast differentiates into chromoplast at about pollination, did not show significantly higher amounts of both transcripts compared to YYBB, in which chromoplast develops early prior to pollination. In the Bicolor Gourds, in which the chromoplast and chloroplast containing tissues lie in juxtaposition on the same fruit, showed little differences in rbcL and psbA transcripts between the two tissues, if any the chromoplast containing tissue contained more of both transcripts than the chloroplast containing tissue. In Fordhook Zucchini fruits, where the chloroplast containing tissue developed early prior to pollination and was maintained, the steady-state amounts of rbcL transcripts increased to a maximum at 3 days post-pollination and levelled at 14 and 21 days post-pollination. In contrast, in Fordhook Zucchini fruits, the psbA transcript increased gradually up to 21 days post-pollination. In Fordhook Zucchini, the apparent ratios of psbA transcripts versus rbcL transcripts ranged from 2.5 to 3.9, at day 3 to 21 post-pollination, while in Bicolor Gourds were 2.9 and 4.5 at days 14 and 21 post-pollination. The two photosynthetic genes, psbA and rbcL were developmentally regulated and differentially expressed. However, their expression in chloroplast containing fruit tissues was not higher than in the chromoplast containing fruit tissues.

Blotting, Northern↗
PubMed1995

Alternative oxidase from mango (Mangifera indica, L.) is differentially regulated during fruit ripening.

Alternative oxidase is a respiratory-chain component of higher plants and fungi that catalyzes cyanide-resistant oxygen consumption. The activity of a alternative oxidase has been detected during ripening in several climacteric fruit including mango (Mangifera indica L.). Synthetic oligonucleotides, corresponding to conserved regions of the Sauromatum guttatum and Arabidopsis thaliana nucleotide sequences, were used as primers for polymerase chain reaction to amplify genomic DNA extracted from mango leaves. The 623-bp fragment was found to encode an open reading frame of 207 amino acids showing high identity to the S. guttatum enzyme. Using this fragment to screen a ripe mango mesocarp cDNA library, one full-length cDNA clone, designated pAOMI.1, was obtained that contained an open reading frame encoding a polypeptide of 318 amino acids. The predicted amino-acid sequence exhibited 62, 64 and 68% identity to the S. guttatum, soybean, and A. thaliana enzymes respectively, indicating that this cDNA encodes a mango homologue of the alternative oxidase. Gel blot hybridization showed that pAOMI.1 is likely to be encoded by a single-copy gene. The 1.6 kb-transcript was induced during mango fruit ripening although the transcript was clearly detectable in unripe and developing fruit. Antibodies raised against the S. guttatum enzyme recognized three bands of approximately 27, approximately 33 and approximately 36 kDa from mitochondrial mango proteins. Two of the bands were detectable before ripening and increase in ripe fruit, the other band (27 kDa) was barely present in unripe fruit but accumulated during ripening. The clone pAOMI.1 was able to complement an Escherichia coli hemA mutant deficient in cytochrome-mediated aerobic respiration. This is the first report on the analysis of alternative oxidase at the molecular level during the ripening of a climacteric fruit.

Amino Acid Sequence↗
PubMed2002

Characterization of a fertilization-induced and developmentally regulated plasma-membrane aquaporin expressed in reproductive tissues, in the wild potato Solanum chacoense Bitt.

Fertilization triggers a unique and complex developmental program leading to embryogenesis and seed set. Growth and differentiation are accompanied by rapid metabolic changes as well as massive cellular reorganization inside the fertilized ovule. Using differential display to isolate genes involved in pollen-pistil interactions and early fertilization events, we isolated from Solanum chacoense Bitt. a fertilization-induced plasma-membrane intrinsic protein of the PIP2 family that is predominantly expressed in pistil and anthers tissues. Major intrinsic proteins (MIPs) consist of a large family of highly conserved membrane-spanning proteins that are mainly represented in plants by the aquaporins. Aquaporins, mainly of the PIP and TIP type, have been shown in many species to serve as water channels. In S. chacoense the ScPIP2a mRNA is developmentally regulated during anthesis, with mRNA levels gradually decreasing as the pistil reaches maturity. In flowers, strongest expression was observed in elongating styles, in stamens, and transiently in ovules following fertilization. In styles, maximal expression levels correlated with phases of rapid style elongation and with the formation of epidermal papillae. ScPIP2a mRNA was also strongly expressed in developing fruit, consistent with a role in cell expansion during maturation and development.

Amino Acid Sequence↗
PubMed2001

Auxin modification of the incompatibility response in Theobroma cacao.

The time course and control of floral abscission and fruit set in Theobroma cacao were studied after spray application of growth regulators. 1-Naphthaleneacetic acid (NAA) prevented flower abscission in a concentration dependent manner and induced the early stages of fruit development. The cytokinin benzylaminopurine (BAP) counteracted NAA but resulted in longer fruit retention. Measurements of endogenous levels of indole-3-acetic acid showed an inverse correlation between the number of flowers per plant and auxin content. The results suggest that the genetic control of self-incompatibility in T. cacao may be modulated by the hormonal content of the flower.

Journal Article↗
PubMed1995

Requirement for Drosophila cytoplasmic tropomyosin in oskar mRNA localization.

The localization of oskar (osk) RNA to the posterior pole of the developing fruit fly (Drosophila) oocyte induces the assembly of pole plasm, causing development of the abdomen and germ line. Failure to localize oskar RNA results in embryos that lack abdomen and germ cells. Conversely, mis-targeting of oskar RNA to the anterior of the oocyte causes formation of ectopic abdomen and germ cells at the anterior pole. Maternal mutants that have reduced pole plasm activity produce sterile adults with normal abdominal development, suggesting that germ cells are more sensitive than abdomen to defects in pole plasm assembly. Thus mutations in genes that reduce oskar RNA localization or activity can be recovered as viable sterile adults. In a screen for mutants defective in germ cell formation, we isolated nine alleles of the tropomyosin II gene. Here we show that mutations in tropomyosin II (TmII) virtually abolish oskar RNA localization to the posterior pole, suggesting an involvement of the actin network in oskar RNA localization.

Actins↗
PubMed2002

Tobacco to tomatoes: a phylogenetic perspective on fruit diversity in the Solanaceae.

The Solanaceae contains many species of agricultural importance. Several of these are cultivated for their fruits, such as the tomato, the pepper and the aubergine. The family is very diverse in fruit type with capsules, drupes, pyrenes, berries, and several sorts of dehiscent non-capsular fruits occurring in the 90+ genera. In this paper, recent work on fruit type evolution in angiosperms is reviewed in relation to dispersal agents and habitat ecology. Defining fruit types in the Solanaceae in a simple five state system, then mapping them onto a previously published molecular phylogeny based on chloroplast DNA allows discussion of the evolution of these fruit types in a phylogenetic framework. Capsules are plesiomorphic in the family, and although berries are a synapomorphy (shared derived character) for a large clade including the genus Solanum (tomatoes and aubergines), they have arisen several times in the family as a whole. Problems with homology of drupes and pyrenes are discussed, and areas for future investigation of fruit structure homology identified. The distribution of fruit types in the large and diverse genus Solanum is also discussed in the light of monophyletic groups identified using chloroplast gene sequences. This variety is related to recent advances in the understanding of the molecular biology of fruit development. Finally, several key areas of future comparative, phylogenetic investigation into fruit type evolution in the family are highlighted.

Atropa belladonna↗
PubMed2004

Pectin esterase gene family in strawberry fruit: study of FaPE1, a ripening-specific isoform.

Pectin esterases (PE, EC 3.1.1.11) catalyse the demethylation of pectin. As a result of its activity, structural interactions among cell wall components during cell wall turnover and loosening are affected. In plants, PEs are typically encoded by a gene family. This family has been studied in strawberry (Fragaria x ananassa Duch.) in order to investigate the role of distinct PE genes during fruit ripening and senescence. By a combination of a PCR-based library screening and RT-PCR four different strawberry PE cDNAs, termed FaPE1 to FaPE4, have been isolated. Differential expression of each FaPE gene in various organs and during fruit development was revealed by northern blot. FaPE1 is specifically expressed in fruit, showing an increasing expression during the ripening process up to a maximum in the turning stage. Concerning hormone regulation, auxin treatment increased FaPE1 mRNA levels in green fruit, whereas exogenous ethylene decreased FaPE1 mRNA levels in ripe and senescing fruits. It is proposed that this repression of FaPE1 expression could be involved in textural changes occurring during fruit senescence.

Amino Acid Sequence↗
PubMed1994

Endo-1,4-[beta]-Glucanase, Xyloglucanase, and Xyloglucan Endo-Transglycosylase Activities Versus Potential Substrates in Ripening Tomatoes.

In ripening fruits of tomato (Lycopersicon esculentum L. var 83-G-38), the amounts of cellulose and xyloglucan (XG) remained constant during tissue softening, but the relative molecular weight (Mr) of XG decreased markedly and the Mr of cellulose declined slightly. These changes could have been due to activities of non-specific endo-1,4-[beta]-glucanases and/or buffer-soluble XG endo-transglycosylase, both of which increased when tissue firmness declined most rapidly. Tomato extracts also reduced the viscosity of XG solutions, especially in the presence of added XG oligosac-charides. This depolymerizing (XGase) capacity differed from [beta]-glucanase and XG transglycosylase activity (a) by being almost entirely buffer insoluble, and (b) by declining precipitously during fruit softening. Although it disappeared from ripe fruit, XGase may have functioned in promoting wall loosening at earlier stages of fruit development when its activity was highest. By contrast, during aging of fruit in the ripening-inhibited mutant rin there was no change in Mr of XG or cellulose, and activities of [beta]-glucanases and XG transglycosylase were lower than in wild-type tomato. Nevertheless, some softening of the fruit did take place over time and XG amounts declined, possibly because high XGase activity was maintained in the mutant, unlike in wild-type fruit.

Journal Article↗
PubMed2000

Fermentative metabolism in grape berries: isolation and characterization of pyruvate decarboxylase cDNA and analysis of its expression throughout berry development.

The involvement of pyruvate decarboxylase (PDC) in the control of alcohol production during ripening of fruit tissues under aerobic conditions has been only partially studied. Enzymological studies showed a significant increase in PDC activity during the ripening of oranges and pears, concurrently with the induction of ethanol production. In tomato, on the other hand, the induction of ethanol production and ADH gene expression after the onset of ripening was not accompanied by induction of PDC activity. The isolation of PDC cDNA from fruits has not yet been reported, nor has its expression pattern during fruit development. We report here the isolation of a cDNA clone encoding for a grape PDC and the characterization of its expression throughout berry development. The pattern of PDC gene expression throughout berry development, combined with earlier findings on constitutive PDC activity in the berry, may suggest that PDC is not the limiting factor for the production of ethanol in the berry, which is induced only after the onset of berry ripening. Alternatively, the induction of ADH gene expression, which occurs only after the onset of ripening in both tomatoes and grape berries, may serve as a regulator of ethanol production in response to a ripening-related cue.

Journal Article↗
PubMed1999

Protective effect of ascorbic acid against the browning developed in apple fruit treated with high hydrostatic pressure.

Apple Reineta variety was used as an apple dessert. The 1-1.5-cm cubes were immersed in a sucrose solution (30% w/v) and subjected to high pressure (HP) of 400 MPa for 30 min at 5 degrees C. Different ascorbic acid concentrations were used to protect the fruit from the browning developed after the HP treatment. After 2 months of storage at 5 degrees C, no brown color was observed in the samples treated with 20 mM ascorbic acid, and they were acceptable to consumers. However, untreated samples presented fermentation, and they were not acceptable to consumers. The electric conductivity and potassium content were found to be good indicators of the metabolites released from the fruit to the solution in samples treated with high pressure. HP did not affect the peroxidase activity but eliminated the microbial population.

Ascorbic Acid↗
PubMed2004

DVL, a novel class of small polypeptides: overexpression alters Arabidopsis development.

Small polypeptides can act as important regulatory molecules that coordinate cellular responses required for differentiation, growth, and development. In a gain-of-function genetic screen for genes that influence fruit development in Arabidopsis, we identified a novel gene -DEVIL1 (DVL1) - encoding a small protein. Overexpression of DVL1 results in pleiotropic phenotypes featured by shortened stature, rounder rosette leaves, clustered inflorescences, shortened pedicles, and siliques with pronged tips. cDNA analysis indicates that DVL1 has a 153-nucleotide (nt) open-reading frame (ORF) encoding a 51-amino acid polypeptide that shares no significant similarity to previously identified proteins. Sequence alignment shows that DVL1 belongs to a family of related genes that are limited to angiosperm plants. Ectopic overexpression of each of the five closely related Arabidopsis DVL genes causes similar phenotypic changes, suggesting overlapping function in the DVL gene family. Point mutations of conserved amino acids in the C-terminal region of the DVL1 polypeptide reveal that these conserved residues are required for DVL1-overexpression phenotypes. Our results show that the DVL family is a novel class of small polypeptides and the overexpression phenotypes suggest that these polypeptides may have a role in plant development.

Amino Acid Sequence↗
PubMed2006

Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10.

Anthocyanin concentration is an important determinant of the colour of many fruits. In apple (Malus x domestica), centuries of breeding have produced numerous varieties in which levels of anthocyanin pigment vary widely and change in response to environmental and developmental stimuli. The apple fruit cortex is usually colourless, although germplasm does exist where the cortex is highly pigmented due to the accumulation of either anthocyanins or carotenoids. From studies in a diverse array of plant species, it is apparent that anthocyanin biosynthesis is controlled at the level of transcription. Here we report the transcript levels of the anthocyanin biosynthetic genes in a red-fleshed apple compared with a white-fleshed cultivar. We also describe an apple MYB transcription factor, MdMYB10, that is similar in sequence to known anthocyanin regulators in other species. We further show that this transcription factor can induce anthocyanin accumulation in both heterologous and homologous systems, generating pigmented patches in transient assays in tobacco leaves and highly pigmented apple plants following stable transformation with constitutively expressed MdMYB10. Efficient induction of anthocyanin biosynthesis in transient assays by MdMYB10 was dependent on the co-expression of two distinct bHLH proteins from apple, MdbHLH3 and MdbHLH33. The strong correlation between the expression of MdMYB10 and apple anthocyanin levels during fruit development suggests that this transcription factor is responsible for controlling anthocyanin biosynthesis in apple fruit; in the red-fleshed cultivar and in the skin of other varieties, there is an induction of MdMYB10 expression concurrent with colour formation during development. Characterization of MdMYB10 has implications for the development of new varieties through classical breeding or a biotechnological approach.

Amino Acid Sequence↗
PubMed1999

Comparative morphology and physiology of fruit and seed development in the two shrubs Rhus aromatica and R. glabra (Anacardiaceae).

Morphology and physiology of fruit and seed development were compared in Rhus aromatica and R. glabra (Anacardiaceae), both of which produce drupes with water-impermeable endocarps. Phenology of flowering/fruiting of the two species at the study site was separated by ∼2 mo. However, they were similar in the timetable and pattern of fruit and seed development; it took ∼2 mo and ∼1.5 mo for flowers of Rhus aromatica and R. glabra, respectively, to develop into mature drupes. The single sigmoidal growth curve for increase in fruit size and in dry mass of these two species differs from the double-sigmoidal one described for typical commercial drupes such as peach and plum. Order of attainment of maximum size was fruit and endocarp (same time), seed coat, and embryo. By the time fruits turned red, the embryo had reached full size and become germinable; moisture content of seed plus endocarp had decreased to ∼40%. The endocarp was the last fruit component to reach physiological maturity, which coincided with development of its impermeability and a seed plus endocarp moisture content of <10%. At this time, ∼50, 37, and 13% of the dry mass of the drupe was allocated to the exocarp plus mesocarp unit, endocarp, and seed, respectively. The time course of fruit and seed development in these two species is much faster than that reported for other Anacardiaceae, including Rhus lancea, Protorhus, and Pistacia.

Journal Article↗
PubMed1991

Regulation of metallocarboxypeptidase inhibitor gene expression in tomato.

Tomato fruits contain a metallocarboxypeptidase inhibitor (MCPI) the sequence of which has already been determined. Here we report the isolation of a tomato cDNA clone that encodes the mature MCPI protein as well as an N-terminal signal peptide for entry into the secretory system and an eight amino acid carboxyterminal extension. MCPI RNA is present at very high levels in anthesis stage ovaries and decreases quite rapidly during fruit development. MCPI protein accumulation reflects the pattern of MCPI RNA accumulation in fruit, consistent with a transcriptional control of MCPI gene activity. In leaves, the levels of MCPI RNA and protein are very low. Wounding of the leaves causes a dramatic (100-fold) increase in steady-state level of MCPI RNA without a concomitant increase in MCPI protein level suggesting a control at the post-transcriptional or translational level of gene expression. Genomic DNA blot hybridization data indicate that MCPI in tomato may be encoded by a single gene.

Amino Acid Sequence↗
PubMed2006

Preferential localization of abscisic acid in primordial and nursing cells of reproductive organs of Arabidopsis and cucumber.

Abscisic acid (ABA) is known to function in plant stress responses and seed dormancy, and much is known about its detailed mechanisms of signal transduction. Recent studies suggest that this hormone may also play important roles in sugar signaling and assimilate distribution during fruit development. However, little is known about the role of ABA in actively growing or differentiating fruits and other plant organs or tissues. To explore whether ABA functions during the early development of reproductive organs, we carried out ABA immunolocalization using monoclonal antibodies. The specific ABA accumulation pattern was verified by gas chromatography-mass spectrometry (GC-MS). ABA was not only detected in primordial cells of flower organs, but was also detected in nursing cells (e.g. tapetum and integuments), which function in supplying nutrition for germ cell development. These findings suggest that, in addition to its well-known function as a 'negative hormone', ABA may play some 'positive' roles during plant development, including possible involvement in the regulation of assimilate distribution.

Abscisic Acid↗