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PubMed2000

A novel alpha-amylase gene is transiently upregulated during low temperature exposure in apple fruit.

An alpha-amylase gene product was isolated from apple fruit by reverse-transcriptase PCR using redundant primers, followed by 5' and 3' RACE. The gene is a member of a small gene family. It encodes a putative 46.9 kDa protein that is most similar to an alpha-amylase gene from potato (GenBank accession M79328). In apple fruit this new gene was expressed at low levels, as detected by reverse-transcriptase PCR, in a number of plant tissues and during fruit development. Highest levels of mRNA for this transcript were observed 3 to 9 days after placing apple fruit at 0.5 degrees C. Phylogenetic analysis of amino acid sequence places the potato and apple proteins as a distinct and separate new subgroup within the plant alpha-amylases, which appears to have diverged prior to the split between monocotyledonous and dicotyledonous plants. These two divergent alpha-amylases lack the standard signal peptide structures found in all other plant alpha-amylases, and have sequence differences within the B-domain and C-domain. However, comparisons with structures of known starch hydrolases suggest that these differences are unlikely to affect the enzymatic alpha-1,4-amylase function of the protein. This is the first report of upregulation of a dicotyledonous alpha-amylase in response to low temperature, and confirms the presence of a new family of alpha-amylases in plants.

Amino Acid Sequence↗

PubMed1972

Translocation and Metabolism of Ricinine in the Castor Bean Plant, Ricinus communis L.

Ricinine-3,5-(14)C (N-methyl-3-cyano-4-methoxy-2-pyridone) administered to senescent leaves of Ricinus communis L. was translocated to all other tissues of the plant. Developing fruit and especially seeds were found to be labeled the most rapidly. Young growing leaves and other developing tissues of the plant imported ricinine from the senescent leaves much more quickly than mature leaves. Relative intensities of the radioactive ricinine imported and deposited in various tissues indicate a possible functional role of ricinine in the castor bean plant. Data on N-demethyl ricinine presented here may stimulate interest in the possible physiological role of the ricinine to N-demethyl ricinine interconversion.

Journal Article↗

PubMed2006

A gene encoding starch branching enzyme I (SBEI) in apple (Malusxdomestica, Rosaceae) and its phylogenetic relationship to Sbe genes from other angiosperms.

An apple starch-branching enzyme SbeI gene (GenBank Accession No. DQ115404) has been isolated, cloned, and sequenced. The SbeI is a single copy gene in the apple genome, consisting of 14 exons and 13 introns, and covering 6075bp. As detected by RT-PCR, the apple SbeI is expressed at very low levels during early stages of fruit development; while, the highest levels of mRNA transcripts are observed at approximately 44 days post-pollination. Besides fruits, the apple SbeI is also expressed in buds and flowers, and very weakly in leaves. The genomic structure of SbeI in apple is strikingly similar to those reported so far in grasses (Poaceae), with exons 4 through 13 being of identical lengths in both apple and grasses. Moreover, structure similarities in exon lengths have also been detected in SbeII genes of both grasses and eudicots. These findings prompted the investigation of the evolutionary process of the Sbe gene family in angiosperms. A total of 26 Sbe sequences, representing an array of monocots and eudicots, are investigated in this study. Phylogenetic analysis has suggested that Sbe genes have duplicated into SbeI and SbeII prior to the divergence of moncots from eudicots. The SbeII gene is further duplicated into SbeIIa and SbeIIb prior to the radiation of grasses; however, it is not yet clear whether this duplication event has occurred before or after the radiation of the eudicots.

1,4-alpha-Glucan Branching Enzyme↗

PubMed2004

Transcriptional programs of early reproductive stages in Arabidopsis.

The life cycle of flowering plants alternates between a diploid sporophytic and a haploid gametophytic generation. After fertilization of each the egg and central cells by one male gamete, the development of both fertilization products occurs coordinated with the maternally derived seed coat and carpel tissues forming the fruit. The reproduction program is likely to involve the concerted activity of many genes. To identify genes with specific functions during reproduction, we have analyzed the expression profile of more than 22,000 genes present on the Arabidopsis ATH1 microarray during three stages of flower and fruit development. We found 1,886 genes regulated during reproductive development and 1,043 genes that were specifically expressed during reproduction. When compared to cells from an Arabidopsis suspension culture, S-phase genes were underrepresented and G2 and M-phase genes were strongly enriched in the set of specific genes, indicating that important functions during reproduction are exerted in the G2 and M phases of the cell cycle. Many potential signaling components, such as receptor-like protein kinases, phosphatases, and transcription factors, were present in both groups of genes. Members of the YABBY, MADS box, and Myb transcription factor families were significantly overrepresented in the group of specific genes, revealing an important role of these families during reproduction. Furthermore, we found a significant enrichment of predicted secreted proteins smaller than 15 kD that could function directly as signaling molecules or as precursors for peptide hormones. Our study provides a basis for targeted reverse-genetic approaches aimed to identify key genes of reproductive development in plants.

Arabidopsis↗

PubMed1978

Carbon Dioxide Fixation in the Carbon Economy of Developing Seeds of Lupinus albus (L.).

The effects of CO(2) concentration and illumination on net gas exchange and the pathway of (14)CO(2) fixation in detached seeds from developing fruits of Lupinus albus (L.) have been studied.Increasing the CO(2) concentration in the surrounding atmosphere (from 0.03 to 3.0% [v/v] in air) decreased CO(2) efflux by detached seeds either exposed to the light flux equivalent to that transmitted by the pod wall (500 to 600 micro-Einsteins per square meter per second) in full sunlight or held in darkness. Above 1% CO(2) detached seeds made a net gain of CO(2) in the light (up to 0.4 milligrams of CO(2) fixed per gram fresh weight per hour) but (14)CO(2) injected into the gas space of intact fruits (containing around 1.5% CO(2) naturally) was fixed mainly by the pod and little by the seeds.Throughout development seeds contained ribulose-1,5-bisphosphate carboxylase activity (EC 4.1.1.39), especially in the embryo (up to 99 micromoles of CO(2) fixed per gram fresh weight per hour) and phosphoenolpyruvate carboxylase (EC 4.1.1.31) in both testa (up to 280 micromoles of CO(2) fixed per gram fresh weight per hour) and embryo (up to 355 micromoles of CO(2) fixed per gram fresh weight per hour).In kinetic experiments the most significant early formed product of (14)CO(2) fixation in both light and dark was malate but in the light phosphoglyceric acid and sugar phosphates were also rapidly labeled. (14)CO(2) fixation in the light was linked to the synthesis of sugars and amino acids but in the dark labeled sugars were not formed.

Journal Article↗

PubMed1988

Sucrose and Malic Acid as the Compounds Exported to the Apical Bud of Pea following CO(2) Labeling of the Fruit : No Evidence for a Senescence Factor.

The G2 line of peas (Pisum sativum L.) displays senescence and death of the apical bud only in long days and in the presence of fruit. As the removal of fruit prevents senescence, one possible mechanism by which fruits induce senescence is that the fruits produce some ;senescence factor' under long day conditions, which is then transported to the apical bud. Allowing developing fruits to photosynthesize in the presence of (14)CO(2) results in the recovery of label in the apical bud. In order to determine the chemical nature of this radiolabeled material, fruits of G2 peas, growing under long days, were exposed to (14)CO(2) at the time when the first senescence symptoms start to appear. The radiolabeled material from apical buds was then extracted, purified, and identified. Using HPLC and GC-MS the major labeled compound found in the apical bud following exposure of pea fruits to (14)CO(2) was identified as sucrose, while malic acid was identified as the major ethyl acetate-soluble compound. These compounds accounted for about 73 and 16%, respectively, of the radioactivity in the apical bud. No other compounds were present in significant amounts. As neither of these chemicals is likely to have any kind of senescence effect, we report no evidence for a senescence factor.

Journal Article↗

PubMed2026

Phytochrome-interacting factor 1b (SlPIF1b) affects the fruit quality of tomato by regulating chloroplast development.

The increased abundance and functionality of fruit chloroplasts could promote the accumulation of nutrients and flavor in the fruit. Tomato fruit has fully developed fruit chloroplasts, whose abundance and functionality have much untapped potential in improving fruit quality by controlling fruit chloroplast development. Previous studies have identified many regulatory factors that specifically regulate fruit chloroplast development in tomatoes, but there are fewer reports on tomato phytochrome-interacting factors (SlPIFs). Arabidopsis AtPIFs have been implicated in chloroplast development and chlorophyll biosynthesis. In this study, we identified and characterized an SlPIF1b mutant in tomato, named GS, which exhibited a dark green fruit shoulder with enhanced chloroplast development. RNA-seq and genotyping analysis identified a - 21 bp (A → T) mutation in the promoter of SlPIF1b, resulting in the absence of the TATA-box core transcriptional element and inhibiting SlPIF1b transcription. The overexpression of SlPIF1b in GS inhibited chloroplast development of fruits, leading to a lighter green shoulder color, decreased chlorophyll content, reduced photosynthetic activity, diminished starch accumulation, and compromised fruit quality upon ripening. Conversely, the down expression of SlPIF1b significantly enhanced fruit chloroplast development and functionality in fruits, resulting in increased chlorophyll and carotenoid accumulation. Further analysis of expression profile and transcriptional activity indicated that SlPIF1b could bind to G/PBE-box elements present in SlGLK2, SlTKN4, SlCAO1a, SlPOR1, SlPOR3, SlCAB1 and SlCAB1b promoters, thereby inhibiting their expression. This study revealed the specific regulatory mechanism by which SlPIF1b modulates chloroplast development and chlorophyll synthesis in tomato fruit and provided valuable genetic resources and a theoretical basis for tomato quality improvement.

Solanum lycopersicum↗

PubMed1988

Interaction of a DNA binding factor with the 5'-flanking region of an ethylene-responsive fruit ripening gene from tomato.

To understand how fruit development is controlled, we have begun experiments to identify DNA sequences and proteins that regulate gene expression during tomato (Lycopersicon esculentum) fruit ripening. We have focused on the E8 gene because its transcription is responsive to ethylene hormone and is activated at the onset of fruit ripening. We report here that sequences required for ethylene-responsive and developmentally regulated E8 gene expression in transgenic tomato plants are contained on a 4.4 kb restriction fragment which includes sequences 2 kb 5' and 0.5 kb 3' to the gene. In addition, we have identified a DNA-binding factor that specifically interacts with DNA sequences that flank the E8 gene. This DNA-binding activity is low in unripe fruit and increases during fruit ripening. This factor also binds to the 5'-flanking region of another ethylene-responsive gene which is coordinately expressed during tomato fruit ripening. These data suggest that the DNA binding-factor may be involved in the regulation of gene expression during fruit ripening.

Base Sequence↗

PubMed1976

Fruiting organs of Cladosporium werneckii.

Submerged mycelia of a strain of Cladosporium werneckii isolated from tinea nigra palmaris, when cultured on enriched corn-meal agar media, developed fruiting bodies resembling perithecia.

Cladosporium↗

PubMed2004

Cheating is not always punished: killer female plants and pollination by deceit in the dwarf palm Chamaerops humilis.

Because the interests of mutualists are not perfectly aligned, conflicts between partners often arise, rendering mutualism unstable by allowing the evolution of cheating. The dwarf palm Chamaerops humilis is engaged in a nursery pollination mutualism with a specific weevil Derelomus chamaeropsis. In exchange for pollen dispersal, dwarf palms provide pollinators with food, shelter and egg-laying sites, but pollinators can develop only within male inflorescences. Here we show that weevils lay eggs in female inflorescences but processes associated with fruit development prevent larval development. The cost imposed by developing larvae probably differs between male and female plants, explaining why only females defend their inflorescences. Female palms thus cheat their pollinating weevil, and pollinators are expected to 'punish' (avoid) them. We found no evidence for such punishment: weevils visit female plants and the duration of visits to male and female inflorescences does not differ. Thus mutualists do not always co-operate and cheating may not be necessarily punished.

Animals↗

PubMed2002

Changes in the phosphorylation state of sucrose synthase during development of Japanese pear fruit.

Changes in the protein level and phosphorylation state of sucrose synthase (SS) were studied throughout the development of Japanese pear fruit. The level of SS protein was high at the young stage, dropped with fruit enlargement and increased again with fruit maturation. Antibody against phospho-Ser reacted with SS from young fruit, but did not react with SS that had been dephosphorylated by alkaline phosphatase (AP). The activities of SS isozymes were separated by ion-exchange chromatography. It was found that the fluctuation in SS activity was caused by two SS isozymes (SSI and SSII); (SSI reacted with antibody against phospho-Ser, while SSII did not. Phosphorylation of SS affected its kinetic parameters, that is, the affinity of phosphorylated SS for UDP was higher than that of dephosphorylated SS, while it was the contrary for UDP-glucose. The reaction of dephosphorylated SS was inclined toward sucrose synthesis more than that of phosphorylated SS. Phosphorylated SS protein was most abundant in young fruit, but decreased with fruit development, while non-phosphorylated SS protein increased in mature fruit. These results suggest that SS isoforms may be affected by post-translational modifications such as phosphorylation, and that the regulation of phosphorylation may potentially control the properties and functions of SS throughout the development of Japanese pear fruit.

Journal Article↗

PubMed2003

Girdling affects carbohydrate-related gene expression in leaves, bark and roots of alternate-bearing citrus trees.

Effects of girdling on carbohydrate status and carbohydrate-related gene expression in citrus trees were investigated. Alternate-bearing 'Murcott' (a Citrus reticulata hybrid of unknown origin) trees were girdled during autumn (25 Sep. 2001) and examined 10 weeks later. Girdling brought about carbohydrate (soluble sugar and starch) accumulation in leaves and shoot bark above the girdle, in trees during their fruitless, 'off' year. Trees during their heavy fruit load, 'on' year did not accumulate carbohydrates above the girdle due to the high demand for carbohydrates by the developing fruit. Girdling caused a strong decline in soluble sugar and starch concentrations in organs below the girdle (roots), in both 'on' and 'off' trees. Expression of STPH-L and STPH-H (two isoforms of starch phosphorylase), Agps (ADP-glucose pyrophosphorylase, small subunit), AATP (plastidic ADP/ATP transporter), PGM-C (phosphoglucomutase) and CitSuS1 (sucrose synthase), all of which are associated with starch accumulation, was studied. It was found that gene expression is related to starch accumulation in all 'off' tree organs. RNA levels of all the genes examined were high in leaves and bark that accumulated high concentrations of starch, and low in roots with declining starch concentrations. It may be hypothesized that changes in specific sugars signal the up- and down-regulation of genes involved in starch synthesis.

Carbohydrate Metabolism↗

PubMed2003

A putative plastidic glucose translocator is expressed in heterotrophic tissues that do not contain starch, during olive (Olea europea L.) fruit ripening.

Metabolite-specific transporters are present in the inner membrane of the plastid envelope allowing transport between the plastid and other cellular compartments. A plastidic glucose translocator (pGlcT) in leaf mesophyll cells transports glucose from chloroplast stroma to the cytosol after amylolytic starch degradation at night. Here we report the cloning of a pGlcT expressed in olive fruits (Olea europea L.). Our results showed high expression of pGlcT in non-green heterotrophic fruit tissues. Expression of pGlcT in olive fruits was somewhat higher compared to leaves, and continued until the black, mature fruit stage. We cloned part of tomato pGlcT and found that it is also expressed throughout fruit development implying a role for pGlcT in heterotrophic tissues. Light and electron microscopic characterization of plastid structural changes during olive fruit ripening revealed the transition of chloroplast-like plastids into starchless, non-green plastids; in mature olive fruits only chromoplasts were present. Together, these findings suggest that olive pGlcT is abundant in chromoplasts during structural changes, and provide evidence that pGlcT may play different physiological roles in ripening fruits and possibly in other non-photosynthetic organs.

Amino Acid Sequence↗

PubMed1992

Comparison of Pch313 (pTOM13 Homolog) RNA Accumulation during Fruit Softening and Wounding of Two Phenotypically Different Peach Cultivars.

Pch313 was isolated as a cDNA whose RNA accumulated during the softening period of peach (Prunus persica L. Batsch) fruit development. To better understand the role of the gene, we compared the amount of pch313-related RNA detected during fruit softening and tissue wounding between cultivars with different softening characteristics. The cultivar that softened faster, "Bailey," had a significantly higher amount of pch313-related RNA accumulate during softening than the slower-softening cultivar, "Suncrest." Pch313 was sequenced and found to be related to a tomato fruit cDNA clone, pTOM13, which has been shown to encode the ethylene-forming enzyme. The derived amino acid sequence of pch313 is 74 to 83% identical to the pTOM13-related sequences. A pch313-3' noncoding region probe was used to demonstrate that pch313 is related to both a wound-induced RNA transcript and the major fruit-softening transcript. The relationship of pch313 RNA accumulation and ethylene evolution was examined upon wounding and appeared to be both tissue and cultivar specific. When leaves were wounded, more pch313-related RNA was detected in Bailey and the rate of ethylene evolved was also higher in Bailey. When fruits were wounded, the levels of ethylene evolved were nearly identical but Suncrest accumulated more pch313-related RNA. Southern analysis of the DNA indicated a small number of related genes.

Journal Article↗

PubMed2000

Characterization of ripening-regulated cDNAs and their expression in ethylene-suppressed charentais melon fruit.

Charentais melons (Cucumis melo cv Reticulatus) are climacteric and undergo extremely rapid ripening. Sixteen cDNAs corresponding to mRNAs whose abundance is ripening regulated were isolated to characterize the changes in gene expression that accompany this very rapid ripening process. Sequence comparisons indicated that eight of these cDNA clones encoded proteins that have been previously characterized, with one corresponding to ACC (1-aminocyclopropane-1-carboxylic acid) oxidase, three to proteins associated with pathogen responses, two to proteins involved in sulfur amino acid biosynthesis, and two having significant homology to a seed storage protein or a yeast secretory protein. The remaining eight cDNA sequences did not reveal significant sequence similarities to previously characterized proteins. The majority of the 16 ripening-regulated cDNAs corresponded to mRNAs that were fruit specific, although three were expressed at low levels in vegetative tissues. When examined in transgenic antisense ACC oxidase melon fruit, three distinct patterns of mRNA accumulation were observed. One group of cDNAs corresponded to mRNAs whose abundance was reduced in transgenic fruit but inducible by ethylene treatment, indicating that these genes are directly regulated by ethylene. A second group of mRNAs was not significantly altered in the transgenic fruit and was unaffected by treatment with ethylene, indicating that these genes are regulated by ethylene-independent developmental cues. The third and largest group of cDNAs showed an unexpected pattern of expression, with levels of mRNA reduced in transgenic fruit and remaining low after exposure to ethylene. Regulation of this third group of genes thus appears to ethylene independent, but may be regulated by developmental cues that require ethylene at a certain stage in fruit development. The results confirm that both ethylene-dependent and ethylene-independent pathways of gene regulation coexist in climacteric fruit.

Cloning, Molecular↗

PubMed1976

[Effects of Flavorseal and Tag waxes on the ripening and texture of Kent mangos].

Mangos KENT at the green maturity stage were waxed with Tag or Flavorseal and were allowed to ripe at 23+/-2 degrees C and 16+/-1 degrees C. The relative humidity was 60-70%. Unwaxed fruits held under the same conditions were used as controls. It was found that both waxes prolonged the shelf life of the mangos. The results also showed that Tag treated fruits developed their internal and external coloration normally, whereas mangos with Falvorseal coating did not develop their external coloration nor their red internal coloration. The content of Total Saluble Solids (T.S.S.) was higher in control fruits than in Tag treated fruits but it was lowest in Flavorseal coated mangos. As evident from the penetration and compression tests, the texture of Tag treated fruits was softer than the texture of the control and Flavorseal treated mangos. However, the texture of the control mangos was also much softer than the texture of the Flavorseal treated mangos. The patterns of the penetration and compression curves were similar in control and TAG treated fruits while in Flavorseal coated mangos the curves were uncharteristic. The significance of the results is discussed and a possible explanation for the differences between the effects of the two waxes is given.

Antioxidants↗

PubMed2006

Downregulation of Solanum americanum genes encoding proteinase inhibitor II causes defective seed development.

Proteinase inhibitor II proteins (PIN2) are serine proteinase inhibitors found in the Solanaceae. Here, we assign functions in seed development to two Solanum americanum genes, SaPIN2a and SaPIN2b, encoding proteinase inhibitor II. Their mRNAs and proteins have been previously localized to the reproductive tissues, including the inner cell layers of ovules in senescent flowers at the beginning of fruit development, suggestive of their endogenous roles in reproductive development. We have employed RNA interference (RNAi)-induced post-transcriptional gene silencing (PTGS) to further investigate the role of SaPIN2a and SaPIN2b during seed development. A SaPIN2a-derived construct that shared 83% nucleotide homology to SaPIN2b was used in PTGS to silence both genes. Northern blot analyses confirmed that the PIN2-RNAi transgenic plants contain small interfering RNAs (siRNAs) and exhibit reduced levels of SaPIN2a and SaPIN2b mRNAs at various stages of floral development. A reduction in seed set due to seed abortion was observed in PIN2-RNAi transgenic lines. Cytological and molecular analyses of these lines showed the lack of SaPIN2a and SaPIN2b mRNAs and proteins at the inner cell layers of the ovules in senescent flowers. Aborted seeds in transgenic fruits had an abnormal endothelium. The anomalous expansion of the endothelium prevented proper development of the endosperm and embryo, leading to seed abortion. Our observations indicate that SaPIN2a and SaPIN2b are essential for seed development and suggest that the endothelium may protect the embryo sac, allowing proper formation of the endosperm and embryo, as a result of its ability to produce proteinase inhibitors.

Base Sequence↗

PubMed2006

Metabolic changes in fruits of the tomato dx mutant.

The tomato DWARF cytochrome P450 protein catalyzes the C-6 oxidation of 6-deoxo-castasterone to castasterone. The d(x) mutant does not produce a functional DWARF enzyme, and d(x) shoots display severe symptoms of brassinosteroid-deficiency. However, fruits express the CYP85A3 protein which compensates for the deficiency of the DWARF protein and produce bioactive brassinosteroids. Here, we report on the metabolic characterization of d(x) fruits. Fruit size, fresh weight, and pigment content were not altered. However, d(x) fruits showed reduced dry mass content. Levels of starch and various sugars were reduced, amino acid levels were elevated. BR application to d(x) leaves partially normalized dry mass content, sugar and amino acid levels in d(x) fruits. The data demonstrate that brassinosteroid in shoots is required for fruit development in tomato.

Amino Acids↗