Biomedical subjects

Ryohei Nakano

Publications and source records attributed to Ryohei Nakano.

8 recordsLinked to original sources

PubMed2006

Differential feedback regulation of ethylene biosynthesis in pulp and peel tissues of banana fruit.

The feedback regulation of ethylene biosynthesis in banana [Musa sp. (AAA group, Cavendish subgroup) cv. Grand Nain] fruit was investigated in an attempt to clarify the opposite effect of 1-methylcyclopropene (1-MCP), an ethylene action inhibitor, before and after the onset of ripening. 1-MCP pre-treatment completely prevented the ripening-induced effect of propylene in pre-climacteric banana fruit, whereas treatment after the onset of ripening stimulated ethylene production. In pre-climacteric fruit, higher concentrations of propylene suppressed ethylene production more strongly, despite their earlier ethylene-inducing effect. Exposure of the fruit ripened by propylene to 1-MCP increased ethylene production concomitantly with an increase in 1-aminocyclopropane-1-carboxylate (ACC) synthase activity and ACC content, and prevented a transient decrease in MA-ACS1 transcripts in the pulp tissues. In contrast, in the peel of ripening fruit, 1-MCP prevented the increase in ethylene production and subsequently the ripening process by reduction of the increase in MA-ACS1 and MA-ACO1 transcripts and of ACC synthase and ACC oxidase activities. These results suggest that ethylene biosynthesis in ripening banana fruit may be controlled negatively in the pulp tissue and positively in the peel tissue. This differential regulation by ethylene in pulp and peel tissues was also observed for MA-PL, MA-Exp, and MA-MADS genes.

Amino Acids, Cyclic↗

PubMed2006

Vacuolar citrate/H+ symporter of citrus juice cells.

We have isolated a cDNA, designated Citrus sinensis citrate transporter 1 CsCit1 encoding a novel vacuolar citrate/symporter. Immunoblots using antibodies raised against CsCit1 showed that the protein is localized to the juice sac cell vacuoles. The highest expression of CsCit1 and the amount of protein in the juice sac cell vacuoles coincided with the developmental stage at which the vacuolar citrate content began declining with the concomitant increase in vacuolar pH. Vacuoles from Sacharomyces cereviseae expressing CsCit1 displayed a citrate-dependent H(+) efflux, and our results clearly demonstrate that CsCit1 is able to mediate the electroneutral co-transport of H(+) and citrate ions, since the citrate-dependent H(+) fluxes are not affected by changing the electrical potential difference across the tonoplast. The roles of CsCit1 in mediating citrate efflux from the vacuole and on citric acid homoestasis in Citrus juice sac cells are discussed.

Amino Acid Sequence↗

PubMed2004

European, Chinese and Japanese pear fruits exhibit differential softening characteristics during ripening.

Softening characteristics were investigated in three types of pear fruit, namely, European pear 'La France', Chinese pear 'Yali', and Japanese pear 'Nijisseiki'. 'La France' fruit softened dramatically and developed a melting texture during ripening, while 'Yali' fruit with and without propylene treatment showed no change in flesh firmness and texture during ripening. Non-treated 'Nijisseiki' did not show a detectable decrease in flesh firmness, whereas continuous propylene treatment caused a gradual decrease in firmness resulting in a mealy texture. In 'La France', the analysis of cell wall polysaccharides revealed distinct solubilization and depolymerization of pectin and hemicellulose during fruit softening. In 'Nijisseiki', propylene treatment led to the solubilization and depolymerization of pectic polysaccharides to a limited extent, but not of hemicellulose. In 'Yali', hemicellulose polysaccharides were depolymerized during ripening, but there was hardly any change in pectic polysaccharides except in the water-soluble fraction. PC-PG1 and PC-PG2, two polygalacturonase (PG) genes, were expressed in 'La France' fruit during ripening, while only PC-PG2 was expressed in 'Nijisseiki' and neither PC-PG1 or PC-PG2 was expressed in 'Yali'. The expression pattern of PC-XET1 was constitutive during ripening in all three pear types. PG activity measured by the reducing sugar assay increased in all three pears during ripening. However, viscometric measurements showed that the levels of endo-PG activity were high in 'La France', low in 'Nijisseiki', and undetectable in 'Yali' fruits. These results suggest that, in pears, cell wall degradation is correlated with a decrease in firmness during ripening and the modification of both pectin and hemicellulose are essential for the development of a melting texture. Furthermore, the data suggest that different softening behaviours during ripening among the three pear fruits may be caused by different endo-PG activity and different expression of PG genes.

China↗

PubMed2003

Differential expression of seven alpha-expansin genes during growth and ripening of pear fruit.

Seven cDNAs, designated PcExp1 to PcExp7, encoding expansin homologues, were isolated from mature pear fruit and their expression profiles were investigated in ripening fruit and other tissues, and in response to ethylene. Accumulation of PcExp2, -3, -5 and -6 mRNA increased markedly with fruit softening and then declined at the over-ripe stage. Treatment of fruit at an early ripening stage with 1-methylcyclopropene (MCP), an inhibitor of ethylene action, suppressed ethylene biosynthesis, fruit softening and the accumulation of the expansin mRNAs. Conversely, propylene treatment at the preclimacteric stage induced accumulation of the same four expansin genes, as well as ethylene production and fruit softening. The expression patterns correlated with alteration in the rate and extent of fruit softening. The abundance of PcExp1 mRNA increased at the late expanding phase of fruit development and further increased during ripening, whereas PcExp4 mRNA levels were constant throughout fruit growth and ripening. The MCP and propylene treatments had little effect on PcExp1 and PcExp4 expression. PcExp7 was expressed in young but not mature fruit. PcExp4 and PcExp6 mRNA was also detected in flowers. The accumulation of PcExp4, -5, -6 and -7 mRNA was more abundant in young growing tissues, but not in fully expanded tissues, suggesting roles for these genes in cell expansion. These results demonstrate that characteristically, multiple expansin genes show differential expression and hormonal regulation during pear fruit development and at least six expansins show overlapping expression during ripening.

Journal Article↗

PubMed2003

Ethylene is required for both the initiation and progression of softening in pear (Pyrus communis L.) fruit.

In order to investigate the physiological role of ethylene in the initiation and subsequent progression of softening, pear fruit were treated with propylene, an analogue of ethylene or 1-methylcyclopropene (1-MCP), a gaseous inhibitor of ethylene action at the preclimacteric or ripening stages. The propylene treatment at the pre-ripe stage stimulated ethylene production and flesh softening while the 1-MCP treatment at the same stage markedly retarded the initiation of the ripening-related events. Moreover, 1-MCP treatment after the initiation of ripening markedly suppressed the subsequent flesh softening and ethylene production. These results clearly indicate that ethylene is not merely a by-product, but plays a crucial role in both the initiation and maintenance of regulating the softening process during ripening. The observations also suggest that ethylene in ripening is regulated entirely in an autocatalytic manner. The mRNA accumulation of pear polygalacturonases (PG) genes, PC-PG1 and PC-PG2, was in parallel with the pattern of fruit softening in both propylene and 1-MCP treatments. However, the expression pattern of pear endo-1,4-beta-D-glucanases (EGase) genes, PC-EG1 and PC-EG2, was not affected in both treatments. The results suggest that ethylene is required for PGs expression even in the late ripening stage, but not for EGases.

Alkenes↗

PubMed2003

Characterization of a novel tomato EIN3-like gene (LeEIL4).

A novel cDNA clone encoding a putative EIN3-like protein (LeEIL4) was identified from ripening tomato (Lycopersicon esculentum) fruit. The predicted amino acid sequence contained conserved domains of EIN3-like proteins in the N-terminal half. In phylogenetic analysis, LeEIL4 was classified into the cluster consisting of EIN3 and EIN3-like proteins known to be involved in ethylene signal transduction. LeEIL4 transcripts accumulated in all tissues examined, with higher levels in ripening fruit.

Amino Acid Sequence↗

PubMed2003

Ethylene biosynthesis in detached young persimmon fruit is initiated in calyx and modulated by water loss from the fruit.

Persimmon (Diospyros kaki Thunb.) fruit are usually classified as climacteric fruit; however, unlike typical climacteric fruits, persimmon fruit exhibit a unique characteristic in that the younger the stage of fruit detached, the greater the level of ethylene produced. To investigate ethylene induction mechanisms in detached young persimmon fruit, we cloned three cDNAs encoding 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (DK-ACS1, 2, and -3) and two encoding ACC oxidase (DK-ACO1 and -2) genes involved in ethylene biosynthesis, and we analyzed their expression in various fruit tissues. Ethylene production was induced within a few days of detachment in all fruit tissues tested, accompanied by temporally and spatially coordinated expression of all the DK-ACS and DK-ACO genes. In all tissues except the calyx, treatment with 1-methylcyclopropene, an inhibitor of ethylene action, suppressed ethylene production and ethylene biosynthesis-related gene expression. In the calyx, one ACC synthase gene (DK-ACS2) exhibited increased mRNA accumulation accompanied by a large quantity of ethylene production, and treatment of the fruit with 1-methylcyclopropene did not prevent either the accumulation of DK-ACS2 transcripts or ethylene induction. Furthermore, the alleviation of water loss from the fruit significantly delayed the onset of ethylene production and the expression of DK-ACS2 in the calyx. These results indicate that ethylene biosynthesis in detached young persimmon fruit is initially induced in calyx and is modulated by water loss through transcriptional activation of DK-ACS2. The ethylene produced in the calyx subsequently diffuses to other fruit tissues and acts as a secondary signal that stimulates autocatalytic ethylene biosynthesis in these tissues, leading to a burst of ethylene production.

Amino Acid Oxidoreductases↗

PubMed2002

Extracting regression rules from neural networks.

This paper proposes a new framework and method for extracting regression rules from neural networks trained with multivariate data containing both nominal and numeric variables. Each regression rule is expressed as a pair of a logical formula on the conditional part over nominal variables and a polynomial equation on the action part over numeric variables. The proposed extraction method first generates one such regression rule for each training sample, then utilizes the kappa-means algorithm to generate a much smaller set of rules having more general conditions, where the number of distinct polynomial equations is determined through cross-validation. Finally, this method invokes decision-tree induction to form logical formulae of nominal conditions as conditional parts of final regression rules. Experiments using four data sets show that our method works well in extracting quite accurate and interesting regression rules.

Algorithms↗