Despite its simple two-carbon structure, the olefin ethylene is a potent modulator of plant growth and development ([Ecker, 1995][1]). The plant hormone ethylene is involved in many aspects of the plant life cycle, including seed germination, root hair development, root nodulation, flower senescence

Ethylene biosynthesis and signaling networks.

1-Methylcyclopropene: a review

Elucidating the mechanisms involved in ripening of climacteric fruit and the role that ethylene plays in the process are key to understanding fruit production and quality. In this review, which is based largely on research in tomato, particular attention is paid to the role of specific isoforms of ACC synthase and ACC oxidase in controlling ethylene synthesis during the initiation and subsequent autocatalytic phase of ethylene production during ripening. Recent information on the structure and role of six different putative ethylene receptors in tomato is discussed, including evidence supporting the receptor inhibition model for ripening, possible differences in histidine kinase activity between receptors, and the importance of receptor LeETR4 in ripening. A number of ethylene-regulated ripening-related genes are discussed, including those involved in ethylene synthesis, fruit texture, and aroma volatile production, as well as experiments designed to elucidate the ethylene signalling pathway from receptor through intermediate components similar to those found in Arabidopsis, leading to transcription factors predicted to control the expression of ethylene-regulated genes.

Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening

The development and maturation of fruits has received considerable scientific scrutiny because of both the uniqueness of such processes to the biology of plants and the importance of fruit as a significant component of the human diet. Molecular and genetic analysis of fruit development, and especially ripening of fleshy fruits, has resulted in significant gains in knowledge over recent years. Great strides have been made in the areas of ethylene biosynthesis and response, cell wall metabolism, and environmental factors, such as light, that impact ripening. Discoveries made in Arabidopsis in terms of general mechanisms for signal transduction, in addition to specific mechanisms of carpel development, have assisted discovery in more traditional models such as tomato. This review attempts to coalesce recent findings in the areas of fruit development and ripening.

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Molecular biology of fruit maturation and ripening.

The use of 1-methylcyclopropene (1-MCP) on fruits and vegetables.

We investigated the feedback regulation of ethylene biosynthesis in tomato (Lycopersicon esculentum) fruit with respect to the transition from system 1 to system 2 ethylene production. The abundance of LE-ACS2, LE-ACS4, and NR mRNAs increased in the ripening fruit concomitant with a burst in ethylene production. These increases in mRNAs with ripening were prevented to a large extent by treatment with 1-methylcyclopropene (MCP), an ethylene action inhibitor. Transcripts for the LE-ACS6 gene, which accumulated in preclimacteric fruit but not in untreated ripening fruit, did accumulate in ripening fruit treated with MCP. Treatment of young fruit with propylene prevented the accumulation of transcripts for this gene. LE-ACS1A, LE-ACS3, and TAE1 genes were expressed constitutively in the fruit throughout development and ripening irrespective of whether the fruit was treated with MCP or propylene. The transcripts for LE-ACO1 and LE-ACO4 genes already existed in preclimacteric fruit and increased greatly when ripening commenced. These increases in LE-ACO mRNA with ripening were also prevented by treatment with MCP. The results suggest that in tomato fruit the preclimacteric system 1 ethylene is possibly mediated via constitutively expressed LE-ACS1A and LE-ACS3 and negatively feedback-regulated LE-ACS6 genes with preexisting LE-ACO1 and LE-ACO4 mRNAs. At the onset of the climacteric stage, it shifts to system 2 ethylene, with a large accumulation of LE-ACS2, LE-ACS4, LE-ACO1, and LE-ACO4 mRNAs as a result of a positive feedback regulation. This transition from system 1 to system 2 ethylene production might be related to the accumulated level of NR mRNA.

Differential Expression and Internal Feedback Regulation of 1-Aminocyclopropane-1-Carboxylate Synthase, 1-Aminocyclopropane-1-Carboxylate Oxidase, and Ethylene Receptor Genes in Tomato Fruit during Development and Ripening

We investigated the characteristics of ethylene biosynthesis associated with ripening in banana (Musa sp. [AAA group, Cavendish subgroup] cv Grand Nain) fruit. MA-ACS1 encoding 1-aminocyclopropane-1-carboxylic acid (ACC) synthase in banana fruit was the gene related to the ripening process and was inducible by exogenous ethylene. At the onset of the climacteric period in naturally ripened fruit, ethylene production increased greatly, with a sharp peak concomitant with an increase in the accumulation of MA-ACS1 mRNA, and then decreased rapidly. At the onset of ripening, the in vivo ACC oxidase activity was enhanced greatly, followed by an immediate and rapid decrease. Expression of the MA-ACO1 gene encoding banana ACC oxidase was detectable at the preclimacteric stage, increased when ripening commenced, and then remained high throughout the later ripening stage despite of a rapid reduction in the ACC oxidase activity. This discrepancy between enzyme activity and gene expression of ACC oxidase could be, at least in part, due to reduced contents of ascorbate and iron, cofactors for the enzyme, during ripening. Addition of these cofactors to the incubation medium greatly stimulated the in vivo ACC oxidase activity during late ripening stages. The results suggest that ethylene production in banana fruit is regulated by transcription of MA-ACS1 until climacteric rise and by reduction of ACC oxidase activity possibly through limited in situ availability of its cofactors once ripening has commenced, which in turn characterizes the sharp peak of ethylene production.

Characterization of ethylene biosynthesis associated with ripening in banana fruit.

We have examined whether or not a positive feedback regulation of gene expression for 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase also operates in ripening tomato (Lycopersicon esculentum) fruit during the burst of ethylene production. Two cDNA fragments for ACC synthase and one for ACC oxidase were cloned with high homology to already known genes involved in ethylene biosynthesis in ripening tomato fruit. Accumulation of mRNAs which hybridize to these cDNA probes were induced in mature green fruit within two days by treatment with propylene. In the fruit ripened from the turning stage, red color development, ethylene production, ACC content, and activities of ACC synthase and ACC oxidase increased as maturity progressed. The abundance of two ACC synthase and one ACC oxidase mRNAs in the fruit increased from the turning to pink stage and were followed by a slight decline towards the red stage. These increases in mRNAs abundance with ripening were prevented to a large extent by treatment with the ethylene action inhibitor, 1-methylcyclopropene (MCP). This was most pronounced in the fruit treated with MCP at the turning stage, in which the accumulation of ACC synthase and ACC oxidase transcripts was almost completely eliminated in the first two d, precisely the same stage at which the control fruit had the greatest level of each mRNA accumulation. The inhibition of transcript accumulation recovered to the control level within two to four d. MCP also decreased ethylene biosynthetic activity, although this decrease did not reflect the reduction in the mRNAs accumulation. These results suggest that a strong positive feedback regulation is involved in ethylene biosynthesis at the gene transcriptional level in tomato fruit, even at the stage with a burst of ethylene production.

Expression and Internal Feedback Regulation of ACC Synthase and ACC Oxidase Genes in Ripening Tomato Fruit

We cloned and sequenced three cDNA homologs of 1-aminocyclopropane-1-carboxylate (ACC) synthase (pCS-ACS1, pCS-ACS2 and pCS-ACS3) and two of ACC oxidase (pCS-ACO1 and pCS-ACO2) from cucumber (Cucumis sativus L.) fruit using RT-PCR (reverse transcription and polymerase chain reaction) and RACE (rapid amplification of cDNA ends) PCR. Conserved amino acid sequences reported for other plant tissues were found in all the cloned ACC synthase and ACC oxidase cDNAs. Among the ACC synthase and ACC oxidase cDNA clones, pCS-ACS1, pCS-ACS3, and pCS-ACO1 were highly homologous to ACC synthase and ACC oxidase genes cloned from other Cucurbitaceae family at both the nucleotide and the amino acid levels. Northern blot analysis showed that wounding induced the accumulation of CS-ACS1, CS-ACS2, and CS-ACO1 mRNAs, whereas IAA treatment induced expression of all the three ACC synthase genes and CS-ACO1. These expressions paralleled ethylene production in both treatments. Transcripts for CS-ACO2 were constitutively expressed and were not affected by wounding and IAA treatments. From the expression pattern and the sequence similarities of these genes, we conclude that CS-ACS1 could be a wound-inducible gene and that CS-ACS3 is an auxin-inducible one. These results strongly suggest that in cucumber fruit these genes are differentially expressed by wounding and auxin and are involved in the regulation of ethylene biosynthesis at the transcriptional level.

Akira Nakatsuka

Papers

Differential Expression and Internal Feedback Regulation of 1-Aminocyclopropane-1-Carboxylate Synthase, 1-Aminocyclopropane-1-Carboxylate Oxidase, and Ethylene Receptor Genes in Tomato Fruit during Development and Ripening

Characterization of ethylene biosynthesis associated with ripening in banana fruit.

Expression and Internal Feedback Regulation of ACC Synthase and ACC Oxidase Genes in Ripening Tomato Fruit

cDNA cloning of ACC synthase and ACC oxidase genes in cucumber fruit and their differential expression by wounding and auxin

Effectiveness of back and foot pressures for assessing drowsiness of drivers