Volatile aroma compounds produced by apple, banana, and tomato are produced throughout development, however, those associated with ripening and edible quality are dependent upon ethylene action. In apple and banana, characteristic aroma is, in large part, dependent upon the formation of volatile esters. In tomato, many of the characteristic aromas are dependent upon tissue disruption and result from aldehydes and alcohols following lipid degradation. For apple and banana, the enzyme alcohol acyl-CoA transferase (AAT, EC 22.214.171.124) is the enzyme responsible for the final reaction in the pathway for ester formation and catalyzes the union of an alcohol and the CoA derivative of fatty acids. In both tissues, AAT gene expression was detected prior to the onset of ester production. In apple, AAT expression was found to be closely tied with the onset of autocatalytic ethylene synthesis. In banana, ethylene synthesis peaked and began to decline well before ester synthesis began. However, the expression of AAT increased as ester production increased for both tissues. Tomato fruit, like apple and banana, produced characteristic aromas following the onset of the ethylene climacteric, suggesting changes in the activity of various components of the lipoxygenase pathway. In all three tissue types, there are continuous, significant shifts in the aroma profile as fruit ripen age, suggesting shifts in specific metabolic pathways associated with precursor synthesis or degradation.
Sastry Jayanty, Mauricio Canoles, Alejandra Ferenczi, Jun Song, and Randolph Beaudry*
Sastry S. Jayanty, Mauricio Cañoles, and Randolph M. Beaudry
We studied the dose-response of `Redchief Delicious' apple [Malus sylvestris (L) Mill. var. domestica (Borkh.) Mansf.] fruit to repeated (weekly) dosages of 0.0, 0.02, 0.1, and 1.0 μL·L-1 1-methylcyclopropene (1-MCP) by measuring fruit firmness and chlorophyll fluorescence throughout an extended storage period at 0, 5, 10, 15, and 20 °C. The rate of firmness loss for nontreated fruit increased with increasing temperature. 1-MCP applied at concentrations of 0.1 and 1.0 μL·L-1 slowed firmness loss. The 1-MCP dose-response curve for the rate of firmness loss was essentially the same for all five temperatures. A concentration of 1.0 μL·L-1 1-MCP prevented firmness loss at all temperatures for the duration of the study; however, after holding fruit for an additional 7 days at room temperature, the fruit stored at 10 °C softened with increasing storage duration, whereas fruit at stored at higher and lower temperatures did not. The influence of 1-MCP on chlorophyll fluorescence (Fo and Fm) was markedly affected by temperature; Fo increased during storage at higher storage temperatures and this increase was enhanced by 1-MCP. Conversely, Fm decreased during storage and the rate of decline was much greater at the higher storage temperatures; the rate of decline was reduced by 1-MCP, but only at the higher storage temperatures. Photochemical efficiency (Fv/Fm) of nontreated fruit declined with time for all storage temperatures. Treatment with 0.1 and 1.0 μL·L-1 1-MCP only marginally reduced the rate of decline of photochemical efficiency. Sample loss due to decay increased with temperature, but was reduced by 1-MCP at all temperatures.
Zhifang Gao, Sastry Jayanty, Randolph Beaudry, and Wayne Loescher
In apple (Malus ×domestica Borkh.), where sorbitol is a primary photosynthetic product that is translocated throughout the plant, accumulation of sorbitol in sink cells appears to require an active carrier-mediated membrane transport step. Recent progress in isolation and characterization of genes for sorbitol transporters in sour cherry (Prunus cerasus L.) and mannitol transporters in celery (Apium graveolens L.) suggested that similar transporters may be present in apple tissues. A defect in these transporters could also explain the occurrence of the fruit disorder watercore, characterized by the accumulation of fluids and sorbitol in the apoplasmic free space. Our objectives therefore included isolation and characterization of genes for sorbitol transporters in apple tissues and comparisons of expression of transporter genes, especially in various sink tissues including watercored and non-watercored fruit tissues. We have isolated and characterized two sorbitol transporter genes, MdSOT1 and MdSOT2. Sequence analyses indicated that these are members of the major facilitator transporter superfamily that gives rise to highly hydrophobic integral membrane proteins. Heterologous expression and measurement of sorbitol uptake in yeast indicated that these are specific and with high affinities for sorbitol, with Kms for sorbitol of 1.0 and 7.8 mm for MdSOT1 and MdSOT2, respectively. Sorbitol transporter expression was evident in all sink tissues tested with the exception of watercore-affected fruit tissues. Sorbitol accumulation in apple sink tissues thus involves an apoplasmic active membrane transport step and watercore results from a defect in that process.
Wayne Loescher, Tad Johnson, Randolph Beaudry, and Sastry Jayanty
Sorbitol is the major carbohydrate translocated into apple fruit where it is normally metabolized to fructose. In watercored apple fruit tissues, however, the intercellular spaces become flooded and sorbitol content is consistently higher than in nonwatercored apples, suggesting a defect in sugar alcohol metabolism or transport. Our previous results have identified and characterized two sorbitol transporters, MsSOT1 and MsSOT2, in apple fruit tissues. Sorbitol transporter gene expression has been implicated in development of watercore with MsSOT expression diminished or absent in certain watercored fruit tissues. To explore this further, we have investigated the relationships between watercore, fruit maturation, fruit composition, and MsSOT expression in a number of apple cultivars that differ in watercore susceptibility. We also compared transporter expression between affected (watercored) and healthy parts of the same fruit and between watercored and nonwatercored fruits throughout the maturation and ripening processes. The MsSOT expression was often dramatically reduced in fruit tissues exhibiting watercore. Thus, in susceptible cultivars, maturing (ripening) fruit parenchyma cells lose the ability to transport sorbitol, and this in turn leads to sorbitol accumulation in the apoplastic free space and subsequent flooding of these spaces. These results are consistent with a relationship between watercore and sorbitol transport and also with a genetic susceptibility to the disorder.
Sastry Jayanty, Jun Song, Nicole M. Rubinstein, Andrés Chong, and Randolph M. Beaudry
The temporal relationship between changes in ethylene production, respiration, skin color, chlorophyll fluorescence, volatile ester biosynthesis, and expression of ACC oxidase (ACO) and alcohol acyl-CoA transferase (AAT) in ripening banana (Musa L. spp., AAA group, Cavendish subgroup. `Valery') fruit was investigated at 22 °C. Ethylene production rose to a peak a few hours after the onset of its logarithmic phase; the peak in production coincided with maximal ACO expression. The respiratory rise began as ethylene production increased, reaching its maximum ≈30 to 40 hours after ethylene production had peaked. Green skin coloration and photochemical efficiency, as measured by chlorophyll fluorescence, declined simultaneously after the peak in ethylene biosynthesis. Natural ester biosynthesis began 40 to 50 hours after the peak in ethylene biosynthesis, reaching maximal levels 3 to 4 days later. While AAT expression was detected throughout, the maximum level of expression was detected at the onset of natural ester biosynthesis. The synthesis of unsaturated esters began 100 hours after the peak in ethylene and increased with time, suggesting the lipoxygenase pathway be a source of ester substrates late in ripening. Incorporation of exogenously supplied ester precursors (1-butanol, butyric acid, and 3-methyl-1-butanol) in the vapor phase into esters was maturity-dependent. The pattern of induced esters and expression data for AAT suggested that banana fruit have the capacity to synthesize esters over 100 hours before the onset of natural ester biosynthesis. We hypothesize the primary limiting factor in ester biosynthesis before natural production is precursor availability, but, as ester biosynthesis is engaged, the activity of alcohol acyl-CoA transferase the enzyme responsible for ester biosynthesis, exerts a major influence.