Fruit ripening in Vitis vinifera: light intensity before and not during ripening determines the concentration of 2-methoxy-3-isobutylpyrazine in Cabernet Sauvignon berries.

The roles of light and temperature in the accumulation of the vegetal impact compound 2-methoxy-3-isobutylpyrazine (MIBP) in grape (Vitis vinifera L.) berries were determined. Individual clusters were exposed to various light intensities using neutral density shade cloth before ripening, during ripening or throughout the season in three growing seasons. A recently developed method using headspace solid-phase microextraction combined with GC-MS in the selected ion-monitoring mode was employed to measure MIBP in berries. Berry MIBP concentration increased subsequent to berry set, reached a maximum prior to onset of ripening, and then decreased thereafter until harvest. Complete shading of clusters increased the concentration of MIBP more than 100% compared to unshaded controls in 2 out of 3 years. Light increasingly inhibited MIBP concentrations up to 25-50% of ambient light intensities (1500 µmol photons m(-2) s(-1) ). However, only changes in light intensity before ripening had any effect on MIBP accumulation or final MIBP concentration. Analyses of weather data showed that the 1 year in which shading was ineffective was unusually warm, warm early in the season, and had more hot days and higher early season degree days than the other 2 years. In controlled environment experiments, warm growth conditions reduced MIBP concentrations in fruit about as much as light exposure reduced MIBP concentrations in the field experiments. The results indicate that both light and temperature significantly affect MIBP in harvested fruit, but that the light environment during ripening does not significantly affect MIBP concentrations in the berries at harvest.

2-Methoxy-3-isobutylpyrazine in grape berries and its dependence on genotype.

2-Methoxy-3-isobutylpyrazine (MIBP) contributes a bell pepper aroma to many grape cultivars and has a reported aroma threshold of ∼2 ng L(-1) in water. The purpose of this study was twofold: (1) develop a procedure using headspace solid phase micro-extraction combined with GC-MS in the selected ion monitoring mode (HS-SPME-GC-MS-SIM) for analysis of MIBP in grape berries, and (2) determine the location of MIBP biosynthesis in grapevines by approach grafting clusters of Vitis vinifera L. cvs Cabernet Sauvignon and Muscat blanc onto each other. The soluble solids and pH of the grape juice/homogenate matrix from different grape berry developmental stages influenced the method precision; therefore, quantification via the method of standard addition was used. Using our developed method, the limit of detection (LOD) and limit of quantitation (LOQ) of MIBP were 0.1 ng L(-1) and 2 ng L(-1), respectively, measured in a model juice and non-MIBP containing Chardonnay juice. Spiked recoveries averaged between 91% and 112% in Cabernet Sauvignon and Pinot noir homogenates and the overall relative standard deviation was less than 10%. The method was used to analyze MIBP in 29 grape cultivars and in fruit from clusters grafted to Cabernet Sauvignon or Muscat vines. Quantifiable levels were found only in Cabernet franc, Cabernet Sauvignon, Merlot, Sauvignon blanc and Semillon, providing information on the genetic connection for the occurrence of MIBP in grapes. No MIBP was detected in the berries of Muscat blanc clusters grafted onto Cabernet Sauvignon vines when sampled at fruit maturity. MIBP was detected in all berries of Cabernet Sauvignon regardless the graft configuration. The data indicate that MIBP or its precursors originate in the berry and its formation depends upon grape genotype.