Insights into the Uptake, Distribution, and Metabolism of 3-Isobutyl-2-hydroxypyrazine in Grapevine Using a Stable Isotope Tracer.

Methoxypyrazines (MPs) are potent aroma compounds that have been predominately studied in grape berries but can also be detected in other vine tissues. The synthesis of MPs in berries from hydroxypyrazines by VvOMT3 is well established, but the origin of MPs in vine tissues that have negligible VvOMT3 gene expression is unknown. This research gap was addressed through the application of stable isotope tracer 3-isobutyl-2-hydroxy-[2H2]-pyrazine (d2-IBHP) to the roots of Pinot Meunier L1 microvines and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) quantification of HPs from grapevine tissues following a novel solid-phase extraction method. Four weeks post-application, d2-IBHP and its O-methylated product 3-isobutyl-2-methoxy-[2H2]-pyrazine (d2-IBMP) were present in excised cane, berry, leaf, root, and rachis material. Translocation of d2-IBHP and d2-IBMP was investigated, but results were inconclusive. Nonetheless, knowledge that d2-IBHP, and potentially d2-IBMP, are translocated from roots to other vine organs, including the berries, could provide opportunities for controlling MP accumulation in grapevine tissues pertinent to winemaking.

Methoxypyrazine concentrations in the grape bunch rachis of Vitis vinifera L. Cv Shiraz: Influence of rootstock, region and light.

Vitis vinifera L. cv Shiraz appears unable to synthesise 3-alkyl-2-methoxypyrazines (MPs) in the berry, but can still produce significant concentrations in rachis. MPs are readily extracted from rachis during fermentation, producing Shiraz wines with uncharacteristic "green" flavours. Recently, rootstocks were shown to significantly alter MP concentrations in Cabernet Sauvignon rachis compared to own-rooted varieties, but whether Shiraz followed a similar trend required investigation. This study considered the effect of thirteen rootstocks on the concentrations of 3-isobutyl-2-methoxypyrazine (IBMP), 3-isopropyl-2-methoxypyrazine (IPMP), and 3-sec-butyl-2-methoxypyrazine (SBMP) in the rachis of Shiraz bunches sampled during multiple vintages across several Australian growing regions. Although IBMP was the most abundant, all measured MP concentrations were significantly affected by vintage, rootstock, and region. In addition, vine vigour showed positive correlations with IBMP, which were attributed to changes in canopy coverage impacting rachis light exposure. This hypothesis was explored with light exclusion trials, which significantly increased rachis IBMP concentrations.

Rootstock, Vine Vigor, and Light Mediate Methoxypyrazine Concentrations in the Grape Bunch Rachis of Vitis vinifera L. cv. Cabernet Sauvignon.

Ramsey rootstock has previously been implicated in an approximate 8-fold increase of 3-isobutyl-2-methoxypyrazine (IBMP) levels in the rachis (grape bunch stem) of Vitis vinifera L. cv. Shiraz scions over own-rooted Shiraz vines at harvest. IBMP extracted from rachis during red wine fermentation can contribute potent "green" flavors. Methoxypyrazines (MPs) are normally present in Cabernet Sauvignon grapes, rachis, and wines, but it is unknown whether rootstocks can influence the MP concentration in the rachis. This study considered the effect of eight rootstocks including Ramsey and own roots on the concentrations of IBMP, 3-isopropyl-2-methoxypyrazine (IPMP), and 3-sec-butyl-2-methoxypyrazine (SBMP) in the rachis and grapes of Cabernet Sauvignon over two seasons. IBMP predominated, and its concentration in rachis and berries at harvest was significantly affected by rootstock and growing season. In the 2020 vintage, light exclusion, vine vigor, and spatial variation in vine vigor were shown to significantly affect MP concentrations in rachis.