A One-Step Polyphenol Removal Approach for Detection of Multiple Phytohormones from Grape Berry

Phytohormones play an important role in regulating the maturation process and the quality-related metabolite accumulation of fruits, and their concentration variation has always been concerned during fruit development. However, berry fruits, such as grape berries, are rich in a large number of secondary metabolites, which brings great challenges to the isolation and determination of hormones. In this work, we used grapes as experimental materials and proposed a solid-phase extraction (SPE) protocol to efficiently isolate multiple hormones from phenol-rich matrix using a mixture of dichloromethane, methanol and formic acid as eluent. A highly sensitive method based on ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) was developed to quantify a total of 11 plant growth regulators, including the recognized phytohormones, in grape pericarp and seed. The established method showed satisfactory precision (RSD < 11.3%) and linearity (R2 > 0.9980). The limits of detection (LOD) and the limit of quantification (LOQ) were 0.001–0.75 ng/mL and 0.004–2.5 ng/mL, respectively. The recovery for the three levels of analytes spiked ranged from 63% to 118%, and the matrix effect was between 73% and 119%. Finally, the proposed method was applied to investigate the dynamic hormone concentration in Vitis vinifera L. cv. Cabernet Sauvignon berries from different vineyards, and assess the changes in endogenous hormones in grapes after treatment with exogenous growth regulators. We found that the contents of IP, ABA and IAA in pericarp and IP, IAA, IBA and SA in seed were significantly down-regulated after 10 days of treatment with NAA concentrations of 10 mg/L and 40 mg/L. In conclusion, this method helps to elucidate the role played by phytohormones in the maturation process and the accumulation of quality-related metabolites in phenol-rich fruits

A One-Step Polyphenol Removal Approach for Detection of Multiple Phytohormones from Grape Berry

Phytohormones play an important role in regulating the maturation process and the quality-related metabolite accumulation of fruits, and their concentration variation has always been concerned during fruit development. However, berry fruits, such as grape berries, are rich in a large number of secondary metabolites, which brings great challenges to the isolation and determination of hormones. In this work, we used grapes as experimental materials and proposed a solid-phase extraction (SPE) protocol to efficiently isolate multiple hormones from phenol-rich matrix using a mixture of dichloromethane, methanol and formic acid as eluent. A highly sensitive method based on ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) was developed to quantify a total of 11 plant growth regulators, including the recognized phytohormones, in grape pericarp and seed. The established method showed satisfactory precision (RSD 2 > 0.9980). The limits of detection (LOD) and the limit of quantification (LOQ) were 0.001–0.75 ng/mL and 0.004–2.5 ng/mL, respectively. The recovery for the three levels of analytes spiked ranged from 63% to 118%, and the matrix effect was between 73% and 119%. Finally, the proposed method was applied to investigate the dynamic hormone concentration in Vitis vinifera L. cv. Cabernet Sauvignon berries from different vineyards, and assess the changes in endogenous hormones in grapes after treatment with exogenous growth regulators. We found that the contents of IP, ABA and IAA in pericarp and IP, IAA, IBA and SA in seed were significantly down-regulated after 10 days of treatment with NAA concentrations of 10 mg/L and 40 mg/L. In conclusion, this method helps to elucidate the role played by phytohormones in the maturation process and the accumulation of quality-related metabolites in phenol-rich fruits

Evolution of Seed-Soluble and Insoluble Tannins during Grape Berry Maturation

Condensed tannins (CT) in wine are derived from the seeds and skins of grapes, and their composition and content contribute to the bitterness/astringency characteristics and ageing potential of the wine. Global warming has accelerated the ripening process of grape berries, making them out of sync with seed ripening. To understand the influence of berry ripening on the seed CT composition and content, we analyzed the changes in the soluble and insoluble CT in the seeds of ‘Cabernet Sauvignon’ grapes from two vineyards over two years. The results showed that the seed-soluble CT presented a slight downward trend in fluctuation during grape berry development, while the insoluble CT increased continuously before the véraison and remained at a high level afterwards. Relatively speaking, a lower sugar increment in developing grape berries favored the conversion of seed CT towards a higher degree of polymerization. The terminal unit of soluble CT was dominated by epigallocatechin gallate, the content of which decreased as the seeds matured. It is suggested that the seeds should be fully matured to reduce this bitter component in tannins. This study provides a reference for us to control the grape ripening process and produce high-quality grapes for wine making

Dissecting Seed Proanthocyandin Composition and Accumulation under Different Berry Ripening Process in Wine Grapes

Grape berry proanthocyandin (PA) mainly exists in the skin and seeds. Its content and composition determine the intensity of bitterness and astringency. Affected by global warming, the world’s wine-producing regions, in particular in dry-hot regions such as western China, are facing the problem of unsynchronized berry ripening and seed ripening. Therefore, it is urgent to understand the influence of berry ripening progression on the composition and accumulation of seed PA, ultimately providing strategies for grape harvest decisions. In this paper, Vitis vinfera L. cv. Cabernet sauvignon and Marselan grapes from four sub-regions with different maturation processes were used as experimental materials to study the changes of soluble and insoluble PA contents as well as differences in their composition and mean degree of polymers (mDP) in seeds. The results showed that compared with ‘Cabernet sauvignon’ seeds, the mDP of soluble and insoluble PA were higher in ‘Marselan’ seeds. Both varieties showed that the grape berry, with the fastest sugar accumulation, had relatively high soluble PA content in seeds and a high content of (-)-epigallocatechin-3-gallate and (-)-epicatechin in the seed PA composition units. In contrast, the ‘Cabernet sauvignon’ grapes from the YQ vineyard exhibited the slowest sugar accumulation speed among the four studied vineyards, and their seed PA had the highest mDP and the lowest proportion of (-)-epigallocatechin-3-gallate in the composition units when commercially harvested. According to the results, it is suggested that a faster maturation process would bring about higher levels of bitterness composition, such as (-)-epigallocatechin-3-gallate in seed PA, which is not conducive to the formation of good-tasting tannins

Effect of Cluster-Zone Leaf Removal at Different Stages on Cabernet Sauvignon and Marselan (<i>Vitis vinifera</i> L.) Grape Phenolic and Volatile Profiles

This study investigated the effect of leaf removal at three stages of grape development on the phenolic and volatile profiles of Cabernet Sauvignon and Marselan grapevines for two consecutive years in the Jieshi Mountain region, an area of eastern China with high summer rainfall. The results indicated that cluster-zone leaf removal generally reduced the titratable acidity of both varieties, but did not affect the total soluble solids of grape berries. Leaf-removal treatments increased the anthocyanin and flavonol content of berries in both varieties. However, in Cabernet Sauvignon, leaf removal negatively affected the norisoprenoid compounds, with a more pronounced impact observed when the leaf removal was conducted at an early stage. This negative effect may be related to a decrease in the levels of violaxanthin and neoxanthin, potential precursors of vitisprine and β-damascenone. In contrast, the removal of leaves had no effect on the norisoprenoid aroma of Marselan grapes

A Novel Position-Specific Encoding Algorithm (SeqPose) of Nucleotide Sequences and Its Application for Detecting Enhancers

Enhancers are short genomic regions exerting tissue-specific regulatory roles, usually for remote coding regions. Enhancers are observed in both prokaryotic and eukaryotic genomes, and their detections facilitate a better understanding of the transcriptional regulation mechanism. The accurate detection and transcriptional regulation strength evaluation of the enhancers remain a major bioinformatics challenge. Most of the current studies utilized the statistical features of short fixed-length nucleotide sequences. This study introduces the location information of each k-mer (SeqPose) into the encoding strategy of a DNA sequence and employs the attention mechanism in the two-layer bi-directional long-short term memory (BD-LSTM) model (spEnhancer) for the enhancer detection problem. The first layer of the delivered classifier discriminates between enhancers and non-enhancers, and the second layer evaluates the transcriptional regulation strength of the detected enhancer. The SeqPose-encoded features are selected by the Chi-squared test, and 45 positions are removed from further analysis. The existing studies may focus on selecting the statistical DNA sequence descriptors with large contributions to the prediction models. This study does not utilize these statistical DNA sequence descriptors. Then the word vector of the SeqPose-encoded features is obtained by using the word embedding layer. This study hypothesizes that different word vector features may contribute differently to the enhancer detection model, and assigns different weights to these word vectors through the attention mechanism in the BD-LSTM model. The previous study generously provided the training and independent test datasets, and the proposed spEnhancer is compared with the three existing state-of-the-art studies using the same experimental procedure. The leave-one-out validation data on the training dataset shows that the proposed spEnhancer achieves similar detection performances as the three existing studies. While spEnhancer achieves the best overall performance metric MCC for both of the two binary classification problems on the independent test dataset. The experimental data shows that the strategy of removing redundant positions (SeqPose) may help improve the DNA sequence-based prediction models. spEnhancer may serve well as a complementary model to the existing studies, especially for the novel query enhancers that are not included in the training dataset