Lipoxygenase Hydroperoxide Lyase Research Articles

Persicaria minor F-box Gene Pmf-box1 Indirectly Affects Arabidopsis thaliana LOX-HPL Pathway for Green Leaf Volatile Production

Green leaf volatiles (GLVs) play an essential role in plant defence, plant-plant interaction and plant-insect interaction. The plant releases GLVs and inhibits the growth and propagation of plant pathogens. In this study, overexpression of PmF-box1 in wild type A. thaliana showed the downregulation of genes involved in the lipoxygenase-hydroperoxide lyase (LOX-HPL) pathway, which contributes to the biosynthesis of GLVs. It resulted in a marked reduction of hexanal production in the PmF-box1-overexpressing plant. The expression pattern of LOX-HPL branch genes in the kelch-repeat modified PmF-box1 (KMF)-overexpressing plant showed a pattern much closer to the expression of LOX-HPL branch genes in the vector control (VC) plant. It was shown that the functional KMF protein sequence was not responsible for the significant reduction of all GLVs including hexanal, 1-hexanol, (Z)-3-hexen-1-ol, and the carbon 5 (C5) volatile, 1-penten-3-ol, in plants overexpressing KMF. Furthermore, this study also showed that the relative proportion of production of 1-penten-3-ol to hexanal was higher in the PmF-box1-overexpressing plant. Based on the current comparative literature search, PmF-box1 does not appear to interact directly with the proteins or transcription factors of the LOX-HPL pathway. On the other hand, PmF-box1 interacts with SAMS1, which subsequently influences the HPL pathway enzyme genes. Thus, this study highlights the potential roles of PmF-box1 in the manipulation of GLV productions.

Identification of the key genes contributing to the LOX-HPL volatile aldehyde biosynthesis pathway in jujube fruit

Jujube (Ziziphus jujuba Mill.) is a traditional popular fruit widely grown in China. The volatiles in jujube determine its unique flavor and the high fruit quality required by consumers. However, the biosynthesis of volatiles in jujube were remain unknown. By using gas chromatography–mass spectrometry, there were 46 volatile compounds were identified and determined from three representative jujube fruit types at six developmental stages, including the dry-used (Z. jujuba cv. ‘Junzao’), the fresh-used (Z. jujuba cv. ‘Dongzao’), and wild jujube (Z. jujuba var. spinosa Hu. cv. ‘Qingjiansuanzao’). The aldehydes were identified as major volatile contributors to flavor, of which (E)-2-hexenal was the primary volatile in jujube fruit. Then LOX and HPL gene family were identified in jujube, which were involved in aldehyde biosynthesis through the lipoxygenase-hydroperoxide lyase (LOX-HPL) pathway. Gene expression analysis suggested that ZjLOX3, ZjLOX4, and ZjHPL1 were highly correlated with the accumulation of (E)-2-hexenal, and their proteins were localized to the nucleus and cytoplasm. Transient over-expression of ZjLOX3, ZjLOX4, and ZjHPL1 in jujube fruit significantly enhanced the accumulation of (E)-2-hexenal. Our study provides valuable information on the major volatiles and their biosynthesis in different types of jujube fruit. These results will help determine flavor improvements for future breeding.

Metabolic Flow of C6 Volatile Compounds From LOX-HPL Pathway Based on Airflow During the Post-harvest Process of Oolong Tea.

Aroma is an essential quality indicator of oolong tea, a tea derived from the Camellia sinensis L. plant. Carboxylic 6 (C6) acids and their derivative esters are important components of fatty acid (FA)-derived volatiles in oolong tea. However, the formation and regulation mechanism of C6 acid during postharvest processing of oolong tea remains unclear. To gain better insight into the molecular and biochemical mechanisms of C6 compounds in oolong tea, a combined analysis of alcohol dehydrogenase (ADH) activity, CsADH2 key gene expression, and the FA-derived metabolome during postharvest processing of oolong tea was performed for the first time, complemented by CsHIP (hypoxia-induced protein conserved region) gene expression analysis. Volatile fatty acid derivative (VFAD)-targeted metabolomics analysis using headspace solid-phase microextraction–gas chromatography time-of-flight mass spectrometry (HS-SPEM-GC-TOF-MS) showed that the (Z)-3-hexen-1-ol content increased after each turnover, while the hexanoic acid content showed the opposite trend. The results further showed that both the ADH activity and CsADH gene expression level in oxygen-deficit-turnover tea leaves (ODT) were higher than those of oxygen-turnover tea leaves (OT). The C6-alcohol-derived ester content of OT was significantly higher than that of ODT, while C6-acid-derived ester content showed the opposite trend. Furthermore, the HIP gene family was screened and analyzed, showing that ODT treatment significantly promoted the upregulation of CsHIG4 and CsHIG6 gene expression. These results showed that the formation mechanism of oolong tea aroma quality is mediated by airflow in the lipoxygenase–hydroperoxide lyase (LOX-HPL) pathway, which provided a theoretical reference for future quality control in the postharvest processing of oolong tea.

Profiling Analysis of Volatile and Non-volatile Compounds in Vitis Vinifera Berries (cv. Chardonnay) and Spontaneous Bud Mutation

A novel clonal variety of Vitis vinifera was identified from “Chardonnay” using inter-simple sequence repeat (ISSR) markers and called “bud mutation. ” The metabolomic profiles in Chardonnay and bud mutation berries indicated essential differences in the expression of key genes in the pathways of 2-C-methyl-D-erythritol-4-phosphate (MEP) and lipoxygenase-hydroperoxide lyase (LOX-HPL). Bud mutation fruits also matured 10 days earlier than Chardonnay and have higher carotenoid, sugar, and acidic compound contents. Furthermore, the gene expression was examined in the biosynthetic pathways of two ripening-associated hormones, abscisic acid (ABA) and jasmonic acid (JA), which significantly increased in bud mutation compared with the Chardonnay fruit. The synthesis and metabolism of amino acids, terpenes, fatty acids, volatile components, and specialized metabolites significantly increased in bud mutation. Therefore, in comparison with Chardonnay, bud mutation is considered a highly aroma-producing grape variety for an improvement in the beverage industry.

Effects of different color paper bags on aroma development of Kyoho grape berries

This study investigated the influence of red, green, blue, and white paper bags on the free volatile compound development of Kyoho grape berries from green to harvest. Seven functional groups of volatiles were identified during the development of Kyoho grape berries including esters, aldehydes, alcohols, terpenes, ketones, acids, and hydrocarbons. Esters and aldehydes were abundant in Kyoho grape berries, mainly represented by ethyl acetate, ethyl butyrate, and (E)-2-hexenal. They accumulated quickly after veraison and slightly decreased toward maturation. Red, green, blue, and white paper bags promoted the accumulation of esters and inhibited the accumulation of aldehydes, also inhibited the accumulation of alcohols, tepenes, ketones, and acids. Their effect from strong to weak was green, blue, red, and white paper bags. The expression profiles of genes in the lipoxygenase-hydroperoxide lyase (LOX-HPL) pathway were also analyzed and the results indicated that the regulation of red, green, blue, and white paper bags on aldehydes, alcohols, and esters volatile aromas was at transcriptional level. The results expanded our comprehension in grape aroma biosynthesis and berry bagging technique in table grape cultivation.

Differential Expression of VvLOXA Diversifies C6 Volatile Profiles in Some Vitis vinifera Table Grape Cultivars.

C6 volatiles are synthesized through lipoxygenase-hydroperoxide lyase (LOX-HPL) pathway and these volatiles play important roles in the aromatic quality of grape berries. This study investigated the evolution of both C6 volatiles and the key genes in the LOX-HPL pathway in different table grape cultivars during the berry development period, and further assessed the correlation between the accumulation of C6 volatiles and the expression of these genes in these cultivars. Results showed that hexanal, (E)-2-hexenal, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol were found to be the dominant C6 volatiles in these ripened grape cultivars under two consecutive vintages, and their flavor notes were incorporated in the overall aroma of these cultivars. The cultivar “Xiangfei” showed the most abundant level of C6 aldehydes and C6 acid, whereas the cultivar “Tamina” and “Moldova” possessed the highest C6 alcohol content. The “Muscat of Alexandria” cultivar was found to contain the highest level of C6 esters. C6 volatiles were grouped into three evolutionary patterns in these cultivars during berry development, and their evolution was consistent with the evolution of the LOX-HPL pathway genes’ expression. Pearson’s correlation analysis indicated that the LOX-HPL-pathway-related genes were correlated to the accumulation of C6 volatiles in these cultivars, and VvLOXA appeared to be an important gene that regulated the synthesis of all C6 volatiles.

Ozone Improves the Aromatic Fingerprint of White Grapes

Ozone, a powerful oxidative stressor, has been recently used in wine industry as sanitizing agent to reduce spoilage microflora on grapes. In this study, we evaluated ozone-induced metabolic and molecular responses during postharvest grape dehydration. Ozone increased the contents of total volatile organic compounds (VOCs), which have a great impact on the organoleptic properties of grapes and wines. Among terpenes, responsible for floral and fruity aroma, linalool, geraniol and nerol were the major aromatic markers of Moscato bianco grapes. They were significantly affected by the long-term ozone treatment, increasing their concentration in the last phases of dehydration (>20% weight loss). At molecular level, our results demonstrated that both postharvest dehydration and ozone exposure induce the biosynthesis of monoterpenes via methylerythritol phosphate (MEP) pathway and of aldehydes from lipoxygenase-hydroperoxide lyase (LOX-HPL) pathway. Therefore, transcriptional changes occurred and promoted the over-production of many important volatile compounds for the quality of white grapes.

Lipoxygenase Involvement in Ripening Strawberry

The enzymatic activity, subcellular localization, and immunolocalization of plant lipoxygenase (LOX) in strawberry fruits (Fragaria x ananassa, Duch) were investigated. Chemical and enzymatic properties of LOX have been characterized, and the LOX capability of oxygenating free and esterified unsaturated fatty acids into C6 volatile aldehydes has been confirmed. Fruits at unripe, turning, and ripe stages were analyzed for LOX activity and protein localization by Western blots, two-dimensional electrophoresis, and immunolocalization analyses. The ability of strawberry tissues to in vivo metabolize linolenic acid or linoleic acid into C6 volatile aldehydes and the LOX products was also analyzed. Analysis of strawberry proteins showed that a number of LOX forms, corresponding to at least two mobility groups of approximately 100 and 98 kDa and pI values ranging between 4.4 and 6.5, were present. Confocal and electron microscopy analyses support the idea that LOX proteins are associated to lipid-protein aggregates. Both exogenously supplied linoleate and linolenate were converted into hexanal and trans-2-hexenal at the three fruit-ripening stages. Our experiments suggest the presence of different LOX isoforms in strawberry fruits and that the lipoxygenase-hydroperoxide lyase pathway plays a role in converting lipids to C6 volatiles during ripening.

Lipoxygenase and hydroperoxide lyase activities in ripening strawberry fruits.

The enzymes lipoxygenase and hydroperoxide lyase have been identified in strawberry (Fragariax ananassa Duch.) var. Camarosa. Their subcellular localization, substrate preference, and product specificity were determined in mature strawberry fruits. The activity of both enzymes was located mainly in the microsomal fraction. Linolenic acid was the preferred substrate for strawberry lipoxygenase, forming 13- and 9-hydroperoxides of this acid in the proportion 70:30. The strawberry hydroperoxide lyase cleaves 13-hydroperoxide of linoleic (13% relative activity) and linolenic (100% relative activity) acids to form hexanal and (3Z)-hexenal, respectively. Both enzyme activities and endogenous content of volatile aldehydes formed by sequential action of lipoxygenase-hydroperoxide lyase were evaluated during strawberry development and ripening. A sequential enzymatic pathway for the formation of green odor compounds in strawberry is proposed.