3fd6c029a2cece6245c294c6a1f8b830 25TV51_HorticRes_Monfort_pub.pdf 2fe0835b46a640027d50c1920ba449babd2f2fe5 25TV51_HorticRes_Monfort_pub.pdf f30560a7d339dfcadfdef0ccc1222e359e560fa69ec30d4dbff53292309e58aa 25TV51_HorticRes_Monfort_pub.pdf Title: The role of Fragaria vesca homolog of a (Z)-3:(E)-2-hexenal isomerase in the development of green-leafy fruit aroma Subject: DOI: 10.1093/hr/uhaf163, Horticulture Research, 12, 00, 1 10 2025. Abstract: The green leaf volatiles (Z)-3-hexenal and (E)-2-hexenal are key components of the characteristic strawberry aroma, an important determinant of consumer preferences. Green leaf volatiles (GLVs) are C6 compounds that impart fresh, green notes and are involved in plant wounding responses. GLV biosynthesis requires several enzymatic steps to convert polyunsaturated fatty acids to C6-aldehydes, alcohols, and esters, respectively. However, the biosynthesis of GLVs in strawberries, such as the isomerization of (Z)-3-hexenal to (E)-2-hexenal, remains poorly understood. In this study, we identified a (Z)-3:(E)-2-hexenal isomerase gene (FvHI) using phylogenetic analysis, characterized its expression in different tissues, and characterized its function using stable transformation. Volatile analysis by gas chromatography–mass spectrometry (GC–MS) of fruits from a Fragaria vesca near-isogenic line (NIL) collection revealed a distinct ratio of (Z)-3-hexenal and (E)-2-hexenal in lines containing Fragaria bucharica introgressions in the distal end of linkage group 5. Consequently, FvHI was located within this genomic region. The coding sequence of FvHI was nearly identical between the recurrent parent and a selected NIL individual containing an introgression in the distal end linkage group 5, indicating that the contrasting ratio of (Z)-3 and (E)-2 GLV isomers may be attributed to transcriptional differences. Accordingly, FvHI expression in ripe fruits was lower in the selected NIL individual than in the recurrent parent. Lastly, FvHI overexpression decreased (Z)-3-hexenal accumulation and increased (E)-2-hexenal accumulation in the recurrent parent and the selected NIL individual. These results suggest that FvHI plays a role in producing the characteristic strawberry aroma by converting (Z)-3-hexenal to (E)-2-hexenal. Keywords: Author: Creator: LaTeX with hyperref package Producer: Acrobat Distiller 24.0 (Windows); modified using iTextSharp.LGPLv2.Core 3.7.4.0 CreationDate: Tue Jul 8 02:15:38 2025 CEST ModDate: Thu Oct 16 14:52:51 2025 CEST Custom Metadata: yes Metadata Stream: yes Tagged: yes UserProperties: no Suspects: no Form: none JavaScript: no Pages: 9 Encrypted: no Page size: 595.26 x 782.337 pts Page rot: 0 File size: 1047609 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- DCNAJF+CaeciliaLTStd-Bold Type 1C Custom yes yes yes 67 0 DCNAJG+CaeciliaLTStd-Roman Type 1C Custom yes yes yes 1356 0 DCNAPF+CaeciliaLTStd-Italic Type 1C Custom yes yes yes 1357 0 DCNBAF+CaeciliaLTStd-BoldItalic Type 1C Custom yes yes yes 68 0 DCNBHE+CaeciliaLTStd-Light Type 1C Custom yes yes yes 69 0 DCNBKD+CaeciliaLTStd-Heavy Type 1C Custom yes yes yes 70 0 DCNBND+CaeciliaLTStd-LightItalic Type 1C Custom yes yes yes 71 0 JLIZMH+ArialMT TrueType WinAnsi yes yes no 1 0 DCNDFL+MTGU Type 1C Custom yes yes yes 72 0 DCNLDN+MTMI Type 1C Custom yes yes yes 119 0 DCNLDM+MTSYN Type 1C Custom yes yes yes 120 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-11-28 03:10:51 CET RepresentationInformation: 25TV51_HorticRes_Monfort_pub.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-11-27 14:24:09 CET Size: 1047609 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf Profile: Tagged PDF PDFMetadata: Objects: 1400 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: true NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseOutlines Language: en Outlines: Item: Title: The role of Fragaria vesca homolog of a Z-3:E -2-hexenal isomerase in the development of green-leafy fruit aroma Destination: toc.chapter Children: Item: Title: Introduction Destination: section.12.1.1 Item: Title: Results Destination: section.19.1.2 Item: Title: Discussion Destination: section.24.1.3 Item: Title: Materials and methods Destination: section.25.1.4 Item: Title: Acknowledgements Destination: section.32.1.5 Item: Title: Author contributions Destination: section.33.1.6 Item: Title: Data availability Destination: section.34.1.7 Item: Title: Conflict of interest statement Destination: section.35.1.8 Item: Title: Supplementary data Destination: section.36.1.9 Info: Title: The role of Fragaria vesca homolog of a (Z)-3:(E)-2-hexenal isomerase in the development of green-leafy fruit aroma Author: Subject: DOI: 10.1093/hr/uhaf163, Horticulture Research, 12, 00, 1 10 2025. Abstract: The green leaf volatiles (Z)-3-hexenal and (E)-2-hexenal are key components of the characteristic strawberry aroma, an important determinant of consumer preferences. Green leaf volatiles (GLVs) are C6 compounds that impart fresh, green notes and are involved in plant wounding responses. GLV biosynthesis requires several enzymatic steps to convert polyunsaturated fatty acids to C6-aldehydes, alcohols, and esters, respectively. However, the biosynthesis of GLVs in strawberries, such as the isomerization of (Z)-3-hexenal to (E)-2-hexenal, remains poorly understood. In this study, we identified a (Z)-3:(E)-2-hexenal isomerase gene (FvHI) using phylogenetic analysis, characterized its expression in different tissues, and characterized its function using stable transformation. Volatile analysis by gas chromatography mass spectrometry (GC MS) of fruits from a Fragaria vesca near-isogenic line (NIL) collection revealed a distinct ratio of (Z)-3-hexenal and (E)-2-hexenal in lines containing Fragaria bucharica introgressions in the distal end of linkage group 5. Consequently, FvHI was located within this genomic region. The coding sequence of FvHI was nearly identical between the recurrent parent and a selected NIL individual containing an introgression in the distal end linkage group 5, indicating that the contrasting ratio of (Z)-3 and (E)-2 GLV isomers may be attributed to transcriptional differences. Accordingly, FvHI expression in ripe fruits was lower in the selected NIL individual than in the recurrent parent. Lastly, FvHI overexpression decreased (Z)-3-hexenal accumulation and increased (E)-2-hexenal accumulation in the recurrent parent and the selected NIL individual. These results suggest that FvHI plays a role in producing the characteristic strawberry aroma by converting (Z)-3-hexenal to (E)-2-hexenal. Keywords: Creator: LaTeX with hyperref package Producer: Acrobat Distiller 24.0 (Windows); modified using iTextSharp.LGPLv2.Core 3.7.4.0 CreationDate: Tue Jul 08 02:15:38 CEST 2025 ModDate: Thu Oct 16 14:52:51 CEST 2025 ID: 0x0c44f2d1de88f1f7943893de0c7845cf, 0x5b9921ec991d61611afb37c612deae5b Filters: FilterPipeline: FlateDecode FilterPipeline: DCTDecode Images: Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 1800 ImageHeight: 2333 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 2075 ImageHeight: 1474 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 950 ImageHeight: 1212 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 2075 ImageHeight: 1649 ColorSpace: RGB BitsPerSample: 8 BitsPerSampleUnit: integer Fonts: Type1: Font: BaseFont: DCNAJF+CaeciliaLTStd-Bold FontSubset: true FirstChar: 40 LastChar: 121 FontDescriptor: FontName: DCNAJF+CaeciliaLTStd-Bold Flags: Serif, Symbolic, ForceBold FontBBox: -166, -273, 1141, 938 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNBAF+CaeciliaLTStd-BoldItalic FontSubset: true FirstChar: 46 LastChar: 118 FontDescriptor: FontName: DCNBAF+CaeciliaLTStd-BoldItalic Flags: Serif, Symbolic, Italic, ForceBold FontBBox: -169, -273, 1082, 932 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNBHE+CaeciliaLTStd-Light FontSubset: true FirstChar: 37 LastChar: 250 FontDescriptor: FontName: DCNBHE+CaeciliaLTStd-Light Flags: Serif, Symbolic FontBBox: -164, -266, 1130, 908 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNBKD+CaeciliaLTStd-Heavy FontSubset: true FirstChar: 46 LastChar: 124 FontDescriptor: FontName: DCNBKD+CaeciliaLTStd-Heavy Flags: Serif, Symbolic, ForceBold FontBBox: -170, -277, 1146, 970 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNBND+CaeciliaLTStd-LightItalic FontSubset: true FirstChar: 41 LastChar: 252 FontDescriptor: FontName: DCNBND+CaeciliaLTStd-LightItalic Flags: Serif, Symbolic, Italic FontBBox: -166, -267, 1033, 909 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNLDN+MTMI FontSubset: true FirstChar: 60 LastChar: 60 FontDescriptor: FontName: DCNLDN+MTMI Flags: Serif, Symbolic, Italic FontBBox: 0, -213, 987, 680 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNDFL+MTGU FontSubset: true FirstChar: 2 LastChar: 181 FontDescriptor: FontName: DCNDFL+MTGU Flags: Symbolic FontBBox: 24, -213, 839, 683 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNLDM+MTSYN FontSubset: true FirstChar: 2 LastChar: 215 FontDescriptor: FontName: DCNLDM+MTSYN Flags: Symbolic FontBBox: 0, -954, 1043, 900 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNAJG+CaeciliaLTStd-Roman FontSubset: true FirstChar: 40 LastChar: 237 FontDescriptor: FontName: DCNAJG+CaeciliaLTStd-Roman Flags: Serif, Symbolic FontBBox: -167, -270, 1136, 926 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: DCNAPF+CaeciliaLTStd-Italic FontSubset: true FirstChar: 70 LastChar: 118 FontDescriptor: FontName: DCNAPF+CaeciliaLTStd-Italic Flags: Serif, Symbolic, Italic FontBBox: -166, -270, 1053, 915 FontFile3: true EncodingDictionary: Differences: true ToUnicode: true TrueType: Font: BaseFont: JLIZMH+ArialMT FontSubset: true FirstChar: 32 LastChar: 121 FontDescriptor: FontName: JLIZMH+ArialMT Flags: Nonsymbolic FontBBox: -664, -324, 2000, 1039 FontFile2: true Encoding: WinAnsiEncoding XMP: application/pdf The role of Fragaria vesca homolog of a (Z)-3:(E)-2-hexenal isomerase in the development of green-leafy fruit aroma Zhang RongNunnally Martínez DylanKoskela Elli A.Monfort Amparo DOI: 10.1093/hr/uhaf163, Horticulture Research, 12, 00, 1 10 2025. Abstract: The green leaf volatiles (Z)-3-hexenal and (E)-2-hexenal are key components of the characteristic strawberry aroma, an important determinant of consumer preferences. Green leaf volatiles (GLVs) are C6 compounds that impart fresh, green notes and are involved in plant wounding responses. GLV biosynthesis requires several enzymatic steps to convert polyunsaturated fatty acids to C6-aldehydes, alcohols, and esters, respectively. However, the biosynthesis of GLVs in strawberries, such as the isomerization of (Z)-3-hexenal to (E)-2-hexenal, remains poorly understood. In this study, we identified a (Z)-3:(E)-2-hexenal isomerase gene (FvHI) using phylogenetic analysis, characterized its expression in different tissues, and characterized its function using stable transformation. Volatile analysis by gas chromatography–mass spectrometry (GC–MS) of fruits from a Fragaria vesca near-isogenic line (NIL) collection revealed a distinct ratio of (Z)-3-hexenal and (E)-2-hexenal in lines containing Fragaria bucharica introgressions in the distal end of linkage group 5. Consequently, FvHI was located within this genomic region. The coding sequence of FvHI was nearly identical between the recurrent parent and a selected NIL individual containing an introgression in the distal end linkage group 5, indicating that the contrasting ratio of (Z)-3 and (E)-2 GLV isomers may be attributed to transcriptional differences. Accordingly, FvHI expression in ripe fruits was lower in the selected NIL individual than in the recurrent parent. Lastly, FvHI overexpression decreased (Z)-3-hexenal accumulation and increased (E)-2-hexenal accumulation in the recurrent parent and the selected NIL individual. These results suggest that FvHI plays a role in producing the characteristic strawberry aroma by converting (Z)-3-hexenal to (E)-2-hexenal. © The Author(s) 2025. Published by Oxford University Press on behalf of Nanjing Agricultural University. 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