Abstract：Green leaf volatiles (GLV), including C6 and C9 aldehydes, alcohols and lipids, are important aroma components in grapes(Vitis vinifera L). and wines. Lipoxygenase (LOX) is a key enzyme for the synthesis of GLVs. In this study, full-length cDNAs of VvLOX1 (KF033130) and VvLOX2 (KF033131) were cloned from Cabernet Sauvignon(V. vinifera) berries. By bioinformatics analysis, these two genes belonged to type Ⅰ 9-LOX and type Ⅱ 13-LOX, respectively. The expression levels of VvLOX1 and VvLOX2 were higher in leaves and roots than those in stems and tendrils. During the development of grape berries, the expression levels of VvLOX1 and VvLOX2 were related to GLV synthesis, but the peaks of two genes expression appeared 2 weeks earlier than those of C6 and C9 contents. After wounding treatment, there was a significant positive correlation between VvLOX2 expression level and C6 content, which indicated that this gene played an important role in plant stress resistance. The results of this study provide the basic data for exploring the function mechanism of LOXs in aroma formation of grape berries.
AGRAWAL G K, TAMOGAMI S, HAN O, et al. 2004. Rice octadecanoid pathway[J]. Biochemical & Biophysical Research Communications, 317(1): 1-15.ALAM M M, NAHAR K, HASANUZZAMAN M, et al. 2014. Exogenous jasmonic acid modulates the physiology, antioxidant defense and glyoxalase systems in imparting drought stress tolerance in different brassica species[J]. Plant Biotechnology Reports, 8(3): 279-293.BANNENBERG G, MARTíNEZ M, HAMBERG M, et al. 2009. Diversity of the enzymatic activity in the lipoxygenase gene family of Arabidopsis thaliana[J]. Lipids, 44(2): 85.BLéE E. 1998. Phytooxylipins and plant defense reactions[J]. Progress in Lipid Research, 37(1): 33-72.CHEHAB E W, PEREA J V, GOPALAN B, et al. 2007. Oxylipin pathway in rice and arabidopsis[J]. Journal of Integrative Plant Biology, 49(1): 43-51.CHEN G, HACKETT R, WALKER D, et al. 2004. Identification of a specific isoform of tomato lipoxygenase (tomloxc) involved in the generation of fatty acid-derived flavor compounds[J]. Plant Physiology, 136(1): 2641-51.CHEN S, ZHANG R, HAO L, et al. 2015. Profiling of volatile compounds and associated gene expression and enzyme activity during fruit development in two cucumber cultivars[J]. Plos One, 10(3): 1-22.FEUSSNER I, WASTERNACK C. 2003. The lipoxygenase pathway[J]. Annual Review of Plant Biology, 53(1): 275-297.FEUSSNER I. 2006. Lipoxygenases: Occurrence, functions and catalysis[J]. Journal of Plant Physiology, 163(3): 348-357.HALITSCHKE R, BALDWIN I T. 2003. Antisense lox expression increases herbivore performance by decreasing defense responses and inhibiting growth-related transcriptional reorganization in nicotiana attenuata[J]. Plant Journal for Cell & Molecular Biology, 36(6): 794-807.HEITZ T, BERGEY D R, RYAN C A. 1997. A gene encoding a chloroplast-targeted lipoxygenase in tomato leaves is transiently induced by wounding, systemin, and methyl jasmonate[J]. Plant Physiology, 114(3): 1085-1093.HOU Y, MENG K, HAN Y, et al. 2015. The persimmon 9-lipoxygenase gene DKLOX3 plays positive roles in both promoting senescence and enhancing tolerance to abiotic stress[J]. Frontiers in Plant Science, 6:1-15.HOWE G A, SCHILMILLER A L. 2002. Oxylipin metabolism in response to stress[J]. Current Opinion in Plant Biology, 5(3): 230-236.JIN H S, VAN K, DONG H K, et al. 2008. The lipoxygenase gene family: A genomic fossil of shared polyploidy between glycine max and medicago truncatula[J]. BMC Plant Biology, 8(1): 133.JU Y L, LIU M, ZHAO H, et al. 2016. Effect of exogenous abscisic acid and methyl jasmonate on anthocyanin composition, fatty acids, and volatile compounds of Cabernet Sauvignon (Vitis Vinifera L.) grape berries[J]. Molecules, 21(10): 1354.KOLOMIETS M V, HANNAPEL D J, CHEN H, et al. 2001. Lipoxygenase is involved in the control of potato tuber development[J]. Plant Cell, 13(3): 613-626.LI M, LI L, DUNWELL J M, et al. 2014. Characterization of the lipoxygenase (LOX) gene family in the chinese white pear (Pyrus Bretschneideri) and comparison with other members of the rosaceae[J]. BMC Genomics, 15(1): 444.LIM C W, HAN S W, HWANG I S, et al. 2015. The pepper lipoxygenase calox1 plays a role in osmotic, drought and high salinity stress response[J]. Plant & Cell Physiology, 56(5): 930-942.LIU S Q, LIU X H, JIANG L W. 2011. Genome-wide identification, phylogeny and expression analysis of the lipoxygenase gene family in cucumber[J]. Genetics & Molecular Research, 10(4): 2613-2636.MATSUI K. 2006. Green leaf volatiles: Hydroperoxide lyase pathway of oxylipin metabolism[J]. Current Opinion in Plant Biology, 9(3): 274-280.OLIVEIRA J M, FARIA M, Sá F, et al. 2006. C6 -alcohols as varietal markers for assessment of wine origin[J]. Analytica Chimica Acta, 563(1–2): 300-309.PILATI S, BRAZZALE D, GUELLA G, et al. 2014. The onset of grapevine berry ripening is characterized by ros accumulation and lipoxygenase-mediated membrane peroxidation in the skin[J]. BMC Plant Biology, 14(1): 1-15.PODOLYAN A, WHITE J, JORDAN B, et al. 2010. Identification of the lipoxygenase gene family from vitis vinifera and biochemical characterisation of two 13-lipoxygenases expressed in grape berries of sauvignon blanc[J]. Functional Plant Biology, 37(8): 767-784.SANTINO A, DE P A, GALLO A, et al. 2003. Biochemical and molecular characterization of hazelnut (Corylus avellana) seed lipoxygenases[J]. European Journal of Biochemistry, 270(21): 4365-4375.SHAH J. 2009. Plants under attack: Systemic signals in defence[J]. Current Opinion in Plant Biology, 12(4): 459-464.VICENTE J, KULASEKARAN S, VELLOSILLO T, et al. 2011. Antagonistic role of 9-lipoxygenase-derived oxylipins and ethylene in the control of oxidative stress, lipid peroxidation and plant defence[J]. Plant Journal, 67(3): 447-458.VOGT J, SCHILLER D, ULRICH D, et al. 2013. Identification of lipoxygenase (lox) genes putatively involved in fruit flavour formation in apple ( malus × domestica )[J]. Tree Genetics & Genomes, 9(6): 1493-1511.XU X Q, CHENG G, DUAN L L, et al. 2015. Effect of training systems on fatty acids and their derived volatiles in cabernet sauvignon grapes and wines of the north foot of mt. Tianshan[J]. Food Chemistry, 181: 198-206.Zhang B, CHEN K S, BOWEN J, et al. 2006. Differential expression within the lox gene family in ripening kiwifruit[J]. Journal of Experimental Botany, 57(14): 3825-3836.ZHANG B, LI X,CHEN K S, et al. 2008. Molecular cloning of lipoxygenase gene family members in kiwifruit based on est database[J]. Acta Horticulturae Sinica, 35(3): 337-342.ZHANG B, SHEN J Y, WEI W W, et al. 2010. Expression of genes associated with aroma formation derived from the fatty acid pathway during peach fruit ripening[J]. Journal of Agricultural & Food Chemistry, 58(10): 6157-6165.ZHANG C, JIN Y, LIU J, et al. 2014. The phylogeny and expression profiles of the lipoxygenase (lox) family genes in the melon ( Cucumis Melo L.) genome[J]. Scientia Horticulturae, 2014, 170(3): 94-102.陈巧, 陈书霞, 王聪颖, 等. 2013. 黄瓜脂氧合酶基因CsLOX2的克隆及表达分析[J]. 中国农业科学, 46(11): 2285-2297. (CHEN Q, CHEN S X, WANG C Y, et al. Cloning and Expression Analysis of Lipoxygenase Gene CsLOX2 in Cucumis sativus (Cucumber) [J]. Scientia Agriculture Sinica, 46(11): 2285-2297.)陈书霞, 陈巧, 王聪颖, 等. 2012. 绿叶挥发物代谢调控及分子机理研究进展[J]. 中国农业科学, 45(8): 1545-1557. (CHEN S X, CHEN Q, WANG C Y, et al. Progress in Research on the Metabolic Regulation and Molecular Mechanism of Green Leave Volatiles (GLVs) [J]. Scientia Agriculture Sinica, 45(8): 1545-1557.)侯力璇, 张京芳，徐洪宇. 基于香气成分含量对酿酒红葡萄品种的分类研究[J]. 西北植物学报, 2012, 32(9): 1819-1828. (HOU L X, ZHANG J F, XU H Y. Classification of the red wine grapes based on the content of aroma components [J]. Acta Botanica Boreali-Occidentalia Sinica, 2012, 32(9): 1819-1828.)姜文广, 李记明, 徐岩, 等. 2011. 4种酿酒红葡萄果实的挥发性香气成分分析[J]. 食品科学, 32(6): 225-229. (JIANG W G, LI J M, XU Y, et al. 2011. Analysis of aroma components in four red grape varieties[J]. Food Science, 32(6): 225-229.)康文怀, 徐岩，崔彦志. 2010. 不同酿酒葡萄品种C6醛、醇风味化合物的比较[J]. 食品科学, 31(8): 252-256. (KANG W H, XU Y, CUI Y Z. Comparative analysis of contents of C6 aldehydes and alcohols responsible for the aroma of different varieties of wine grapes [J]. Food Science, 31(8): 252-256.)杨晓帆, 高媛, 韩梅梅，等. 2014. 云南高原区酿酒葡萄果实香气物质的积累规律[J]. 中国农业科学, 47(12):2405-2416. (YANG X F, GAO Y, HAN M M, et al. Accumulation Characteristics of Volatile Compounds in Wine Grape Berries Grown in High Altitude Regions of Yunnan [J]. Scientia Agriculture Sinica, 47(12):2405-2416.)张今今, 王跃进, 王西平, 等. 2003. 葡萄总RNA提取方法的研究[J]. 果树学报, 20(3): 178-181. (ZHANG J J, WANG Y J, WANG X P, et al. An improved method for rapidly extracting total RNA from Vitis [J]. Journal of Fruit Science, 20(3): 178-181.)张可文, 安钰, 胡增辉, 等. 2005. 脂氧合酶、脱落酸与茉莉酸在合作杨损伤信号传递中的相互关系[J]. 林业科学研究, 18(3): 300-304. (ZHANG K W, A Y, HU Z H, et al. Relationship between Lipoxygenase and ABA and JA in Wounded Signal Transduction of Heathy Populus Seedlings [J]. Forest Research, 18(3): 300-304.)
