二氧化硫(SO_2)熏蒸对葡萄果实结构和花色素苷含量的影响
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摘要
目的:探讨二氧化硫(SO_2)气体熏蒸处理对红地球葡萄果实采后细胞超微结构、花色素苷降解和相关基因表达的影响。方法:红地球葡萄果实经100、500μL/L SO_2气体间歇熏蒸方式处理,采用电镜扫描观察果梗表面结构的变化情况,分析测定果皮中花色素苷含量、花色素苷酶、苯丙氨酸解氨酶和4-香豆酸Co A连接酶活性变化,实时荧光定量PCR测定花色素苷合成相关基因表达的差异。结果:100μL/L SO_2熏蒸处理红地球葡萄降低了果梗气孔导度,抑制了果皮中花色素苷酶活性,诱导PAL、4CL活性增强,上调了PAL、4CL、LDOX基因的表达,但下调了DFR基因表达。结论:适宜浓度SO_2可以通过提高花色素苷合成相关酶的活性和基因表达量来调控花色素苷含量,从而延缓了花色素苷含量的下降,减缓了SO_2对果皮的漂白伤害作用。
Objective:The effects of SO_2fumigation on cell ultrastructure,anthocyanin degradation and related gene expression of the red globe grape were studied.Methods:The red globe grape was fumigated by 100 and 500μL/L SO_2.The surface structure of the stem was observed by scanning electron microscope(SEM).The content of anthocyanin and the activity of anthocyaninase,PAL,4CL in peel were detected by ulfraviolet spectrophotometry.The expression level of anthocyanin synthaserelated genes were determined byreal-time quantitative PCR.Results:100μL/L SO_2fumigation treatment on the red globe grape inhibited the stem of grape stomatal,the activity of anthocyanin in peel and the increasing of PAL,4CL activity.The expression of PAL,4CL and LDOX genes was up-regulated,but the DFR gene was down-regulated by SO_2.Conclusion:The lower concentration of SO_2would through increase the anthocyanin synthesis of enzyme activity and gene expression to control the content of anthocyanins.Thus delayed the decline of the content of anthocyanins and the reduce fade symptoms of the peel.
Objective:The effects of SO_2fumigation on cell ultrastructure,anthocyanin degradation and related gene expression of the red globe grape were studied.Methods:The red globe grape was fumigated by 100 and 500μL/L SO_2.The surface structure of the stem was observed by scanning electron microscope(SEM).The content of anthocyanin and the activity of anthocyaninase,PAL,4CL in peel were detected by ulfraviolet spectrophotometry.The expression level of anthocyanin synthaserelated genes were determined byreal-time quantitative PCR.Results:100μL/L SO_2fumigation treatment on the red globe grape inhibited the stem of grape stomatal,the activity of anthocyanin in peel and the increasing of PAL,4CL activity.The expression of PAL,4CL and LDOX genes was up-regulated,but the DFR gene was down-regulated by SO_2.Conclusion:The lower concentration of SO_2would through increase the anthocyanin synthesis of enzyme activity and gene expression to control the content of anthocyanins.Thus delayed the decline of the content of anthocyanins and the reduce fade symptoms of the peel.
引文
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[5]姜卫兵,徐莉莉,翁忙玲,等.环境因子及外源化学物质对植物花色素苷的影响[J].生态环境学报,2009,18(4):1546-1552.
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[9]黄海雄,黄育强.不同剂量SO2保鲜剂对冷处理护色荔枝贮藏安全品质的影响[J].保鲜与加工,2014,(2):7-11.
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[14]刘龙博,安玉艳,熊丽君,等.5-ALA诱导的黄酮醇积累参与调节苹果叶片气孔开度[J].园艺学报,2016,43(5):817-828.
[15]张家荣.负载量及其调控方式对酿酒葡萄果实中花色苷的影响[D].济南:齐鲁工业大学,2013.
[16]Martino A,Pifferi P G,Spagna G.The separation of pectinlyase fromβ-glucosidase in a commercial preparation[J].Journal of Chemical Technology and Biotechnology Biotechnology,2010,61(3):255-260.
[17]曹建康,姜微波,赵玉梅.果蔬采后生理生化实验指导[M].北京:中国轻工业出版社,2007:142-144.
[18]杜红豆.氮素对飞机草(Eupatorium odoratum)4CL酶活性和基因表达影响的研究[D].哈尔滨:哈尔滨师范大学,2012.
[19]周莉.NAA和ABA对葡萄花色苷生物合成相关基因表达的影响[D].哈尔滨:东北林业大学,2009.
[20]高登涛,李秋利,魏志峰,等.植物对二氧化硫胁迫反应与应答机制研究进展[J].广东农业科学,2016,43(11):27-35.
[21]成果,周思泓,黄羽,等.‘桂葡6号’葡萄花色苷组成特点与基因表达规律解析[J].植物生理学报,2017(1):103-114.
[22]Kader F,Irmouli M,Nicolas J P,et al.Proposed mechanism for the degradation of pelargonidin 3-glucoside by caffeic acid o-quinone[J].Food Chemistry,2001,75(2):139-144.
[23]马立娜,惠竹梅,霍珊珊,等.油菜素内酯和脱落酸调控葡萄果实花色苷合成的研究[J].果树学报,2012,29(5):830-836.
[24]范丙友,胡诗宇,陆海,等.毛白杨4-香豆酸:辅酶A连接酶可溶性原核表达及活性检测[J].北京林业大学学报,2006,28(2):1-8.
[25]李颖颖.红掌花PAL基因的克隆与花色苷含量的研究[D].海口:海南大学,2012.
[26]张波,赵志常,高爱平,等.芒果二氢黄酮醇4-还原酶(DFR)基因的克隆及其表达分析[J].分子植物育种,2015,13(4):816-821.
[27]Appelhagen I,Jahns O,Bartelniewoehner L,et al.Leucoanthocyanidin Dioxygenase in Arabidopsis thaliana:Characterization of mutant alleles and regulation by MYB-BHLH-TTG1 transcription factor complexes[J].Gene,2011,484(1-2):61.
[28]Jackman R L,Yada R Y,Tung M A,et al.Anthocyanins as food colorants-a review[J].Journal of Food Biochemistry,1987,11(3):201-247.
[29]Hiratsuka S,Onodera H,Kawai Y,et al.Enzyme activity changes during anthocyanin synthesis in‘Olympia’grape berries[J].Scientia Horticulturae,2001,90(3-4):255-264.
[30]母洪娜,孙陶泽,徐晨,等.桂花(Osmanthus fragrans Lour.)4-香豆酸辅酶A连接酶(4CL)基因克隆与表达分析[J].分子植物育种,2016,14(3):536-541.
[31]Wang H,Fan W,Li H,et al.Functional Characterization of Dihydroflavonol-4-Reductase in Anthocyanin Biosynthesis of Purple Sweet Potato Underlies the Direct Evidence of Anthocyanins Function against Abiotic Stresses[J].Plo S one,2013,8(11):44.
[32]He F,Mu L,Yan G L,et al.Biosynthesis of Anthocyanins and Their Regulation in Colored Grapes[J].Molecules,2010,15(12):9057-9091.
[33]Dai Z W,Ollat N,GomèS E,et al.Ecophysiological,Genetic,and Molecular Causes of Variation in Grape Berry Weight and Composition:A Review[J].American Journal of Enology and Viticulture,2011,62(4):413-425.
[34]Honda C,Kotoda N,Wada M,et al.Anthocyanin biosynthetic genes are coordinately expressed during red coloration in apple skin[J].Plant Physiology and Biochemistry,2002,40(11):955-962.
[35]文习成.桃不同品种花瓣和叶片呈色变化过程中花色苷合成的相关基因表达差异性研究[D].南京:南京农业大学,2012.
[36]孙欣,韩键,房经贵,等.葡萄浆果着色分子机理的重要研究进展[J].植物生理学报,2012,48(4):333-342.
[37]Jeong S T,Gotoyamamoto N,Hashizume K,et al.Expression of the flavonoid 3'-hydroxylase and flavonoid 3',5'-hydroxylase genes and flavonoid composition in grape(Vitis vinifera)[J].Plant Science,2006,170(1):61-69.
[38]Kobayashi S,Ishimaru M,Ding C K,et al.Comparison of UDP-glucose:flavonoid 3-O-glucosyltransferase(UFGT)gene sequences between white grapes(Vitis vinifera)and their sports with red skin[J].Plant Science An International Journal of Experimental Plant Biology,2001,160(3):543-550.
[2]葛毅强,谭敦炎,张维一,等.SO2对鲜葡萄采后熏蒸处理的组织解剖研究[J].西北植物学报,1997,17(4):516-521.
[3]周江.二氧化硫(SO2)间歇熏蒸对红地球葡萄采后品质的影响[D].乌鲁木齐:新疆农业大学,2016.
[4]王小青,韩键,文杨,等.呈色机制不同的桃叶片花色素苷积累及合成相关基因表达的季节性差异[J].南京农业大学学报,2016,39(6):924-931.
[5]姜卫兵,徐莉莉,翁忙玲,等.环境因子及外源化学物质对植物花色素苷的影响[J].生态环境学报,2009,18(4):1546-1552.
[6]刘丽青,王燕楠,仪慧兰,等.二氧化硫对玫瑰香葡萄果实采后保鲜的机理[J].应用与环境生物学报,2016,22(2):338-341.
[7]Cantín C M,Palou L,Bremer V,et al.Evaluation of the use of sulfur dioxide to reduce postharvest losses on dark and green figs[J].Postharvest Biology and Technology,2011,59(2):150-158.
[8]刘文浩.熏硫及脱硫处理对龙眼贮藏生理的影响及转录组研究[D].广州:华南农业大学,2016.
[9]黄海雄,黄育强.不同剂量SO2保鲜剂对冷处理护色荔枝贮藏安全品质的影响[J].保鲜与加工,2014,(2):7-11.
[10]吴斌,张玉丽,程琳琳,等.鲜食葡萄花色苷的研究进展[J].现代食品科技,2012,28(4):441-444.
[11]Hernandez A,Calero A,Barberan T.Quality and enhancement of bioactive phenolics in Cv.Napoleon table grapes exposed to different postharvest gaseous treatments[J].Journal of Agricultural and Food Chemistry,2003,51(18):5290-5295.
[12]张玉萍.新疆葡萄甜瓜保鲜技术研究与应用[D].乌鲁木齐:新疆大学,2006.
[13]赵晓敏,杨玉荣,李建鲲,等.1-MCP处理对库尔勒香梨采后果皮蜡质变化的影响[J].食品科学,2015,36(18):262-266.
[14]刘龙博,安玉艳,熊丽君,等.5-ALA诱导的黄酮醇积累参与调节苹果叶片气孔开度[J].园艺学报,2016,43(5):817-828.
[15]张家荣.负载量及其调控方式对酿酒葡萄果实中花色苷的影响[D].济南:齐鲁工业大学,2013.
[16]Martino A,Pifferi P G,Spagna G.The separation of pectinlyase fromβ-glucosidase in a commercial preparation[J].Journal of Chemical Technology and Biotechnology Biotechnology,2010,61(3):255-260.
[17]曹建康,姜微波,赵玉梅.果蔬采后生理生化实验指导[M].北京:中国轻工业出版社,2007:142-144.
[18]杜红豆.氮素对飞机草(Eupatorium odoratum)4CL酶活性和基因表达影响的研究[D].哈尔滨:哈尔滨师范大学,2012.
[19]周莉.NAA和ABA对葡萄花色苷生物合成相关基因表达的影响[D].哈尔滨:东北林业大学,2009.
[20]高登涛,李秋利,魏志峰,等.植物对二氧化硫胁迫反应与应答机制研究进展[J].广东农业科学,2016,43(11):27-35.
[21]成果,周思泓,黄羽,等.‘桂葡6号’葡萄花色苷组成特点与基因表达规律解析[J].植物生理学报,2017(1):103-114.
[22]Kader F,Irmouli M,Nicolas J P,et al.Proposed mechanism for the degradation of pelargonidin 3-glucoside by caffeic acid o-quinone[J].Food Chemistry,2001,75(2):139-144.
[23]马立娜,惠竹梅,霍珊珊,等.油菜素内酯和脱落酸调控葡萄果实花色苷合成的研究[J].果树学报,2012,29(5):830-836.
[24]范丙友,胡诗宇,陆海,等.毛白杨4-香豆酸:辅酶A连接酶可溶性原核表达及活性检测[J].北京林业大学学报,2006,28(2):1-8.
[25]李颖颖.红掌花PAL基因的克隆与花色苷含量的研究[D].海口:海南大学,2012.
[26]张波,赵志常,高爱平,等.芒果二氢黄酮醇4-还原酶(DFR)基因的克隆及其表达分析[J].分子植物育种,2015,13(4):816-821.
[27]Appelhagen I,Jahns O,Bartelniewoehner L,et al.Leucoanthocyanidin Dioxygenase in Arabidopsis thaliana:Characterization of mutant alleles and regulation by MYB-BHLH-TTG1 transcription factor complexes[J].Gene,2011,484(1-2):61.
[28]Jackman R L,Yada R Y,Tung M A,et al.Anthocyanins as food colorants-a review[J].Journal of Food Biochemistry,1987,11(3):201-247.
[29]Hiratsuka S,Onodera H,Kawai Y,et al.Enzyme activity changes during anthocyanin synthesis in‘Olympia’grape berries[J].Scientia Horticulturae,2001,90(3-4):255-264.
[30]母洪娜,孙陶泽,徐晨,等.桂花(Osmanthus fragrans Lour.)4-香豆酸辅酶A连接酶(4CL)基因克隆与表达分析[J].分子植物育种,2016,14(3):536-541.
[31]Wang H,Fan W,Li H,et al.Functional Characterization of Dihydroflavonol-4-Reductase in Anthocyanin Biosynthesis of Purple Sweet Potato Underlies the Direct Evidence of Anthocyanins Function against Abiotic Stresses[J].Plo S one,2013,8(11):44.
[32]He F,Mu L,Yan G L,et al.Biosynthesis of Anthocyanins and Their Regulation in Colored Grapes[J].Molecules,2010,15(12):9057-9091.
[33]Dai Z W,Ollat N,GomèS E,et al.Ecophysiological,Genetic,and Molecular Causes of Variation in Grape Berry Weight and Composition:A Review[J].American Journal of Enology and Viticulture,2011,62(4):413-425.
[34]Honda C,Kotoda N,Wada M,et al.Anthocyanin biosynthetic genes are coordinately expressed during red coloration in apple skin[J].Plant Physiology and Biochemistry,2002,40(11):955-962.
[35]文习成.桃不同品种花瓣和叶片呈色变化过程中花色苷合成的相关基因表达差异性研究[D].南京:南京农业大学,2012.
[36]孙欣,韩键,房经贵,等.葡萄浆果着色分子机理的重要研究进展[J].植物生理学报,2012,48(4):333-342.
[37]Jeong S T,Gotoyamamoto N,Hashizume K,et al.Expression of the flavonoid 3'-hydroxylase and flavonoid 3',5'-hydroxylase genes and flavonoid composition in grape(Vitis vinifera)[J].Plant Science,2006,170(1):61-69.
[38]Kobayashi S,Ishimaru M,Ding C K,et al.Comparison of UDP-glucose:flavonoid 3-O-glucosyltransferase(UFGT)gene sequences between white grapes(Vitis vinifera)and their sports with red skin[J].Plant Science An International Journal of Experimental Plant Biology,2001,160(3):543-550.