外源6-BA对葡萄果实花色苷含量及相关基因表达的影响
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摘要
【目的】阐明6-BA对葡萄果实花色苷含量及其生物合成相关基因表达的影响,为提高葡萄果实花色苷含量的研究提供参考。【方法】以‘梅洛’葡萄为试材,对其进行不同质量浓度(0,100,200,400mg/L)6-BA处理,研究6-BA对葡萄果实花色苷含量及其合成相关基因表达的影响,分析花色苷含量与其合成相关基因表达量的相关性。【结果】100mg/L 6-BA处理可提高葡萄果实花色苷含量,花后110d达到最大,为5.21mg/g;100mg/L 6-BA处理葡萄果实PAL、CHS1、F3′H、UFGT基因的表达量在花后90d显著提高,200mg/L 6-BA处理F3′5′H基因的表达量显著提高,400mg/L 6-BA处理则相反。相关性分析表明,100mg/L 6-BA处理葡萄果实花色苷含量与CHS1、UF-GT基因相对表达量呈显著正相关,200和400mg/L 6-BA处理组花色苷含量与PAL基因相对表达量呈显著和极显著正相关;100和400mg/L 6-BA处理组MybA1与UFGT的相对表达量呈显著正相关。【结论】低质量浓度6-BA处理有利于提高果实花色苷含量,在果实着色后期可影响花色苷合成相关基因的表达。
【Objective】This study elucidated the effect of 6-BA on anthocyanin content and its biosynthetic gene expression in grape berry to provide reference for the improvement of anthocyanin in grape berry.【Method】‘Merlot'was treated with 6-BA at different concentrations of 0,100,200,and 400 mg/L to investigate its effects on anthocyanin content and expression of related biosynthesis genes in grape berry.The correlation between anthocyanin content and the expression of related genes was also analyzed.【Result】The treatment of 100 mg/L 6-BA increased the content of anthocyanin in grape berry,reaching the maximum of 5.21 mg/g 110 days after anthesis.The expression levels of PAL,CHS1,F3′H and UFGT genes in grape fruits treated with 100 mg/L 6-BA were significantly increased 90 days after flowering,The expression levels of F3′5′H gene in grape fruits treated with 200 mg/L 6-BA were significantly increased,whereas the 400 mg/L 6-BA treatment showed the opposite.The correlation analysis showed that there was a significant positive correlation between the anthocyanin content in 100 mg/L 6-BA treated grape berry and the relative expression levels of CHS1 and UFGT genes.The anthocyanin contents of 200 and 400mg/L 6-BA treatment groups were significantly and positively correlated with the relative expression level of PALgene.There was a significant positive correlation between the relative expressions of MybA1 and UFGTin 100 and 400 mg/L 6-BA treatment groups.【Conclusion】Low-concentration 6-BA treatment was beneficial to increase the content of anthocyanin in grape berries and affected the expression of anthocyanin synthesis related genes in the late stage of fruit coloring.
【Objective】This study elucidated the effect of 6-BA on anthocyanin content and its biosynthetic gene expression in grape berry to provide reference for the improvement of anthocyanin in grape berry.【Method】‘Merlot'was treated with 6-BA at different concentrations of 0,100,200,and 400 mg/L to investigate its effects on anthocyanin content and expression of related biosynthesis genes in grape berry.The correlation between anthocyanin content and the expression of related genes was also analyzed.【Result】The treatment of 100 mg/L 6-BA increased the content of anthocyanin in grape berry,reaching the maximum of 5.21 mg/g 110 days after anthesis.The expression levels of PAL,CHS1,F3′H and UFGT genes in grape fruits treated with 100 mg/L 6-BA were significantly increased 90 days after flowering,The expression levels of F3′5′H gene in grape fruits treated with 200 mg/L 6-BA were significantly increased,whereas the 400 mg/L 6-BA treatment showed the opposite.The correlation analysis showed that there was a significant positive correlation between the anthocyanin content in 100 mg/L 6-BA treated grape berry and the relative expression levels of CHS1 and UFGT genes.The anthocyanin contents of 200 and 400mg/L 6-BA treatment groups were significantly and positively correlated with the relative expression level of PALgene.There was a significant positive correlation between the relative expressions of MybA1 and UFGTin 100 and 400 mg/L 6-BA treatment groups.【Conclusion】Low-concentration 6-BA treatment was beneficial to increase the content of anthocyanin in grape berries and affected the expression of anthocyanin synthesis related genes in the late stage of fruit coloring.
引文
[1]刘双双,王忠华.葡萄花色素苷生物合成调控研究进展[J].科技创新导报,2015,2(6):89-92.Liu S S,Wang Z H.Advances in regulation of anthocyanin biosynthesis in grapevine[J].Science and Technology Innovation Herald,2015,2(6):89-92.
[2]刘笑宏,郭淑华,牛彦杰,等.8个葡萄品种的花色苷组分及含量分析[J].果树学报,2017,34(4):444-453.Liu X H,Guo S H,Niu Y J,et al.Analysis of anthocyanin compositions and contens in eight grape cultiars[J].Journal of Fruit Science,2017,34(4):444-453.
[3]李颖畅,孟宪军,孙靖靖,等.蓝莓花色苷的降血脂和抗氧化作用[J].食品与发酵业,2008,34(10):44-48.Li Y C,Meng X J,Sun J J,et al.Effects of anthocyanins from blueberry on lowing the cholesterol and antioxidation[J].Food and Fermentation Industries,2008,34(10):44-48.
[4]Chang H,Yu B,Yu X,et al.Anticancer activities of anthocyanin-rich extract from black rice against breast cancer cells in vitro and in vivo[J].Nutrition&Cancer,2010,62(8):1128-1136.
[5]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.Sun X,Han J,Fang J G,et al.Important research progress of coloring molecular mechanisms in grape berry[J].Plant Physiology Journal,2012,48(4):333-342.
[6]贾赵东,马佩勇,边小峰,等.植物花青素合成代谢途径及其分子调控[J].西北植物学报,2014,34(7):1496-1506.Jia Z D,Ma P Y,Bian X F,et al.Biosynthesis metabolic pathway and molecular regulation of plants ant hocyanin[J].Acta Botanica Boreali-Occidentalia Sinica,2014,34(7):1496-1506.
[7]Jenong S T,Goto-yamamoto N,Kobayash S,et al.Effects of plant hormones and shading on the accumulation of anthocyanins and the expression of anthocyanin biosynthetic genes in grape berry skins[J].Plant Science,2004,167(2):247-252.
[8]Kobayashi S,Ishimaru M,Hiraoka K,et al.Myb-related genes of the Kyoho grape(Vitis labruscana)regulate anthocyanin biosynthesis[J].Plant,2002,215(6):924-933.
[9]Ageorges A,Fernandez L,Vialet S,et al.Four specific isogenes of the anthocyanin metabolic pathway are systematicall co-expressed with the red colour of grape berries[J].Plant Science,2006,170(2):372-383.
[10]于淼,刘海峰,王军.ABA对葡萄花色苷合成相关基因表达的影响[J].果树学报,2012,29(1):29-35,157.Yu M,Liu H F,Wang J.Effects of ABA on expression of genes related to anthocyanin biosynthesis in grapevine[J].Journal of Fruit Science,2012,29(1):29-35,157.
[11]吴岚芳,钟玉,曾丽萍,等.6-BA对芒果糖分代谢的影响[J].食品工业科技,2010,11(31):335-337.Wu L F,Zhong Y,Zeng L P,et al.Effects of 6-BA on sugar metabolism in mango[J].Science and Technology of Food Industry,2010,11(31):335-337.
[12]王惠聪,黄辉白,黄旭明.妃子笑荔枝果实着色不良原因的研究[J].园艺学报,2002,29(5):408-412.Wang H C,Huang H B,Huang X M.A study on the causative factors retarding pigmentation in the fruit of‘Feizixiao’litch[J].Acta Horticulturae Sinica,2002,29(5):408-412.
[13]赵权,王军.ABA和6-BA对山葡萄果实着色及相关品质的影响[J].江苏农业科学,2010(2):189-190,220.Zhao Q,Wang J.Effects of ABA and 6-BA on berries anthocyanin content and berry quality of Vitis amurensis[J].Jiangsu Agricultural Sciences,2010(2):189-190,220.
[14]刘善军,王萍,范国荣.6-BA对甜柿果皮色素的影响[J].江西农业学报,2004,16(4):24-29.Liu S J,Wang P,Fan G R.Influence of 6-BA on pericarpial pigment in nonastringent persimmon[J].Acta Agriculturae Jiangxi,2004,16(4):24-29.
[15]刘洪海,张晓丽,杜平,等.pH示差法测定烟73葡萄中花青素含量[J].中国调味品,2009,34(4):110-111,117.Liu H H,Zhang X L,Du P,et al.Determination the anthocyanin content in Yan 73grape by pH-differential method[J].China Condiment,2009,34(4):110-111,117.
[16]查倩,奚晓军,蒋爱丽,等.葡萄实时定量PCR中稳定内参基因的筛选[J].果树学报,2016,27(3):268-274.Zha Q,Xi X J,Jiang A L,et al.Identification of appropriate reference geneby Real-time quantitative PCR in grape[J].Journal of Fruit Science,2016,27(3):268-274.
[17]Hashimoto K,Eckert C,Anschutz U,et al.Phosphorylation of calcineurin B-like(CBL)calcium sensorproteins by their CBL-interacting protein kinases(CIPKs)is required for full activity of CBL-CIPK complexes toward their target proteins[J].Journal of Biological Chemistry,2012,287(11):7956.
[18]赵权.葡萄酚类物质及其生物合成相关结构基因表达[D].哈尔滨:东北林业大学,2010.Zhao Q.Phenolic compounds and expression of related biosynthesis structural genes in grapevine[D].Harbin:Northeast Forestry University,2010.
[19]步洪凤,杜云安,顾振华,等.6-BA对月季切花保鲜效果的研究[J].上海农业学报,2016,34(4):66-70.Bu H F,Du Y A,Gu Z H,et al.Effect of 6-BA on preservantion of China rose cut flower[J].Acta Agriculturae Shanghai,2016,34(4):66-70.
[20]郑强卿,郁松林,孟凤.6-BA对葡萄果实发育过程中糖积累及转化酶活性的影响[J].新疆农业科学,2009,46(4):793-799.Zheng Q Q,Yu S L,Meng F.Effects of 6-BA on the sugar accumulation and invertase activities during development of‘Red Globe’greap berries[J].Xinjiang Agricultural Sciences,2009,46(4):793-799.
[21]王博.根域限制促进鲜食葡萄果皮花色苷合成的机制研究[D].上海:上海交通大学,2013.Wang B.Study on the mechanism of improved anthocyanin biosynthesis under root restriction in table grape berry skin[D].Shanghai:Shanghai Jiaotong University,2013.
[22]Azuma,Yakushiji H,Koshita Y,et al.Flavonoid biosynthesisrelated genes in grape skin are differentially regulated by temperature and light conditions[J].Planta,2012,236(4):1-14.
[23]栾丽英.油菜素内酯和脱落酸对酿酒葡萄花色苷调控及葡萄酒品质影响的研究[D].陕西杨凌:西北农林科技大学,2014.Luan L Y.Study on the regulation of anthocyanin synthesis of grape and the quality of wine after brassinolide and abscisic acid treatments[D].Yangling,Shaanxi:Northwest A&F U-niversity,2014.
[24]俞信光,刘双双,冯亚斌,等.有机基质栽培对巨峰葡萄花色苷合成基因表达的影响[J].核农学报,2016,30(11):2133-2143.Yu X G,Liu S S,Feng Y B,et al.Effects of organic substrate cultivation on anthocyanins biosynthetic genes expression in Kyoho grape berries[J].Journal of Nuclear Agricultural Sciences,2016,30(11):2133-2143.
[25]牛生洋.中国野生刺葡萄(Vitis davidii)果皮颜色变异机理研究[D].陕西杨凌:西北农林科技大学,2016.Niu S Y.Studies on the mechanism of color variation in grape berry skin of Chinese Wild Grape(Vitis davidii)[D].Yangling,Shaanxi:Northwest A&F University,2016.
[26]Peppi M C,Walker M A,Fidelibus M W.Application of abscis2016.ic acid rapidly upregulated UFGT gene expression and improved color of grape berries[J].Vitis,2008,47(1):11-14.
[27]孟祥春,彭建宗,王小菁.光和糖对非洲菊花色素苷积累及CHS、DFR基因表达的影响[J].园艺学报,2007,34(1):227-230.Meng X C,Peng J Z,Wang X J.Anthocyan in accumulation and CHS,DFRgene expression regulated by light and sugar in Gerbera jamesonii Bolus ray floret[J].Acta Horticulturae Sinica,2007,34(1):227-230.
[28]Deluc L,Barrieu F,Marchive C,et al.Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway[J].Plant Physiology,2006,140(2):499-511.
[29]Dixon R A,Xie D,Sharma S B.Proanthocyanidins:a final frontier in flavonoid research?[J].New Phytologist,2005,165(1):9-28.
[30]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(Amsterdam,Netherlands),2001,160(3):543-550.
[31]Walker A R,Lee E,Bogs J,et al.White grapes arose through the mutation of two similar and adjacent regulatory genes[J].Plant Journal,2007,49(5):772-785.
[2]刘笑宏,郭淑华,牛彦杰,等.8个葡萄品种的花色苷组分及含量分析[J].果树学报,2017,34(4):444-453.Liu X H,Guo S H,Niu Y J,et al.Analysis of anthocyanin compositions and contens in eight grape cultiars[J].Journal of Fruit Science,2017,34(4):444-453.
[3]李颖畅,孟宪军,孙靖靖,等.蓝莓花色苷的降血脂和抗氧化作用[J].食品与发酵业,2008,34(10):44-48.Li Y C,Meng X J,Sun J J,et al.Effects of anthocyanins from blueberry on lowing the cholesterol and antioxidation[J].Food and Fermentation Industries,2008,34(10):44-48.
[4]Chang H,Yu B,Yu X,et al.Anticancer activities of anthocyanin-rich extract from black rice against breast cancer cells in vitro and in vivo[J].Nutrition&Cancer,2010,62(8):1128-1136.
[5]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.Sun X,Han J,Fang J G,et al.Important research progress of coloring molecular mechanisms in grape berry[J].Plant Physiology Journal,2012,48(4):333-342.
[6]贾赵东,马佩勇,边小峰,等.植物花青素合成代谢途径及其分子调控[J].西北植物学报,2014,34(7):1496-1506.Jia Z D,Ma P Y,Bian X F,et al.Biosynthesis metabolic pathway and molecular regulation of plants ant hocyanin[J].Acta Botanica Boreali-Occidentalia Sinica,2014,34(7):1496-1506.
[7]Jenong S T,Goto-yamamoto N,Kobayash S,et al.Effects of plant hormones and shading on the accumulation of anthocyanins and the expression of anthocyanin biosynthetic genes in grape berry skins[J].Plant Science,2004,167(2):247-252.
[8]Kobayashi S,Ishimaru M,Hiraoka K,et al.Myb-related genes of the Kyoho grape(Vitis labruscana)regulate anthocyanin biosynthesis[J].Plant,2002,215(6):924-933.
[9]Ageorges A,Fernandez L,Vialet S,et al.Four specific isogenes of the anthocyanin metabolic pathway are systematicall co-expressed with the red colour of grape berries[J].Plant Science,2006,170(2):372-383.
[10]于淼,刘海峰,王军.ABA对葡萄花色苷合成相关基因表达的影响[J].果树学报,2012,29(1):29-35,157.Yu M,Liu H F,Wang J.Effects of ABA on expression of genes related to anthocyanin biosynthesis in grapevine[J].Journal of Fruit Science,2012,29(1):29-35,157.
[11]吴岚芳,钟玉,曾丽萍,等.6-BA对芒果糖分代谢的影响[J].食品工业科技,2010,11(31):335-337.Wu L F,Zhong Y,Zeng L P,et al.Effects of 6-BA on sugar metabolism in mango[J].Science and Technology of Food Industry,2010,11(31):335-337.
[12]王惠聪,黄辉白,黄旭明.妃子笑荔枝果实着色不良原因的研究[J].园艺学报,2002,29(5):408-412.Wang H C,Huang H B,Huang X M.A study on the causative factors retarding pigmentation in the fruit of‘Feizixiao’litch[J].Acta Horticulturae Sinica,2002,29(5):408-412.
[13]赵权,王军.ABA和6-BA对山葡萄果实着色及相关品质的影响[J].江苏农业科学,2010(2):189-190,220.Zhao Q,Wang J.Effects of ABA and 6-BA on berries anthocyanin content and berry quality of Vitis amurensis[J].Jiangsu Agricultural Sciences,2010(2):189-190,220.
[14]刘善军,王萍,范国荣.6-BA对甜柿果皮色素的影响[J].江西农业学报,2004,16(4):24-29.Liu S J,Wang P,Fan G R.Influence of 6-BA on pericarpial pigment in nonastringent persimmon[J].Acta Agriculturae Jiangxi,2004,16(4):24-29.
[15]刘洪海,张晓丽,杜平,等.pH示差法测定烟73葡萄中花青素含量[J].中国调味品,2009,34(4):110-111,117.Liu H H,Zhang X L,Du P,et al.Determination the anthocyanin content in Yan 73grape by pH-differential method[J].China Condiment,2009,34(4):110-111,117.
[16]查倩,奚晓军,蒋爱丽,等.葡萄实时定量PCR中稳定内参基因的筛选[J].果树学报,2016,27(3):268-274.Zha Q,Xi X J,Jiang A L,et al.Identification of appropriate reference geneby Real-time quantitative PCR in grape[J].Journal of Fruit Science,2016,27(3):268-274.
[17]Hashimoto K,Eckert C,Anschutz U,et al.Phosphorylation of calcineurin B-like(CBL)calcium sensorproteins by their CBL-interacting protein kinases(CIPKs)is required for full activity of CBL-CIPK complexes toward their target proteins[J].Journal of Biological Chemistry,2012,287(11):7956.
[18]赵权.葡萄酚类物质及其生物合成相关结构基因表达[D].哈尔滨:东北林业大学,2010.Zhao Q.Phenolic compounds and expression of related biosynthesis structural genes in grapevine[D].Harbin:Northeast Forestry University,2010.
[19]步洪凤,杜云安,顾振华,等.6-BA对月季切花保鲜效果的研究[J].上海农业学报,2016,34(4):66-70.Bu H F,Du Y A,Gu Z H,et al.Effect of 6-BA on preservantion of China rose cut flower[J].Acta Agriculturae Shanghai,2016,34(4):66-70.
[20]郑强卿,郁松林,孟凤.6-BA对葡萄果实发育过程中糖积累及转化酶活性的影响[J].新疆农业科学,2009,46(4):793-799.Zheng Q Q,Yu S L,Meng F.Effects of 6-BA on the sugar accumulation and invertase activities during development of‘Red Globe’greap berries[J].Xinjiang Agricultural Sciences,2009,46(4):793-799.
[21]王博.根域限制促进鲜食葡萄果皮花色苷合成的机制研究[D].上海:上海交通大学,2013.Wang B.Study on the mechanism of improved anthocyanin biosynthesis under root restriction in table grape berry skin[D].Shanghai:Shanghai Jiaotong University,2013.
[22]Azuma,Yakushiji H,Koshita Y,et al.Flavonoid biosynthesisrelated genes in grape skin are differentially regulated by temperature and light conditions[J].Planta,2012,236(4):1-14.
[23]栾丽英.油菜素内酯和脱落酸对酿酒葡萄花色苷调控及葡萄酒品质影响的研究[D].陕西杨凌:西北农林科技大学,2014.Luan L Y.Study on the regulation of anthocyanin synthesis of grape and the quality of wine after brassinolide and abscisic acid treatments[D].Yangling,Shaanxi:Northwest A&F U-niversity,2014.
[24]俞信光,刘双双,冯亚斌,等.有机基质栽培对巨峰葡萄花色苷合成基因表达的影响[J].核农学报,2016,30(11):2133-2143.Yu X G,Liu S S,Feng Y B,et al.Effects of organic substrate cultivation on anthocyanins biosynthetic genes expression in Kyoho grape berries[J].Journal of Nuclear Agricultural Sciences,2016,30(11):2133-2143.
[25]牛生洋.中国野生刺葡萄(Vitis davidii)果皮颜色变异机理研究[D].陕西杨凌:西北农林科技大学,2016.Niu S Y.Studies on the mechanism of color variation in grape berry skin of Chinese Wild Grape(Vitis davidii)[D].Yangling,Shaanxi:Northwest A&F University,2016.
[26]Peppi M C,Walker M A,Fidelibus M W.Application of abscis2016.ic acid rapidly upregulated UFGT gene expression and improved color of grape berries[J].Vitis,2008,47(1):11-14.
[27]孟祥春,彭建宗,王小菁.光和糖对非洲菊花色素苷积累及CHS、DFR基因表达的影响[J].园艺学报,2007,34(1):227-230.Meng X C,Peng J Z,Wang X J.Anthocyan in accumulation and CHS,DFRgene expression regulated by light and sugar in Gerbera jamesonii Bolus ray floret[J].Acta Horticulturae Sinica,2007,34(1):227-230.
[28]Deluc L,Barrieu F,Marchive C,et al.Characterization of a grapevine R2R3-MYB transcription factor that regulates the phenylpropanoid pathway[J].Plant Physiology,2006,140(2):499-511.
[29]Dixon R A,Xie D,Sharma S B.Proanthocyanidins:a final frontier in flavonoid research?[J].New Phytologist,2005,165(1):9-28.
[30]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(Amsterdam,Netherlands),2001,160(3):543-550.
[31]Walker A R,Lee E,Bogs J,et al.White grapes arose through the mutation of two similar and adjacent regulatory genes[J].Plant Journal,2007,49(5):772-785.