海州常山花萼转色期生理变化研究
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摘要
【目的】探究海州常山花萼色素的组成及动态变化规律。【方法】以分别从盐城亭湖、镇江句容、连云港云台山、徐州贾汪、泰安泰山和驻马店东官庄引种至南京的6个种源海州常山为试验材料,用显色反应和紫外-可见光谱检测海州常山花萼中色素的组成,运用HPLC法鉴定花青素组分,于盛花初期、盛花中期、花果共存期、结果期、果萼宿存期和落叶期采样测定花萼变色过程中叶绿素、花青素、可溶性糖含量和相关酶活性的变化,并分析花青素与其他指标的相关性。【结果】海州常山花萼中含有叶绿素、花青素和类黄酮化合物,不含类胡萝卜素。从不同发育期海州常山花萼中共检测出3种花青素成分,确定其中一种为矢车菊素-3-O-葡萄糖苷。在花萼变色过程中,叶绿素含量降低,花青素与可溶性糖含量升高,苯丙氨酸解氨酶(PAL)活性在转色后期波动较大,查尔酮异构酶(CHI)活性明显升高。相关性分析发现,花青素含量与叶绿素、可溶性糖含量和PAL、CHI活性明显相关。【结论】转色过程中CHI酶的变化是海州常山花萼转色的主要原因。
【Objective】To explore the composition and dynamic changes of the calyx pigment in Clerodendrum trichotomum.【Method】Six Clerodendrum trichotomum samples(Tinghu,Yancheng;Jurong,Zhenjiang;Yuntaishan,Lianyungang;Jiawang,Xuzhou;Taishan,Tai'an;Dongguanzhuang,Zhumadian)from different provenances introduced into Nanjing were used as experimental materials to detect pigment composition in the calyx through identifying the anthocyanin components using color reaction,UV-VIS spectroscopy and HPLC method.At the six developmental stages(primary full-blooming stage,middle fullblooming stage,flower and fruit coexistence stage,fruit-bearing stage,fruit and calyx persistent stage,defoliating stage)sampling,mesuring the changes of chlorophyll,anthocyanin,soluble sugar and anthocyanidin synthase in the process of calyx discoloration,meanwhile analyzing the correlation between anthocyanins and other indicators.【Result】It showed that there were chlorophyll,anthocyanin and flavonoids,no carotenoids in the Clerodendrum trichotomum calyx.A total of 3 anthocyanin constituents were identified in which the Cyanidin-3-O-glucoside was confirmed.During the process of color change,the content of chlorophyll decreased,the content of anthocyanin and soluble sugar increased,PAL enzyme activity fluctuated greatly at the later color-turning stage and the activity of CHI enzyme increased obviously.Through correlation analysis found that anthocyanin content was significantly correlated with chlorophyll content,soluble sugar content,PAL and CHI enzyme activities.【Conclusion】Change in CHI enzyme during color conversion was the main cause of calyx color change in Clerodendrum trichotomum.
【Objective】To explore the composition and dynamic changes of the calyx pigment in Clerodendrum trichotomum.【Method】Six Clerodendrum trichotomum samples(Tinghu,Yancheng;Jurong,Zhenjiang;Yuntaishan,Lianyungang;Jiawang,Xuzhou;Taishan,Tai'an;Dongguanzhuang,Zhumadian)from different provenances introduced into Nanjing were used as experimental materials to detect pigment composition in the calyx through identifying the anthocyanin components using color reaction,UV-VIS spectroscopy and HPLC method.At the six developmental stages(primary full-blooming stage,middle fullblooming stage,flower and fruit coexistence stage,fruit-bearing stage,fruit and calyx persistent stage,defoliating stage)sampling,mesuring the changes of chlorophyll,anthocyanin,soluble sugar and anthocyanidin synthase in the process of calyx discoloration,meanwhile analyzing the correlation between anthocyanins and other indicators.【Result】It showed that there were chlorophyll,anthocyanin and flavonoids,no carotenoids in the Clerodendrum trichotomum calyx.A total of 3 anthocyanin constituents were identified in which the Cyanidin-3-O-glucoside was confirmed.During the process of color change,the content of chlorophyll decreased,the content of anthocyanin and soluble sugar increased,PAL enzyme activity fluctuated greatly at the later color-turning stage and the activity of CHI enzyme increased obviously.Through correlation analysis found that anthocyanin content was significantly correlated with chlorophyll content,soluble sugar content,PAL and CHI enzyme activities.【Conclusion】Change in CHI enzyme during color conversion was the main cause of calyx color change in Clerodendrum trichotomum.
引文
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[5]郭鸿飞,张延龙,牛立新,等.8种中国野生百合花色素成分分析[J].西北农林科技大学学报(自然科学版),2015,43(3):98-104.Guo H F,Zhang Y L,Niu L X,et al.Petal pigments of eight wild Liliumspecies native to China[J].Journal of Northwest A&F University(Natural Science Edition),2015,43(3):98-104.
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[8]朱满兰,王亮生,张会金,等.耐寒睡莲花瓣中花青素苷组成及其与花色的关系[J].植物学报,2012,47(5):437-453.Zhu M L,Wang L S,Zhang H J,et al.Composition of anthocyanin in hardy Nymphaeapetals and its relationship with flower color[J].Chinese Bulletin of Botany,2012,47(5):437-453.
[9]钱见平,丰震,王长宪,等.元宝枫无性系秋季叶片主要成分含量动态变化[J].中国农学通报,2013,29(7):26-30.Qian J P,Feng Z,Wang C X,et al.The dynamic changes of main component in leaves of Acer truncatum Bunge clones in autumn[J].Chinese Agricultural Science Bulletin,2013,29(7):26-30.
[10]王艺菲,辛秀兰,陈亮,等.pH示差法测定不同种类蓝果忍冬总花色苷含量[J].食品研究与开发,2014,35(7):75-78.Wang Y F,Xin X L,Chen L,et al.Total anthocyanins content in different species of Lonicera cearulea Linn by pH-differential spectrophotometry[J].Food Research and Development,2014,35(7):75-78.
[11]李文砚,韦持章,孔方南,等.蒽酮法测定木奶果果实中可溶性糖含量的研究[J].中国园艺文摘,2015,31(12):7-8,28.Li W Y,Wei C Z,Kong F N,et al.Determination of soluble saccharide contents of Baccaurea ramiflora Lour.usinganthrone colorimetric method[J].Chinese Journal of Horticulture,2015,31(12):7-8,28.
[12]赵宗方,赵勇,吴桂法.果实花青素含量与PAL活性关系的研究[J].园艺学报,1994(2):199-200.Zhao Z F,Zhao Y,Wu G F.Study on the relationship between anthocyanin content and PAL activity in fruits[J].Journal of Horticulture,1994(2):199-200.
[13]王庆菊,李晓磊,王磊,等.紫叶稠李叶片花色苷及其合成相关酶动态[J].林业科学,2008(3):45-49.Wang Q J,Li X L,Wang L,et al.Dynamic changes of anthocyanin and the relevant biosynthesis enzymes in Padus virginiana‘Schubert’leaves[J].Scientia Silvae Sinicae,2008(3):45-49.
[14]Silva V O,Freitas A A,Macanita A L,et al.Chemistry and photochemistry of natural plant pigments:the anthocyanins[J].Journal of Physical Organic Chemistry,2016,29(11):594-599.
[15]Sakata Y,Aoki N,Tsunematsu S,et al.Petal coloration and pigmentation of tree peony bred and selected in Daikon Island(Shimane Prefecture)[J].Engei Gakkai Zasshi,2008,64(2):351-357.
[16]梁娟,蔺银鼎.元宝枫秋叶着色生理指标的研究[J].山西农业大学学报(自然科学版),2009,29(1):37-40.Liang J,Lin Y D.Studies on the physiological indexes of Acer truncatumfall leaves during coloring process[J].Journal of Shanxi Agricultural University(Natural Science Edition),2009,29(1):37-40.
[17]王雅琼,郑伟尉,臧运祥,等.不同品种羽衣甘蓝生长期色素含量变化规律研究[J].中国农学通报,2013,29(10):154-161.Wang Y Q,Zheng W W,Zang Y X,et al.Studies on change of pigment contents during different developmental stages among several flowering kale cultivars[J].Chinese Agricultural Science Bulletin,2013,29(10):154-161.
[18]Escribano-Bailon M T,Santos-Buelga C,Rivas-Gonzalo J C.Anthocyanins in cereals[J].Journal of Chromatography A,2004,1054(1):129-141.
[19]胡静静,沈向,李雪飞,等.黄连木秋季叶色变化与可溶性糖和矿质元素的关系[J].林业科学,2010,46(2):80-86.Hu J J,Shen X,Li X F,et al.Relationships of leaf color changes with soluble sugars and mineral elements in leaves of Pistacia chinensis in autumn[J].Scientia Silvae Sinicae,2010,46(2):80-86.
[20]杨暖,姜琳,姜官恒,等.北美豆梨秋冬叶色与生理变化的关系[J].农学学报,2015,5(11):73-78.Yang N,Jiang L,Jiang G H,et al.The relationship between leaf color in autumn and physiological changes of Pyrus calleryana Decne[J].Journal of Agriculture,2015,5(11):73-78.
[21]曹丽春,徐翔宇,李金平,等.不同色泽无花果果皮色素变化特征及其与果实糖分累积的关系[J].中国农业大学学报,2017,22(2):54-60.Cao L C,Xu X Y,Li J P,et al.Peel pigment change during the development of figs with different colors and the correlation with soluble sugar accumulation[J].Journal of China Agricultural University,2017,22(2):54-60.
[22]李云萍,郭晋雅,高峰.紫心甘薯花青素积累与PAL活性的关系[J].西南大学学报(自然科学版),2010,32(2):68-72.Li Y P,Guo J Y,Gao F.Relationship between anthocyanin accumulation and PAL activity in Ipomoea batatas Lam[J].Journal of Southwest University(Natural Science Edition),2010,32(2):68-72.
[23]Mori T,Sakurai M,Sakuta M.Changes in PAL,CHS,DAHPsynthase(DS-Co and Ds-Mn)activity during anthocyanin synthesis in suspension culture of Fragaria ananassa[J].Plant Cell Tissue&Organ Culture,2000,62(2):135-139.
[24]王惠聪,黄旭明,胡桂兵,等.荔枝果皮花青苷合成与相关酶的关系研究[J].中国农业科学,2004,37(12):2028-2032.Wang H C,Huang X M,Hu G B,et al.Studies on the relationship between anthocyanin biosynthesis and related enzymes in Litchi pericarp[J].Scientia Agricultura Sinica,2004,37(12):2028-2032.
[25]陈继卫,沈朝栋,贾玉芳,等.鸡爪槭转色期叶色变化生理研究[J].核农学报,2010,24(1):171-175,180.Chen J W,Shen Z D,Jia Y F,et al.Physiological study on leaf color change during the color change in Acer palmatum[J].Journal of Nuclear Agricultural Sciences,2010,24(1):171-175,180.
[26]朱书香,王中华,李彦慧,等.紫叶矮樱叶片色素理化性质的研究[J].河北农业大学学报,2010,33(3):33-36,45.Zhu S X,Wang Z H,Li Y H,et al.Study on the physicalchemical properties of leaf pigment of Prunus×cistenena‘Purpleleaf and Chevry’[J].Journal of Agricultural University of Hebei,2010,33(3):33-36,45.
[27]于晓南,张启翔.光强与光质对“美人梅”叶色的影响[J].北京林业大学学报,2001,23(S2):36-38.Yu X N,Zhang Q X.Effects of light intensity and light quality on leaf coler expression of Prunus blireiana[J].Journal of Beijing Forestry University,2001,23(S2):36-38.
[2]杨秀莲,林燕青,王良桂.海州常山花粉低温贮藏及其生理生化特征研究[J].西北植物学报,2015,35(9):1808-1814.Yang X L,Lin Y Q,Wang L G.Cold storage and physio-biochemical characteristics of Clerodendrum trichotomum Thumb pollen[J].Acta Botanica Boreali-Occidentalia Sinica,2015,35(9):1808-1814.
[3]李红光.色彩丰富的观赏植物:海州常山[N].中国绿色时报,2009-07-14(B2).Li H G.Colorful ornamental plants:Clerodendrum trichotomum Thumb[N].China Green Times,2009-07-14(B2).
[4]崔会平.海州常山:观花观果两相宜[N].中国绿色时报,2007-11-20(B4).Cui H P.Clerodendrum trichotomum Thumb:Ornamental flowers and enjoy the fruit[N].China Green Times,2007-11-20(B4).
[5]郭鸿飞,张延龙,牛立新,等.8种中国野生百合花色素成分分析[J].西北农林科技大学学报(自然科学版),2015,43(3):98-104.Guo H F,Zhang Y L,Niu L X,et al.Petal pigments of eight wild Liliumspecies native to China[J].Journal of Northwest A&F University(Natural Science Edition),2015,43(3):98-104.
[6]安田齐.花色的生理生物化学[M].北京:中国林业出版社,1989:15-54.An T Q.Physiological biochemistry[M].Beijing:China Forestry Publishing House,1989:15-54.
[7]白新祥,胡可,戴思兰,等.不同花色菊花品种花色素成分的初步分析[J].北京林业大学学报,2006(5):84-89.Bai X X,Hu K,Dai S L,et al.Components of flower pigments in the petals of different color Chrysanthemum morifolium Ramat cultivars[J].Journal of Beijing Forestry University,2006(5):84-89.
[8]朱满兰,王亮生,张会金,等.耐寒睡莲花瓣中花青素苷组成及其与花色的关系[J].植物学报,2012,47(5):437-453.Zhu M L,Wang L S,Zhang H J,et al.Composition of anthocyanin in hardy Nymphaeapetals and its relationship with flower color[J].Chinese Bulletin of Botany,2012,47(5):437-453.
[9]钱见平,丰震,王长宪,等.元宝枫无性系秋季叶片主要成分含量动态变化[J].中国农学通报,2013,29(7):26-30.Qian J P,Feng Z,Wang C X,et al.The dynamic changes of main component in leaves of Acer truncatum Bunge clones in autumn[J].Chinese Agricultural Science Bulletin,2013,29(7):26-30.
[10]王艺菲,辛秀兰,陈亮,等.pH示差法测定不同种类蓝果忍冬总花色苷含量[J].食品研究与开发,2014,35(7):75-78.Wang Y F,Xin X L,Chen L,et al.Total anthocyanins content in different species of Lonicera cearulea Linn by pH-differential spectrophotometry[J].Food Research and Development,2014,35(7):75-78.
[11]李文砚,韦持章,孔方南,等.蒽酮法测定木奶果果实中可溶性糖含量的研究[J].中国园艺文摘,2015,31(12):7-8,28.Li W Y,Wei C Z,Kong F N,et al.Determination of soluble saccharide contents of Baccaurea ramiflora Lour.usinganthrone colorimetric method[J].Chinese Journal of Horticulture,2015,31(12):7-8,28.
[12]赵宗方,赵勇,吴桂法.果实花青素含量与PAL活性关系的研究[J].园艺学报,1994(2):199-200.Zhao Z F,Zhao Y,Wu G F.Study on the relationship between anthocyanin content and PAL activity in fruits[J].Journal of Horticulture,1994(2):199-200.
[13]王庆菊,李晓磊,王磊,等.紫叶稠李叶片花色苷及其合成相关酶动态[J].林业科学,2008(3):45-49.Wang Q J,Li X L,Wang L,et al.Dynamic changes of anthocyanin and the relevant biosynthesis enzymes in Padus virginiana‘Schubert’leaves[J].Scientia Silvae Sinicae,2008(3):45-49.
[14]Silva V O,Freitas A A,Macanita A L,et al.Chemistry and photochemistry of natural plant pigments:the anthocyanins[J].Journal of Physical Organic Chemistry,2016,29(11):594-599.
[15]Sakata Y,Aoki N,Tsunematsu S,et al.Petal coloration and pigmentation of tree peony bred and selected in Daikon Island(Shimane Prefecture)[J].Engei Gakkai Zasshi,2008,64(2):351-357.
[16]梁娟,蔺银鼎.元宝枫秋叶着色生理指标的研究[J].山西农业大学学报(自然科学版),2009,29(1):37-40.Liang J,Lin Y D.Studies on the physiological indexes of Acer truncatumfall leaves during coloring process[J].Journal of Shanxi Agricultural University(Natural Science Edition),2009,29(1):37-40.
[17]王雅琼,郑伟尉,臧运祥,等.不同品种羽衣甘蓝生长期色素含量变化规律研究[J].中国农学通报,2013,29(10):154-161.Wang Y Q,Zheng W W,Zang Y X,et al.Studies on change of pigment contents during different developmental stages among several flowering kale cultivars[J].Chinese Agricultural Science Bulletin,2013,29(10):154-161.
[18]Escribano-Bailon M T,Santos-Buelga C,Rivas-Gonzalo J C.Anthocyanins in cereals[J].Journal of Chromatography A,2004,1054(1):129-141.
[19]胡静静,沈向,李雪飞,等.黄连木秋季叶色变化与可溶性糖和矿质元素的关系[J].林业科学,2010,46(2):80-86.Hu J J,Shen X,Li X F,et al.Relationships of leaf color changes with soluble sugars and mineral elements in leaves of Pistacia chinensis in autumn[J].Scientia Silvae Sinicae,2010,46(2):80-86.
[20]杨暖,姜琳,姜官恒,等.北美豆梨秋冬叶色与生理变化的关系[J].农学学报,2015,5(11):73-78.Yang N,Jiang L,Jiang G H,et al.The relationship between leaf color in autumn and physiological changes of Pyrus calleryana Decne[J].Journal of Agriculture,2015,5(11):73-78.
[21]曹丽春,徐翔宇,李金平,等.不同色泽无花果果皮色素变化特征及其与果实糖分累积的关系[J].中国农业大学学报,2017,22(2):54-60.Cao L C,Xu X Y,Li J P,et al.Peel pigment change during the development of figs with different colors and the correlation with soluble sugar accumulation[J].Journal of China Agricultural University,2017,22(2):54-60.
[22]李云萍,郭晋雅,高峰.紫心甘薯花青素积累与PAL活性的关系[J].西南大学学报(自然科学版),2010,32(2):68-72.Li Y P,Guo J Y,Gao F.Relationship between anthocyanin accumulation and PAL activity in Ipomoea batatas Lam[J].Journal of Southwest University(Natural Science Edition),2010,32(2):68-72.
[23]Mori T,Sakurai M,Sakuta M.Changes in PAL,CHS,DAHPsynthase(DS-Co and Ds-Mn)activity during anthocyanin synthesis in suspension culture of Fragaria ananassa[J].Plant Cell Tissue&Organ Culture,2000,62(2):135-139.
[24]王惠聪,黄旭明,胡桂兵,等.荔枝果皮花青苷合成与相关酶的关系研究[J].中国农业科学,2004,37(12):2028-2032.Wang H C,Huang X M,Hu G B,et al.Studies on the relationship between anthocyanin biosynthesis and related enzymes in Litchi pericarp[J].Scientia Agricultura Sinica,2004,37(12):2028-2032.
[25]陈继卫,沈朝栋,贾玉芳,等.鸡爪槭转色期叶色变化生理研究[J].核农学报,2010,24(1):171-175,180.Chen J W,Shen Z D,Jia Y F,et al.Physiological study on leaf color change during the color change in Acer palmatum[J].Journal of Nuclear Agricultural Sciences,2010,24(1):171-175,180.
[26]朱书香,王中华,李彦慧,等.紫叶矮樱叶片色素理化性质的研究[J].河北农业大学学报,2010,33(3):33-36,45.Zhu S X,Wang Z H,Li Y H,et al.Study on the physicalchemical properties of leaf pigment of Prunus×cistenena‘Purpleleaf and Chevry’[J].Journal of Agricultural University of Hebei,2010,33(3):33-36,45.
[27]于晓南,张启翔.光强与光质对“美人梅”叶色的影响[J].北京林业大学学报,2001,23(S2):36-38.Yu X N,Zhang Q X.Effects of light intensity and light quality on leaf coler expression of Prunus blireiana[J].Journal of Beijing Forestry University,2001,23(S2):36-38.