茉莉酸甲酯对滇龙胆叶中4种裂环烯醚萜类物质含量的影响
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摘要
目的:研究外源性茉莉酸甲酯对滇龙胆叶中4种裂环烯醚萜类物质(龙胆苦苷、獐牙菜苦苷、獐牙菜苷和苦龙胆酯苷)含量及生理生化指标的影响,为茉莉酸甲酯在滇龙胆规范化种植中的应用提供科学依据。方法:分别以10、50、100、200 mg/L茉莉酸甲酯喷施滇龙胆植株2、4、6、8、10 d(喷施量均为500 m L)后,收集药材叶为样品材料,并收集相应时段未喷施处理的药材叶为对照。采用高效液相色谱(HPLC)法测定4个质量浓度下茉莉酸甲酯喷施10 d后滇龙胆叶中4种裂环烯醚萜类物质含量,优选茉莉酸甲酯的最佳喷施质量浓度(有效成分含量最高)。采用HPLC法测定以最佳喷施质量浓度茉莉酸甲酯喷施不同时间后滇龙胆叶4种裂环烯醚萜类物质的含量,并测定其中相关生理生化指标(超氧化物歧化酶、过氧化物酶、过氧化氢酶和丙二醛)水平。结果:以10、50、100、200 mg/L茉莉酸甲酯喷施滇龙胆植株10 d后,叶中4种裂环烯醚萜类物质的含量较未处理叶均不同程度增加,其中以100 mg/L茉莉酸甲酯喷施后效果最好,此时叶中龙胆苦苷、獐牙菜苦苷和獐牙菜苷的含量分别为未处理叶的1.88、2.36和1.87倍(P<0.05)。以100 mg/L茉莉酸甲酯喷施2、4、6、8、10 d后,叶中4种裂环烯醚萜类物质的含量均不同程度高于同时段未处理叶;在喷施4 d后叶中龙胆苦苷含量开始出现显著差异(P<0.05),在喷施6 d后叶中4种裂环烯醚萜类物质的含量总体较高,此时叶中龙胆苦苷、獐牙菜苦苷、獐牙菜苷和苦龙胆酯苷含量分别为同时段未处理叶的1.88、1.88、1.47、1.82倍(P<0.05);并自喷施4 d起,叶中各生理生化指标水平较未处理叶均显著升高(P<0.05)。结论:以100 mg/L茉莉酸甲酯喷施6 d后滇龙胆叶中4种裂环烯醚萜类物质含量升高最明显,这可能与其提高了叶中相关生理生化指标的水平有关。
OBJECTIVE:To study the effects of exogenous methyl jasmonate on the contents of 4 kinds of secoiridoidsubstance(gentiopicrin,swertiamain,sweroside and amarogentin) and biochemical indexes in the leaves of Gentiana rigescens,and to provide scientific evidence for the application of methyl jasmonate in standardized planting of G. rigescens. METHODS:2,4,6,8,10 d after spraying 10,50,100,200 mg/L methyl jasmonate(spraying amount was 500 mL)for G. rigescens strain,theleaves of medicinal material were collected as sample,and the other leaves without spraying methyl jasmonate were collected ascontrol. HPLC method was used to determine the contents of 4 kinds of secoiridoid substance in the leaves of G. rigescens afterspraying 4 kinds of concentrations of methyl jasmonate for 10 d. The concentration of methyl jasmonate was optimized(the contentof effective component was the highest). HPLC method was adopted to determine the contents of 4 kinds of secoiridoid substancein the leaves of G. rigescens after spraying the optimal concentration of methyl jasmonate for different time. The levels of relevantbiochemical indexes(SOD,POD,CAT,MDA)were determined. RESULTS:10 d after spraying 10,50,100,200 mg/L methyljasmonate for G. rigescens strains,the contents of 4 kinds of secoiridoid substance in the leaves were increased to different extent.Compared with untreated leaves,100 mg/L methyl jasmonate had the best effect after spraying,and the contents of gentiopicrin,swertiamain and sweroside in treated leaves were 1.88,2.36 and 1.87 times of those in untreated leaves,respectively(P<0.05).2,4,6,8,10 d after spraying 100 mg/L methyl jasmonate,the contents of 4 kinds of secoiridoid substance in treated leaves werehigher than untreated leaves at corresponding stage;the content of secoiridoid had significant difference after spraying for 4 d(P<0.05). The contents of active components in general wererelatively high after spraying for 6 d, and the contents ofgentiopicrin, swertiamain, sweroside and amarogentin intreated leaves were 1.88,1.88,1.47,1.82 times of those inuntreated leaves, respectively(P<0.05). The levels ofrelevant biochemical indexes in treated leaves were increasedsignificantly since 4 d of spraying,compared with untreated leaves(P<0.05). CONCLUSIONS:After spraying 100 mg/L methyljasmonate for 6 d,the contents increase of 4 kinds of secoiridoid substance in the leaves of G. rigescens are most obvious,whichmay be associated with improving the levels of related biochemical indexes.
OBJECTIVE:To study the effects of exogenous methyl jasmonate on the contents of 4 kinds of secoiridoidsubstance(gentiopicrin,swertiamain,sweroside and amarogentin) and biochemical indexes in the leaves of Gentiana rigescens,and to provide scientific evidence for the application of methyl jasmonate in standardized planting of G. rigescens. METHODS:2,4,6,8,10 d after spraying 10,50,100,200 mg/L methyl jasmonate(spraying amount was 500 mL)for G. rigescens strain,theleaves of medicinal material were collected as sample,and the other leaves without spraying methyl jasmonate were collected ascontrol. HPLC method was used to determine the contents of 4 kinds of secoiridoid substance in the leaves of G. rigescens afterspraying 4 kinds of concentrations of methyl jasmonate for 10 d. The concentration of methyl jasmonate was optimized(the contentof effective component was the highest). HPLC method was adopted to determine the contents of 4 kinds of secoiridoid substancein the leaves of G. rigescens after spraying the optimal concentration of methyl jasmonate for different time. The levels of relevantbiochemical indexes(SOD,POD,CAT,MDA)were determined. RESULTS:10 d after spraying 10,50,100,200 mg/L methyljasmonate for G. rigescens strains,the contents of 4 kinds of secoiridoid substance in the leaves were increased to different extent.Compared with untreated leaves,100 mg/L methyl jasmonate had the best effect after spraying,and the contents of gentiopicrin,swertiamain and sweroside in treated leaves were 1.88,2.36 and 1.87 times of those in untreated leaves,respectively(P<0.05).2,4,6,8,10 d after spraying 100 mg/L methyl jasmonate,the contents of 4 kinds of secoiridoid substance in treated leaves werehigher than untreated leaves at corresponding stage;the content of secoiridoid had significant difference after spraying for 4 d(P<0.05). The contents of active components in general wererelatively high after spraying for 6 d, and the contents ofgentiopicrin, swertiamain, sweroside and amarogentin intreated leaves were 1.88,1.88,1.47,1.82 times of those inuntreated leaves, respectively(P<0.05). The levels ofrelevant biochemical indexes in treated leaves were increasedsignificantly since 4 d of spraying,compared with untreated leaves(P<0.05). CONCLUSIONS:After spraying 100 mg/L methyljasmonate for 6 d,the contents increase of 4 kinds of secoiridoid substance in the leaves of G. rigescens are most obvious,whichmay be associated with improving the levels of related biochemical indexes.
引文
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[8]朱宏涛,李江,李元,等.茉莉酸甲酯对三七组培苗中总皂苷含量的影响[J].西部林业科学,2014,43(2):72-78.
[9]陈林,崔培梧,鲁耀邦,等.茉莉酸甲酯对茯苓三萜生物合成的调控研究[J].湖南中医药大学学报,2017,37(6):606-610.
[10]谢阳姣,何志鹏,林伟,等.茉莉酸甲酯对苦玄参生长及苦玄参苷积累的影响研究[J].作物杂志,2013(2):80-84.
[11]王焕,杨锦芬,邓可,等.茉莉酸甲酯影响阳春砂挥发性萜类代谢和基因转录[J].世界科学技术-中医药现代化,2014,16(7):1528-1536.
[12]李嘉丰,许嘉,任跃英,等.不同生长年限北五味子植株黑斑病发病规律及生理指标的变化[J].河南农业科学,2018,47(9):88-92.
[13]王学奎.植物生理生化实验原理和技术[M]. 2版.北京:高等教育出版社,2006:167-281.
[14]韩多,赵志莲,刘卫红,等.坚龙胆主要次生代谢产物及分子差异的研究[J].中国实验方剂学杂志,2016,22(18):35-39.
[15]王逸文,胡高升,贾景明. 2-氨基磷酸茚和茉莉酸甲酯对高山红景天愈伤组织生长和红景天苷积累的影响[J].沈阳药科大学学报,2012,29(9):718-723.
[16]从春蕾,郅军锐,廖启荣,等.蓟马取食、机械损伤以及外源水杨酸甲酯和茉莉酸对菜豆叶片防御酶活性的影响[J].昆虫学报,2014,57(5):564-571.
[17]张梅,林棋,娄本勇,等.茉莉酸甲酯对锦绣杜鹃黄酮的累积及相关酶活性的影响[J].甘肃农业大学学报,2015,50(4):165-169.
[18]熊茜,王春幸,王凯凯,等.苋菜芽生长中相关酶及抗氧化活性的变化[J].食品工业科技,2017,38(24):10-15.
[2]国家药典委员会.中华人民共和国药典:一部[S]. 2015年版.北京:中国医药科技出版社,2015:96.
[3]赵志莲,张琳,李海峰,等.大理白族自治州不同居群坚龙胆叶中裂环烯醚萜类物质含量的研究[J].中国药房,2018,29(5):647-651.
[4]王丽,杨雁,方艳,等.中药材滇龙胆资源调查和分析研究[J].西南农业学报,2017,30(2):267-272.
[5]韩多,赵志莲,刘卫红,等. 3种龙胆属药用植物叶中主要有效成分的积累[J].中国医药工业杂志,2016,47(7):865-869.
[6]赵志莲,张琳,刘卫红,等.滇龙胆裂环烯醚萜类活性成分积累规律的研究[J].时珍国医国药,2016,27(9):2073-2075.
[7]滕红梅,谢旭蓉,王玲丽,等.外源物质对远志根中皂苷含量的影响[J].南方农业学报,2014,42(12):2143-2147.
[8]朱宏涛,李江,李元,等.茉莉酸甲酯对三七组培苗中总皂苷含量的影响[J].西部林业科学,2014,43(2):72-78.
[9]陈林,崔培梧,鲁耀邦,等.茉莉酸甲酯对茯苓三萜生物合成的调控研究[J].湖南中医药大学学报,2017,37(6):606-610.
[10]谢阳姣,何志鹏,林伟,等.茉莉酸甲酯对苦玄参生长及苦玄参苷积累的影响研究[J].作物杂志,2013(2):80-84.
[11]王焕,杨锦芬,邓可,等.茉莉酸甲酯影响阳春砂挥发性萜类代谢和基因转录[J].世界科学技术-中医药现代化,2014,16(7):1528-1536.
[12]李嘉丰,许嘉,任跃英,等.不同生长年限北五味子植株黑斑病发病规律及生理指标的变化[J].河南农业科学,2018,47(9):88-92.
[13]王学奎.植物生理生化实验原理和技术[M]. 2版.北京:高等教育出版社,2006:167-281.
[14]韩多,赵志莲,刘卫红,等.坚龙胆主要次生代谢产物及分子差异的研究[J].中国实验方剂学杂志,2016,22(18):35-39.
[15]王逸文,胡高升,贾景明. 2-氨基磷酸茚和茉莉酸甲酯对高山红景天愈伤组织生长和红景天苷积累的影响[J].沈阳药科大学学报,2012,29(9):718-723.
[16]从春蕾,郅军锐,廖启荣,等.蓟马取食、机械损伤以及外源水杨酸甲酯和茉莉酸对菜豆叶片防御酶活性的影响[J].昆虫学报,2014,57(5):564-571.
[17]张梅,林棋,娄本勇,等.茉莉酸甲酯对锦绣杜鹃黄酮的累积及相关酶活性的影响[J].甘肃农业大学学报,2015,50(4):165-169.
[18]熊茜,王春幸,王凯凯,等.苋菜芽生长中相关酶及抗氧化活性的变化[J].食品工业科技,2017,38(24):10-15.