生态因子及关键酶基因表达对秋季黄芩采收期主要药效成分合成的影响
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摘要
目的研究黄芩秋季采收期药效成分的动态变化及生态因子和关键酶表达对其的影响。方法以人工栽培的一年生黄芩为研究对象,采用实时荧光定量PCR(qRT-PCR)法测定秋季连续时间黄芩根组织中9种关键酶基因(PAL、C4H、4CL、CHS、CHI、FNS、F6H、UBGAT、GUS)的表达量;采用HPLC法测定黄芩根中4种主要黄酮类化合物(黄芩苷、汉黄芩苷、黄芩素、汉黄芩素)的含量;以生态气象站对黄芩样地气象数据进行采集;采用SPSS统计软件和DPS统计软件进行数据分析。结果 4种黄酮类化合物含量在一年生黄芩秋季呈缓慢下降的趋势,因此黄芩秋季最佳采收时间应在9月初;灰色关联度分析结果表明影响4种黄酮类药效成分较大的生态因子为土壤含水量、最高空气温度、空气湿度、光合有效辐射;C4H、UBGAT基因的表达对黄芩秋季根部黄酮类化合物的积累具有重要影响;最大降雨强度可能通过对关键酶基因表达的影响来间接影响黄芩药效成分的积累。结论明确了一年生黄芩秋季采收期4种主要黄酮类药效成分的动态变化和黄酮类化合物关键酶基因的表达,为黄芩黄酮类化合物合成生理生态机制的明晰和黄芩药材质量的提高提供了理论依据。
Objective To study the dynamic changes of the pharmacodynamic components of Scutellaria baicalensis in the harvest period and the effects of ecological factors and key enzyme expression on it. Methods The artificial cultivated annual S. baicalensis was studied and the expression of nine key enzyme genes(PAL, C4 H, 4 CL, CHS, CHI, FNS, F6 H, UBGAT, and GUS) in the roots of S. baicalensis were determined by real-time quantitative PCR. The content of four main flavonoids(baicalin, wogonoside, baicalein, and wogonin) in the roots was determined by HPLC. The meteorological data of S. baicalensis were collected by the ecological meteorological station. SPSS statistical software and DPS statistical software were used for data analysis. Results The content of flavonoids of four monomers of annual S. baicalensis decreased slowly in autumn, so the best harvest time of S. baicalensis was in early September. The results of grey correlation analysis showed that the ecological factors affecting the four flavonoids were SWC, Max Ta, RH, and PAR. The expression of C4 H and UBGAT genes had an important effect on the accumulation of flavonoids in the roots of S. baicalensis in autumn. Maximum rainfall intensity may indirectly affect the accumulation of the pharmacodynamic components of S.baicalensis by affecting the gene expression of key enzymes. Conclusion The dynamic changes of four main flavonoids of annual S. baicalensis in autumn and the expression of key enzyme genes of S. baicalensis annual are clarified, providing the theoretical basis for the clarification of the physiological and ecological mechanism of the biosynthesis of S. baicalensis and the improvement of the quality of S. baicalensis.
Objective To study the dynamic changes of the pharmacodynamic components of Scutellaria baicalensis in the harvest period and the effects of ecological factors and key enzyme expression on it. Methods The artificial cultivated annual S. baicalensis was studied and the expression of nine key enzyme genes(PAL, C4 H, 4 CL, CHS, CHI, FNS, F6 H, UBGAT, and GUS) in the roots of S. baicalensis were determined by real-time quantitative PCR. The content of four main flavonoids(baicalin, wogonoside, baicalein, and wogonin) in the roots was determined by HPLC. The meteorological data of S. baicalensis were collected by the ecological meteorological station. SPSS statistical software and DPS statistical software were used for data analysis. Results The content of flavonoids of four monomers of annual S. baicalensis decreased slowly in autumn, so the best harvest time of S. baicalensis was in early September. The results of grey correlation analysis showed that the ecological factors affecting the four flavonoids were SWC, Max Ta, RH, and PAR. The expression of C4 H and UBGAT genes had an important effect on the accumulation of flavonoids in the roots of S. baicalensis in autumn. Maximum rainfall intensity may indirectly affect the accumulation of the pharmacodynamic components of S.baicalensis by affecting the gene expression of key enzymes. Conclusion The dynamic changes of four main flavonoids of annual S. baicalensis in autumn and the expression of key enzyme genes of S. baicalensis annual are clarified, providing the theoretical basis for the clarification of the physiological and ecological mechanism of the biosynthesis of S. baicalensis and the improvement of the quality of S. baicalensis.
引文
[1]中国药典[S].一部.2015.
[2]付国辉,马香芹.黄芩的化学成分及药理作用研究进展[J].中国当代医药,2015,22(22):18-20.
[3]雷桅,税晓容,胡侃.黄芩苷生物合成途径与生物技术研究进展[J].北方园艺,2014(22):185-189.
[4]Shen J,He C N,Liu H T,et al.Simultaneous determination of 15 flavonoids from different parts of Scutellaria baicalensis and its chemometrics analysis[J].Chin Herb Med,2019,11(1):20-27.
[5]庞力贤,张文岭,吕维红.黄芩药材质量影响因素的研究进展[J].山东中医药大学学报,2015,39(2):195-197.
[6]Yuan Q J,Zhang Z Y,Hu J,et al.Impacts of recent cultivation on genetic diversity pattern of a medicinal plant,Scutellaria baicalensis(Lamiaceae)[J].BMCGenetics,2010,11(1):29.
[7]Wang W,Chen J K,Zhou T S.CHS and UBGATExpression and baicalin accumulation in the roots of Scutellaria baicalensis Georgi during cultivation seasons[J].复旦学报(自然科学版),2006,45(5):674-678.
[8]安瑜,王旭鹏,赵建军.宁夏六盘山栽培黄芩适宜采收期研究[J].中国实验方剂学杂志,2013,19(13):161-164.
[9]靳维荣,石俊英,张小伟.不同采收期黄芩中六种黄酮类成分的动态研究[J].山东中医杂志,2008,1(4):268-271.
[10]张媛,宋双红,王喆之.栽培黄芩中黄酮类成分的动态积累研究[J].中草药,2009,40(9):1478-1480.
[11]雷燕妮,张小斌.不同采收季节黄芩苷含量比较[J].浙江农业科学,2013(8):964-965.
[12]缪晓素,宋国虎,刘容秀,等.不同栽培年限及采收期对黄芩药材有效成分含量的影响研究[J].中国现代中药,2015,17(8):836-839.
[13]Cheng L,Han M,Yang L,et al.Changes in the physiological characteristics and baicalin biosynthesis metabolism of Scutellaria baicalensis Georgi.under drought stress[J].Industrial Crops Prod,2018,122:473-482.
[14]程林.干旱胁迫黄芩生理生态变化及其黄芩苷生物合成的分子生态机制[D].长春:吉林农业大学,2018.
[15]苏文华,张光飞,李秀华,等.植物药材次生代谢产物的积累与环境的关系[J].中草药,2005,36(9):1415-1418.
[16]杨利民,张永刚,林红梅,等.中药材质量形成理论与控制技术研究进展[J].吉林农业大学学报,2012,34(2):119-124.
[17]陈顺钦,袁媛,罗毓健,等.光照对黄芩黄酮类活性成分积累及其相关基因表达的影响[J].中国中药杂志,2010,35(6):682-685.
[18]颜永刚,王红艳,邓翀,等.生长年限、海拔和光照对大黄中8种成分量的影响研究[J].中草药,2017,48(11):2285-2291.
[19]邵玺文,韩梅,韩忠明,等.不同生境条件下黄芩光合日变化与环境因子的关系[J].生态学报,2009,29(3):1470-1477.
[20]曾燕,郭兰萍,王继永,等.植物黄酮类化合物的化学生态学功能及中药材定向栽培的构想[J].中国现代中药,2015,17(8):776-790.
[21]邵玺文,韩梅,韩忠明,等.水分供给量对黄芩生长与光合特性的影响[J].生态学报,2006,(10):3214-3220.
[22]Ji-Chao H U,Cao W X,Zhang J B,et al.Quantifying responses of winter wheat physiological processes to soil water stress for use in growth simulation modeling[J].Pedosphere,2004,14(4):509-518.
[23]张永刚,韩梅,姜雪,等.黄芩对干旱复水的生理生态响应[J].中国中药杂志,2013,38(22):3845-3850.
[24]杨金艳,杨万勤,王开运,等.木本植物对CO2浓度和温度升高的相互作用的响应[J].植物生态学报,2003,27(3):304-310.
[25]Jarvi M P,Burton A J.Acclimation and soil moisture constrain sugar maple root respiration in experimentally warmed soil[J].Tree Physiol,2013,33(9):949-959.
[26]刘金花.环境因子对黄芩植株代谢的影响[D].济南:山东中医药大学,2011.
[27]Yang D,Ma P,Liang X,et al.PEG and ABA trigger methyl jasmonate accumulation to induce the MEP pathway and increase tanshinone production in Salvia miltiorrhiza hairy roots[J].Physiol Plantarum,2012,146(2):173-183.
[28]邵玺文.黄芩高产栽培及质量调控生理生态机制研究[D].长春:吉林农业大学,2008.
[29]李帅.黄芩种子及药材最佳采收期研究[D].长春:吉林农业大学,2011.
[30]李化.黄芩采收规律及其数学模型的研究[D].北京:中国中医科学院,2008.
[2]付国辉,马香芹.黄芩的化学成分及药理作用研究进展[J].中国当代医药,2015,22(22):18-20.
[3]雷桅,税晓容,胡侃.黄芩苷生物合成途径与生物技术研究进展[J].北方园艺,2014(22):185-189.
[4]Shen J,He C N,Liu H T,et al.Simultaneous determination of 15 flavonoids from different parts of Scutellaria baicalensis and its chemometrics analysis[J].Chin Herb Med,2019,11(1):20-27.
[5]庞力贤,张文岭,吕维红.黄芩药材质量影响因素的研究进展[J].山东中医药大学学报,2015,39(2):195-197.
[6]Yuan Q J,Zhang Z Y,Hu J,et al.Impacts of recent cultivation on genetic diversity pattern of a medicinal plant,Scutellaria baicalensis(Lamiaceae)[J].BMCGenetics,2010,11(1):29.
[7]Wang W,Chen J K,Zhou T S.CHS and UBGATExpression and baicalin accumulation in the roots of Scutellaria baicalensis Georgi during cultivation seasons[J].复旦学报(自然科学版),2006,45(5):674-678.
[8]安瑜,王旭鹏,赵建军.宁夏六盘山栽培黄芩适宜采收期研究[J].中国实验方剂学杂志,2013,19(13):161-164.
[9]靳维荣,石俊英,张小伟.不同采收期黄芩中六种黄酮类成分的动态研究[J].山东中医杂志,2008,1(4):268-271.
[10]张媛,宋双红,王喆之.栽培黄芩中黄酮类成分的动态积累研究[J].中草药,2009,40(9):1478-1480.
[11]雷燕妮,张小斌.不同采收季节黄芩苷含量比较[J].浙江农业科学,2013(8):964-965.
[12]缪晓素,宋国虎,刘容秀,等.不同栽培年限及采收期对黄芩药材有效成分含量的影响研究[J].中国现代中药,2015,17(8):836-839.
[13]Cheng L,Han M,Yang L,et al.Changes in the physiological characteristics and baicalin biosynthesis metabolism of Scutellaria baicalensis Georgi.under drought stress[J].Industrial Crops Prod,2018,122:473-482.
[14]程林.干旱胁迫黄芩生理生态变化及其黄芩苷生物合成的分子生态机制[D].长春:吉林农业大学,2018.
[15]苏文华,张光飞,李秀华,等.植物药材次生代谢产物的积累与环境的关系[J].中草药,2005,36(9):1415-1418.
[16]杨利民,张永刚,林红梅,等.中药材质量形成理论与控制技术研究进展[J].吉林农业大学学报,2012,34(2):119-124.
[17]陈顺钦,袁媛,罗毓健,等.光照对黄芩黄酮类活性成分积累及其相关基因表达的影响[J].中国中药杂志,2010,35(6):682-685.
[18]颜永刚,王红艳,邓翀,等.生长年限、海拔和光照对大黄中8种成分量的影响研究[J].中草药,2017,48(11):2285-2291.
[19]邵玺文,韩梅,韩忠明,等.不同生境条件下黄芩光合日变化与环境因子的关系[J].生态学报,2009,29(3):1470-1477.
[20]曾燕,郭兰萍,王继永,等.植物黄酮类化合物的化学生态学功能及中药材定向栽培的构想[J].中国现代中药,2015,17(8):776-790.
[21]邵玺文,韩梅,韩忠明,等.水分供给量对黄芩生长与光合特性的影响[J].生态学报,2006,(10):3214-3220.
[22]Ji-Chao H U,Cao W X,Zhang J B,et al.Quantifying responses of winter wheat physiological processes to soil water stress for use in growth simulation modeling[J].Pedosphere,2004,14(4):509-518.
[23]张永刚,韩梅,姜雪,等.黄芩对干旱复水的生理生态响应[J].中国中药杂志,2013,38(22):3845-3850.
[24]杨金艳,杨万勤,王开运,等.木本植物对CO2浓度和温度升高的相互作用的响应[J].植物生态学报,2003,27(3):304-310.
[25]Jarvi M P,Burton A J.Acclimation and soil moisture constrain sugar maple root respiration in experimentally warmed soil[J].Tree Physiol,2013,33(9):949-959.
[26]刘金花.环境因子对黄芩植株代谢的影响[D].济南:山东中医药大学,2011.
[27]Yang D,Ma P,Liang X,et al.PEG and ABA trigger methyl jasmonate accumulation to induce the MEP pathway and increase tanshinone production in Salvia miltiorrhiza hairy roots[J].Physiol Plantarum,2012,146(2):173-183.
[28]邵玺文.黄芩高产栽培及质量调控生理生态机制研究[D].长春:吉林农业大学,2008.
[29]李帅.黄芩种子及药材最佳采收期研究[D].长春:吉林农业大学,2011.
[30]李化.黄芩采收规律及其数学模型的研究[D].北京:中国中医科学院,2008.