基于遗传-化学-生态特征的西洋参品质生态型研究
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摘要
我国不同生态型的同种药材品质差异显著,本课题组前期发现国产西洋参根据人参皂苷的特征存在关外型与关内型两大生态型,然而引起西洋参品质变异的遗传生态机制尚不清楚。本文基于遗传-化学-生态策略,运用转录组学技术、HPLC技术,并进行西洋参转录组数据、人参皂苷含量及环境气候生态因子的相关性分析。转录组结果显示人参皂苷生物合成关键基因如羟甲基戊二酰辅酶A还原酶(hydroxymethylglutaryl-CoA reductase,HMGR)、β-氨基酸合成酶(beta-amyrin synthase, AS)和法呢基焦磷酸合成酶(farnesyl diphosphate synthase, FPS)等在关内型均显著下调。HPLC结果表明关外型西洋参中人参皂苷含量高于关内型西洋参。相关性分析结果显示西洋参人参皂苷的含量与甲羟戊酸二磷酸脱羧酶(mevalonate kinase, MK)、羟甲基戊二酰辅酶A合酶(hydroxymethylglutaryl-CoA synthase, HMGS)、HMGR和AS等人参皂苷生物合成关键基因的表达成正相关,而与糖基转移酶GT的表达成负相关,西洋参的人参皂苷含量与两大生态型所处环境的温度、日照等气候因子成负相关,与水分成正相关。本文为阐释西洋参两大生态型品质变异的形成机制提供了新思路,也为道地药材品质生态学理论研究提供科学依据。
Medicinal materials in China differ in quality by different ecological types. Our research group found that there were two ecotypes of domestic Panax quinquefolium L. according to the characteristics of ginsenosides, inside versus outside Shanhaiguan. The genetic and ecological mechanisms of quality variation of Panax quinquefolium L. is unknown. Based on the genetic-chemical-ecological strategy, transcriptome and HPLC technology were used for comprehensive correlation analyses of transcriptomic data, ginsenoside content and environmental climate ecological factors. The transcriptomic results showed that key genes of ginsenoside biosynthesis, such as HMGR, AS and FPS, were significantly down-regulated in the inside Shanhaiguan ecotype. HPLC results showed that the quality of outside Shanhaiguan ecotype Panax quinquefolium L. was higher than that of the inside ecotype,with the content of ginsenosides in outside Panax quinquefolium L. was higher than that of inside ecotype except Rb2. Correlation analyses revealed that content of Panax quinquefolium L. ginsenoside is positively related to the expression levels of ginsenoside biosynthesis key genes(MK, HMGS, HMGR, and AS), and negatively related to the expression of glycosyl transferase(GT). The content of ginsenosides is negative related with climate factors,such as temperature, sunshine, and is positively related with moisture in both ecological environments. This study has provided a new mechanistic insight into the quality variations of two ecotypes for Panax quinquefolium L. and established a scientific basis for studying the ecological factors for the quality of traditional Chinese medicine.
Medicinal materials in China differ in quality by different ecological types. Our research group found that there were two ecotypes of domestic Panax quinquefolium L. according to the characteristics of ginsenosides, inside versus outside Shanhaiguan. The genetic and ecological mechanisms of quality variation of Panax quinquefolium L. is unknown. Based on the genetic-chemical-ecological strategy, transcriptome and HPLC technology were used for comprehensive correlation analyses of transcriptomic data, ginsenoside content and environmental climate ecological factors. The transcriptomic results showed that key genes of ginsenoside biosynthesis, such as HMGR, AS and FPS, were significantly down-regulated in the inside Shanhaiguan ecotype. HPLC results showed that the quality of outside Shanhaiguan ecotype Panax quinquefolium L. was higher than that of the inside ecotype,with the content of ginsenosides in outside Panax quinquefolium L. was higher than that of inside ecotype except Rb2. Correlation analyses revealed that content of Panax quinquefolium L. ginsenoside is positively related to the expression levels of ginsenoside biosynthesis key genes(MK, HMGS, HMGR, and AS), and negatively related to the expression of glycosyl transferase(GT). The content of ginsenosides is negative related with climate factors,such as temperature, sunshine, and is positively related with moisture in both ecological environments. This study has provided a new mechanistic insight into the quality variations of two ecotypes for Panax quinquefolium L. and established a scientific basis for studying the ecological factors for the quality of traditional Chinese medicine.
引文
[1]Cheng YQ,Wang CX,Xu T,et al.Progress in quality evaluation of American ginseng medicinal materials[J].Shandong Sci(山东科学),2018,4:39-43,49.
[2]Lee SH,Jung BH,Kim SY,et al.The antistress effect of ginseng total saponin and ginsenoside Rg3and Rb1evaluated by brain polyamine level under immobilization stress[J].Pharmacol Res,2006,54:46-49.
[3]Lee J,Jung E,Lee J,et al.Panax ginseng induces human Type Icollagen synthesis through activation of Smad signaling[J].JEthnopharmacol,2007,109:29-34.
[4]Chen QQ,Wu QQ,Gao XJ,et al.Research progress on fungal biotransformation of ginseng saponins in American ginseng[J].Food Res Devel(食品研究与开发),2019,40:220-224.
[5]Wang J,Li J,Li J,et al.Transcriptome profiling shows gene regulation patterns in ginsenoside pathway in response to methyl jasmonate in Panax quinquefolium adventitious root[J].Sci Rep,2016,6:37263.
[6]Huang LF,Suo FM,Song JY,et al.Quality variation and ecotype division of Panax quinquefolium in China[J].Acta Pharm Sin(药学学报),2013,48:580-589.
[7]Huang LF,Chen SL.Quality ecology of traditional Chinese medicine:an emerging interdisciplinary subject[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2017,23:1-11.
[8]Huang LF,Fu J,Chen SL.Academic discussion on ecological variation of Chinese medicinal materials[J].Chin Tradit Herb Drugs(中草药),2012,43:1249-1258.
[9]Cheng RZ,Wu ML,Shen L,et al.Analysis of ecological suitability of global producing area of traditional Chinese medicine[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2017,23:19-24.
[10]Xu J,Shen L,Wang Y,et al.Analysis of global ecological suitable production area of Ginkgo biloba based on GMPGIS[J].World Chin Med(世界中医药),2017,12:969-973.
[11]Jiang XY,Sun HB,Qin JH,et al.Study on the functional zoning of Fritillaria cirrhosa based on growth suitability and quality suitability[J].China J Chin Mater Med(中国中药杂志),2016,41:3194-3201.
[12]Zheng SH,Jiang X,Huang LF,et al.Chemical and genetic dis‐crimination of Cistanches Herba based on UPLCQTOF/MS and DNA barcoding[J].PLoS One,2014,9:e98061.
[13]Lin L,Josef BA,Bing L,et al.Three-dimensional evaluation on ecotypic diversity of traditional Chinese medicine:a case study of Artemisia annua L.[J].Front Plant Sci,2017,8:1225.
[14]Li L,Zheng SH,Brinckmann JA,et al.Chemical and genetic diversity of Astragalus mongholicus grown in different ecoclimatic regions[J].PLoS One,2017,12:e0184791.
[15]Li L,Zheng SH,Yang QZ,et al.Distinguishing Astragalus mongholicus and its planting soil samples from different regions by ICP-AES[J].Molecules.2016,21:482.
[16]Wang ZH.Study on the Quality and Origin Variation of American Ginseng and Screening of EGFR Kinase Inhibitors for 49 Medicinal Extracts(西洋参品质和产地变异研究及49种药材提取物EGFR激酶抑制剂筛选)[D].Beijing:Peking Union Medical College,2015.
[17]Yang JL,Gao LL,Zhu P.Advances in the biosynthesis research of ginsenosides[J].Acta Pharm Sin(药学学报),2013,48:170-178.
[18]He DT,Wang B,Chen JM.Research progress on pharmacological effects of ginsenoside[J].J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2012,14:118-121.
[19]Christensen JC.Ginsenosides chemistry,biosynthesis,analysis,and potential health effects[J].Adv Food Nutr Res,2009,55:1-99.
[20]Wu Q,Sun C,Chen SL.Identification and expression analysis of a 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from American ginseng[J].Plant OMICS:J Plant Mol Biol Omics,2012,5:414-420.
[21]Kim OT,Bang KH,Jung SJ,et al.Molecular characterization of ginseng farnesyl diphosphate synthase gene and its up-regulation by methyl jasmonate[J].Biol Plant,2010,54:47-53.
[22]Lee MH,Jeong JH,Seo JW,et al.Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene[J].Plant Cell Physiol,2004,45:976-984.
[23]Han JY,In JG,Kwon YS,et al.Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxi‐dase gene in Panax ginseng[J].Phytochemistry,2010,71:36-46.
[24]Lee MH,Han JY,Kim HJ,et al.Dammarenediol-II production confers TMV Tolerance in transgenic tobacco expressing Panax ginseng dammarenediol-II synthase[J].Plant Cell Physiol,2012,53:173-182.
[25]Yue CJ,Zhong JJ.Purification and characterization of UDPG:ginsenoside Rd glucosyltransferase from suspended cells of Panax notoginseng[J].Proc Biochem,2005,40:3742-3748.
[26]Xing N,Liu LL,Yin SG,et al.The association between property of Chinese medicinal herbs and chemical components,ecologi‐cal environment[J].J Chin Med Mater(中药材),2011,34:324-325.
[2]Lee SH,Jung BH,Kim SY,et al.The antistress effect of ginseng total saponin and ginsenoside Rg3and Rb1evaluated by brain polyamine level under immobilization stress[J].Pharmacol Res,2006,54:46-49.
[3]Lee J,Jung E,Lee J,et al.Panax ginseng induces human Type Icollagen synthesis through activation of Smad signaling[J].JEthnopharmacol,2007,109:29-34.
[4]Chen QQ,Wu QQ,Gao XJ,et al.Research progress on fungal biotransformation of ginseng saponins in American ginseng[J].Food Res Devel(食品研究与开发),2019,40:220-224.
[5]Wang J,Li J,Li J,et al.Transcriptome profiling shows gene regulation patterns in ginsenoside pathway in response to methyl jasmonate in Panax quinquefolium adventitious root[J].Sci Rep,2016,6:37263.
[6]Huang LF,Suo FM,Song JY,et al.Quality variation and ecotype division of Panax quinquefolium in China[J].Acta Pharm Sin(药学学报),2013,48:580-589.
[7]Huang LF,Chen SL.Quality ecology of traditional Chinese medicine:an emerging interdisciplinary subject[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2017,23:1-11.
[8]Huang LF,Fu J,Chen SL.Academic discussion on ecological variation of Chinese medicinal materials[J].Chin Tradit Herb Drugs(中草药),2012,43:1249-1258.
[9]Cheng RZ,Wu ML,Shen L,et al.Analysis of ecological suitability of global producing area of traditional Chinese medicine[J].Chin J Exp Tradit Med Form(中国实验方剂学杂志),2017,23:19-24.
[10]Xu J,Shen L,Wang Y,et al.Analysis of global ecological suitable production area of Ginkgo biloba based on GMPGIS[J].World Chin Med(世界中医药),2017,12:969-973.
[11]Jiang XY,Sun HB,Qin JH,et al.Study on the functional zoning of Fritillaria cirrhosa based on growth suitability and quality suitability[J].China J Chin Mater Med(中国中药杂志),2016,41:3194-3201.
[12]Zheng SH,Jiang X,Huang LF,et al.Chemical and genetic dis‐crimination of Cistanches Herba based on UPLCQTOF/MS and DNA barcoding[J].PLoS One,2014,9:e98061.
[13]Lin L,Josef BA,Bing L,et al.Three-dimensional evaluation on ecotypic diversity of traditional Chinese medicine:a case study of Artemisia annua L.[J].Front Plant Sci,2017,8:1225.
[14]Li L,Zheng SH,Brinckmann JA,et al.Chemical and genetic diversity of Astragalus mongholicus grown in different ecoclimatic regions[J].PLoS One,2017,12:e0184791.
[15]Li L,Zheng SH,Yang QZ,et al.Distinguishing Astragalus mongholicus and its planting soil samples from different regions by ICP-AES[J].Molecules.2016,21:482.
[16]Wang ZH.Study on the Quality and Origin Variation of American Ginseng and Screening of EGFR Kinase Inhibitors for 49 Medicinal Extracts(西洋参品质和产地变异研究及49种药材提取物EGFR激酶抑制剂筛选)[D].Beijing:Peking Union Medical College,2015.
[17]Yang JL,Gao LL,Zhu P.Advances in the biosynthesis research of ginsenosides[J].Acta Pharm Sin(药学学报),2013,48:170-178.
[18]He DT,Wang B,Chen JM.Research progress on pharmacological effects of ginsenoside[J].J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2012,14:118-121.
[19]Christensen JC.Ginsenosides chemistry,biosynthesis,analysis,and potential health effects[J].Adv Food Nutr Res,2009,55:1-99.
[20]Wu Q,Sun C,Chen SL.Identification and expression analysis of a 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from American ginseng[J].Plant OMICS:J Plant Mol Biol Omics,2012,5:414-420.
[21]Kim OT,Bang KH,Jung SJ,et al.Molecular characterization of ginseng farnesyl diphosphate synthase gene and its up-regulation by methyl jasmonate[J].Biol Plant,2010,54:47-53.
[22]Lee MH,Jeong JH,Seo JW,et al.Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene[J].Plant Cell Physiol,2004,45:976-984.
[23]Han JY,In JG,Kwon YS,et al.Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxi‐dase gene in Panax ginseng[J].Phytochemistry,2010,71:36-46.
[24]Lee MH,Han JY,Kim HJ,et al.Dammarenediol-II production confers TMV Tolerance in transgenic tobacco expressing Panax ginseng dammarenediol-II synthase[J].Plant Cell Physiol,2012,53:173-182.
[25]Yue CJ,Zhong JJ.Purification and characterization of UDPG:ginsenoside Rd glucosyltransferase from suspended cells of Panax notoginseng[J].Proc Biochem,2005,40:3742-3748.
[26]Xing N,Liu LL,Yin SG,et al.The association between property of Chinese medicinal herbs and chemical components,ecologi‐cal environment[J].J Chin Med Mater(中药材),2011,34:324-325.