两种氮素形态对药用植物三叶青中黄酮类活性成分积累的影响及相关分子机制的初步探讨  

Preliminary Study on the Effects of Two Different Nitrogen Forms on Flavonoids Accumulation and Related Molecular Mechanism in Medicinal Plant Tetrastigma hemsleyanum

作　　者：张强[1,2] 刘一 吴晓敏 束良佐[1,2] 宋运贤 陈楚[1,2] 龙民慧 ZHANG Qiang;LIU Yi;WU Xiao-Min;SHU Liang-Zuo;SONG Yun-Xian;CHEN Chu;LONG Min-Hui(School of Life Sciences/Information College,Huaibei Normal University, Huaibei 235000;Key Laboratory of Plant Resources and Biology of Anhui Province, Huaibei 235000)

机构地区：[1]淮北师范大学生命科学学院/信息学院,淮北235000 [2]资源植物生物学安徽省重点实验室,淮北235000

出　　处：《植物研究》2018年第3期367-376,共10页Bulletin of Botanical Research

基　　金：国家自然科学基金(31500594;31572202);安徽省教育厅重点项目(KJ2014A227;KJ2015A302);安徽高校优秀青年人才支持计划重点项目(gxyq ZD2016415)~~

摘　　要：以珍稀濒危药用植物三叶青(Tetrastigma hemsleyanum Diels et Gilg)为材料,通过遮荫条件下(30%自然光)的盆栽试验,研究了硝态氮(NO_3^--N)和铵态氮(NH_4^+-N)两种不同氮素形态对其根和叶中黄酮类活性成分含量的影响并探讨其机制。结果表明:NO_3^--N处理后的三叶青叶中原花青素B1、芦丁、山柰酚-3-O-芸香糖苷、槲皮素、异槲皮苷、山柰酚的含量比NH_4^+-N处理的高;其根中的原花青素B1、芦丁、山柰酚-3-O-芸香糖苷、槲皮素、异槲皮苷、山柰酚的含量比NH_4^+-N处理的高。进一步的研究发现NO_3^--N处理过的三叶青其根、叶中与黄酮类化合物合成相关的酶苯丙氨酸解氨酶(PAL)、查尔酮合成酶(CHS)转录与表达水平高于NH_4^+-N处理的三叶青。此外,NO_3^--N处理的三叶青叶、根中的PAL和CHS酶活性也高于NH_4^+-N处理。利用PAL特异性抑制剂AOA处理后,AOA能显著抑制PAL酶活性(P<0.05)并导致黄酮类成分含量极显著下降(P<0.01)。综上所述,施用NO_3^--N更有益于三叶青黄酮类化合物的生物合成,其主要是通过增加PAL代谢途径关键酶的基因表达量进而提高其酶活性来增强黄酮类化合物的合成。 The effects of two different nitrogen（N） forms（nitrate-N and ammonium-N） on the accumulation of flavonoids（procyanidin B1,rutin,kaempferol-3-O-rutinoside,quercetin,isoquercitrin and kaempferol） in the endangered medicinal plant Tetrastigma hemsleyanum （from Zhejiang Province） were examined through potted experiment under shady condition （30% natural light）,and its influence mechanism was also discussed.The flavonoids content was relatively higher in the root and leaf of T.hemsleyanum under nitrate-N supply than ammonium-N supply.The levels of transcription and expression of phenylalanine ammonia-lyase （PAL） and chalcone synthase（CHS） which are related to flavonoids synthesis were relatively higher in the root and leaf of T.hemsleyanum under nitrate-N supply than ammonium-N supply.Besides,the enzyme activities of PAL and CHS were also higher in the root and leaf of T.hemsleyanum under nitrate-N supply than ammonium-N supply.The PAL-specific inhibitor（AOA） could significantly inhibit the enzyme activity of PAL（P 〈0.05） and lead to a very significant decrease in flavonoids content（P 〈0.01）.In conclusion,nitrate-N supply is more beneficial to flavonoids synthesis in T.hemsleyanum,which mainly enhances the flavonoids synthesis by increasing the gene expression and activity of key enzymes in PAL metabolic pathway.

关 键 词：氮素形态 氮同化 黄酮类 三叶青 

分 类 号：Q945[生物学—植物学]