Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (23): 4772-4780.doi: 10.3864/j.issn.0578-1752.2011.23.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Effect of Exogenous L-Phentermine and UV-C on the Accumulation of Rutin Compounds and the Expression of Rutin Biosynthesis Genes in Fagopyrum tartaricum

 SUN  Chao-Xia, HOU  Si-Yu, YANG  Wu-De   

  1. 1.山西农业大学旱作农业工程研究所,山西太谷 030801
  • Received:2011-06-03 Online:2011-12-01 Published:2011-08-31

Abstract: 【Objective】 The objective of this study is to investigate the content of rutin and the expression of the related genes in response to L-phentermine (Phe) and UV-C treatment, and to provide molecular information for high rutin content buckwheat breeding. 【Method】 Rutin biosynthesis-related genes were cloned from the leaves of F. tataricum by 3′RACE. Rutin content in leaves under Phe treatment (1, 2 and 4 mg?L-1, 1 to 5 d) was compared with that of untreated buckwheat by high performance liquid chromatography (HPLC). The gene expression level was analyzed by qRT-PCR. 【Result】 Three partial CDS of rutin biosynthesis-related genes were cloned. They were chalcone synthase (FtCHS), flavonol synthase-like (FtFLS-like) and flavone 3-hydroxylase (FtF3H). Seedlings were cultured in solution containing Phe (4 mg×L-1) for 1 d, rutin content was two times more than the control (12.55 mg×gFW-1), reached 26.15 mg×gFW-1. The higher rutin content (17.36 mg×gFW-1) was found in leaves at 3 h under UV-C treatment. Differential expression patterns of FtCHS, FtFLS-like and FtF3H were observed in different Phe concentration treatments in leaves of F. tartaricum. Seedlings were cultured in solution containing Phe (1 mg×L-1) for 4 d, FtCHS expression  level was 4.84 fold higher than the control. The higher expression levels of FtF3H and FtFLS-like were observed after 4 and 2 mg×L-1 Phe treatment for 5 d, they were 1.06 and 3.90 fold higher than the control, respectively. UV-C radiation for 1-3 h, the expression      of FtCHS, FtFLS-like and FtF3H was higher than control. In particular, the expression level of FtFLS-like at 1 h under UV-C radiation was 127.71 fold higher than the control. 【Conclusion】 Rutin content could be enhanced to a great extent by Phe treatment and UV-C radiation, and gene expression patterns of rutin biosysthesis-related genes were regulated by UV-C radiation. It can be used to analyze the pathway of rutin biosynthesis. Further study should be focused on the influential factors of rutin metabolic process.

Key words: Fagopyrum tartaricum, rutin, L-phenylalanine, UV-C, gene expression

[1]陈庆富. 五个中国荞麦 (Fagopyrum) 种的核型分析. 广西植物, 2001, 21(2): 107-110.

Chen Q F. Karyotype analysis of five Fagopyrum species native to China. Guihaia, 2001, 21(2): 107-110. (in Chinese)

[2]刘光德, 李名扬, 祝钦泷, 李艳冬, 眭顺照. 资源植物野生金荞麦的研究进展. 农业资源与环境科学, 2006, 22(10): 380-389.

Liu G D, Li M Y, Zhu Q L, Li Y D, Sui S Z. The research advance on resource plant Fagopyrum dibotrys. Chinese Agricultural Science Bulletin, 2006, 22(10): 380-389. (in Chinese)

[3]Kim S J, Kawaharda C, Suzuki T, Saito K, Hashimoto N, Takigawa S, Noda T, Matsuura-Endo C, Yamauchi H. Effect of natural light periods on rutin, free amino acid and vitamin C contents in the sprouts of common (Fagopyrum esculentum Moench) and tartary (F. tataricum Gaertn) buckwheats. Food Science and Technology Research, 2006, 12(3): 199-205.

[4]Kreft I, Fabjan N, Yasumoto K. Rutin content in buckwheat (Fagopyrum esculentum Moench) food materials and products. Food Chemistry, 2006, 98: 508-512.

[5]Metha R G, Murillo G, Naithani R, Peng X J. Cancer chemoprevention by natural products: how far have we come? Pharmaceutical Research, 2010, 27(16): 950-961.

[6]Jiang P, Burczynski F, Campbell C, Pierce G, Austria J A, Briggs C J. Rutin and flavonoid contents in three buckwheat species Fagopyrum esculentum, F. tataricum, and F. homotropicum and their protective effects against lipid peroxidation. Food Research International, 2007, 40: 356-364.

[7]张美莉, 胡小松. 荞麦生物活性物质及其功能研究进展. 杂粮作物, 2004, 24(1): 26-29.

Zhang M L, Hu X S. Research advance of buckwheat biological active substance and fuction. Rain Fed Crops, 2004, 24(1): 26-29. (in Chinese)

[8]文  平, 陈进红. 荞麦芦丁的研究进展. 中国粮油学报, 2006, 21(3): 107-111.

Wen P, Chen J H. Research progress on rutin in buekwheat. Journal of the Chinese Cereals and Oil Association, 2006, 21(3): 107-111. (in Chinese)

[9]Suzuki T, Honda Y, Mukasa Y. Effects of UV-B radiation, cold and desiccation stress on rutin concentration and rutin glucosidase activity in tartary buckwheat (Fagopyrum tataricum) leaves. Plant Science, 2005, 168: 1303-1307.

[10]Kreft S, Strukelj B, Gaberscik A, Kreft I. Rutin in buckwheat herb grown at different UV-B radiation levlels: comparison of two UV spectrophotometric and HPLC method. Journal of Experimental Botany, 2002, 53(375): 1801-1804.

[11]González-Aguilar G A, Zavaleta-Gatica R, Tiznado-Hernández M E. Improving postharvest quality of mango ‘Haden’ by UV-C treatment. Postharvest Biology and Technology, 2007, 45: 108-116.

[12]程水源, 顾曼如, 束怀瑞. 银杏叶黄酮研究进展. 林业科学, 2000, 36(6): 110-115.

Cheng S Y, Gu M R, Shu H R. Advances in research on flavonoids in ginkgo biloba leaf. Scientia Silvae Sinicae, 2000, 36(6): 110-115. (in Chinese)

[13]吕东平, 赵德修, 黄  艳, 赵  乔. 前体对水母雪莲悬浮培养细胞黄酮合成的影响. 云南植物研究, 2001, 23(4): 497-503.

Lü D P, Zhao D X, Huang Y, Zhao Q. The effect of precursor feeding on flavonoids biosynthesis in cell suspension cultures of Saussurea medusa. Acta Botanica Yunnanica, 2001, 23(4): 497-503. (in Chinese)

[14]Edahiro J I, Nakamura M, Seki M, Furusaki S. Enhanced accumulation of anthocyanin in cultured strawberry cells by repetitive feeding of L-phenylalanine into the medium. Journal of Bioscience and Bioengineering, 2005, 99(1): 43-47.

[15]Li X, Park N I, Xu H, Woo S H, Park C H, Park S U. Differential expression of flavonoid biosynthesis genes and accumulation of phenolic compounds in common buckwheat (Fagopyrum esculentum). Journal of Agricultural Food Chemistry, 2010, 58: 12176-12181.

[16]蒙  华, 李成磊, 吴  琦, 邵继荣, 陈  惠. 金荞麦查尔酮合成酶基因CHS的克隆及序列分析. 草业学报, 2010, 19(3): 162-169.

Meng H, Li C L, Wu Q, Shao J R, Chen H. Cloning and sequence analysis of the chalcone synthase gene (CHS) from Fagopyrum dibotrys. Acta Pratacul Turae Sinica, 2010, 19(3): 162-169. (in Chinese)

[17]马  婧, 祝钦泷, 郭铁英, 刘光德, 眭顺照, 李名扬. 金荞麦MYB转录因子基因FdMYBP1的克隆及分子特征分析. 中国中药杂志, 2009, 34(17): 2155-2159.

Ma J, Zhu Q L, Guo T Y, Liu G D, Sui S Z, Li M Y. Cloning and molecular characteristics analysis of a MYB gene from Fagopyrum dibotrys. China Journal of Chinese Materia Medica, 2009, 34(17): 2155-2159. (in Chinese)

[18]刘光德, 雷兴华, 祝钦泷, 钟光驰, 郭铁英, 李艳冬, 眭顺照, 李名扬. 金荞麦二氢黄酮醇4-还原酶基因 (FdDFR1) 的克隆及序列分析. 中国农业科学, 2009, 42(1): 55-63.

Liu G D, Lei X H, Zhu Q L, Zhong G C, Guo T Y, Li Y D, Sui S Z, Li M Y. Cloning and sequence analysis of a DFR gene from Fagopyrum dibotrys (D. Don) Hara. Scientia Agricultura Sinica, 2009, 42(1): 55-63. (in Chinese)

[19]王  岚, 李玉英, 蔡桂红, 张  政, 王转花. 重组苦荞麦过敏蛋白TBa的原核表达及其免疫活性鉴定. 中国生物化学与分子生物学报, 2006, 22(4): 308-312.

Wang L, Li Y Y, Cai G H, Zhang Z, Wang Z H. Prokaryotic expression and immunological identification of tartary buckwheat allergenic protein (TBa). Chinese Journal of Biochemistry and Molecular Biology, 2006, 22(4): 308-312. (in Chinese)

[20]高  丽, 李玉英, 张  政, 王转花, 王宏伟, 张  丽, 朱  镭. 重组荞麦胰蛋白酶抑制剂对HL-60细胞的促凋亡作用. 中国实验血液学杂志, 2007, 15(1): 59-62.

Gao L, Li Y Y, Zhang Z, Wang Z H, Wang H W, Zhang L, Zhu L. Apoptosis of HL-60 cells induced by recombinant common buckwheat trypsin inhibitor. Journal of Experimental Hematology, 2007, 15(1): 59-62. (in Chinese)

[21]Winkel-Shirley B. Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiology, 2001, 126: 485-493.

[22]侯思宇, 张  锐, 郭三堆. 棉花育性相关基因GhPG2的克隆与表达分析. 作物学报, 2009, 35(11): 2008-2014.

Hou S Y, Zhang R, Guo S D. Cloning and expression profile analysis of GhPG2 gene associated with fertility in cotton (Gossypium hirsutum L.). Acta Agronomica Sinica, 2009, 35(11): 2008-2014. (in Chinese)

[23]邹  勇, 尹礼国, 贾雪峰, 钟  耕. HPLC法测定苦荞叶中芦丁的含量. 粮油食品科技, 2007, 15(3): 57-58.

Zou Y, Yin L G, Jia X F, Zhong G. Determination of rutin in buckwheat leaves by HPLC. Science and Technology of Cereals, Oils and Foods, 2007, 15(3): 57-58. (in Chinese)

[24]杨  英, 何  峰, 季家兴, 雷  晶, 陈雪红, 余龙江. 四种前体对胀果甘草细胞悬浮培养生产甘草黄酮的调控效果评价. 武汉植物学研究, 2007, 25(5): 484-489.

Yang Y, He F, Ji J X, Lei J, Chen X H, Yu L J. The effect of precursor feeding on flavonoids biosynthesis in cell suspension cultures of Glycyrrhiza inflata Bat.. Journal of Wuhan Botanical Research, 2007, 25(5): 484-489. (in Chinese)

[25]Masoumian M, Arbakariya A, Syahida A, Maziah M. Effect of precursors on flavonoid production by Hydrocotyle bonariensis callus tissues. African Journal of Biotechnology, 2011, 10(32): 6021-6029.

[26]Cominelli E, Gusmaroli G, Allegra D, Galbiati M, Wade H K, Jenkins G I, Tonelli C. Expression analysis of anthocyanin regulatory genes in response to different light qualities in Arabidopsis thaliana. Journal of Plant Physiology, 2008, 165: 886-894.

[27]Maccarrone M, Melino G, Finazzi-Agrò A. Lipoxygenases and their involvement in programmed cell death. Cell Death and Differentiation, 2001, 8: 776-784.
[1] ZHANG KeKun,CHEN KeQin,LI WanPing,QIAO HaoRong,ZHANG JunXia,LIU FengZhi,FANG YuLin,WANG HaiBo. Effects of Irrigation Amount on Berry Development and Aroma Components Accumulation of Shine Muscat Grape in Root-Restricted Cultivation [J]. Scientia Agricultura Sinica, 2023, 56(1): 129-143.
[2] GU LiDan,LIU Yang,LI FangXiang,CHENG WeiNing. Cloning of Small Heat Shock Protein Gene Hsp21.9 in Sitodiplosis mosellana and Its Expression Characteristics During Diapause and Under Temperature Stresses [J]. Scientia Agricultura Sinica, 2023, 56(1): 79-89.
[3] LAI ChunWang, ZHOU XiaoJuan, CHEN Yan, LIU MengYu, XUE XiaoDong, XIAO XueChen, LIN WenZhong, LAI ZhongXiong, LIN YuLing. Identification of Ethylene Synthesis Pathway Genes in Longan and Its Response to ACC Treatment [J]. Scientia Agricultura Sinica, 2022, 55(3): 558-574.
[4] SHU JingTing,SHAN YanJu,JI GaiGe,ZHANG Ming,TU YunJie,LIU YiFan,JU XiaoJun,SHENG ZhongWei,TANG YanFei,LI Hua,ZOU JianMin. Relationship Between Expression Levels of Guangxi Partridge Chicken m6A Methyltransferase Genes, Myofiber Types and Myogenic Differentiation [J]. Scientia Agricultura Sinica, 2022, 55(3): 589-601.
[5] GUO ShaoLei,XU JianLan,WANG XiaoJun,SU ZiWen,ZHANG BinBin,MA RuiJuan,YU MingLiang. Genome-Wide Identification and Expression Analysis of XTH Gene Family in Peach Fruit During Storage [J]. Scientia Agricultura Sinica, 2022, 55(23): 4702-4716.
[6] KANG Chen,ZHAO XueFang,LI YaDong,TIAN ZheJuan,WANG Peng,WU ZhiMing. Genome-Wide Identification and Analysis of CC-NBS-LRR Family in Response to Downy Mildew and Powdery Mildew in Cucumis sativus [J]. Scientia Agricultura Sinica, 2022, 55(19): 3751-3766.
[7] YuXia WEN,Jian ZHANG,Qin WANG,Jing WANG,YueHong PEI,ShaoRui TIAN,GuangJin FAN,XiaoZhou MA,XianChao SUN. Cloning, Expression and Anti-TMV Function Analysis of Nicotiana benthamiana NbMBF1c [J]. Scientia Agricultura Sinica, 2022, 55(18): 3543-3555.
[8] JIN MengJiao,LIU Bo,WANG KangKang,ZHANG GuangZhong,QIAN WanQiang,WAN FangHao. Light Energy Utilization and Response of Chlorophyll Synthesis Under Different Light Intensities in Mikania micrantha [J]. Scientia Agricultura Sinica, 2022, 55(12): 2347-2359.
[9] YUAN JingLi,ZHENG HongLi,LIANG XianLi,MEI Jun,YU DongLiang,SUN YuQiang,KE LiPing. Influence of Anthocyanin Biosynthesis on Leaf and Fiber Color of Gossypium hirsutum L. [J]. Scientia Agricultura Sinica, 2021, 54(9): 1846-1855.
[10] SHU JingTing,JI GaiGe,SHAN YanJu,ZHANG Ming,JU XiaoJun,LIU YiFan,TU YunJie,SHENG ZhongWei,TANG YanFei,JIANG HuaLian,ZOU JianMin. Expression Analysis of IGF1-PI3K-Akt-Dependent Pathway Genes in Skeletal Muscle and Liver Tissue of Yellow Feather Broilers [J]. Scientia Agricultura Sinica, 2021, 54(9): 2027-2038.
[11] ZHAO Ke,ZHENG Lin,DU MeiXia,LONG JunHong,HE YongRui,CHEN ShanChun,ZOU XiuPing. Response Characteristics of Plant SAR and Its Signaling Gene CsSABP2 to Huanglongbing Infection in Citrus [J]. Scientia Agricultura Sinica, 2021, 54(8): 1638-1652.
[12] ZHAO Le,YANG HaiLi,LI JiaLu,YANG YongHeng,ZHANG Rong,CHENG WenQiang,CHENG Lei,ZHAO YongJu. Expression Patterns of TETs and Programmed Cell Death Related Genes in Oviduct and Uterus of Early Pregnancy Goats [J]. Scientia Agricultura Sinica, 2021, 54(4): 845-854.
[13] ZHU FangFang,DONG YaHui,REN ZhenZhen,WANG ZhiYong,SU HuiHui,KU LiXia,CHEN YanHui. Over-expression of ZmIBH1-1 to Improve Drought Resistance in Maize Seedlings [J]. Scientia Agricultura Sinica, 2021, 54(21): 4500-4513.
[14] YUE YingXiao,HE JinGang,ZHAO JiangLi,YAN ZiRu,CHENG YuDou,WU XiaoQi,WANG YongXia,GUAN JunFeng. Comparison Analysis on Volatile Compound and Related Gene Expression in Yali Pear During Cellar and Cold Storage Condition [J]. Scientia Agricultura Sinica, 2021, 54(21): 4635-4649.
[15] LIU ChangYun,LI XinYu,TIAN ShaoRui,WANG Jing,PEI YueHong,MA XiaoZhou,FAN GuangJin,WANG DaiBin,SUN XianChao. Cloning, Expression and Anti-Virus Function Analysis of Solanum lycopersicum SlN-like [J]. Scientia Agricultura Sinica, 2021, 54(20): 4348-4357.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!