木瓜抗烟草花叶病毒活性及其有效成分分离

doi:10.3864/j.issn.0578-1752.2013.17.005

摘要/Abstract

摘要： 【目的】测定木瓜（Chaenomeles sinensis）抗烟草花叶病毒（Tobacco mosaic virus，TMV）的活性，并对其中的有效成分进行研究，为发现新的植物源抗病毒物质提供依据。【方法】以心叶烟（Nicotiana glutinosa）和普通烟（N. tabacum）为材料，结合活性追踪法，对木瓜中抗TMV成分进行分离和结构鉴定，并采用半叶枯斑法和室内盆栽试验对粗提物及单体化合物抗TMV的活性进行测定。【结果】木瓜乙醇提取物对TMV表现出较强的钝化作用（10 mg干样/mL的抑制率达97.76%），对TMV系统侵染具有预防作用（10 mg干样/mL的防效为49.98%）；乙醇浸膏的乙酸乙酯、正丁醇和水萃取物对TMV具较强的体外钝化活性，其抑制率分别为89.02%、82.84%和84.74%；从木瓜乙醇提取物的乙酸乙酯萃取相中分离得到邻苯二甲酸二戊酯（WZ-31，Dipentyl phthalate）、β-香树素（WZ-32，β-Amyrin）、（8E）-十六烯酸（L3，8E-hexadecenoic acid）和（8E）-十九烯酸（L4，8E-nonadecenoic acid）共4个化合物；β-香树素、（8E）-十六烯酸和（8E）-十九烯酸在浓度为1.0 mg•mL-1时对TMV表现出较好的体外钝化活性，其抑制率分别为27.51%、33.33%和41.08%，相当于或显著高于1.0 mg•mL-1 20%盐酸吗啉胍•乙铜可湿性粉剂（MCA）。【结论】木瓜提取物具有较好的抗TMV活性，具备进一步开发和应用的潜力，β-香树素、（8E）-十六烯酸和（8E）-十九烯酸是其抗TMV有效成分。

关键词: 木瓜 , 抗TMV活性成分 , β-香树素 , （8E）-十六烯酸 , （8E）-十九烯酸

Abstract: 【Objective】In order to find new botanical antiviral compounds, antiviral ingredients of Chaenomeles sinensis (Thouin) Koehne was isolated and evaluated against Tobacco mosaic virus (TMV). 【Method】Anti-TMV ingredients were isolated with bioassay-guided fractionation methods. Bioassay against TMV infection and replication of raw extracts and monomeric compound were determined using local lesion and potted plant methods on Nicotiana glutinosa and N. tabacum. 【Result】 Ethanol extract of C. sinensis displayed a significant inactivation effect on TMV in N. glutinosa with inhibition ratio of 97.76% and protection effect on TMV infection on N. tabacum var. K326 with efficiency of 49.98% at the concentration of 10 mg•mL-1. Ethyl acetate extract, n-butyl alcohol extract and water extract from ethanol extractum of C. sinensis all inactivated significantly TMV and the inhibition ratios were 89.02%, 82.84% and 84.74%. Dipentyl phthalate, β-amyrin, (8E)-hexadecenoic acid and (8E)-nonadecenoic acid were isolated and identified through comparison with data obtained via spectroscopic methods, and from values reported in the literature. Among them, β-Amyrin, (8E)-hexadecenoic acid and (8E)-nonadecenoic acid showed similar or better anti-MTV activities than 20% moroxydine hydrochloride•copper acetate WP, and the inhibition ratios were 27.51%, 33.33% and 41.08% at the concentration of 1.0 mg•mL-1. 【Conclusion】C. sinensis can be used as a biological antiviral agent in tobacco production and β-amyrin, (8E)-hexadecenoic acid and (8E)-nonadecenoic acid are its antiviral ingredients.

Key words: Chaenomeles sinensis , anti-TMV ingredients , β-amyrin , (8E)-hexadecenoic acid , (8E)-nonadecenoic acid

梁红艳1, 戎谞1, 陈丕庭1, 马志卿12, 张兴12. 木瓜抗烟草花叶病毒活性及其有效成分分离[J]. 中国农业科学, 2013, 46(17): 3571-3579.

LIANG Hong-Yan-1, RONG Xu-1, CHEN Pi-Ting-1, MA Zhi-Qing-12, ZHANG Xing-12. Anti-TMV Activity and Isolation of Active Ingredients in Fruit from Chaenomeles sinensis[J]. Scientia Agricultura Sinica, 2013, 46(17): 3571-3579.

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参考文献

[1]Bos L. Crop losses caused by viruses. Crop Protection, 1982, 1(3): 263-282.

[2]Ritzenthaler C. Resistance to plant viruses: old issue, new answers? Current Opinion in Biotechnology, 2005, 16: 118-122.

[3]翟熙伦, 杨金广, 申莉莉, 钱玉梅, 王盼, 孙丽萍, 王惠卿, 赵洪东, 王永, 王凤龙. 一株对TMV和PVY具有拮抗活性生防菌的筛选与鉴定.中国农业科学, 2012, 45(11): 2180-2188.

Zhai X L, Yang J G, Shen L L, Qian Y M, Wang P, Sun L P, Wang H Q, Zhao H D, Wang Y, Wang F L. Selection and identification of a biocontrol bacteria strain with inhibitory activity against TMV and PVY. Scientia Agricultura Sinica, 2012, 45(11): 2180-2188. (in Chinese)

[4]Verma H N, Baranwal V K, Srivastava S. Antiviral substances of plant origin//Hadidi A, Khetarpal R K, Koganezawa H. Plant Virus Disease Control. St Paul, MN, USA: APS Press, 1998: 154-162.

[5]Chen J, Yan X H, Dong J H, Sang P, Fang X, Di Y T, Zhang Z K, Hao X J. Tobacco mosaic virus (TMV) inhibitors from Picrasma quassioides Benn. Journal of Agricultural and Food Chemistry, 2009, 57(15): 6590-6595.

[6]Sancheti S, Sancheti S, Bafna M, Seo S Y. Antihyperglycemic, antihyperlipidemic, and antioxidant effects of Chaenomeles sinensis fruit extract in streptozotocin-induced diabetic rats. European Food Research and Technology, 2010, 231: 415-421.

[7]Gao H, Wu L, Kuroyanagi M, Harada K, Kawahara N, Nakane T, Umehara K, Hirasawa A, Nakamura Y. Antitumor-promoting constituents from Chaenomeles sinensis KOEHNE and their activities in JB6 mouse epidermal cells. Chemical & Pharmaceutical Bulletin (Tokyo), 2003, 51(11): 1318-1321.

[8]Oku H, Ueda Y, Ishiguro K. Antipruritic effects of the fruits of Chaenomeles sinensis. Biological & Pharmaceutical Bulletin, 2003, 26(7): 1031-1034.

[9]Sawai R, Kuroda K, Shibata T, Gomyou R, Osawa K, Shimizu K. Anti-influenza virus activity of Chaenomeles sinensis. Journal of Ethnopharmacology, 2008, 118(1): 108-112.

[10]刘厚佳, 胡晋红, 孙莲娜, 蔡溱, 石晶, 刘涛. 木瓜中齐墩果酸抗乙型肝炎病毒研究. 解放军药学学报, 2002, 18(5): 272-274.

Liu H J, Hu J H, Sun L N, Cai Q, Shi J, Liu T. Inhibition effects of oleanolic acid from Chaenomeles lagenaria on Hepatitis B virus in vitro. Pharmaceutical Journal of Chinese People’s Liberation Army, 2002, 18(5): 272-274. (in Chinese)

[11]Kostova I, Iossifova T. Chemical components of Fraxinus species. Fitoterapia, 2007, 78(1): 85-106.

[12]王学贵, 沈丽淘, 田永清, 徐汉虹. 岩木瓜化学成分的分离、结构鉴定与杀虫活性. 昆虫学报, 2010, 53(5): 525-530.

Wang X G, Shen L T, Tian Y Q, Xu H H. Isolation, structure identification and insecticidal activity of compounds from Ficus tsiangii Merr. ex Corner. Acta Entomologica Sinica, 2010, 53(5): 525-530. (in Chinese)

[13]Jing B N, Ma Z Q, Feng J T, Liang H Y, Li C, Zhang X. Evaluation of the antiviral activity of extracts from plants grown in the Qinling Region of China against infection by Tobacco mosaic virus (TMV). Journal of Phytopathology, 2012, 160(4): 181-186.

[14]Gooding G V, Hebert T T. A simple technique for purification of tobacco mosaic virus in large quantities. Phytopathology, 1967, 57(11): 1285.

[15]田波, 裴美云. 植物病毒研究方法. 北京: 科学出版社, 1987.

Tian B, Pei M Y. Research Technique of Plant Virus. Beijing: Science Press, 1987. (in Chinese)

[16]吴惠勤, 朱志鑫, 黄晓兰, 林晓珊, 黄芳, 马叶芬, 罗辉泰, 邓欣, 潘亮君. 不同类别食品中21种邻苯二甲酸酯的气相色谱-质谱测定及其分布情况研究. 分析测试学报, 2011, 30(10): 1079-1087.

Wu H Q, Zhu Z X, Huang X L, Lin X S, Huang F, Ma Y F, Luo H T, Deng X, Pan L J. Study on determination of 21 phthalate acid esters and their distributions in different types of foods by gas chromatography-mass spectrometry. Journal of Instrumental Analysis, 2011, 30(10): 1079-1087. (in Chinese)

[17]Anjaneyulu V, Babu J S, Babu B H, Reddy M V. Terpenoids from a mangrove, Excoecaria agallocha. Acta Ciencia Indica Chemistry, 1993, 19(4): 123-126.

[18]Chandler R F, Chung A P K, Safe L M, Hooper S N. Constituents of Furcellaria fastigiata. Canadian Journal of Pharmaceutical Sciences, 1977, 12(4): 92-94.

[19]Busarova N G, Isai S V, Revtsova I Y. Lipids from the marine worm Urechis unicinctus. Chemistry of Natural Compounds, 2011, 47(4): 515-518.

[20]张冬松, 高慧媛, 吴立军. 光皮木瓜的化学成分药理活性及临床研究进展. 沈阳药科大学学报, 2007, 24(11): 721-726.

Zhang D S, Gao H Y, Wu L J. Advances in the study of Chaenomeles sinensis. Journal of Shenyang Pharmaceutical University, 2007, 24(11): 721-726. (in Chinese)

[21]席忠新, 王燕, 赵贵钧, 李霞, 吴志军, 孙连娜. 鼠曲草石油醚部位的化学成分. 第二军医大学学报, 2011, 32(3): 311-313.

Xi Z X, Wang Y, Zhao G J, Li X, Wu Z J, Sun L N. Chemical constituents of petroleum ether fractions of Gnaphalium affine D. Don. Academic Journal of Second Military Medical University, 2011, 32(3): 311-313. (in Chinese)

[22]白玉华, 于辉, 常乃丹, 安东政義. 日本苦苣菜的化学成分. 中国药科大学学报, 2008, 39(3): 279- 281.

Bai Y H, Yu H, Chang N D, An D Z X. Chemical constituents of Japanese herb Sonchus oleraceus. Journal of China Pharmaceutical University, 2008, 39(3): 279-281. (in Chinese)

[23]刘慧灵, 马国需, 杨峻山, 许旭东. 苦石莲的抗肿瘤化学成分研究. 中草药, 2012, 43(10): 1901-1904.

Liu H L, Ma G X, Yang J S, Xu X D. Antitumor constituents from seeds of Caesalpinia minax. Chinese Traditional and Herbal Drugs, 2012, 43(10): 1901-1904. (in Chinese)

[24]Vitor C E, Figueiredo C P, Hara D B, Bento A F, Mazzuco T L, Calixto J B. Therapeutic action and underlying mechanisms of a combination of two pentacyclic triterpenes, α- and β-amyrin, in a mouse model of colitis. British Journal of Pharmacology, 2009, 157(6): 1034-1044.

[25]Chen J C, Ho T Y, Chang Y S, Wu S L, Li C C, Hsiang C Y. Identification of Escherichia coli enterotoxin inhibitors from traditional medicinal herbs by in silico, in vitro, and in vivo analyses. Journal of Ethnopharmacology, 2009, 121(3): 372-378.

[26]陈庆园, 胡玉录, 王发鹏. 抑制烟草病毒病植物源活性物质的研究进展. 中国烟草科学, 2006(3): 16-19.

Chen Q Y, Hu Y L, Wang F P. Research advances in plant source active substances of inhibition tobacco virus diseases. Chinese Tobacco Science, 2006(3): 16-19. (in Chinese)

[27] 张冰涛, 赵婷, 邹艳敏, 邹烨, 郑大恒, 曲洪媛, 李峰, 仰榴青. 玫瑰茄花萼中脂肪酸成分的GC-MS分析. 江苏大学学报: 医学版, 2012, 22(2): 147-150.

Zhang B T, Zhao T, Zou Y M, Zou Y, Zhen D H, Qu H Y, Li F, Yang L Q. Analysis of fatty acids from Hibiscus sabdariffa L. calyx by GC-MS. Journal of Jiangsu University: Medicine Edition, 2012, 22(2): 147-150. (in Chinese)

[28]祝红, 祝玲, 叶曼红. 番木瓜种子油提取方法及成分的研究. 中药材, 2007, 30(7): 857-860.

Zhu H, Zhu L, Ye M H. Study on the extraction method and constituents of papaw seed oil. Journal of Chinese Medicinal Materials, 2007, 30(7): 857-860. (in Chinese)

[29]沈建国, 张正坤, 吴祖建, 谢联辉, 林奇英. 臭椿抗烟草花叶病毒活性物质的提取及其初步分离. 中国生物防治, 2007, 23(4): 348-352.

Shen J G, Zhang Z K, Wu Z J, Xie L H, Lin Q Y. Extraction and preliminary isolation of antiviral substances from Ailanthus altissima against TMV. Chinese Journal of Biological Control, 2007, 23(4): 348-352. (in Chinese)

[30]刘国坤, 谢联辉, 林奇英, 吴祖建，陈启建. 15种植物的单宁提取物对TMV的抑制作用. 植物病理学报, 2003, 3(33): 279-283.

Liu G K, Xie L H, Lin Q Y, Wu Z J, Chen Q J. Inhibition of the infectivity of TMV by tannins. Acta Phytopathologica Sinica, 2003, 3(33): 279-283. (in Chinese)

[31]马志卿, 李威, 王海鹏, 张兴. 植物源病毒抑制剂VFB对烟草抗烟草花叶病毒的诱导作用研究. 植物病理学报, 2010, 40(4): 419-425.

Ma Z Q, Li W, Wang H P, Zhang X. Induced resistance to tobacco mosaic virus (TMV) on tobacco by botanical-derived VFB. Acta Phytopathologica Sinica, 2010, 40(4): 419-425. (in Chinese)