木瓜抗烟草花叶病毒活性及其有效成分分离

Anti-TMV Activity and Isolation of Active Ingredients in Fruit from Chaenomeles sinensis

【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-120%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。

[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 compoundweredeterminedusinglocal lesionand 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% andprotection effect on TMV infection on N. tabacum var. K326 with e^ciency of 49.98% at the concentration of 10 mg.mL^-1. Ethylacetate 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, （SE）-hexadecenoic acid and （SE）-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, （SE）-hexadecenoic acid and （SE）-nonadecenoic acid showed similar orbetter 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.