云南烤烟中新颖的酚类化合物及其抗烟草花叶病毒活性研究

The secondary metabolites play an important role for the plant to defense the stimulus of environment. The tobacco plant would make a defense against the virus and generated the antiviral secondary metabolites when they inffected with the tobacco mosaic virus. In previous work, we disclose that the artificially infecting the tobacco mosaic virus to the tobacco plant can cause the plant to generate more antiviral phenolic compounds. The types and content of phenolic compounds in antiviral plant are high than those of in the less resistance plants. Based on this, to stimulate the tobacco generating more antiviral phenolic compounds, the artificially infecting virus to flue-cure tobacco of different varieties will be carried out at difference place. The representative samples will be selected out form the antiviral plant of these samples by HPLC analysis and the bioactivity screening of their extract. The chemical constitutions of the slected samples will be carefully studied by modern separation technology. As a result, more than thirty new phenolic compounds (incloding 3-5 compounds with new skeleton) were be isolated from these samples.The anti-tobacco mosaic virus activities of the isolated compounds will be carefully screened, and 3-5 compounds with notable antiviral activities will be selected out. These will provide the leading compounds for the development of efficient and safe virus inhibitor to plants diseases. Furthermore, the antiviral mechanism for some compounds with high activities will be studied; the mechanisms of antiviral activities, the relationships of phenolic structures and antiviral activities, the biosynthetic pathway of the compounds will be discussed; and the defense mechanism of tobacco plant under the forceof tobacco mosaic virus will also be clarify in this project.

次生代谢产物在植物防御外界环境刺激过程中发挥重要作用，当烟草受到花叶病毒浸染时会对病毒做出防御，产生抗病毒活性的次生代谢产物。我们前期研究发现烟草抗花叶病毒植株酚类化合物的种类和含量都比不抗病毒植株高，可通过人工浸染病毒的方法诱导烟草产生更多的抗花叶病毒活性酚类化合物。基于这一发现，本项目将对不同产地、品种的云南烤烟进行人工浸染病毒诱导，刺激烟草植株产生抗病毒活性的酚类化合物；再通过HPLC分析和提取物活性筛选的方法从中选出代表性样品，用现代分离技术对样品中抗病毒活性酚类化合物进行深入研究。预期将从3个选定样品中发现30个以上的新酚类化合物，包括3-5个新骨架，并筛选出3-5活性显著的化合物结构，为高效、安全的植物病毒抑制剂开发提供先导化合物。同时，项目还将对部分活性好的化合物进行抗病毒机理研究，探讨其构效关系和生源途径，并初步阐明烟草在花叶病毒胁迫下的防御机制。

项目组紧扣计划任务书要求，用现代分离技术对样品中抗病毒活性酚类化合物进行深入研究。共采集了15个不同产地、品种的样品，其中包括8个花叶病田间浸染病毒实验中筛选出的高抗病毒植株样品。通过植物化学分离的方法，对样品中的酚类化合物进行了分离鉴定。此外，扩展性的对5个香料烟、地方晾晒烟样品，以及其它植物样品的抗烟草花叶病毒活性成分进行了研究。项目组总共分离到酚类化合物263个，包括106个新酚类化合物，其中6个为新骨架分子；所有化合物均通过核磁共振、质谱、红外、紫外光谱等波谱技术对结构进行准确鉴定。项目组还完成了148个化合物的抗烟草花叶病毒活性筛选，发现了10个对烟草花叶病毒抑制率显著超过对照(宁南霉素)的化合物结构，并选择活性好的化合物对其抗病毒机理进行了研究。项目成果共在SCI收录期刊发表研究论文43篇(两篇论文在SCI一区期刊发表)，所有发表论文均标准了基金资助号；还获得了29件发明专利授权。超额完成了计划任务书规定的任务。