Ethylene signalling is essential for the resistance of Nicotiana attenuata against Alternaria alternata and phytoalexin scopoletin biosynthesis

The phytohormone ethylene plays an important role in plant defence responses to pathogen attack. When infected by the necrotrophic fungal pathogen Alternaria alternata (tobacco pathotype), which causes severe diseases in Nicotiana species, the wild tobacco plant Nicotiana attenuata accumulates a high amount of the jasmonate (JA)‐dependent phytoalexin scopoletin to defend itself against this fungal pathogen. However, it is still not known whether ethylene signalling is also involved in scopoletin biosynthesis and the resistance of N. attenuata. After infection, ethylene biosynthetic genes were highly elicited. Furthermore, plants strongly impaired in ethylene biosynthesis or perception had dramatically decreased scopoletin levels, and these plants became more susceptible to the fungus, while A. alternata‐elicited JA levels were increased, indicating that the decreased defence responses were not due to lower JA levels. Thus, it is concluded that after infection, ethylene signalling is activated together with JA signalling in N. attenuata plants and this subsequently regulates scopoletin biosynthesis and plant resistance.     

东莨菪内酯在水中的光解特性研究

实验室条件下,采用高效液相色谱法研究了东莨菪内酯在水溶液中的光解特性以及初始质量浓度、光源、pH值和温度等因素对其光解的影响。结果表明,东莨菪内酯在水溶液中的光解均符合一级动力学方程。在初始质量浓度为5~40 mg/L范围内,其光解速率随初始质量浓度的增大而降低;在不同光源条件下,其光解速率从大到小依次是:500 W高压汞灯18 W紫外灯500 W氙灯;在pH值为5~9的缓冲溶液中,其光解半衰期随pH值的升高而降低,pH为9时的光解半衰期最短,为4.81 h;在5~65℃范围内,其光解速率随温度升高呈先升高后降低趋势,于15℃时达到最大值。研究表明,东莨菪内酯在水溶液中的光解与其初始质量浓度、光源、pH值和温度相关。所得结果可为其环境风险评价提供参考。     

氯化钙与东莨菪内酯联用对朱砂叶螨的杀螨毒力及其Ca2+-ATP酶活性的影响

为了明确钙离子与东莨菪内酯联合作用的杀螨效果,进而为东莨菪内酯的开发利用提供参考,采用玻片浸渍法测定了钙离子(Ca2+)与东莨菪内酯混用对朱砂叶螨Tetranychus cinnabarinus雌成螨的杀螨毒力,并测定了活体和离体条件下对螨体内Ca2+-ATP酶活性的影响,对Ca2+的增效作用机理进行了初步分析。结果表明:Ca2+与东莨菪内酯联合使用能显著增强东莨菪内酯的杀螨效果,其中联合作用24和48 h的LC50值分别比东莨菪内酯单独处理降低20%和45%;对朱砂叶螨Ca2+-ATP酶活性的测定结果表明,无论在活体还是离体条件下,Ca2+与东莨菪内酯联用均能显著增强对Ca2+-ATP酶的抑制作用,而相同浓度的Ca2+单独作用则对Ca2+-ATP酶活性无影响,这也在一定程度上证明了Ca2+-ATP酶是东莨菪内酯的重要作用靶标之一。     

瑞香狼毒杀螨活性物质的分离与鉴定

 采用活性跟踪法,对瑞香狼毒（Stellera chamaejasme L.）中的杀螨活性成分进行研究。从瑞香狼毒乙酸乙酯提取物中分离出对朱砂叶螨（Tetranychus cinnabarinus）具有较强触杀和内吸活性的化合物Ⅳ,通过核磁共振（1H NMR,13C NMR）、红外色谱（IR）等方法对化合物Ⅳ进行鉴定,确定该化合物为东莨菪内酯（scopoletin）。杀螨活性的测定结果表明,东莨菪内酯可能是瑞香狼毒中重要的杀螨活性物质。东莨菪内酯对朱砂叶螨触杀活性的半致死浓度（LC50）值为1.267 mg · mL^-1,内吸活性的LC50值为7.388 mg · mL^-1。     

Inhibition of growth ofErwinia carotovora in vitro by phenolics

Summary The phenolic acids caffeic, cinnamic, ferulic, salicylic, sinapic, and vanillic, together with scopoletin and coniferyl alcohol, at concentrations of ≤1 mg/ml, variously inhibited growth ofErwinia carotovora in Nutrient Broth buffered to pH 6 by 0.2 M phosphate buffer. Chlorogenic acid at 1 mg/ml did not inhibit growth but it did so in agar diffusion bioassays at high concentrations (3–10 mg/well) probably by lowering of the pH of the agar.     

东莨菪素对朱砂叶螨的毒力及杀螨机理初探

采用玻片浸渍法测定东莨菪素对朱砂叶螨的触杀毒力,观察东莨菪素对朱砂叶螨的致毒症状,并测定其对朱砂叶螨神经系统靶标酶(乙酰胆碱酯酶、单胺氧化酶、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶)活性的影响。结果表明,东莨菪素对朱砂叶螨24 h的触杀LC50值为1.24 mg/mL;中毒试螨表现出神经毒剂的致毒症状,如兴奋、痉挛等。在活体和离体试验中,东莨菪素均显著抑制试螨乙酰胆碱酯酶、单胺氧化酶、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶的活性。研究还发现东莨菪素会引起试螨乙酰胆碱含量增加,可能是东莨菪素显著抑制试螨乙酰胆碱酯酶活性引起。据此推测东莨菪素对朱砂叶螨可能有神经毒性。     

核黄素诱导烟草悬浮细胞酚类物质和木质素的积累

 【目的】阐明核黄素对烟草悬浮细胞苯丙氨酸解氨酶(phenylalanine ammonia-lyase,PAL)、过氧化物酶(peroxidase,POD)活性以及抗病相关的酚类物质、木质素等次生代谢产物含量的影响。【方法】1 mmol?L-1的核黄素处理烟草悬浮细胞,用生物化学、细胞化学和高效液相色谱(high performance liquid chromatography,HPLC)等方法测定苯丙烷代谢有关的生理和生化指标。【结果】核黄素处理后,PAL和POD活性明显升高,处理12 h 和24 h后酶活性分别达到峰值;荧光观察细胞壁积累了更多的酚类物质,胞内和胞外scopoletin(7-羟基-6-甲氧基香豆素)含量明显升高,在处理12 h后都达到峰值,分别是对照的3.6和3.9倍;细胞染色和含量测定表明,核黄素处理后烟草悬浮细胞沉积了更多的木质素,处理24 h和48 h后,木质素含量分别为对照的1.5和1.8倍。【结论】核黄素处理后提高了烟草悬浮细胞PAL、POD活性,积累了更多与抗病有关的次生代谢产物。     

4CL基因在海巴戟叶片莨菪亭累积过程中的功能研究

为明确海巴戟叶片中莨菪亭累积规律,探究4CL基因在其累积过程中起到的作用。利用高效液相色谱仪对离体的海巴戟成熟叶0～48 h内莨菪亭含量进行测定,并测定经不同浓度的阿魏酸处理后莨菪亭含量的变化。以阿魏酸为底物,利用紫外分光光度计进行酶活反应的4CL总酶活性测定。对RNA-seq结果中7个4CL基因进行qPCR,筛选出一个表达趋势与莨菪亭含量及酶活变化趋势一致的4CL基因作为关键候选基因,进行全长克隆以及生物信息学分析,最后用qRT-PCR研究其表达特性。海巴戟叶片在采摘后48 h内,莨菪亭含量在12 h时最高,达0.35 mg/g,48 h最低,为0.18mg/g。4CL总酶活性变化与莨菪亭含量趋势相近。阿魏酸处理后的叶片中4CL总酶活增加。通过qRT-PCR筛选到4CL(DN15707)基因,克隆、测序后发现其长度为1632 bp,具备完整开放阅读框,在NCBI进行序列比对及系统进化树分析,确定该基因为4CL基因家族中的4CL2基因,命名为Mc4CL2。Mc4CL2基因可能是莨菪亭累积过程中的关键基因,它能启动4CL酶活,充分利用阿魏酸底物进而导致莨菪亭累积。     

Coumarin compounds in cassava diets: 2 health implications of scopoletin in gari

Scopoletin has been isolated and identified in gari, a cassava food consumed in Nigeria (West Africa). Its levels in gari and cassava flour is not altered by post processing treatments such as sundrying, refrigeration and storage. Scopoletin has also been identified as an active principle in the traditional herbal infusion of the fruit ofTetrapleura tetraptera TAUB used in the ethnopharmacology of West Africa. It is a potent hypotensive and non-specific spasmolytic agent. These pharmacological effects of Scopoletin are probably the underlying factors in the slowly developing tropical neuropathy characterised by optic atrophy, nerve deafness and ataxia endemic among populations subsisting on cassava diets such as gari. Hitherto, these toxicities were attributed to cyanogenic glucosides (cyanide) present in cassava.     

马铃薯块茎中的6-羟基-7-甲氧基香豆素的分离和鉴定

用甲醇将６－羟基－７－甲氧基香豆素从马铃薯粉中萃取出来后，采用薄层层析法进行分离和纯化，并且用紫外光谱和质谱对其进行了鉴定。