长角血蜱性信息素的主要成分及其功能

发布时间：2018-03-16 08:59

本文选题：长角血蜱　切入点：性信息素　出处：《河北师范大学》2007年硕士论文　论文类型：学位论文

【摘要】： 蜱类为专性吸血的外寄生动物,是节肢动物中的一个特殊类群,传播多种人、畜和野生动物疾病,对人类健康和畜牧业造成极大危害。蜱类性信息素由吸血后雌蜱释放,刺激并吸引性活动的雄蜱,从而完成寻找配偶和交配过程,对生殖起关键作用。因此,对蜱类性信息素的研究对于控制其种群数量,减小危害有重要意义。本研究以广泛分布于我国的长角血蜱(Haemaphysalis longicornis Neumann)为研究对象,运用固相萃取、薄层层析、气相色谱和气相色谱-质谱联用等现代生物学及交叉学科的研究方法对多种收集方法得到的性信息素样品进行分析,鉴定其中主要化学物质的种类和含量,并通过生物测定确定其活性成分和生物学作用。本研究深入阐明长角血蜱性信息素的主要成分和生物学功能,对于蜱类防治有重要理论和实际应用意义。主要结果如下: 长角血蜱交配期雄蜱对气流收集法、腺体浸提法和整蜱浸提法三种方法得到的样品均有一系列大致相同的反应:开始是第一对附肢在空中挥舞表现出积极的寻找行为;在15min内多数雄蜱会陆续停止爬行并停留在一定区域;最后将附肢收拢在躯体下面,保持静止不动的状态。气流收集法得到的样品对雄蜱吸引率最高。气流收集法、腺体浸提法和整蜱浸提法得到的性信息素样品对交配期雄蜱吸引率依次为:88㳠、76㳠和59㳠。性信息素样品提取方法的比较研究发现,不同收集方法样品成分种类和含量均由较大差异。气流收集法对保留时间较小的物质收集效率极高。这类物质挥发性很强,并且在结构上有共同点,均携带有数目不等的氯原子(六氯乙烷、五氯丙烷、八氯丙烷、六氯丙烷和七氯丙烷)。这类物质在长角血蜱中的生物学作用尚未确定。腺体收集法能够增加性信息素的提取量,在样品中检测到微量2,6-二氯酚(2,6-DCP)和含量较大的胆甾醇。整蜱浸提法对于保留时间小于12min的物质提取效率很低,而在14min-21min色谱峰比较集中,这种方法能够较好提取长角血蜱体表物质,如几种不同的长链脂肪酸和胆甾醇酯类等。本研究采用气相色谱-质谱联用方法在长角血蜱腺体提取物中鉴定出微量2,6-DCP,为吸引性信息素主要成分。气相色谱分析长角血蜱交配期雌蜱挥发物中可能存在另外一种结构类似的物质——2,4-DCP,与2,6-DCP的比例为1∶8.74,在生物测定中对交配期雄蜱吸引率达到62㳠,与2,6-DCP并无显著差异,2,4-DCP在长角血蜱交配中也有重要作用。本研究通过气相色谱-质谱联用方法鉴定出长角血蜱交配期雌蜱性信息素样品中存在胆甾醇、胆甾醇油酸酯和胆甾二醇乙酸酯。通过体表点滴法进行生物测定,发现胆甾醇、胆甾醇油酸酯和胆甾二醇二乙酸酯类似物胆甾醇乙酸酯三种标准样品均可使长角血蜱交配成功率显著恢复,可确定胆甾醇和胆甾醇油酸酯为长角血蜱MSP主要成分。本研究还首次在长角血蜱交配期雌蜱样品中发现长链饱和脂肪酸——十四烷酸、十八烷酸,这两种物质是长角血蜱GSP可能成分。
[Abstract]:Foreign animal parasitic ticks, is a special group of arthropod animal in the spread of a variety of people, livestock and wild animal diseases, causing great harm to human health and animal husbandry. Tick pheromone released by a female ticks after feeding, stimulate and attract activity of the male ticks, thus completing the looking for a spouse and mating process, plays a key role in reproduction. Therefore, the tick sex pheromone for the control of the population, have important significance to reduce the harm.
In this study, widely distributed in China and haemaphysalislongicornis (Haemaphysalis longicornis Neumann) as the research object, using solid phase extraction, thin-layer chromatography, gas chromatography and gas chromatography-mass spectrometry method in the research of modern biology and interdisciplinary method to collect a variety of sex pheromone samples analysis, type and content the main chemical substances identified, and the biological determination of active ingredients and determine its biological role. This study further clarify the information of haemaphysalislongicornis main components and biological elements of the function, has important theoretical and practical significance for the prevention and control of ticks. The main results are as follows:
Haemaphysalis longicornis mating male ticks on the air collecting method, gland extraction and extraction from the whole tick three methods of samples have a series of roughly the same reaction: the beginning is the first pair of appendages waving in the air show actively seeking behavior; most male ticks will continue to stop crawling and stay in a certain area in 15min; the appendage folded in the body below the stationary state. The sample collection method of air flow rate was the highest. The male ticks attract air collecting method, gland extraction and whole tick extracted sex pheromone samples of mating male ticks attract rate as follows: 88, 76?? and 59?. a comparative study of the sex pheromone sample extraction method, different methods of collecting sample component and the content are quite different. The retention time of air collecting method small matter collection efficiency is very high. This kind of material is highly volatile, and at the junction Have a common structure, are carrying the number of chlorine atoms (hexachloroethane, five epichlorohydrin, eight epichlorohydrin, hexachloropropane and seven chloro propane). The biological effects of these substances in Haemaphysalis longicornis in glands has not been determined. Collection method can increase the amount of information extraction, to trace 2,6- two detection of chlorophenol in the sample (2,6-DCP) and high content of cholesterol. The whole tick extraction for material retention time is less than 12min the extraction efficiency is very low, and more concentrated in the 14min-21min peak, this method can better extract haemaphysalislongicornis surface material, such as different kinds of long chain fatty acids and cholesterol esters.
This research uses gas chromatography - mass spectrometry identification of trace 2,6-DCP in haemaphysalislongicornis gland extracts with methods, in order to attract major sex pheromone components. Gas chromatographic analysis of haemaphysalislongicornis mating female ticks were there may be another kind of structural similar material -- 2,4-DCP from 2,6-DCP, and the ratio of 1 8.74, in the bioassay of mating male ticks to attract up to 62?, and 2,6-DCP had no significant difference, 2,4-DCP also plays an important role in haemaphysalislongicornis mating.
This study by gas chromatography-mass spectrometry method for identifying Haemaphysalis longicornis mating exist cholesterol period female ticks of sex pheromone in the sample, cholesteryl oleate and cholesteryl acetate. Glycol bioassay was carried out through the surface drip method found that cholesterol, cholesteryl oleate and cholesteryl ester analogues of two alcohol two cholesteryl acetate three standard samples can make the longicornis mating success rate recovered significantly, can determine cholesterol and cholesteryl oleate as the main component of haemaphysalislongicornis MSP.
In this study, for the first time, long chain saturated fatty acids - fourteen alkanic acid and eighteen alkanic acid were found in the female ticks of the ticks, and these two substances were GSP components of the tick.

【学位授予单位】：河北师范大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R384;S852.7

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