论文标题:蝶蛹金小蜂和丽蝇蛹集金小蜂毒液的生化特性与生理功能
Biochemical Characters and Physiological Functions of Venoms from Two Peromalid Wasps, Pteromalus Puparum and Nasonia Vitripennis (Hymenoptera: Pteromalidae)
论文作者 张忠
论文导师 胡萃；叶恭银,论文学位 博士,论文专业 农业昆虫与害虫防治
论文单位 浙江大学,点击次数 151,论文页数 202页File Size18192k
2005-05-01论文网 http://www.lw23.com/lunwen_4163382/ 蝶蛹金小蜂;丽蝇蛹集金小蜂;毒液;电泳;质谱分析;菜粉蝶;柑桔风蝶;血细胞;延展;存活;包囊反应;非免疫系统组织;非寄主昆虫;昆虫细胞系;表皮蛋白;表皮可溶性蛋白;表皮碳氢化合物;表皮脂类;脂肪体;血淋巴;蛋白质;脂类;氨基酸
Pteromalus puparum;Nasonia vitripennis; Venom; Electrophoresis; MALDI-TOF analysis; Pieris rapae; Papilio xuthus; Hemocyte; Spreading; Viability; Encapsulation capability;Non-immune system; Non-target insects; Insect cell lines; Cuticular protein; Soluble protein in cuticle; Cuticular hydrocarbon; Cuticular Lipid; Fat body; Protein; Lipid; Amino acid
本论文针对国内外有关寄生蜂毒液性质与功能研究主要限于姬蜂科和茧蜂科,而有关其它科寄生蜂研究甚少之现状,特对隶属金小蜂科的蝶蛹金小蜂和丽蝇蛹集金小蜂毒液的性质和功能进行了系统研究,主要结果概括如下: 1 蝶蛹金小蜂成蜂生殖系统抑制寄主细胞免疫因子的确定 蝶蛹金小蜂卵巢、卵子、输卵管、睾丸和雄性附腺提取物处理其寄主菜粉蝶蛹和柑桔凤蝶蛹血细胞后,其细胞延展率、死亡率及对Sephadex A-50微珠的包囊率与对照相比均无显著差异;而雌蜂毒液则可显著抑制血细胞延展及对Sephadex A-50微珠的包囊,并引起血细胞的显著死亡。可见,蝶蛹金小蜂毒液是抑制其寄主血细胞免疫的主要因子。此蜂毒液的功能显然不同于具多分DNA病毒(PDV)的茧蜂或姬蜂。 2 两种金小蜂毒器官发育动态及其与卵子发育的关系 饲以蜂蜜的蝶蛹金小蜂和丽蝇蛹集金小蜂雌蜂发育过程中,毒腺长度变化不大,毒囊变化明显,雌蜂刚羽化时,毒囊形态较瘪,随雌蜂的发育毒囊体积迅速膨大,蛋白含量迅速增加,3d后稳定于一定水平,并持续至死亡。未喂食蜂蜜水的蝶蛹金小蜂发育过程中,毒腺长度变化不明显,毒囊发育相对较慢,雌蜂寿命明显缩短。交配和喂食蜂蜜水对蝶蛹金小蜂和丽蝇蛹集金小蜂雌毒腺长度均无显著影响,交配不影响两种金小蜂雌蜂毒囊大小和毒液蛋白含量,但喂食蜂蜜水对此可产生显著正面影响。 喂食蜂蜜水并正常交配的蝶蛹金小蜂毒器官各组成部分的发育与其卵巢中成熟卵的数量均存在正相关关系,而与其卵巢中Ⅰ级卵和Ⅱ级卵的数量成负相关关系。毒器官各组成部分之间的发育也呈现相应的线性关系。丽蝇蛹集金小蜂毒囊直径发育与其卵巢中Ⅱ级卵数量存在相关关系,毒器官其它组分与卵巢中各级卵数量均无线性相关关系。毒器官各组成部分之间的发育呈线性关系。 3 两种金小蜂毒液的蛋白组成及分子特性 蝶蛹金小蜂毒液蛋白的梯度PAGE电泳结果显示由12条谱带组成,分子量为17.47～879.67kDa,其中7条带大于100kDa,蛋白含量以分子量为558.64kDa、308kDa和178.17kDa的三条蛋白谱带为主。丽蝇蛹集金小蜂毒液蛋白的梯度PAGE图谱由9条带组成,分子量为17.62～809.94kDa,其中以分子量为246.20kDa蛋白含量为最高。SDS-PAGE分析表
Studies of biochemial characters and physiological founctions of parasitoid were foused on braconid and ichneumonid wasps, but in other families were fewer. So we studied the biochemical characters and physiological functions of venoms from two peromalid wasps, Pteromalus puparum and Nasonia vitripennis. The results were shown as follows.1. Host immune inhibiting factors in the reproductive system of Pteromalus puparumExtracts of ovary, eggs, oviduct from female P. puparum and testes, male accessory gland from male P. puparum had no effects on spreading, mortality and encapsulation capability of hemocytes from P. rapae and P. xuthus pupae. But venom from female P. puparum could inhibit the spreading and encapsulation capability of the hemocytes and elicit significant death of hemocytes. We considered venom is the host immunity inhibiting factor in P. puparum female. Venom functions in the parasitoid was different with venoms from braconid and ichneumonid wasps had polydnavirus.2. Development of venom apparatus from two pteromalid wasps and their relationship with the oogenesisThe length of venom glands from female P. puparum and N. vitripennis fed with 20% honey water had no significantly change with the development of female wasps, but the diameters of venom reservoir and proteins in them increased rapidly in the first 3 days after female wasps" eclosion, and persisted at the levels until the death of female wasps. The length of venom glands from non-fed P. puparum and N. vitripennis female also had no significantly changes, but the development of venom reservoirs were much slower. Mating and feeding had no effects on the length of venom glands from P. puparum and N. vitripennis female, mating had no effects on thediameters and protein content of venom reservoirs, but feeding had significantly beneficial effects on them.Development of venom apparatus from mating and feeding P. puparum had positive relationship with the numbers of mature eggs and negative relationship with the numbers of grade I and grade II eggs from the ovary. But development of venom apparatus from mating and feeding N. vitrirpennis had no correlation with the numbers of mature eggs and immature eggs except that venom reservoir diameter had negative relationship with the numbers of grade II eggs in the ovary. Parts of venom apparatus of two pteromalid wasps also had positive relationships with each other.3. Composition and molecular character of venom proteins from two pteromalid waspsAnalysis of the 5 ~ 15% gradent-PAGE profile revealed the presence of 12 protein bands with molecular weight ranging from 17.47 to 879.67 kDa in the P. puparum venom, of which 558.64 kDa, 308 kDa and 178.17 kDa proteins were the highest three in the venom. The factors in P. puparum venom inhibiting the spreading and inducing the death of hosts" hemocytes were both proteins with native molecular weight above 100 kDa. Conversely, 9 protein bands with molecular weight between 17.62 to 809.94 kDa were detected in the N. vitripennis venom.Analysis of the 10% SDS-PAGE profile revealed the presence of 17 protein bands with molecular weight ranging from 14.11 kDa to 165.18 kDa, of which 9 protein bands between 21.17 ~ 44.41 kDa were relatively abundant, were observed in P. puparum venom. In contrast, 14 protein bands between 14.11 ~ 155.93 kDa, of which 6 protein bands between 21.15 ~ 36.68 kDa were relatively rich, were found in N. vitripennis venom. The factors in P. puparum venom inhibiting the spreading and inducing the death of hosts" hemocytes were both proteins with native molecular weight above 100 kDa.pi of venom proteins from P. puparum were between 4 and 7, and 5 to 10 from N. vitripennis. After analyzed with MALDI-TOF MS for seven higher expression spots from 2-DE gel of P. puparum venom and searched in UCSF with MS-FIT, we found the matching ratio of the checking proteins and proteins from database were very lower, with the highest matching ratio of 35%.4. Physical and chemical factors affected the physiological functions of venomsfrom Pteromalus puparum and N
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