柞蚕蛹血淋巴中凝集素的分离鉴定

在柞蚕蛹血淋巴中可以测得血凝活力,但在大肠杆菌诱导后血淋巴中血凝滴度有很大的升高.本文报道了分离正常柞蚕蛹血淋巴中凝集素的提取步骤.所得的凝集素在SDS垂直板电泳中表现单一条带,免疫扩散呈现单一的沉淀带,分子量为40,000道尔顿.制剂能凝集兔、鸭、豚鼠、羊、马及人的A、B、O和AB型的红细胞.其凝集活力可被半乳糖,乳糖,及N-乙酰半乳糖胺所抑制. 诱导后的柞蚕蛹血淋巴的凝集素成分比较复杂,经亲和层析后的制剂在免疫琼脂双扩散盘中呈现二条沉淀带,在垂直电泳中至少有二条带.     

Population dynamics of Dremomys pernyi and Callosciurus erythraeus in protective and non-protective pine forests at different ages

Four pine forests (6-10,11-15,16-20,and 31-40 year-old)located in the Cangshan Mountain and Erhai Lake National Reserve and 7 pine forests (1-5,6-10,11-15,16-20,21-30,31-40,and more than 50 year-old)located in the non-protective area near the national reserve were selected.Three replications of each forest was set and a total of 33 sites were investigated.At each site,we quantified 6 habitat variables (species richness,abundance,and percentage of grasses and shrubs coverage respectively at the bottom layer of forests)within randomly determined 5 m×5 m areas.One hundred cages were set in five lines at each site to trap small mammals,whose species and numbers were recorded.Dominance of Dremomys pernyi and Callosciurus erythraeus in small mammal communities,time niche breadth,and time niche overlap between the two small mammals were calculated,respectively.Step-wise regression was used to analyze the relationship between small mammals and habitat factors.Our results indicated that D.pernyi occurred earlier than C.erythraeus in protective pine forests.D.pernyi was captured in 6-10 year-old forest initially,and C.erythraeus was captured in 16-20 year-old forest initially.D.pernyi and C.erythraeus were captured in the 31-40 and 21-30 year-oldforests initially in the non-protective area,respectively.Populations of D.pernyi and C.erythraeus in the 31-40 year-old protective forests were 3 and 3.75 times of those in the sameaged non-protective forests,respectively.Shrubs significantly influenced the populations of the two small mammals.The population of D.pernyi was positively correlated with the density of shrubs;the population of C.allosciurus erythraeus was positively correlated with the coverage of shrubs,and negatively correlated with the coverage of grasses.D.remomys pernyi and C.allosciurus erythraeus were important for pine forests to scatter pine seeds.Human activities in the nonprotective pine forests decreased the vegetation heterogeneity at the bottom layer of pine forests,postponed the occurrence of D.pernyi and C.erythraeus,and decreased the populations of the two small mammals.     

利用全长转录本优化柞蚕基因组注释(英文)

【目的】优化柞蚕Antheraea pernyi基因组注释,更好地扩展其在比较基因组学及品种改良研究中的应用。【方法】对柞蚕进行全长转录组测序分析;经全长转录本与参考基因组比对,鉴定新基因及新转录本,并对这些新基因和新转录本进行功能注释及长链非编码RNAs (lncRNAs)预测。利用大量的蛋白质编码转录本和lncRNAs对柞蚕基因组中基因结构进行修订。最后创建矫正后的柞蚕基因组基因注释。【结果】新发现1 997个蛋白编码基因和3 399个lncRNA基因,分别由2 402个和3 574个全长转录本数据支持。发现柞蚕基因组含25 021个基因,其中19 825个基因是蛋白编码基因,包括7个保幼激素酸甲基转移酶基因。【结论】本研究促进了对柞蚕基因组基因注释信息的认识,为柞蚕及相关物种功能基因组及比较基因组学研究提供了很有用的数据资源。     

柞蚕抗菌肽CA1的分离

应用FPLC、HPLC系统配合MALDI TOF MS等技术 ,分离得到一个以灭活的Escherichiacoli诱导产生的具有明显杀菌活性的柞蚕抗菌肽CA₁。自动蛋白质序列分析仪测定其一级结构为WNPFKELERAGSRVRDAIISAGVAVATVAQATAILK ,含有 36个氨基酸残基 ,经联机检索 ,与cecropinD有 88%的同源性 ,仅有 4个氨基酸残基的差异 ,其中铰链区第 2 1～ 2 3位氨基酸为AGV ,与已知柞蚕抗菌肽A、D铰链区AGP有所不同 ,提示抗菌肽存在多态性。     

柞蚕卵繁殖赤眼蜂实验种群生命表的编制与分析

通过解剖柞蚕卵统计赤眼蜂幼期存活率与生殖力数据的方法,编制了松毛虫赤眼蜂在柞蚕卵上繁殖的实验种群生命表,松毛虫赤眼蜂在柞蚕卵上的单雌平均产卵量为86．15粒,赤眼蜂的卵、幼虫、蛹的各期存活率分别为1．00、0．9186、0．8895;羽化率为0．8668;种群趋势指数Ⅰ为54．48可见,大卵(柞蚕卵)繁蜂与小卵(如米蛾卵)繁蜂不同,大卵繁蜂时的幼期死亡率接近30％,是一个不可忽视的因素,文中还讨论了净生殖力R0、Ⅰ、内禀增长率rm3个重要生命表参数计算及其相互关系,认为R0和Ⅰ在生物学意义上相同,由Ⅰ值也可计算rm。     

柞蚕蛹及胚胎发育中几种同工酶基因的表达研究

本文通过聚丙烯酰胺凝胶电泳法研究了柞蚕蛹及胚胎发育过程中酯酶、苹果酸脱氢酶、乳酸脱氢酶和超氧化物歧化酶等几种同工酶的表达模式。结果表明,苹果酸脱氨酶和超氧化物歧化酶在蛹脂肪体、血淋巴、卵母细胞以及胚胎发育中均表达;乳酸脱氢酶只在蛹脂肪体中测到,而在血淋巴和卵母细胞中没有测到,但在胚胎发育晚期获表达;酯酶在脂肪体中获强表达,但在血淋巴和卵母细胞中表达很弱,在胚胎发育过程中仅在晚期得到较充分表达。胚胎发育中未检测到胆碱酯酶,与抗性有关的羧酸酯酶、过氧化物酶和过氧化氢酶在蛹血淋巴、脂肪体及胚胎发育中均未检测到。该研究提供了正常生理状态下柞蚕蛹及胚胎发育过程中几种同工酶基因表达的一般模式,为研究鳞翅目昆虫同工酶积累了资料。     

柞蚕核型多角体病毒(AnpeNPV)三地理株的全基因组比较分析揭示其遗传多样性

[目的]柞蚕核型多角体病毒(Antheraea pernyi nucleopolyhedroviruses,AnpeNPV)能引起柞蚕脓病大面积暴发.尽管之前已完成了两株核型多角体病毒的基因组测序,但基于比较基因组的方法调查AnpeNPV间的遗传变异研究则相对较少.[方法]为了研究AnpeNPV不同地理株系间的遗传多样性,我们测序了1株从中国河南省分离的柞蚕NPV(AnpeNPV-H)基因组,并和之前从辽宁省分离的2株柞蚕NPV(AnpeNPV-L和AnpeNPV-Z)进行了比较基因组学分析.[结果]AnpeNPV-H的全基因组大小为125 605 bp,总共预测了146个开放阅读框(ORF),包括95个功能注释蛋白和51个假定蛋白.我们发现这3株AnpeNPV间的基因组成非常相似.在这3个AnpeNPV株系间,有6个开放阅读框存在碱基变异而导致基因的变短.并鉴定超过200个单核苷酸多态性(single nucleotide variations,SNVs),其中85％位于蛋白编码区.同时,odv-e56,p94-like和egt3个基因的非同义突变较高,表明这3个蛋白质编码基因可能经历了正向选择或纯化选择.我们发现在这3株AnpeNPV间一些基因的氨基酸发生变异,例如编码膜蛋白的基因、抑制凋亡和蜕皮的基因及与DNA复制相关的DNA聚合酶和解旋酶等基因.最后,展示了3株AnpeNPV间遗传重组的证据.[结论]本研究揭示了AnpeNPV种内水平的变异和株系内基因组的动态结构.     

Immunolocalization of pheromone-binding protein and general odorant-binding protein in olfactory sensilla of the silk moths Antheraea and Bombyx

The distribution of odorant-binding proteins among olfactory sensilla of three moth species was studied by immuno-electron microscopy. Two polyclonal antisera were used in a post-embedding labelling protocol on sections of cryo-substituted antennae. The first was directed against the pheromone-binding protein (PBP) of Antheraea polyphemus, the second against the general odorant-binding protein (GOBP) of the same species. Immunoblots showed that these antisera were highly specific; both antisera did, however, cross-react with related proteins in the related species A. pernyi, and in the bombycid moth B. mori. PBP and GOBP were localized only in olfactory sensilla trichodea and sensilla basiconica, the principal site being the sensillum lymph surrounding the sensory dendrites. In the males of all three species, the pheromone-sensitive long sensilla trichodea exclusively contained PBP. the majority of the sensilla basiconica in both sexes in these species contained GOBP; these sensilla are known to respond to plant and other general odours. Some sensilla were not labelled by either antiserum; presumably, these held an odorantbinding protein of a different subfamily. Never were PBP and GOBP co-localized in the same sensillum. Two observations deserve special attention: (1) PBP was also found in a few sensilla in females, and (2) in B. mori, where the long sensilla trichodea have a different functional specificity in males (pheromone) and females (plant odours), the expression of the odorant-binding protein (males: PBP; females: GOBP) is similarly different. The distinct and complex distribution pattern of odorant-binding proteins supports the notion that these proteins participate in stimulus recognition.Dedicated to Professor Ya.A. Vinnikov on the occasion of his 85. birthdayThis work was partly supported by DFG grant ste 501/3-1.     

不同诱导源对柞蚕蛹血淋巴及生殖腺中抗菌物质产生的影响

比较了不同诱导源对诱导柞蚕蛹血淋巴中抗菌活力的动力学变化。各种化学试剂包括生理盐水都能诱导产生抗菌物质。在不引起死亡的刺激量下,刺激量愈大,诱导的抗菌活力愈高。不同金属络合剂所诱导产生的抗菌物质组份可有很大区别。如二氮杂菲使P_(9E)及P_(9A)的组份比例增高,而EDTA则可提高P_(9B)及P_(9D)组份的比例。P_(9E)和P_(9A),P_(9D)和P_(9B)的产生分别有对应关系,讨论了是否由两个不同基因所控制。 大肠杆菌诱导后的柞蚕蛹,除在血淋巴中测得抗菌活力,在雌雄的生殖腺内含物中也能测得。摘除生殖腺后虽然在血淋巴中仍能诱导出抗菌物质,但应答较慢,活力也较小,也可能由于手术创伤较大,同样的刺激量容易引起死亡。     

柞蚕蛹抗菌肽D对大肠杆菌K_(12)D_(31)的作用

本文报道了不同浓度的柞蚕蛹抗菌肽D,对大肠杆菌K_(12)D_(31)的杀灭作用动力学。在LEG培养液中,抗菌从D的浓度在5微克/毫升时显效。浓度在10微克/毫升以上时,其杀菌速度大于细菌的增殖速度。固定抗菌肽浓度为10微克/毫升,细菌浓度在3×10~7个细菌/毫升,培养在磷酸钾盐缓冲液中,4小时后能全部杀灭。同样浓度细菌在LEG培养液中,4小时后细菌数下降到约为10~2个细菌/毫升,但不能全部杀灭。同时还提供了柞蚕蛹抗菌肽D和B对大肠杆菌K_(12)D_(31)作用不同时间的电镜照片。