野蚕黑卵蜂寄主识别利它素的cDNA文库构建

根据家蚕性附腺中含有的利它素能诱导野蚕黑卵蜂寄生涂有该利它素的人造卵及能用家蚕卵繁育野蚕黑卵蜂的特性 ,用化蛹 9天的家蚕雌蛹性附腺分泌部的高丰度的mRNA构建了富含利它素基因的cDNA文库。构建成的未扩增文库的滴度为 3.2 9× 10 4 pfu μL ,重组率为 90 .0 5 % ,扩增后的文库总滴度为 1.5 6× 10 7pfu μL。用PCR方法对总文库和随机挑取的噬菌斑进行扩增 ,表明λZiplox载体中已含有大小为 0 .5～ 8.0kb的cDNA插入片段 ,并在 2 .5kb、1.1kb和 0 .75kb处cDNA插入片段相对集中。所有分析结果表明 ,采用特定的发育时期和特定的组织提取的mRNA构建而成的cDNA表达文库具有较高频率的利它素基因片段和合适的滴度 ,定向插入技术并结合λZiplox噬菌体的表达功能 ,使构建的cDNA文库具有表达外源DNA的功能 ,有利于进行利它素基因的免疫筛选。     

家蚕雌性附腺及其Ng突变体的蛋白质组差异研究

家蚕雌蛾性附腺在化蛾前2到3天开始大量分泌胶状粘性蛋白,其贮存部迅速地膨大,而其Ng突变体的雌蛾性附腺不能正常分泌胶状粘性物质.分别对家蚕(Bombyx mori)的正常及Ng突变体雌蛾性附腺分泌部组织的蛋白质进行提取,并采用双向凝胶电泳和计算机辅助分析方法,对提取的蛋白质混合物进行分离和比较分析,并对主要差异表达的蛋白质用质谱鉴定.实验结果表明,用银染法,平均每张电泳图谱可以分离约700个蛋白质点,其中大部分的蛋白质点分布在pH 4～8范围内,其分子质量主要集中在30～70 ku区域.比较分析发现一些差异表达蛋白,其中No2,3蛋白质点经质谱鉴定为肌动蛋白A3,该蛋白质只在化蛹后期正常雌性附腺组织中特异表达,而Ng突变体中肌动蛋白A3的缺失,暗示了肌动蛋白A3可能与家蚕雌性附腺的胶状粘性物质的胞外分泌有关.     

家蚕蛹-成虫变态期中肠和涎腺超微结构变化与功能

为进一步明确家蚕Bombyx mori变态期间消化系统的生理功能以及溶茧酶的来源器官,通过透射电镜观察和酶活性检测,对家蚕蛹 成虫变态期中肠和涎腺的超微结构、水解酶活力以及中肠内容物的变化进行了观察和分析。结果表明: 蛹第7日到羽化前1日家蚕中肠细胞和刚羽化成虫的涎腺细胞中,均可观察到大量的分泌泡、分泌颗粒、微绒毛等分泌细胞的结构特征以及活跃的分泌现象。潜成虫的中肠和涎腺中都存在活性较高的水解酶活力,其中每毫克中肠组织中蛋白酶活力、酯酶活力和纤溶酶活力分别为726 U、751 U和263 U,每毫克涎腺组织中上述3种酶的活力分别为603 U、523 U和147 U,说明中肠和涎腺可能都具有分泌溶茧酶的功能。家蚕蛹期中肠内容物的主要成分是蛋白质、脂质和糖,三者占内含物总量的95%以上,其中蛋白质含量占78.8%～80.2%。中肠内容物的重量在刚化蛹时为20.1～21.9 mg/头,化蛹后7日内无明显变化,化蛹第9日内容物重量减少63.01%～66.17%,到成虫羽化时已所剩无几,可能是因内容物被消化吸收所致。据此推测,在家蚕变态期中肠还具有贮存和释放营养物质的功能,而溶茧酶的另一个功能可能是分解消化中肠内容物。     

家蚕雌性附腺及其Ng突变体的蛋白质组差异研究(英)

家蚕雌蛾性附腺在化蛾前2到3天开始大量分泌胶状粘性蛋白,其贮存部迅速地膨大,而其Ng突变体的雌蛾性附腺不能正常分泌胶状粘性物质.分别对家蚕（Bombyx mori）的正常及Ng突变体雌蛾性附腺分泌部组织的蛋白质进行提取,并采用双向凝胶电泳和计算机辅助分析方法,对提取的蛋白质混合物进行分离和比较分析,并对主要差异表达的蛋白质用质谱鉴定.实验结果表明,用银染法,平均每张电泳图谱可以分离约700个蛋白质点,其中大部分的蛋白质点分布在pH 4～8 范围内,其分子质量主要集中在30～70 ku区域.比较分析发现一些差异表达蛋白,其中No2, 3蛋白质点经质谱鉴定为肌动蛋白A3,该蛋白质只在化蛹后期正常雌性附腺组织中特异表达,而Ng突变体中肌动蛋白A3的缺失,暗示了肌动蛋白A3可能与家蚕雌性附腺的胶状粘性物质的胞外分泌有关.     

野蚕黑卵蜂寄主识别利它素的纯化及氨基酸组成分析

针对利它素特殊的物理及化学性质,采用特定的温度分离方法,成功地从野蚕Theophila mandarina和家蚕Bombyx mori的雌蛾性附腺中分别获得了纯度较高的野蚕黑卵蜂Telenomus theophilae寄主识别利它素。对这两种利它素氨基酸组成的分析表明,来自家蚕和野蚕雌蛾性附腺的利它素在氨基酸的组成和含量方面非常相似,在检测到的15种氨基酸中,甘氨酸、谷氨酸和天冬氨酸的克分子百分数在10%以上。从家蚕得到的分别为28.1%、18.5%和12.6%。从野蚕得到的分别为24.4%、18.1%和10.1%。这3种氨基酸之和在家蚕和野蚕中均超过50%以上。用凝胰乳蛋白酶对这两种利它素进行水解时发现溶液中均产生非水溶性沉淀,电泳表明沉淀的多肽蛋白分子量在10 Kd左右。这显示在野蚕和家蚕利它素结构中,有着与家蚕丝蛋白相类似的结构,存在着结晶区域（沉淀）和无定形区域（上清）。     

野蚕黑卵蜂寄主识别利它素的纯化及氨基酸组成分析

针对利它素特殊的物理及化学性质 ,采用特定的温度分离方法 ,成功地从野蚕Theophilamandarina和家蚕Bombyxmori的雌蛾性附腺中分别获得了纯度较高的野蚕黑卵蜂Telenomustheophilae寄主识别利它素。对这两种利它素氨基酸组成的分析表明 ,来自家蚕和野蚕雌蛾性附腺的利它素在氨基酸的组成和含量方面非常相似 ,在检测到的 15种氨基酸中 ,甘氨酸、谷氨酸和天冬氨酸的克分子百分数在 10 %以上。从家蚕得到的分别为 2 8 1%、 18 5 %和 12 6 %。从野蚕得到的分别为 2 4 4%、18 1%和 10 1%。这 3种氨基酸之和在家蚕和野蚕中均超过 5 0 %以上。用凝胰乳蛋白酶对这两种利它素进行水解时发现溶液中均产生非水溶性沉淀 ,电泳表明沉淀的多肽蛋白分子量在 10kD左右。这显示在野蚕和家蚕利它素结构中 ,有着与家蚕丝蛋白相类似的结构 ,存在着结晶区域 (沉淀 )和无定形区域 (上清 )。     

长颚斗蟋长翅和短翅型雄成虫营养物质的积累与分配差异

【目的】翅多型雄虫在繁殖方面的能量投入与雌虫相异,这种差异可能会导致雄虫飞行与繁殖权衡的生理机制发生改变。因此,本研究旨在探究翅二型长颚斗蟋Velarifictorus aspersus雄成虫在营养物质积累与分配方面是否存在飞行与繁殖的权衡关系。【方法】选取长颚斗蟋V. aspersus头幅相近的长翅和短翅型雄成虫,对羽化后7 d内两型雄成虫的体重的动态变化进行了比较,利用分光光度计法测定了两型雄成虫虫体及飞行和繁殖器官内蛋白质、总脂和糖原含量。【结果】长颚斗蟋V. aspersus长翅雄成虫在羽化后前7 d内体重保持不变,而短翅雄成虫的体重显著增加。羽化当日长、短翅雄成虫体内蛋白质含量分别为46.5±2.7和42.3±1.9 mg,二者差异显著,且长翅雄成虫羽化后体内蛋白质含量的增加速率明显高于短翅雄成虫;而羽化当日两型雄成虫体内总脂含量无显著差异,但羽化后第5和7天短翅雄成虫体内总脂含量均显著高于长翅雄成虫。成虫羽化后7 d内长翅雄成虫飞行肌内蛋白质和总脂含量均显著高于短翅雄成虫,羽化当日两型雄成虫附腺内的蛋白质与总脂含量无显著差异,但从第3天开始,短翅雄成虫附腺内蛋白质和总脂含量均显著高于长翅雄成虫。【结论】 结果说明翅二型长颚斗蟋V. aspersus雄成虫体内营养物质的积累与分配方面存在飞行与繁殖的权衡关系。     

小地老虎雄性附腺细微结构和功能及高温的影响

本文通过光镜、电镜和生化分析等方法,研究了小地老虎Agrotis ypsilon(Rottemberg)雄性附腺的细微结构和功能,结果表明：(1)雄性附腺是一对管状腺,基段粉红色,中段桔红色,端段乳白色。形态分化在蛹前期完成,分泌功能在羽化后4天进入旺盛期;(2)附腺细胞属蛋白质合成型,具有旺盛的合成蛋白质的能力,胞内含有致密的粗面内质网和游离核糖体颗粒,大量的分泌液泡均匀地分布在细胞质中;(3)顶浆分泌和局部分泌是腺体的二种主要分泌方式, 前者分泌的颗粒物为糖蛋白性质(Pas阳性),后者吩泌的网状物为非糖蛋白性质(Pas阴性),二者在腺腔内呈有规则的放射状排列“4”雄性附腺分泌物具有种的特异性,小地老虎、棉铃虫和粘虫等夜蛾科昆虫分泌物的蛋白电泳谱带存在明显的种间差异,高温(32℃)抑制了雄性附腺分泌某些蛋白质的能力,从而改变精液的成分。     

水稻强弱势籽粒核酸和蛋白质含量的差异

水稻抽穗当天,强势粒(上部一次枝梗籽粒)子房中总RNA、mRNA含量都高于弱势粒(下部二次枝梗籽粒),蛋白质含量却低于弱势粒.抽穗后第5天,强势粒子房中总RNA含量仍然高于弱势粒,mRNA含量两者差异不大,蛋白质含量依然低于弱势粒.抽穗后0～5d,强势粒子房中总RNA、mRNA含量变化不大,蛋白质含量下降显著;弱势粒子房中总RNA含量上升显著,mRNA含量、蛋白质含量变化不大.抽穗当天和抽穗后第5天,强势粒子房中有一些小分子rRNA带出现;弱势粒则相反,小分子rRNA基本上看不见.     

天蚕卵黄原蛋白的合成、运转与沉积

系统测定了天蚕Antheraea yamamai吐丝结茧至成虫期脂肪体、血淋巴和卵巢中卵黄蛋白和可溶性蛋白总含量的动态变化。结果表明,脂肪体是卵黄原蛋白(Vg)合成场所,Vg合成始于吐丝结茧后第4天;脂肪体、血淋巴中Vg滴度在吐丝结茧后第4天开始上升,化蛹后第6天或第8天达高峰,成虫羽化第1天则明显下降。卵巢对Vg摄取始于化蛹第1天,此后随蛹日龄逐渐上升,并渐趋平稳。同一卵巢管中卵黄蛋白(Vt)含量自顶端至基端随卵室增大而逐渐升高,不同日龄蛹中相应序号卵室的Vt含量以日龄大者为高;卵室中Vt含量与卵室体积大小呈正线性关系。电镜观察表明,Vg被卵母细胞摄入后以卵黄体形式存在,不同发育阶段卵巢中卵母细胞内卵黄体大小不同,以早期者为小;同一卵巢管中不同卵母细胞内卵黄体以顶端为小,基端明显增大,且卵黄体呈网状。     

Snythesis of mucoprotein secretion in the salivary glands of Drosophila melanogaster IIIrd instar larvae]

The process of mucoprotein (glue) accumulation is described. This substance is secreted by salivary glands of the III instar larvae of Drosophila melanogaster. Granules of the secretory substance appear in the glandular cells 90 hrs after egg laying. Its secretion takes place within 120 hrs. The template RNA of glue proteins appears to be synthesized within the range of 72-85 hrs since actinomycin treatment of larvae at this time suppresses the formation of secretory substance. The portion of secretory proteins amounts to 23-32% of the total gland protein. In a mutant, 13tl, deficient by the absence of pupation, no PAS-positive, staining of glands or secretory granules identified under the phase-contrast microscope was found. A possible participation of fat body in the formation of mucoprotein secretory substance is discussed.     

Venom gland of the ichneumonid Diadromus collaris： morphology, ultrastructure and age-related changes

Females of the solitary parasitoid Diadromus collaris （Insecta： Hymenoptera： Ichneumonidae） lay eggs in the pupae of Plutella xylostella （Lepidoptera： Plutellidae）, and the venom is synchronously injected into hosts. The venom apparatus consists of two glandular tubules terminating in a common reservoir, A ductule connects the reservoir with the sting apparatus, by which the reservoir content enters the latter. Secretory units line the two glandular tubules. All secretory cells belong to dermal gland type Ⅲ. Dermal gland cells in glandular tubules are more abundant and developed than those in the reservoir. There are extensive rough endoplasmic reticulum and electrondense vesicles, and the microvilli are well developed. By the cuticle-lined central funnel secretion products of secretory units reach the reservoir. Moreover, the secretory apparatus undergoes age-related changes. The secretory units in the venom gland are better developed and more vigorous 7 days after eclosion than those 1 day after eclosion; autolytic processes occur 15 days after eclosion, and the tissue of the reservoir is more prostrate 15 day after eclosion than those 1 day after eclosion. The ovipostion peak of this parasitoid, about 3-7 days after eclosion, corresponds with the period when the venom gland is highly developed in the life span of the wasp.     

Patterns of leucine incorporation in the spermathecal accessory glands of the post-ecdysial adult female Tenebrio molitor

The spermathecal accessory glands of female mealworm beetles become fully active by six days after pupal-adult ecdysis. Coincident with the morphological maturation, increases occur in protein and RNA content and in total leucine incorporation. Three classes of differentiation-specific proteins are increasingly prominent and show heightened rates of leucine incorporation by the sixth day.     

Simultaneous determination of molting and juvenile hormone titers of the greater wax moth

A simple and rapid extraction procedure was developed to determine simultaneously the molting hormone (MH) and juvenile hormone (JH) activity in a single insect tissue sample. From the onset of the last larval stage to adult eclosion of the greater wax moth, Galleria mellonella, three JH peaks were noted: at the time of the sixth larval ecdysis, 1 day before the seventh larval ecdysis, and at the time of adult eclosion. Three MH peaks were recorded for the male: at 1 day before the sixth larval ecdysis, 1 day before the seventh larval ecdysis, and 2 days after pupation. In the female, a fourth peak was shown at the time of adult eclosion. This fourth peak exhibits the highest molting hormone activity of all samples, 1600 Musca units/g of fresh tissue or an equivalent of 5.6 μg/g of ecdysterone. Eighty per cent of this MH accumulated in the ovary. The significance of MH and JH titers as related to the endocrine regulation of development is discussed in the light of this finding.     

DEVELOPMENT OF THE ACINOUS AND ACCESSORY SALIVARY GLANDS IN NUCELLA LAPILLUS: (NEOGASTROPODA: MURICOIDEA)

The acinous and accessory salivary glands in Nucella lapillusare derived from two distinctly separate sites; the acinoussalivary glands evaginate from the walls of the buccal cavity,whilst the accessory salivary glands arise as paired invaginationsof the epithelium of the ventral lip of the mouth. During thedevelopment of the oesophagus, the acinous salivary glands growposteriorly and come to lie behind the nerve ring, but are pulledanteriorly through it when the proboscis elongates during development.The ducts of the accessory salivary glands fuse to form a singleduct with paired tubular glands during proboscis formation.The secretory cells in both pairs of salivary glands differentiateprior to the crawlaway's emergence from the egg capsule. Theontogeny of the salivary glands in Nucella shows that the accessorysalivary glands cannot be homologdus with the acinous salivaryglands or venom apparatus of the Conoidea. (Received 13 September 1996; accepted 25 November 1996)     

空心莲子草叶甲室内大量繁殖研究

为了大量繁殖供环境释放的空心莲子草叶甲Agasicles hygrophila以实现空心莲子草区域减灾, 我们探索出室内大量饲养与繁殖空心莲子草叶甲的方法与流程, 包括用叶片法或苗水培法孵化卵粒、用盒养法饲养各龄幼虫与成虫并供成虫产卵、用栽培活苗笼养法化蛹羽化。在室内成虫终日均能取食、交配、产卵, 产卵前期约4～5 d, 产卵高峰期在羽化后第7～24 天, 每雌平均产卵21.08块, 约570粒。盒养法叶片平均可着卵4.28块, 叶背与叶面着卵量相近; 笼养法叶片平均着卵为1.46块, 卵主要产于叶背。盒养法与笼养法得到的卵孵化率分别为94.02%与92.50%。空心莲子草叶甲除化蛹需在栽培活苗上完成外, 各龄幼虫与成虫均可用离体新鲜苗盒养法密集饲养。初孵1龄幼虫转株（叶）期、3龄老熟幼虫转化蛹苗期是室内大量饲养与繁殖空心莲子草叶甲的关键时期, 高密度成功饲养与繁殖空心莲子草叶甲的最适化蛹接虫量是每株8头, 产卵期雌虫的最适密度是每株5头。     

The architecture of the accessory reproductive glands of the male desert locust. 5: ultrastructure during maturation

The accessory reproductive glands of the male desert locust were studied with the electron microscope from the time of adult emergence until full sexual maturity was attained (10 days). Observations on the changes in the nucleus and the cytoplasm of the glandular epithelium, particularly those organelles involved in the elaboration of cell secretions, were made during the maturation of the accessory glands. Cells of gland 16 (the functional seminal vesicle) exhibit numerous ribosomal aggregates, have a concentration of mitochondria along their luminal surface, and show some secretory activity, even at the time of adult emergence. On the other hand, all other accessory glands show very little development of the secretory apparatus (rough-surfaced endoplasmic reticulum, polyribosomes, Golgi elements, elaborate mitochondria, etc.) until after the fifth adult day. These changes parallel changes in the secretory activity of the adult corpus allatum; and it is suggested that the corpus allatum hormone regulates the maturation of the truly glandular accessory glands of the locust, but not that of the functional vesicle.     

Ultrastructure and nature of secretory proteins in the male accessory gland of Drosophila funebris

The ultrastructural differentiation of the secretory cells and the nature of secretory proteins in the male accessory gland of Drosophila funebris have been studied by electron-microscopic and immunological methods. (1) In the pupae at $\text{1}\text{1}\text{2}$ days before eclosion, secretory products can be detected in the lumen, even though most glandular cells are at the initial phase of differentiation. At the time of eclosion both main and secondary cells are fully differentiated, but the whole set of five immunologically active proteins are detectable only on the second to third day of adult life. (2) The secondary cells contain giant protein granules, the so-called filamentous bodies, which become partially fused and the filaments assume a twisted form. Randomly dispersed filaments and closely packed filament bundles are also visible in the gland lumen. Antigenic labelling of ultrathin sections and immunoreplica electrophoresis yielded no evidence for the microtubular nature of these filaments. The secretion stored in the lumen contains in addition a large quantity of flocculent proteins which have their origin in the main cells. (3) During the period of high secretory activity in the 7-day-old male flies no vacuolization and disintegration of either the main or secondary cells have been observed. We conclude that both types of cells have the merocrine secretory mechanism. (4) Ultrastructural alterations in the glandular cells confirmed our previous observation that copulation stimulates RNA and protein synthesis.