家蚕蜕皮激素合成相关的细胞色素P450基因的鉴定分析

【目的】昆虫变态发育的启动主要受前胸腺合成分泌的蜕皮激素所调控,而蜕皮激素的合成是由细胞色素P450基因催化完成。家蚕（Bombyx mori）是一种产丝昆虫,蚕业生产中如果能阻断或延迟家蚕蛹变态发育进程,将有利于改进蚕茧处理工艺,提高蚕丝品质。论文旨在鉴定参与家蚕蜕皮激素合成的细胞色素P450基因,从而为人为遗传调节家蚕变态发育提供靶基因。【方法】基于序列同源性比对,筛选家蚕及其他昆虫中参与蜕皮激素合成的P450基因。利用ClustalW软件,分析昆虫蜕皮激素合成相关P450基因的遗传发生关系。通过全基因组表达芯片数据分析及RT-PCR验证,调查家蚕蜕皮激素合成相关P450基因的时空表达特征。利用RNAi技术分析Cyp314a1表达下调对家蚕变态发育的影响。【结果】家蚕基因组中有4个参与家蚕蜕皮激素合成的P450基因,即Cyp306a1、Cyp302a1、Cyp315a1和Cyp314a1。比较分析显示,这4个蜕皮激素合成相关的P450基因在家蚕及其他昆虫中都是单拷贝,而且每个基因的同源体在遗传发生树上能很好地聚成一类,表明昆虫蜕皮激素合成相关的P450基因及其负责的蜕皮激素合成通路非常保守。时空表达谱分析显示,在家蚕幼虫5龄第3天,Cyp302a1、Cyp315a1和Cyp314a1主要在家蚕幼虫卵巢、精巢和头部等组织器官中高表达;在幼虫-蛹-成虫变态发育进程中,Cyp302a1、Cyp315a1和Cyp314a1主要在蛹变态发育后期表达,其中Cyp314a1分别在上簇、化蛹及羽化前高表达,这与蜕皮激素滴度高峰出现的时期基本一致。Cyp314a1的RNAi导致家蚕不能正常化蛹及雌蛾卵巢发育异常,且降低了蜕皮激素信号通路关键基因HR3及Ftz-f1的表达。【结论】家蚕蜕皮激素合成相关的P450基因在进化上非常保守,其表达水平降低能引起家蚕蛹变态发育受阻,暗示蜕皮激素合成相关P450基因可以用作家蚕变态发育控制的靶标基因之一。     

风信子鹿角珊瑚的胚胎和幼虫发育研究

采集异株风信子鹿角珊瑚Acropora hyacinthus自然排放的精卵,进行人工授精,并在显微镜下连续观察其胚胎和幼虫的发育过程。结果表明：风信子鹿角珊瑚是雌雄同体,体外受精,在月圆后2~3d同期排卵;其胚胎发育分为6个阶段,即受精卵期、卵裂期、囊胚期、原肠期、浮浪幼虫期、珊瑚幼体;受精卵经过48 h发育为浮浪幼虫,再经7 d着床变形发育为珊瑚幼体;其浮浪幼虫附着在附着基上后逐渐膨大,由圆形变成蝶形完全附着在基底上,口盘朝上,随后幼虫发生附着变形,长出触手,形成珊瑚幼体。本研究为国内对文昌云龙湾风信子鹿角珊瑚的胚胎和幼虫发育过程的首次报道,研究结果可为珊瑚礁的生态修复提供参考依据。     

半滑舌鳎的育苗效果及生物学特征的观察

在水温为21．0～23．7℃,盐度为33．5的条件下,用约7．8×10^5粒受精卵孵出仔鱼约55万尾,孵化率达70％左右,再经过79d的培育,培育出幼鱼2．287×10^5尾,成活率达到41．5％．初孵仔鱼平均全长（3．16±0．22）mm;初孵仔鱼至孵化后3日龄为前期仔鱼,冠状幼鳍形成,大部分仔鱼已开口,逐渐建立外源性摄食关系;4日龄仔鱼进入后期仔鱼阶段,冠状幼鳍继续增高,出现鳍条;5日龄,冠状幼鳍加长;13日龄,仔鱼明显集群,变态开始,进入稚鱼期;22日龄左右,大部分稚鱼右眼已经完全转到左侧,变态基本完成．在35日龄,有的个体发育进入幼鱼期,鳞片完全长齐,体色开始加深．育苗过程中,白化率低于0．5％．全长（LT）与日龄（D）的关系式为LT=6．480e0.033d,体重（W）与日龄的关系式为彬=0．747e 0.117D,全长与体重的回归方程为W形=0．0022LT 32.76。     

Distribution, larval stage duration and growth of the sea lamprey ammocoetes, Petromyzon marinus L., in a highly modified river basin

Abstract  – Larval stage duration of the sea lamprey, Petromyzon marinus , in the River Mondego is estimated to last for 4 years. The number of annuli provides reliable age estimates when compared with length–frequency distributions analysis. The growth rate of the sea lamprey ammocoetes displays strong seasonal patterns, and reaches its highest value during the first 2 years of larval stage. About 69% of the length increment between hatching and metamorphosis is attained at the end of the second year. There is a longitudinal gradient associated with ammocoete distribution along the river. Relative abundance of ammocoetes decreases downstream from the Açude-Ponte dam, the first obstruction encountered by the adult sea lampreys in their upstream spawning migration along the River Mondego.     

绵羊粪便中毛圆线虫卵培养和感染性幼虫分离法的比较

目的　为比较毛圆线虫卵培养和感染性幼虫分离的几种方法。方法　采用培养中添加锯末,蛭石培养羊粪及分离幼虫用纱布袋浸泡、贝尔曼分虫法、广口罐分离法。结果　在绵羊粪便毛圆线虫卵的培养中,用锯末作为添加物较蛭石作添加物,幼虫的孵化率提高1.8倍,两者差异显著(P≤0.05);用纱布袋浸泡法进行粪便培养物中感染性幼虫的分离,较贝尔曼分虫法提高3.7倍;较广口罐分离法提高2.6倍。结论　以上方法分离的感染性幼虫再经蔗糖垫层离心提取后,可得到纯净、高密度的活性虫体。     

Larval development and salinity tolerance of Japanese flounder (Paralichthys olivaceus) from hatching to juvenile settlement

Salinity tolerance and growth of Japanese flounder Paralichthys olivaceus at different developmental stages were evaluated, including newly hatched larvae (nhl), yolk sac larvae (ysl), oil droplet larvae (odl), post oil droplet larvae (podl), premetamorphic larvae (preml) and prometamorphic larvae (proml), at 11 salinities from 5 to 55 g L^?1 for 96 h. The ontogenesis during the early life of P. olivaceus was investigated under hatchery salinity 35 g L^?1. The results showed that suitable salinities for nhl, ysl, odl, podl, preml and proml larvae were 10 to 25 g L^?1, 10 to 30 g L^?1, 20 to 30 g L^?1, 30 g L^?1, 10 to 30 g L^?1, 15 g L^?1, respectively, demonstrating an ontogenetic variation of salinity tolerance. The salinity tolerance of nhl, ysl, preml was higher than that of odl, podl and proml. The ysl and preml larvae displayed wide salinity tolerances. The present findings demonstrate that the suitable salinity for larviculture of P. olivaceus is 20–25 g L^?1 before the depletion of oil droplet; after that, higher salinity (30 g L^?1) should be ensured for the post‐oil droplet larvae; the premetamorphic larvae can be cultured at a wide salinity range (10–30 g L^?1), and the metamorphosed larvae should be reared at salinity about 15 g L^?1.     

短须裂腹鱼胚胎与仔鱼早期发育特性研究

为积累短须裂腹鱼的基础生物学资料,并对规模化人工繁殖提供理论指导,利用养殖条件下的短须裂腹鱼亲鱼,通过干法授精获得受精卵,对其胚胎发育和仔鱼早期发育全过程进行连续观察。短须裂腹鱼受精卵的平均卵径为2.36 mm,吸水膨胀后为3.68 mm,沉性卵、弱粘性、卵黄丰富。在水温(14±1)℃的条件下,受精后6 h 30 min进入卵裂期,20 h 55 min进入囊胚期,60 h 28 min进入原肠期,70 h 4 min进入原肠中期,77 h 52 min进入神经胚期,142 h肌肉开始收缩,177 h 46 min进入心动期,254 h 40 min开始出膜。孵化全过程所需积温为3 565.3℃·h。初孵仔鱼全长8.7 mm。出膜后第2~9天,胸鳍、鳃、口腔、眼色素、体内血管等器官相继发育完全;第10天仔鱼全长达15.15mm,鳔充气,鱼苗开始平游和觅食。孵化过程中应重点防控水霉病。早期仔鱼可以投喂轮虫、蛋黄或者豆浆等,待仔鱼捕食能力变强之后,可以投喂更加适口的枝角类、嫩口丰年虫等。     

A larval specific OBP able to bind the major female sex pheromone component in Spodoptera exigua(Hübner)

Odorant binding proteins(OBPs) in insects are postulated to solubilize and transport the hydrophobic odorants across the hydrophilic antennal lymph to the olfactory receptors(ORs) located on the dendrite membrane of the sensory neurons. OBPs in adult insects have been intensively reported, but those in larvae are rarely addressed. In our study, a full-length OBP c DNA, namely Sexi OBP13, was cloned by RT-PCR and RACE strategy from the heads of Spodoptera exigua larvae. The quantitative real-time PCR(q PCR) measurement indicated that Sexi OBP13 was highly expressed in larval head, but very low in other parts of larva and was not detected in any tissues of adult. The binding affinities of Sexi OBP13 to plant volatiles and female sex pheromone components were measured by competitive binding assays. Interestingly, Sexi OBP13 displayed a high binding affinity(Ki=3.82 μmol L–1) to Z9,E12–14:Ac, the major sex pheromone component of S. exigua, while low affinities to the tested host plant volatiles(Ki27 μmol L–1). The behavioral tests further confirmed that Z9,E12–14:Ac was indeed active to elicit the behavioral activity of the third instar larvae of S. exigua. Taken together, our results suggest that Sexi OBP13 may play a role in reception of female sex pheromone in S. exigua larvae. The ecological significance of the larvae preference to the adult female sex pheromone was discussed.     

不同饵料对泥鳅苗种前期培育效果的研究

本文研究了蛋黄、轮虫、豆浆、脱脂奶粉、鳗鱼饲料、鱼粉和酵母粉等7种饵料对泥鳅苗种前期培育生长和存活的影响。经过21 d培育,结果显示,泥鳅在不同饵料条件下的生长速度和存活率存在显著差异(P0.05),轮虫组生长速度较快,平均日增全长达到0.31 mm,最终存活率最高,达到39.5%;其次是蛋黄组,达到25.5%;其他组别最终成活率都较低。因此,在本试验条件下,轮虫作为泥鳅开口饵料效果最为理想。Bertalanffy、Gompertz和Logistic三种非线性生长模型均能很好地拟合泥鳅苗种生长曲线(R2=1.000),该期生长拐点在7~8日龄间。     

Larval fish distribution and retention in the Canary Current system during the weak upwelling season

The spatial distribution of fish larvae was studied in the Canaries‐African Coastal Transition Zone, outside the strong upwelling season. An onshore–offshore transition in the larval fish community structure was observed, from a coastal assemblage dominated by small pelagics (sardine, anchovy, mackerel), bounded by the upwelling front, to an offshore assemblage dominated by mesopelagic species (mainly Myctophidae, Phosichthydae, Gonostomatidae). Distribution of the neritic larvae was deeply influenced by the intense mesoscale activity found in the area, both horizontally (larvae were advected offshore but were always retained within the upwelling area) and vertically (larvae were deepened in the vicinity of two anticyclonic eddies). A combined effect of the upwelling front and a cyclonic–anticyclonic eddy dipole is likely the successful retention mechanism for these larvae. These results support the current belief that retention may be higher than previously thought in upwelling areas. Oceanic larvae were also collected in higher abundances near the front and an anticyclonic eddy. Neritic and oceanic larvae frequently showed a differentiated position in the water column, although they sometimes coexisted. Finally, larval connectivity between Islands within the Canary archipelago is suggested. The present study thus contributes to the understanding of the complex dispersal and retention processes in the Canaries‐African Coastal Transition Zone. However, results also highlight the poor knowledge of this region compared with the other three main Eastern Boundary Upwelling Systems in terms of ichthyoplankton dynamics. The importance of routine monitoring programs of commercial and non‐commercial species in the area is emphasized.