海葵毒素Av3对德国小蠊的毒性及其作用机制

摘要: 【目的】Ⅲ型海葵毒素(Av3)对昆虫具有显著的选择性毒性，对其作用机制的研究对新型高选择性毒性杀虫剂的设计研究具有重要意义。【方法】反向高效液相色谱及电喷雾质谱用于鉴定化学合成的Av3野生型(Av3 wild type, Av3-WT)及其突变体的纯度和分子量；生物活性测定检测Av3-WT及其突变体对德国小蠊Blattella germanica成虫的毒力；双电极电压钳技术检测Av3-WT及其突变体对德国小蠊钠通道BgNav1-1a失活的抑制作用。构建基于BgNav1-1a与大鼠钠通道rNav1.2a的重组嵌合体，通过双电极电压钳技术确定BgNav1-1a上参与Av3-WT选择性毒性的关键区域。【结果】芳香族氨基酸Y7, W8和Y18分别突变后形成的Av3-WT突变体Y7A, W8A和Y18A对德国小蠊成虫的毒力显著降低，半数击倒剂量(KD50)与Av3-WT相比均增加了超过10倍；与Av3-WT对通道失活62%的抑制率相比，250 nmol/L的毒素突变体Y7A, W8A和Y18A对BgNav1-1a失活的抑制作用也显著降低，通道失活的抑制率分别降低到12%, 23%和8%；以rNav1.2a胞外环DI/SS5-S6替换BgNav1-1a的相应序列获得的重组嵌合体钠通道对Av3-WT毒素的敏感性几乎丧失，仅3.6%的通道在1 μmol/L Av3-WT作用下失活被抑制。BgNav1-1a胞外环 DI/SS2-S6上的His404突变为Tyr后几乎丧失对毒素的敏感性，仅6%的通道在1 μmol/L Av3-WT作用下失活被抑制。【结论】芳香族氨基酸Tyr7, Trp8和Tyr18参与到构成Av3-WT分子的生物活性表面；钠离子通道胞外环DI/SS2-S6是影响Av3-WT毒素发挥选择性的关键结合区域，BgNav1-1a的DI/SS2-S6上的His404则是影响Av3-WT选择性毒性的关键氨基酸。     

Pb污染对马蔺生长、体内重金属元素积累以及叶绿体超微结构的影响

通过土壤单因子Pb胁迫盆栽试验和铅锌尾矿污染土壤掺比试验研究了不同土培条件下Pb对马蔺（Iris lactea var chinensis）生长、体内重金属元素积累以及叶绿体超微结构的影响。结果表明：2种不同土壤Pb胁迫下马蔺株高、根长、叶数以及地上、地下部生物量均随Pb浓度的增加呈先增后降的趋势,其中,土壤单Pb胁迫下,低于500 mg?kg-1Pb胁迫（T1—T3）均增加了马蔺的株高和根长,仅800mg?kg-1高Pb浓度处理时马蔺生物量才出现轻微下降（P＞0.05）;与单一土壤Pb胁迫相比,铅锌尾矿污染土壤掺比试验中铅锌尾矿含量低于一定浓度下对马蔺株高、根长生长同样也具有促进效应,只有100%尾矿污染土壤处理下马蔺地上部和根系生物量下降显著（P＜0.05）。马蔺地上部和根系Pb含量均随土壤中Pb浓度和尾矿含量的增加而增加,800mg?kg-1土壤单Pb胁迫下为340.6、1700.02 mg?kg-1,100%尾矿污染土壤处理下为126.9、1725 mg?kg-1。Pb单独胁迫下马蔺地上部Zn含量随Pb含量的增加逐渐下降, Zn和Pb的吸收表现出一定的拮抗效应;而不同掺比污染土壤胁迫下马蔺地上部和根系Zn积累同Pb一样,均随土壤中Pb、Zn浓度的增加而增加,Zn和Pb的吸收表现为一种协同效应。而2种不同土壤Pb胁迫下马蔺对Cu、Cd的吸收均相对较少。本研究中,800mg?kg-1Pb胁迫处理和50%尾砂土壤胁迫处理下马蔺叶片叶绿体双层被膜及内部基粒和类囊体结构与对照相比变化不大。综上表明,马蔺对重金属Pb有较强的耐受性,具备修复Pb污染土壤的潜在能力。     

黄菖蒲和美人蕉对水深梯度的响应差异

通过野外观测研究了黄菖蒲和美人蕉对水深梯度(10、30、50和70cm)的适应性。结果表明:随着水深的增加(1)黄菖蒲和美人蕉的分蘖数显著减少(P<0.05),但对株高、叶长和叶宽的影响不大;(2)2种植物叶片的叶绿素含量和根系活力均逐渐降低,叶片的丙二醛含量则逐渐升高,其中美人蕉的叶绿素含量、根系活力和叶片丙二醛含量的差异显著(P<0.05),而黄菖蒲仅根系活力的差异显著(P<0.05);(3)叶绿素荧光参数中,PSⅡ的最大光能转换效率(Fv/Fm)、表观电子传递速率(ETR)和光化学淬灭(qP)均呈显著下降的趋势(P<0.05),非光化学淬灭(qN)则显著增加(P<0.05);相对电子传递速率(rETR)及潜在最大电子传递速率(rETRm)、对光能的利用效率(α)和对强光的耐受能力(Ik)也都表现出明显的下降;美人蕉在10cm水深条件下各指标均高于黄菖蒲,但在水深较大的条件下总体上低于黄菖蒲。可见,水深较大时对黄菖蒲和美人蕉的生长产生了抑制作用,且对后者的抑制程度要大于前者。因此,在选择黄菖蒲和美人蕉进行湿地植物修复时,应保持浅水条件以利于其生长,其中黄菖蒲适应的水深(30～70cm)大于美人蕉(10cm...     

Cytological features of developing embryogenic callus and somatic embryos of Iris pumila L. and Iris setosa Pall

Abstract

Cytological examination during somatic embryogenesis in Iris pumila L. and Iris setosa Pall, were performed using light and electron microscopy. The first sign of the cellular differentiation in the initial embryogenic callus (EC; stage 1) of both Iris species was the formation of short and elongated cell types. After the onset of embryogenesis, short cells divided producing a mass of densely packed meristematic cells, closely connected with numerous plasmodesmata. Further differentiation into globular embryos (GE) led to a loss of plasmodesmata and cell separation. In vacuolated elongated cells, cytoplasm was located near the wall and around the nucleus. In both cell types amyloplasts and small mitochondria with poorly developed crystae were abundant.

Cell of GE (stage 2) contained an increased number of mitochondria and plastids comparing to those from stage 1, indicating further differentiation. Thylakoids and starch grains were observed within the plastids, while the number of cristae within the mitochondria was increasing.

In cells of embryos with coleoptile (ECl) (stage 3), plastids differentiated into chloroplasts with thylakoids. In all stages of cell differentiation, short and long cisternae of endoplasmic reticulum with ribosomes were seen. Activity of dictyosomes was increased in stages 1 and 2, then reduced in stage 3.

Ultrastructure of EC cells was identical to that of proembryogenic cells, i.e. of early GE. Ultrastructural appearance of GE cells was identical in both Iris species, but evident, and increasing, differences in mitochondria and plastids were observed between GE and ECl embryos.The presence of bi-, three- and eight- cell proembryos demonstrates that they originate from a single cell in both Iris species.     

Odour-mediated foraging by yellowjacket wasps (Hymenoptera: Vespidae): predation on leks of pheromone-calling Mediterranean fruit fly males (Diptera: Tephritidae)

Predation is probably the most important male mortality factor in insect species with courtship displays that render males performing them conspicuous targets of predators. Sexually active Mediterranean fruit fly males, Ceratitis capitata (Wied.), aggregate in leks, where they participate in agonistic encounters and engage in visual, acoustic and pheromone-calling displays to attract receptive females. The objective of this study was to assess: a) whether sexually displaying C. capitata males in leks inside host and non-host foliage are subject to predation by the most prominent predators yellow-jacket wasps, Vespula germanica (F.), and if so, b) whether olfactory, visual or auditive stimuli are used by foraging wasps in locating male C. capitata prey. Studies were carried out in a citrus orchard and surroundings on the island of Chios, Greece. Observations were conducted using perforated containers hung within mulberry, fig or citrus foliage. Living C. capitata flies of different sex and either mature or immature were placed inside. Our results show that the yellowjacket wasps have learned to associate the presence of sexually active medfly males aggregated in leks with their prey's pheromone (kairomone). Foraging wasps, flying through the crowns of host trees, responded to the odour source of C. capitata male pheromone by approaching from downwind. Even inside dense citrus tree foliage, wasps keyed in on aggregations of pheromone-calling males using olfactory stimuli. Stimuli of visual and acoustic male signalling were only used at close range, after having followed the pheromone plume close to its source. Visual cues played a greater role in directing wasp foraging under more open and exposed host foliage conditions. Odour-based foraging of wasps inside host foliage in the mid-morning hours, when medfly male lekking activities peak, shifted gradually to a more visual-based host fruit patrolling in the afternoons to capture ovipositing and feeding medfly females. On ripe fruit, particularly fig, V. germanica visual prey hunting also included the capture of feeding medfly males, other feeding Diptera, as well as medfly larvae extracted from wasp-made perforations in the fruit.     

Structure of yolk granules in oocytes and eggs of Blattella germanica and their interaction with vitellophages and endosymbiotic bacteria during granule degradation

Oocytes and embryos of the cockroach Blattella germanica were examined by optical and electron microscopy to study yolk granule degradation during embryo development. During vitellogenesis, progressively larger yolk granules are formed in the ooplasm and by chorionogenesis, the mature granules are packed so tightly that their shape is highly distorted. Throughout ovarian development, endosymbiotic bacteria lie at the follicle cell/oocyte interface. Just prior to chorionogenesis the endosymbionts transit the oocyte plasma membrane and cluster at the periphery. Bacteria become more numerous over the ventral region of the egg by day 1 postovulation and begin to invade the interior of the yolk mass from the ventral periphery. At that time, lysis of the nearby yolk granules occurs while those in the central ooplasm remain intact and free of bacteria up to day 4. Vitellophages become evident by day 2 postovulation. These cells are also distributed over the egg's periphery but are most numerous in the ventral region. Vitellophages, in association with the endosymbionts, protrude toward the yolk granules and extend filo- and lamellipodia over the granule surface. Portions of the yolk granules are then engulfed and sequestered as large vacuoles in the vitellophage's cytoplasm. The vacuoles then become vesiculated. As embryo development proceeds, the vesiculated portions partition into smaller multivesicular bodies. This study describes the dynamics of yolk granule-vitellophage interaction in embryos of B. germanica and suggests that yolk utilization entails the cooperative efforts of both vitellophages and endosymbiont bacteria.     

Phenotypic plasticity in response to environmental heterogeneity contributes to fluctuating asymmetry in plants: first empirical evidence

Fluctuating asymmetry (FA) is widely used to quantify developmental instability (DI) in ecological and evolutionary studies. It has long been recognized that FA may not exclusively originate from DI for sessile organisms such as plants, because phenotypic plasticity in response to heterogeneities in the environment might also produce FA. This study provides the first empirical evidence for this hypothesis. We reasoned that solar irradiance, which is greater on the southern side than on the northern side of plants growing in the temperate zone of the Northern Hemisphere, would cause systematic morphological differences and asymmetry associated with the orientation of plant parts. We used geometric morphometrics to characterize the size and shape of flower parts in Iris pumila grown in a common garden. The size of floral organs was not significantly affected by orientation. Shape and particularly its asymmetric component differed significantly according to orientation for three different floral parts. Orientation accounted for 10.4% of the total shape asymmetry within flowers in the falls, for 11.4% in the standards and for 2.2% in the style branches. This indicates that phenotypic plasticity in response to a directed environmental factor, most likely solar irradiance, contributes to FA of flowers under natural conditions. That FA partly results from phenotypic plasticity and not just from DI needs to be considered by studies of FA in plants and other sessile organisms.     

Laboratory and field evaluation of Beauveria bassiana bait against two cockroach species (Dictyoptera: Blattellidae,Blattidae) in Jinan City,East China

A Beauveria bassiana bait formulation was evaluated for its efficacy in the laboratory and field against German cockroaches, Blattella germanica (L.), and American cockroaches, Periplaneta americana (L.). The LT₅₀ values were 46 and 119?h, respectively, for B. germanica and P. americana. The cockroach species were lured and killed effectively when linear distance was about 5?m between B. bassiana bait and fresh mouse food. In the field, more than 90% relative decline in density was found 14?d after treatment, which lasted for 90?d. In conclusion, laboratory and field studies demonstrated the high efficacy of a B. bassiana bait formulation against common cockroach species.