Pheromone-induced phosphorylation of antennal proteins from insects

Protein kinase C inhibitors, such a calphostin C, abolish the transient nature of pheromone-induced rapid inositol 1,4,5-triphosphate (IP₃) responses, suggesting that pheromone signalling is terminated by phosphorylation of specific proteins. Challenging antennal preparations fromHeliothis virescens with species-specific pheromones in the presence of [³²P]--ATP led to a rapid, stimulus-dependent incorporation of³²P_i into antennal proteins. Pheromone-induced phosphorylation was completely abolished by a blockade of protein kinase C. Electrophoretic analysis revealed that upon stimulation with a pheromone blend two polypeptide bands were labelled; stimulation solely with the major compound (Z-11-hexadecenal) resulted in only a single labelled band. The data indicate that pheromones cause phosphorylation of specific antennal proteins which may be receptors for pheromones.Abbreviations ATP adenosine 5-triphosphate - DMSO dimethylsulphoxide - DPM disintigrations per minute - DTT dithiothreitol - EDTA ethylenediaminetetra-acetic acid - EGTA ethyleneglycol-bis(-aminoethyl ether)N,N,N,N-tetra-acetic acid - GTP guanosine 5-triphosphate - IP₃ inositol 1,4,5-trisphosphate - MOPS 3-(N-morpholino)propanesulphinic acid - P_i inorganic phosphate - PDBu phorbol-dibutyrate - SDS sodium dodecyl sulphate     

Probing the events of photoinhibition by altering electron-transport activity and light-harvesting capacity in chloroplast thylakoids

Abstract. The effect of photoinhibition on the activity of photosystem II (PSII) in spinach chloroplasts was investigated. Direct light-induced absorbance change measurements at 320 nm (Δ A ₃₂₀) provided a measure of the PSII charge separation reaction and revealed that photoinhibition prevented the stable photoreduction of the primary quinone acceptor Q_A. Sensitivity to photoinhibition was substantially enhanced by treatment of thylakoids with NH₂OH which extracts manganese from the H₂O-splitting enzyme and prevents electron donation to the reaction centre. Incubation with 3-(3,4,-dichlorophenyl)-1,1-dimethylurea (DCMU) during light exposure did not affect the extent of photoinhibitory damage. The chlorophyll (Chl) b -less chlorina (2 mutant of barley displayed a significantly smaller light-harvesting antenna size of PSII (about 20% of that in wild type chloroplasts) and, simultaneously, a lower sensitivity to photoinhibition. These observations suggest that photoinhibition depends on the amount of light absorbed by PSII and that the process of photoinhibition is accelerated when electron donation to the reaction centre is prevented. It is postulated that the probability of photoinhibition is greater when excitation energy is trapped by P680⁺, the oxidized form of the PSII reaction centre. The results are discussed in terms of the D1/D2 heterodimer which contains the functional PSII components P680, pheophytin, Q_A and Q_B.     

Phycobiliproteins and phycobilisomes: the early observations

The purpose of this minireview is to highlight the early observations that led to the discovery of the physico-chemical properties of the phycobiliproteins, their structure and function, and to their architectural organization in supramolecular complexes, the phycobilisomes. Generally attached on the stromal surface of the thylakoid membranes in both prokaryotic (cyanobacteria) and eukaryotic cells (cyanelles, red algae and cryptomonads), these complexes represent the most abundant soluble proteins and the major light-harvesting antennae for photosynthesis. This review mainly focuses on the years prior to the development of the molecular biology of cyanobacteria that flourished in the 1980s. We refer the reader to the comprehensive and excellent review by Sidler (1994) for more recent discoveries and more detailed literature on this topic. [-pt] `It would be difficult to find another series of colouring matters of greater beauty or with such remarkable and instructive chemical and physical peculiarities'. H. Sorby, 1877. This revised version was published online in August 2006 with corrections to the Cover Date.     

Response of <Emphasis Type="Italic">Trichogramma chilonis</Emphasis> to infochemicals: an SEM and electrophysiological investigation

The external morphology and peripheral olfactory responses of antennal chemoreceptors of Trichogramma chilonis Ishii (Hymenoptera:Trichogrammatidae) to host related stimuli were examined by SEM and EAG technique respectively. The antennae of male and female T. chilonis reveal strong sexual dimorphism in structure as well as in types, density and distribution of individual sensillar types. The female antennae are characterized by the presence of a single uniporous sensilla on the apical tip of the dome and numerous multiporous pitted sensilla. Sensilla common to both males and females include the multiporous plate sensilla, the peg sensilla and trichoid sensilla. Differences between males and females occur with respect to their EAG profiles to 22 chemicals commonly found in plants viz., hexanal, -pinene, citronellal, terpinene, citronellol, phytol, caryophyllene, R-(+)-limonene, linalool, carvacrol, cis-3-hexen-1-ol, ethanol, -pinene, trans-2-hexanal, -phellandrene, amyl acetate, cineole, quinolene, S-(–)limonene, -myrcene, carvone, terpineol; hexane leaf extracts of three host plants of Helicoverpa armigera (cotton, chickpea and pigeon pea) and hexane scale extracts of adult males and females of H. armigera. Individually, the largest peak amplitudes in females were obtained with citronellal, phytol, caryophyllene, R-(+)-limonene, linalool, carvacrol and citronellol while in males citronellol, caryophyllene, linalool, R-(+)-limonene and amyl acetate caused the highest response. Among the different chemical classes, the largest peak amplitudes in females were obtained with oxygenated monoterpenes, while in males, fatty acid derivatives caused the highest response. In both males and females, the hydrocarbon monoterpenes caused the least response. Thus, the wide spectrum response of Trichogramma chilonis, to a wide group of chemicals belonging to different chemical classes, though varying, nevertheless suggests that using plant information enhances its parasitization efficiency.     

Low-vacuum electron microscopy of carabid chemoreceptors: a new tool for the identification of live and valuable museum specimens

High-vacuum scanning electron microscopy, following coating of specimens with gold, produces high quality images that have proved invaluable for the study of insect sensilla. Unfortunately, the technique is essentially destructive, and cannot be used on live or valuable museum specimens. In particular, high-vacuum scanning usually causes the collapse of the tips of the palps, interfering with any examination of the sensilla in this area. A new low-vacuum technique is described that avoids these problems. Insect cuticle does not need to be coated with gold, thus avoiding damage to important specimens. Examples are given of scans of the palp tips of live carabid beetles, anaesthetised with CO₂. It was shown that the technique could consistently display these tips in their natural convex state. In all, four types of sensilla were identified by low-vacuum scans of the maxillary palps, and four further types on the terminal segment of the antennae, plus glandular openings. The antennae revealed a type of sensilla that has not previously been described on carabids. These sensilla showed clear structural differences between the two species studied, Pterostichus melanarius Illiger and P. niger Schaller (Coleoptera: Carabidae), and can be used as a diagnostic character for both fresh and dried specimens. The low-vacuum technique can be recommended for examining valuable type specimens without risk of damage.     

cDNA cloning and recombinant expression of the general odorant binding protein II from Spodoptera litura

A cDNA encoding the general odorant binding protein II (GOBP II) was isolated from the antennae of Spodoptera litura (SlGOBP II, GenBank Accession No. EU086371) by homologous cloning and rapid amplification of cDNA ends (RACE). Sequencing and structural analyses revealed that the open reading frame (ORF) of SlGOBP II was 489 bp, encoding 162 amino acids with a predicted MW of 18.2 kD and pI of 5.72. SlGOPB II shared typical structural features of odorant binding proteins with other insects, including the six conservative cysteine residues. The deduced amino acid sequence of SlGOPB II shared significant identity with the GOBP II from S. frugiperda and S. exigua. RT-PCR and Northern blot analyses showed that SlGOBP II was specifically expressed in the antennae. cDNA encoding SlGOBP II was constructed into the pET-32a vector and the recombinant protein was highly expressed in Es-cherichia coli BL21 (DE3) after induction with IPTG. SDS electrophoresis and Western blot analysis confirmed the molecular weight of the recombinant SIGOBPII i.e, 32 kD, which has a 6×His tag at the N-terminus. The recombinant SlGOBP II was purified by single-step Ni-NTA affinity chromatography and used to raise antiserum in rabbits. ELISA showed that the titer of antiserum was 1︰12800, while Western blot analysis showed that the recombinant SlGOBP II was recognized as anti-SlGOBP II an-tiserum.     

圆柏大痣小蜂雌成虫触角、下颚须及产卵器感器超微结构观察

圆柏大痣小蜂Megastigmus sabinae Xu et He是危害祁连圆柏球果种仁的寡食性害虫,其幼虫在球果内蛀食胚乳,并随球果同步发育,致使种子成熟后有果无仁,严重影响祁连圆柏种子生产。观察和分析圆柏大痣小蜂雌成虫触角、下颚须及产卵器感受器形态特征,旨在为圆柏大痣小蜂进化和行为策略研究提供基础。运用扫描电镜技术对圆柏大痣小蜂雌成虫的触角、下颚须及产卵器上的感受器进行观察,并对各感受器的形态特征进行分析。圆柏大痣小蜂触角为膝状,由支角突、柄节、梗节、环节、索节(7节)和棒节(3节)共14亚节组成,触角总长1 797.56±24.29μm,表面有B?hm氏鬃毛、板形感受器、毛形感受器、刺形感受器、锥形乳头状感器5种感器;下颚须由5节组成,第1节粗长,其余4节较细,表面有毛形感受器、刺形感受器、锥形感受器和栓锥形感受器4种感器;外产卵器由产卵鞘和产卵针组成,有毛形感受器、栓锥形感受器、半球形感受器和腔锥形感受器,产卵鞘长3.17±0.22 mm,表面具2类毛形感器,产卵针长5.01±0.41 mm,产卵针从基部开始,分为3个产卵瓣,外侧端部具"八"字形缺刻及2个腔锥形感器。圆柏大痣小蜂雌成虫触角、下颚须、产卵器上的感受器类型多样,形态变化丰富。不同感受器类型间存在一定差异,是圆柏大痣小蜂为适应生存而进化产生的适应性形态,不同感器形态与圆柏大痣小蜂寄主选择、产卵及交配策略具有一定的内在联系。本文首次对圆柏大痣小蜂的触角、下颚须、产卵器的感受器形态进行研究报道,为理解圆柏大痣小蜂的形态特征、进化和行为策略研究提供理了理论依据。     

锈色粒肩天牛触角感器的扫描电镜观察

利用扫描电镜对锈色粒肩天牛Apriona swainsoni雌、雄成虫触角上的感器结构、分布及数量进行观察和研究。结果表明:锈色粒肩天牛成虫触角鞭节状,由柄节,梗节和9节鞭节组成。在雌、雄成虫触角上分布着4类8种感器,分别是2种毛形感器,2种刺形感器,3种锥形感器和1种芽形感器。而发锥形感器和短刺形感器只分布于雄虫的触角上,不同类型感器在雌、雄个体上的长度及分布数量不同。     

Continuous single sensillum recording as a detection method for moth pheromone components in the effluent of a gas chromatograph

ABSTRACT. Single sensillum recordings were made from male antennal pheromone receptors of Agrotis segetum (Schiff.) (Noctuidae) and Adoxophyes orana (F.v.R.) (Tortricidae). A gas chromatograph (GC) was used to prepare and to deliver the odour stimuli. Samples of female abdominal gland extracts were injected into the column of the GC and the responses of the receptor cells to the effluent were recorded continuously. The receptor cells responded with an increase of their action potential frequency during elution of the major pheromone components. The pheromone receptors of Agrotis showed a much higher sensitivity than those of Adoxophyes. In the extract of female glands from Agrotis , compounds were detected which have not been identified in previous studies of the sex pheromone of this species. It is suggested that the combination of GC techniques with direct single sensillum recording may serve as a valuable supplement to electro-antennographic techniques.     

Active antennal movements in Drosophila can tune wind encoding

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