植食性昆虫唾液调控昆虫-植物互作的研究进展

在与寄主植物长期共同进化的过程中,为了更好地适应和利用寄主,植食性昆虫进化出了多种取食方式。同时,为了应对植食性昆虫的取食,植物进化出了多种防御策略,包括直接防御和间接防御。在整个昆虫-植物互作过程中,昆虫唾液起着重要作用。一方面,昆虫唾液中一些组分可以被植物识别并诱导植物防御反应;另一方面,昆虫通过分泌唾液到植物中调控寄主防御反应。该文从昆虫-植物互作的角度出发介绍植食性昆虫唾液的成分与功能,着重对昆虫唾液激发子和效应子的研究进展进行了综述,并对未来唾液的研究及其在害虫防治中的应用进行展望。     

蚜虫传播非持久性病毒的取食行为调控机制

蚜虫能够传播上百种植物病毒,是最重要的农业介体昆虫之一。蚜虫在刺探和取食植物过程中,唾液组分会连同附着在口针中的病毒粒子一同被分泌进入植物内,在调节植物诱导抗性、病毒侵染扩散、介体昆虫行为等过程中均有重要作用。本文围绕蚜虫传播病毒和获取病毒2个关键过程,总结分析了蚜虫独特的刺吸取食行为与传毒效率和获毒效率之间的联系;针对取食活动中关键的唾液蛋白在调控植物免疫抗性、帮助病毒侵染过程中的功能,阐述了蚜虫高效传播非持久病毒的分子基础;针对蚜虫的获毒过程,综述了病毒侵染植物间接调控蚜虫趋向和行为的作用方式。这些研究的开展将为解释蚜虫和病毒协同侵染的分子机制以及有效开展基于蚜虫取食行为调控的病虫害防控新技术提供思路。     

植物寄生线虫效应蛋白调控寄主防卫反应分子机制研究进展

 在植物寄生线虫与寄主互作过程中,线虫分泌器官如食道腺细胞分泌的效应蛋白在寄主细胞壁修饰和调控寄主免疫反应以及取食位点形成和维护中发挥着关键作用。解析植物寄生线虫关键效应蛋白的功能及其与寄主互作机制将为探索植物寄生线虫防控新策略提供重要的理论基础。本文从效应蛋白降解寄主细胞壁、调控寄主基础免疫反应、诱导免疫反应机制和介导翻译后修饰调控寄主免疫反应以及植物激素代谢途径的调控机制等方面进行了概述。     

植物寄生线虫效应蛋白调控寄主防卫反应分子机制研究进展

 在植物寄生线虫与寄主互作过程中,线虫分泌器官如食道腺细胞分泌的效应蛋白在寄主细胞壁修饰和调控寄主免疫反应以及取食位点形成和维护中发挥着关键作用。解析植物寄生线虫关键效应蛋白的功能及其与寄主互作机制将为探索植物寄生线虫防控新策略提供重要的理论基础。本文从效应蛋白降解寄主细胞壁、调控寄主基础免疫反应、诱导免疫反应机制和介导翻译后修饰调控寄主免疫反应以及植物激素代谢途径的调控机制等方面进行了概述。     

Modulation of human lymphocyte proliferation by salivary gland extracts of ixodid ticks (Acari: Ixodidae): effect of feeding stage and sex

Ixodid ticks remain attached to their hosts for several days to weeks. During this extended feeding process new proteins involved in the modulation of host immune responses are expressed in tick salivary glands. In our study a stimulatory or inhibitory effect of salivary gland extracts (SGE) of unfed and partially fed female Ixodes ricinus (Linnaeus, 1758), female and male Amblyomma variegatum (Fabricius, 1794) and Rhipicephalus appendiculatus Neumann, 1901 ticks on human lymphocyte proliferation induced by Concanavalin A (ConA) and phytohaemagglutinin (PHA), respectively, was investigated. SGE of all female ticks examined suppressed proliferation of ConA-induced lymphocytes; highly significant suppression was observed in the presence of unfed I. ricinus and 9-day fed A. variegatum SGE. SGE of partially fed A. variegatum and I. ricinus females suppressed PHA responses of lymphocytes. Lymphocytes showed reduced PHA and ConA responses in the presence of SGE of unfed and 2-day fed R. appendiculatus females, while SGE of 6-day fed females enhanced PHA responses, but reduced their ConA responses; generally SGE of 2-day fed females displayed the strongest inhibition. Amblyomma variegatum male SGE slightly enhanced PHA, but significantly reduced ConA responses of lymphocytes and their inhibitory effect increased during feeding. SGE of unfed and 2-day fed R. appendiculatus males enhanced PHA and ConA responses and those of 6-day fed males suppressed lymphocyte proliferation. The results suggest that (i) species- and sex-specific differences exist in the effects of tick salivary gland antigens on human lymphocyte proliferation and (ii) effect of SGE on human lymphocyte responses to mitogens varies depending on the tick feeding status.     

棉铃虫唾液凝集素的初步研究

昆虫唾液在成功取食寄主中发挥关键作用.而有关植食性昆虫唾液(反吐液)的研究多集中在与寄主植物的互作关系上,鲜有涉及唾液免疫因子的研究.凝集素作为模式识别受体,在病原物的识别中发挥重要作用,是昆虫重要的免疫因子.棉铃虫幼虫血淋巴中已报道的凝集素种类至少有8种,但其唾液中是否存在凝集素却未见报道.本文通过与鸡血红细胞的血凝...     

Virulent and attenuated lines of Leishmania major: DNA karyotypes and differences in metalloproteinase GP63

The Leishmania metalloproteinase GP63 has been reported to play important roles mainly in resistance of promastigotes to complement-mediated lysis and in interaction with macrophage receptors. On the other hand, its function in insect vectors is still unclear. We compared the structure and dosage of gp63 genes and the activity of GP63 in Leishmania major Yakimoff et Schokhor strains and lines differing in virulence for mice and ability to develop in sand flies. The results demonstrate considerable variability in amount and proteolytical activity of GP63 among L. major strains although genomic changes in the gp63 locus were not found. Attenuated LV561/AV line showed low amount and low enzymatic activity of GP63. Serial passages of attenuated parasites through either Phlebotomus duboscqi Neveu-Lemaire or through mice led to a recovery of GP63 proteolytical activity to the level present in virulent LV561/V line. Overexpression of GP63 was found in two L major strains (L119, Neal) with defective lipophosphoglycan (LPG); both these strains were capable to cause mice infection but unable to survive and multiply in sand flies. Differences were found also in karyotypes and in amount of minichromosomes amplified in some lines of the LV561 strain. The results suggest that parasite virulence is not simply correlated with the activity of GP63; however, this enzyme plays a significant role in association with other surface molecules, especially LPG. Overexpression of GP63 can compensate LPG defect in the vertebrate host but in sand flies both molecules fulfil quite different functions and the defect in LPG is lethal for the parasite. On the other hand, linear minichromosomes of about 200 kb found in some lines of the LV561 strain are associated with development in vitro and in the vector but they are not essential for the infection of the vertebrate host.     

Effect of tick salivary gland extract on the cytokine production by mouse epidermal cells

Previous studies have demonstrated that both tick saliva and Borrelia burgdorferi sensu lato antigens modulate the cytokine response of the host. In this paper, the effect of salivary gland extract (SGE) from Ixodes ricinus (L., 1758) ticks on cytokine production by primary cultures of mouse epidermal cells stimulated with Borrelia afzelii Canica, Nato, du Merle, Mazie, Baranton et Postic, 1993 spirochetes was analysed. Epidermal cells were derived from C3H/HeN mice, susceptible to Lyme disease, and BALB/c mice, which are resistant. In cultures from C3H/HeN mice, SGE down regulated production of tumour necrosis factor alpha (TNF-alpha) and up regulated Th2 cytokine, interleukin 4 (IL-4). Cultures from BALB/c mice produced higher basal levels of monitored cytokines, but their production was affected by SGE a different way. While Th2 cytokines IL-6 and IL-10 were down regulated, the effect on TNF-alpha and IL-4 was ambiguous. These results indicate that the effect of tick saliva on the epidermal cells of Lyme disease-susceptible C3H/HeN mice mirrors its effect on other cells of the immune system.     

植物病毒经介体昆虫唾液腺水平传播至寄主韧皮部机制的研究进展及展望

很多植物病毒经介体昆虫以持久循回型的方式水平传播至寄主韧皮部致病，而唾液腺是介体昆虫持久传毒的重要器官，也是植物病毒在介体昆虫内循回需要克服的最后一道防线。持久性植物病毒要完成水平传播，必须突破昆虫唾液腺屏障的阻碍，因此病毒和介体昆虫间形成了“攻”与“守”的较量与对决。揭示持久性植物病毒克服昆虫唾液腺屏障，实现水平传播的机制，对病害控制具有重要意义。该文着眼于介体昆虫唾液腺在持久传毒过程中的重要功能，回顾了虫传植物病毒突破介体昆虫唾液腺侵入屏障和释放屏障的分子机制，探讨了昆虫唾液蛋白通过调节植物或昆虫的适应性和行为促进或抑制病毒水平传播的功能，为制定阻断介体昆虫传播植物病毒途径的防控策略提供理论依据。     

Evolutionary strategies and adaptations for survival between mosquito-parasitic microsporidia and their intermediate copepod hosts: a comparative examination of Amblyospora connecticus and Hyalinocysta chapmani (Microsporidia: Amblyosporidae)

The epizootiology, transmission dynamics, and survival strategies employed by two mosquito-parasitic microsporidia that utilize copepods as intermediate hosts are examined in relation to the biological attributes of their hosts and the environments in which they inhabit. Amblyospora connecticus Andreadis, 1988, a parasite of Ochlerotatus cantator (Coquillett) and Acanthocyclops vernalis (Fischer) is found in an unstable salt marsh environment that is subject to periodic flooding and drying. Both hosts have distinct non-overlapping generations. A. connecticus exhibits a well-defined seasonal transmission cycle that relies heavily on maternal-mediated transovarial transmission by female O. cantator during the summer, and horizontal transmission via the copepod host during the spring (copepod to mosquito) and fall (mosquito to copepod). Its survival strategies include: delayed virulence, low pathogenicity and high tissue specificity that allow for transstadial transmission of horizontally acquired infections and maximum spore production, reliance on living hosts throughout most of its life cycle with overwintering in the copepod, polymorphic development that is well synchronized with host physiology, and production and dissemination of infectious spores that are coincident with the seasonal occurrence of susceptible stages in each host. Hyalinocysta chapmani Hazard et Oldacre, 1975, a parasite of Culiseta melanura (Coquillett) and Orthocyclops modestus (Herrick) is found in a comparatively stable, subterranean habitat that is inundated with water throughout the year. Copepods are omnipresent and C. melanura has overlapping broods. H. chapmani is maintained in a continuous cycle of horizontal transmission between each host throughout the summer and fall but lacks a developmental sequence leading to transovarial transmission in the mosquito host. It relies on living hosts for most of its life cycle and overwinters in diapausing mosquito larvae. Transstadial transmission does not occur and there is no dimorphic development in the mosquito host. The spatial and temporal overlap of both mosquito and copepod hosts during the summer and fall affords abundant opportunity for continuous horizontal transmission and increases the likelihood that H. chapmani will find a target host, thus negating the need for a transovarial route. It is hypothesized that natural selection has favoured the production of meiospores in larval female mosquitoes rather than congenital transfer of infection to progeny via ovarian infection as a strategy for achieving greater transmission success. Analysis of the molecular phylogeny data suggest that (1) transovarial transmission and the developmental sequence leading to ovarian infection have been secondarily lost in H. chapmani, as they occur in all other closely related genera, (2) the ancestral state included complex life cycles involving transovarial transmission and an intermediate host, and (3) mosquito-parasitic microsporidia are adjusting their life cycles to accommodate host ecological conditions.     

Effect of fast protein liquid chromatography fractionated salivary gland extracts from different ixodid tick species on interleukin-8 binding to its cell receptors

Interleukin-8 plays a critical role in inflammatory processes. Hence generation of molecules with anti-IL-8 activity is likely to be important for successful feeding and for survival of the ticks. Anti-IL-8 activity was studied in saliva of three ixodid tick species--Dermacentor reticulatus (Fabricius, 1794), Rhipicephalus appendiculatus Neumann, 1901, and Amblyomma variegatum (Fabricius, 1794). The greatest activity was shown in saliva prepared from D. reticulatus. The activity was attributed to tick salivary gland molecules that bind to IL-8, preventing binding of the chemokine to its specific receptor, rather than to occupation of the IL-8 cell receptor by the tick molecules. The distribution of anti-IL-8 activity in fast protein liquid chromatography (FPLC) fractions of salivary gland extracts (SGE) derived from adult female D. reticulatus, R. appendiculatus and A. variegatum was compared directly by both ELISA and receptor-binding inhibition assays. The correspondence in results with fractions of SGE from ELISA is consistent with detection of tick molecules that inhibit IL-8 binding to its receptor. As IL-8 is an important chemoattractant and activator of neutrophils, the presence of an anti-IL-8 activity in tick saliva indicates that neutrophils play an important role in the host response to parasitism by ticks.     

蚜虫唾液主要成分及其在寄主和害虫互作中的作用

刺吸式害虫唾液里的某些成分通常是植物诱导反应中一些共有的或特异性诱导信号物质。本文综述了蚜虫取食、唾液的主要成分和取食诱发的植物防御信号传导途径,例如水杨酸途径、茉莉酸/乙烯途径,以及唾液诱导使植物防御因子上调效应的分子检测方法;同时还展望了蚜虫唾液研究的应用前景。     

Pathology of cortical invasion by Plasmodiophora brassicae in clubroot resistant and susceptible Brassica oleracea hosts

The differences in physiological response of Brassica oleracea to infection and growth of Plasmodiophora brassicae in infected tissue were studied in clubroot resistant and susceptible hosts, grown in sand solution culture artificially inoculated with spores of P. brassicae (10⁸ spores mL⁻¹). Primary (root hair) and secondary (cortical) stages of P. brassicae occurred in both resistant and susceptible hosts. Symptoms of cortical invasion by P. brassicae in resistant and susceptible hosts included cell wall breaks, presence of vesicles or inclusion bodies within the cell walls, cell wall thickening in association with plasmodesmata and enlarged and/or disorganized host nuclei. The main difference between the resistant and susceptible host reaction was the absence of degradation of the secondary thickening and cell walls of the xylem in the resistant host. This study supports the existence of an amoeboid form of the pathogen in addition to the recognized two-phase life history of P. brassicae. Furthermore, it suggests that resistance in B. oleracea does not prevent the development of this amoeboid form. A reduced number of cell wall breakages suggests movement of the amoeboid form, may have been restricted but not prevented in the resistant host.     

植物病毒与媒介昆虫互作促进其传播的研究进展

近年来植物病毒病频发,严重制约着农作物的产量与品质。绝大多数植物病毒依赖媒介昆虫进行传播,而传播的关键是病毒如何突破昆虫的肠屏障、唾液腺屏障和卵屏障等多个生物屏障。植物病毒一方面利用其外壳蛋白或非结构蛋白突破媒介昆虫的中肠屏障和唾液腺屏障;另一方面则与昆虫体内卵黄原蛋白、共生菌以及精子表面蛋白发生特异性互作,促进病毒跨越卵障碍,最终实现病毒在昆虫体内复制。此外,植物病毒还能通过侵染寄主植物影响其防御性状,间接改变媒介昆虫生理及其行为反应,促进病毒在植物间的传播。该研究对植物病毒突破昆虫生物屏障的分子机制,以及植物病毒-植物-媒介昆虫互作对于病毒传播的影响进行了综述,并对阻断病毒传播的方法进行展望。     

昆虫与弹状病毒互作的研究进展

近年来宏基因组学研究的普及大大丰富了人们对RNA病毒多样性的认识,但对这些新发现病毒的生物学特性却所知甚少。本文围绕RNA病毒中一类重要的负单链RNA病毒--弹状病毒与其昆虫寄主互作的研究进行综述,总结已发现的弹状病毒及其昆虫寄主类型,共有20个属144种弹状病毒可以感染14个属的昆虫;根据已有的系统进化研究对弹状病毒的寄主起源进行推测;并以感染黑腹果蝇Drosophila melanogaster的sigma病毒(Drosophila melanogaster sigma virus,DMelSV)为主要对象,就弹状病毒引起的CO2麻痹致死症状以及昆虫寄主对其的免疫反应研究进行总结,而在对黑腹果蝇的研究中发现很多非经典免疫通路中的新抗病毒基因,暗示存在新的抗病毒免疫通路;通过飞虱、叶蝉与其传播的植物弹状病毒以及长须罗蛉Lutzomyia longipalpis与其传播的脊椎动物病毒的互作研究,发现Toll、IMD信号通路、细胞自噬及小RNA干扰(small interfering RNA,siRNA)通路等可能与昆虫对弹状病毒的免疫反应相关。昆虫是弹状病毒主要的寄主和媒介,也是病毒遗传多样性的储主,因此更好地研究和了解昆虫寄主与弹状病毒的相互关系,有助于病毒致病和传播机制以及昆虫抗病毒免疫机理的深入研究。     

斑痣悬茧蜂对感染细菌寄主的寄生选择和传播病菌作用

本研究旨在探究斑痣悬茧蜂是否寄生被短稳杆菌Empedobacter brevis感染的寄主,以及在寄生感病与健康寄主中是否传播病菌。在斜纹夜蛾幼虫为寄主的非选择性寄生试验中,将感染不同浓度短稳杆菌液及其感染后不同时间（1～5 d）的寄主分别供斑痣悬茧蜂寄生,结果表明,当细菌浓度较高时（5.0×107和1.25×107 cfu/mL）,寄生率随感染后间隔时间延长而具有较大波动（10%～20%）,当细菌浓度较低时（3.13×106 cfu/mL）,寄生率波动较小（10%～15%）,均表现二次曲线关系;但在对感染后不同时间寄主的选择性寄生试验中,当将感染细菌后1d分别与感染2、3、4或5d的寄主幼虫同时供斑痣悬茧蜂寄生时,斑痣悬茧蜂在感菌后1与3d、1与4d的寄主之间,偏好感菌后1d的寄主幼虫,寄生的寄主数量分别是感菌后3 d的2.5倍、4 d的2.0倍。在病菌传播试验中,分别以斜纹夜蛾和黏虫为寄主,结果表明,斑痣悬茧蜂寄生斜纹夜蛾幼虫后的传染率仅约5%;但寄生感菌黏虫寄主后能够传播病菌,而且传播概率随感染后时间的增加而下降,从起初的40%逐渐降至约10%。本研究结果说明,斑痣悬茧蜂对感染短稳杆菌寄主幼虫存在一定程度的寄生选择性,并通过寄生在一定程度上可传播该病菌。     

Passive transfer of humoral resistance against adults of the tsetse fly, Glossina palpalis palpalis (Diptera, Glossinidae), in rabbits

The possibility of passive transfer of rabbit humoral immunity against tsetse fly bites was investigated for the first time. Partial immunity of recipient animals was achieved after two intravenous injections of 15 ml of serum from immunized (donor) rabbits during 48 hrs. This treatment induced an apparent increase of resistance in the passively immunized group of rabbits expressed as direct mortality ("killing effect") of sucking flies within the following 72 hr period. The immunological state of immune sera of both donors and recipients was examined by ELISA, using the water-soluble proteins of tsetse salivary glands as antigen. No direct correlation between the titre of antibodies and the killing of Glossina was detected. These results indicate that these antibodies were not the only humoral factor responsible for tsetse mortality since their titre did not substantially change in the course of 7 days while the "killing effect" had disappeared from the recipient's blood within 72 hrs.     

多堆柄锈菌侵染玉米的细胞学及超微结构特征

为明确玉米对多堆柄锈菌Puccinia polysora侵染后病理反应的细胞学特征,利用扫描和透射电镜技术分析了玉米自交系与多堆柄锈菌互作中二者的细胞变化过程。多堆柄锈菌对玉米的侵染主要以直接穿透叶片表皮侵入为主,少量可从气孔和细胞间隙侵入。接种后,病菌夏孢子在感病自交系叶片上快速并大量萌发,在叶表生长蔓延并侵入表皮组织细胞,7 d后形成夏孢子堆;在抗病自交系上,病菌萌发、菌丝生长均受到明显抑制,少量入侵的病菌也由于寄主细胞死亡而导致菌丝和夏孢子干瘪死亡。侵染早期在感病寄主细胞间隙出现菌丝并穿透细胞壁,在胞内产生分枝菌丝,此时寄主细胞结构正常;随着菌丝进一步扩展,叶绿体等结构发生紊乱,被侵染细胞逐渐死亡。在抗病自交系上,接菌24 h后寄主即出现过敏性坏死反应,侵入位点与周围细胞快速坏死,抑制菌丝生长蔓延;叶绿体中清晰可见深色颗粒状物质;72 h后细胞壁外侧产生大量致密的深色结晶体,应为与抗病反应相关的酚类物质。表明抗多堆柄锈菌的玉米材料可能存在2种抗病途径,即寄主与病菌互作中由分子识别引起的免疫反应和病菌侵入后的系统防卫反应。     

Immune response of the long-tailed field mouse (Apodemus sylvaticus) to tick-borne encephalitis virus infection.

The immune response following infection with a virulent strain of Central European encephalitis (CEE) virus in a natural host, long-tailed field mouse (Apodemus sylvaticus L.) and white laboratory-bread ICR mouse, was compared. Viraemia was demonstrated in ICR mice after intraperitoneal infection with a dose of 10(5) LD50/0.5 ml. The virus titres were high in the spleen and, particularly, in the brain. In A. sylvaticus the virus was detected in the blood and spleen, but not in the brain. CEE virus multiplied in peritoneal macrophages from ICR mice, but not from A. sylvaticus. The infection induced a strong interferon response in both hosts. The natural killer (NK) cell activity increase was twice as high in A. sylvaticus compared to ICR mice. The neutralization antibodies appeared sooner in A. sylvaticus and reached higher titres in the early phases of infection.     

Comparison of the surface coat proteins of the pine wood nematode appeared during host pine infection and in vitro culture by a proteomic approach

Pine wilt disease, caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, has become of worldwide quarantine concern in recent years. Here, we disclosed the surface coat (SC) proteins of the PWN which are thought to be one of the key components in pine wilt development. This is the first report that focused on the SC proteins and thoroughly identified those proteins of a plant-parasitic nematode using the proteomic approach. In this study, SC protein profiles were compared for PWNs grown on the fungus Botrytis cinerea and in host pine seedlings. The results demonstrated that the gross amount of PWN SC proteins drastically increased during infection of the host pine. Thirty-seven protein bands showed significant quantity differences between fungus-grown and host-origin PWNs, and were used for identification by matrix-assisted laser desorption ionization time of flight mass spectrometry analysis. These included several proteins that are presumed to be involved in the host immune response; for example, regulators of reactive oxygen species (ROS) and a ROS scavenger. These results might suggest that the PWN SC proteins are crucial in modulating or evading host immune response. Our data provide a new insight into the mechanism of pine wilt disease and the biological role of the SC proteins of plant-parasitic nematodes.