禾谷缢管蚜在三个小麦品种上取食行为的EPG比较

应用刺探电位图谱（EPG）技术对禾谷缢管蚜Rhopalosiphum padi在不同小麦品种(Ww2730、小偃22和Batis)苗期的取食行为进行了比较研究。结果表明：在小偃22上蚜虫开始取食的第1次刺探时间最晚,且持续时间最短;在Ww2730上取食受到机械阻力的个体最多,且出现F波的几率和持续时间最长;两品种上蚜虫在木质部主动摄取汁液（G波）花费的时间最长。 在Batis上,蚜虫口针第1次到达韧皮部时需要分泌较多水溶性唾液(E1波),但随后只需分泌较少的水溶性唾液就可以成功取食,而且被动吸食韧皮部汁液的时间（E2波）最长。蚜虫口针在到达小偃22韧皮部取食之前,出现多次的口针试探、回撤,并且多次、多量分泌水溶性唾液（E1波）;虽然蚜虫在小偃22上口针最先到达韧皮部,但被动吸食韧皮部汁液的时间（E2波）最短。由此得出结论：小偃22表皮部、韧皮部存在阻碍禾谷缢管蚜取食的物理和生化因素; 禾谷缢管蚜在Ww2730取食遇到更多的是细胞间机械阻力; Batis是较感蚜的品种。     

园艺矿物油乳剂对亚洲柑橘木虱成虫刺吸行为的影响

目的】亚洲柑橘木虱Diaphorina citri是传播柑橘最重要病害黄龙病(huanglongbing, Candidatus Liberibacter asiaticus)的媒介昆虫,树冠喷施园艺矿物油(horticultural mineral oil, HMO)可以减少其在柑橘上的取食和产卵。本研究旨在探索植物组织内残留的园艺矿物油如何影响亚洲柑橘木虱成虫的取食行为。【方法】采用直流型刺吸电位仪(DC-EPG Giga-8)记录亚洲柑橘木虱成虫12 h内在喷施不同浓度(0.25%, 0.5%, 1%和2%, v/v)HMO(nC24)乳剂和清水(对照)时柑橘嫩叶上的刺探和取食行为并转换为波形信号,分析比较各处理的波形参数。【结果】不同浓度HMO乳剂处理显著提高了亚洲柑橘木虱成虫在柑橘叶片上的C波(路径波)持续时间百分比,显著减少了E2波(韧皮部吸食汁液)持续时间百分比,而对D(口针第一次接触韧皮部组织), E1(韧皮部出现唾液分泌) 和G(木质部吸食汁液) 波持续时间百分比没有显著影响。HMO处理组C波的每成虫波形累积时间(WDI)显著长于对照组,而各浓度处理之间没有显著差异,但D波WDI明显短于对照组。随着HMO浓度的增加, E1, E2和G波WDI显著下降,2% HMO处理组E1, E2和G波WDI明显低于其他浓度处理组。HMO乳剂处理明显减少了刺探次数;在HMO处理中,每成虫刺探次数和每成虫各波形次数均显著少于对照组,其中以1%和2% HMO乳剂处理最少。HMO乳剂处理组np波 (非刺探波)、D波、E2波的每成虫单次波形持续时间(WDE)均短于对照组,其中2% HMO乳剂处理后,D, E1和E2波WDE显著短于其他浓度处理和对照。同时,2% HMO乳剂处理推迟了从口器接触叶片到第1次开始刺探的时间和从口器接触叶片到第1次刺探到韧皮部的时间。【结论】结果表明,2% HMO乳剂显著减少亚洲柑橘木虱成虫在柑橘叶片上的取食次数和有效取食时间,同时还缩短了其在韧皮部分泌唾液和吸食汁液时间,因此可推荐矿物油用于柑橘黄龙病和亚洲柑橘木虱的综合防治体系。同时初步推断矿物油的作用机制可能是其进入植物气孔阻止植物挥发性物质的释放,抑制或掩盖柑橘叶片表面的挥发性物质从而减少了亚洲柑橘木虱在寄主上取食。     

番茄黄化曲叶病毒对烟粉虱MED隐种取食行为的影响

为探究番茄黄化曲叶病毒（TYLCV）对烟粉虱刺探取食行为的影响并研究其与烟粉虱之间的互作关系。利用EPG记录了感染TYLCV的MED隐种在番茄寄主上的取食波形,并与未感染TYLCV的MED隐种的取食波形进行了比较研究。结果表明,烟粉虱开始取食行为后,带毒MED和未带毒MED平均刺探次数和非刺吸时间占总取食时间的比例方面没有显著差异。带毒MED从口针刺探开始至到达韧皮部的平均时间为46 min,从记录开始至口针第一次到达韧皮部的平均时间为135 min,未带毒MED相对应的平均时间分别为19 min和25 min,两者结果差异显著;在烟粉虱取食番茄韧皮部阶段,带毒和未带毒MED的唾液分泌总时间分别为9 min、3 min,第一次韧皮部持续吸食平均时间分别为143 min、26 min,带毒MED显著高于未带毒MED。带毒MED在第一次持续吸食至第一次取食结束的平均时间为15 min,显著高于未带毒MED的4 min。综上所述,带毒MED完成整个刺探取食行为的平均时间较未带毒MED显著增加,特别是唾液的分泌时间以及第一次在韧皮部的持续吸食时间。表明,TYLCV能促进MED加强唾液分泌和韧皮部取食,从而促进病毒的传播。     

吡虫啉对3种体色型烟蚜取食行为的影响

为探究吡虫啉对红色型、绿色型和褐色型3种体色型烟蚜取食行为的影响,利用刺探电位图谱(EPG)技术比较3种体色型烟蚜在吡虫啉处理后的烟苗上的取食行为。结果表明:33.3 mg/L吡虫啉处理烟苗后,3种体色型烟蚜的非刺探总时间(np波)均长于清水处理的绿色型烟蚜,其中褐色型烟蚜最长,且与清水处理的绿色型烟蚜存在显著性差异(P0.05);刺探过程中,3种体色型烟蚜在烟苗上口针遇到阻力(F波)的时间均长于清水处理的绿色型烟蚜,但处理间差异不显著;木质部取食阶段,褐色型烟蚜的G波平均持续时间最长;韧皮部取食阶段,3种体色型烟蚜口针在韧皮部分泌唾液(E1波)和被动吸食汁液(E2波)的总持续时间均短于清水处理的绿色型烟蚜,其中被动吸食汁液的时间红色型为15.87±9.33 min,绿色型为29.89±8.79 min,而褐色型仅为6.38±2.15 min。因此,吡虫啉处理烟苗后能明显抑制烟蚜对烟草韧皮部的取食。     

灰飞虱取食行为刺吸电位波形的初步研究

利用刺吸电位技术(EPG)研究了灰飞虱Laodelphax striatellus(Fallén)EPG波形与其取食行为之间的对应关系,并对不同虫龄、性别灰飞虱的取食行为进行EPG特征参数的比较。结果表明,灰飞虱取食行为的EPG波形可分为NP、N1、N2-a、N2-b、N3、N4和N5 7种类型,波谱的形状和频率与已报道的褐飞虱Nilaparvata lugens(Stl)的EPG取食波形基本相同。首次发现灰飞虱在口针刺探水稻维管束进行唾液分泌时可明显分为两个阶段即N2-a和N2-b。N2-a波表示口针移动并伴随唾液的分泌;N2-b波与灰飞虱持续分泌唾液的行为有关。经对比,不同虫龄、性别灰飞虱的EPG取食波形基本相同,但EPG特征参数存在差异。雌虫刺探次数显著多于雄虫和低龄若虫;成虫口针在临近韧皮部细胞外移动的时间显著长于若虫。     

烟粉虱和温室粉虱在甘蓝上的刺探取食行为比较

利用刺吸电波图技术研究B型、ZHJ_1型烟粉虱Bemisia tabaci(Gennadius)和温室粉虱Trialeurodes vaporariorum(Westwood)在甘蓝上的取食行为,将3种粉虱的电波图进行比较,其中,B型和ZHJ_1型烟粉虱记录到np,C,pd,E1,E2,F和G波7种波形,温室粉虱只记录到刺探波形,少有取食波形。B型的20个记录中只有1个没有持续吸食波形;ZHJ_1型的25个记录中有10个记录没有持续吸食波形;温室粉虱没有持续吸食记录。甘蓝叶片韧皮部的结构或汁液的化学成分与温室粉虱的抗性密切相关。     

番石榴精油对柑橘木虱刺吸取食行为的影响

利用刺探电位技术(electrical penetration graph,EPG).比较测定了柑橘木虱Diaphorina citri Kuwayama成虫在经番石榴精油处理和对照柑橘叶片上的刺吸取食行为.结果表明,在正常(对照)叶片上,柑橘木虱成虫的刺吸取食有8种不同波形,表现为迅速进入取食过程,非刺探活动时间和唾液分泌较少,为韧皮部取食,没有木质部取食.而柑橘叶片经1μw/mL番石榴精油处理后,柑橘木虱成虫刺吸取食行为显著改变,表现为非刺探活动和唾液分泌增加,韧皮部取食时间极显著下降,下降率为62.12%,并出现木质部取食.不同时段柑橘木虱的有效取食时间动态结果显示,番石榴精油在第1 h使柑橘木虱完全进入木质部取食,但随后逐步恢复韧皮部取食.     

假眼小绿叶蝉（Empoasca vitis Gothe）在不同品种茶树上的取食行为

应用可视DC-EPG方法研究了假眼小绿叶蝉(Empoasca vitis Gothe)在9个品种茶苗上的口针刺探行为,共发现并初步确定了7种主要波型,即A波、S波、C波、E波、F波和R波、以及非刺探波NP波。A波为刺探波,S波为口针向韧皮部刺探和进入韧皮部中的分泌唾液波,C波为口针到达韧皮部之前的主动取食波,E波和F波为口针在韧皮部中吸收波,R波为取食间歇波。以该叶蝉在不同品种茶树上的平均刺探次数和各波形平均持续时间为指标,或者以其在不同品种茶树上含有各波形的单次刺探平均持续时间为指标,分别进行聚类分析,均将9个品种分为3个不同的组。S、E和F波对应着假眼小绿叶蝉在茶树上的主要取食活动,可能与茶树抗假眼小绿叶蝉的取食密切相关。以S、E和F各波的平均持续时间、以及含有各波形的单次刺探的平均持续时间为指标,对品种的抗性强弱排序,评判9个茶树品种抗叶蝉取食能力由强至弱的顺序为:龙井长叶、黄旦、政和大白茶、黔湄601、红芽佛手、中茶102、中茶302、龙井43和安吉白茶。该顺序与田间查得的9个品种茶树上假眼小绿叶蝉种群密度由小到大的顺序一致,表明DC-EPG方法简捷、可信度高,可作为检测茶树品种对叶蝉抗性的有效手段之一。     

褐飞虱取食人工饲料的刺吸电位波形鉴别

昆虫刺吸电位技术(Electrical penetration graph,EPG)与人工饲料体系相结合,能为昆虫与植物相互关系中的行为、生理/毒理和分子机制等多个领域提供定量的研究手段。由于人工饲料体系与植物组织在结构和成分方面有较大差异,以现有的植物EPG波形作为参照标准的研究方法存在局限性。本研究应用EPG技术记录褐飞虱Nilaparvata lugens(Stl)在人工饲料上的取食行为,结合刺孔检测和饲料染色等方法,鉴定各波形的生物学意义。研究发现,人工饲料EPG波形与褐飞虱取食水稻产生的7种EPG波形相比存在差异。人工饲料EPG波形包括4种基本波形:np波,非刺探波;P波,口针刺入饲囊膜;S波,口针分泌唾液,并在人工饲料中移动;I波,吸食人工饲料。研究结果可为建立准确的人工饲料-刺吸电位技术体系提供基础。     

植物和刺吸式口器昆虫的诱导防御与反防御研究进展

刺吸式口器昆虫在长期的进化过程中形成特殊的口针结构,用于专门吸食植物韧皮部筛管细胞的汁液成分.以蚜虫为例,它们在取食过程中分泌的胶状唾液和水状唾液将有效的降低植物防御反应,其中水状唾液包含的大量酶类不仅可以帮助蚜虫穿刺植物韧皮部,刺探到筛管细胞,同时也是植物感受蚜虫为害的激发因子,诱导出植物防御反应和相关抗性基因的表达...     

Identification of phloem sieve elements as the site of resistance to silverleaf whitefly in resistant alfalfa genotypes

Experiments were conducted to locate the plant tissue where resistance is expressed against silverleaf whitefly, Bemisia argentifolii Bellows and Perring (Homoptera: Aleyrodidae), in alfalfa, Medicago sativa L. (Fabaceae), genotypes previously shown to have high levels of resistance against this pest. Previous work demonstrated that resistance in the resistant alfalfa genotypes was expressed primarily as high first‐instar mortality; consequently this study focused on first‐instar nymphs. Examination of stylets in cleared leaf tissue indicated that first‐instar nymphs located vascular bundles with equal success on resistant and susceptible alfalfa genotypes. Furthermore, direct current electrical penetration graphs (DC‐EPG) indicated that sieve elements were penetrated and phloem ingestion behavior was initiated with equal success on resistant and susceptible genotypes. Thus, the mechanism of resistance does not reside in tissues encountered by the stylets prior to penetrating a phloem sieve element. Honeydew production (as a proxy for ingestion) was greatly reduced on two resistant genotypes compared to the two susceptible genotypes. The frequency distribution of honeydew production was bimodal, indicating that most individuals on the resistant genotypes produced little or no honeydew while some produced as much honeydew as whiteflies on the susceptible genotypes. This indicates that expression of resistance is an all‐or‐nothing phenomenon; an individual nymph either encounters resistance and cannot sustain ingestion or it does not encounter resistance and ingests just as well as on a susceptible plant. Intermediates are rare. DC‐EPGs indicate that phloem ingestion behavior is significantly reduced on two of the resistant genotypes compared to the susceptible genotypes. The primary reason for this appears to be more frequent termination of phloem ingestion behavior on at least one of the resistant genotypes. On one of the resistant genotypes, the productivity of EPG‐measured phloem ingestion behavior (honeydew produced per min of phloem ingestion behavior) was reduced compared to a susceptible control.     

Leafhoppers on leaves: An analysis of feeding behavior using conditional probabilities

Feeding behavior of four deltocephaline leafhoppers,Graminella nigrifrons, G. oquaka, Amblysellus grex, andDalbulus maidis on maize and johnsongrass was analyzed using an electronic monitoring device. Five distinct waveform patterns were identified: secretion of sheath saliva (salivation), nonvascular probing, nonsieve element ingestion, x-waveform, and phloem ingestion. Waveforms were associated with feeding activities by correlation with light microscopic examination of salivary sheath termination points in leaf tissue and analysis of honeydew excreted by monitored leafhoppers. In previous studies x-waveforms have been reported to occur only when the stylets of homopterans are in contact with the phloem; the function of x-waveforms is poorly understood. There were no differences in time spent salivating or ingesting from nonsieve elements amongG. nigrifrons, G. oquaka andA. grex on either plant.D. maidis differed from other species in phloem probing and feeding behavior; only a small proportion produced x-waveforms, although those that did spent significantly more time in this behavior than other species. Also,D. maidis spent more time than other leafhoppers ingesting from tissues other than sieve elements. Kinetic diagrams of transition probabilities show that probing activities of all species were not random regarding the sequence of behaviors culminating in phloem ingestion. Thirty-five percent ofG. nigrifrons x-waveforms were followed by nonsieve element ingestion. This was consistent with observations showing that salivary sheaths of leafhoppers producing x-waveforms sometimes do not terminate in the phloem, but rather in nearby cells. Phloem ingestion was always preceeded by x-waveforms. The quantitative differences in probing behavior are discussed in relation to ability of these leafhoppers to transmit the phloem-associated maize chlorotic dwarf waikavirus.     

Aphid activities during sieve element punctures

Aphid salivation in sieve elements and phloem sap ingestion were linked to waveforms in the Electrical Penetration Graph (EPG). Non-viruliferousRhopalosiphum padi (L.) (Hemiptera, Aphididae) on barley yellow dwarf virus (BYDV) infected wheat could acquire the virus, which was used as an indication for phloem sap ingestion, whereas virus inoculation by viruliferous aphids on healthy plants was associated with salivation in sieve elements or other phloem cells. Probing was monitored and the waveforms recorded were related to ELISA results of test plants. The EPG patterns A, B, and C are indicative of the stylet pathway phase, whereas patterns E1 and E2 reflect the phloem (sieve element) phase with an unknown activity (E1) or with ingestion and concurrent salivation (E2). Aphids showing pathway and E1 rarely acquired virus, suggesting that little or no phloem sap ingestion can occur during these patterns, whereas those showing additionally pattern E2 did so substantially, indicating phloem sap ingestion. The main pattern related to virus inoculation was E1, although some aphids were able to inoculate plants during pathway. Pattern E1 clearly reflects the most important salivation into sieve elements. Pattern E2 had no clear contribution to virus inoculation, supporting the present hypothesis that during this pattern the saliva is mixed with the phloem sap in the single canal at the stylet tips and ingested immediately, without reaching the plant tissue. Sustained sap ingestion did not affect virus inoculation. So, BYDV inoculation mainly occurs during the first period of a sieve element puncture which is always formed by E1. Implications on persistent virus transmission are discussed.     

Salivation into sieve elements in relation to plant chemistry: the case of the aphid Sitobion fragariae and the wheat, Triticum aestivum

Extended sieve element salivation (E1 waveform in the electrical penetration graph) is a characteristic activity during early sieve element punctures, particularly in resistant plants. In order to explore a chemically-mediated mechanism of resistance associated with sieve element salivation, we compared the pattern of feeding behaviour of the aphid, Sitobion fragariae (Walker), on two cultivars of the wheat Triticum aestivum L., with different concentrations of hydroxamic acids (Hx). During 24 h of electronic monitoring, aphids dedicated over 50% of the total time to phloem ingestion from the sieve elements. Total time allocated to E1 in the experiment, time to first E1 within the experiment, time allocated to E1 before a sustained phloem ingestion (E2) and the contribution of sieve element salivation to the phloem phase (E1/[E1+E2]) were significantly higher in the high-Hx cultivar. The increased salivation in plants with higher contents of Hx suggests the existence, at least in this system, of a chemically-mediated sieve element constraint.     

Mi‐mediated aphid resistance in tomato: tissue localization and impact on the feeding behavior of two potato aphid clones with differing levels of virulence

The Mi‐1.2 gene in tomato, Solanum lycopersicum L. (Solanaceae), confers resistance against several herbivores, including the potato aphid, Macrosiphum euphorbiae (Thomas) (Hemiptera: Sternorrhyncha: Aphididae) and the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Sternorrhyncha: Aleyrodidae). Previous studies on the tissue localization of resistance have given varying results; whitefly resistance was attributed to factors localized in the mesophyll or epidermis, whereas aphid resistance was attributed to factors localized in the phloem. Our study utilizes the direct current electrical penetration graph (DC‐EPG) technique to compare aphid feeding behavior on resistant (Mi‐1.2+) and susceptible (Mi‐1.2?) tomato plants. This study also compares the impact of resistance on the feeding behavior of two aphid clones that vary in their virulence, or their ability to survive and reproduce on resistant plants. Previous work had shown that the avirulent WU11 clone is almost completely inhibited by resistance, whereas the semi‐virulent WU12 clone can colonize resistant hosts. Here, DC‐EPG analysis shows that both aphid clones take longer to initiate cell sampling and to establish a confirmed sieve element phase on resistant plants than on susceptible hosts, and have shorter ingestion periods on resistant plants. However, the magnitude of these deterrent effects is far less for the semi‐virulent clone than for the avirulent aphids. In particular, the WU12 clone is less sensitive to factors that limit sieve element ingestion, showing shorter non‐probe duration and rapidly establishing sustained phloem ingestion on resistant plants when compared to the WU11 clone. We conclude that, in addition to previously described factors in the phloem that inhibit ingestion, Mi‐mediated aphid resistance also involves factors (possibly in the mesophyll and/or epidermis) that delay initiation of phloem salivation, and that act in the intercellular spaces to deter the first cell sampling. Furthermore, the relative effectiveness of these components of resistance differs among insect populations.     

Foraging ofRhagoletis pomonella flies in relation to interactive food and fruit resources

Feeding behaviour of the lettuce root aphidPemphigus bursarius was monitored electronically on six lettuce varieties. Aphids, monitored for a minimum of 5 h on lettuce roots, performed recognizable feeding behaviour on the susceptible varieties, Webbs Wonderful and Borough Wonder including non probing, pattern A start of new penetration, C (stylet pathway activities) sometimes including clear B-waves (salivary sheath secretions), potential drops (intracellular penetrations), intracellular E (phloem ingestion) distinguishing E₁ and E₂ (Tjallingii, 1990) and occasionally xylem ingestion. Although aphid probing was recorded on the resistant varieties Lakeland and Grand Rapids continued penetration to the phloem elements was deterred and the time spent ingesting phloem was short. Aphids rarely probed the resistant varieties Avoncrisp and Avondefiance and all attempted penetrations quickly terminated.     

Analysis by DC–EPG of the resistance to Bemisia tabaci on an Mi-tomato line

The tomato Mi gene confers resistance to nematodes, Meloidogyne spp., and to the potato aphid, Macrosiphum euphorbiae (Thomas). Previous greenhouse choice assays with Bemisia tabaci (Gennadius) showed that tomato commercial varieties carrying this gene had significantly lower values of host suitability and whitefly reproduction than varieties lacking Mi. This indicated that Mi, or another gene in its region, could regulate partial resistance. In order to characterise this resistance, probing and feeding behaviour of Bemisia tabaci B-biotype was studied with DC Electrical Penetration Graph (EPG) technique on the near-isogenic tomato lines Moneymaker (without Mi) and Motelle (carrying Mi). Significant differences (P < 0.05) between tomato lines were found in EPG parameters related to epidermis and/or mesophyll tissues. On Motelle, a lower percentage of whiteflies achieved phloem phase and they made more probes before attaining first phloem phase, had a higher ratio (number of probes before first phloem phase)/(total number of probes), had a longer total duration of non-probing time, and a longer time before making the first intracellular puncture and before making the first phloem phase. In contrast, most of the parameters related to phloem phase were found not to differ significantly between these near-isogenic lines. The behavioural data strongly suggest that the partial resistance in the variety Motelle is due to factors in the epidermis and/or mesophyll that inhibit the whiteflies from reaching phloem sieve elements. However, once the stylets reach a sieve element, whitefly behaviour did not differ between the two varieties. Thus, phloem sap of the two varieties appears to be equally acceptable to the whiteflies. Further studies are necessary to provide a better understanding of these mechanisms of resistance to whiteflies in tomatoes.     

Ultrastructure of compatible and incompatible interactions in phloem sieve elements during the stylet penetration by cotton aphids in melon

Resistance of the melon line TGR‐1551 to the aphid Aphis gossypii is based on preventing aphids from ingesting phloem sap. In electrical penetration graphs (EPGs), this resistance has been characterized with A. gossypii showing unusually long phloem salivation periods (waveform E1) mostly followed by pathway activities (waveform C) or if followed by phloem ingestion (waveform E2), ingestion was not sustained for more than 10 min. Stylectomy with aphids on susceptible and resistant plants was performed during EPG recording while the stylet tips were phloem inserted. This was followed by dissection of the penetrated leaf section, plant tissue fixation, resin embedding, and ultrathin sectioning for transmission electron microscopic observation in order to study the resistance mechanism in the TGR. The most obvious aspect appeared to be the coagulation of phloem proteins inside the stylet canals and the punctured sieve elements. Stylets of 5 aphids per genotype were amputated during sieve element (SE) salivation (E1) and SE ingestion (E2). Cross‐sections of stylet bundles in susceptible melon plants showed that the contents of the stylet canals were totally clear and also, no coagulated phloem proteins occurred in their punctured sieve elements. In contrast, electron‐dense coagulations were found in both locations in the resistant plants. Due to calcium binding, aphid saliva has been hypothesized to play an essential role in preventing/suppressing such coagulations that cause occlusion of sieves plate and in the food canal of the aphid's stylets. Doubts about this role of E1 salivation are discussed on the basis of our results.     

On the Protein Inclusions in Sieve Elements of Sterculia urens

Protein inclusions in the sieve tubes of Sterculia urens havebeen examined by light microscopy. Conspicuous inclusions wereobserved in both primary and secondary phloem of young branches,but not in the main stem, where smaller inconspicuous spheroidswere found. It is suggested that spheroidal protein inclusionsmay have previously gone unnoticed in other plants. phloem, evolution of phloem, sieve element, sieve element inclusion