应用EPG技术分析不同品种苜蓿对苜蓿斑蚜的抗性

【目的】明确苜蓿斑蚜Therioaphis trifolii对不同品种苜蓿Medicago sativa的取食行为,探明抗蚜因子和抗虫位点,筛选抗蚜苜蓿品种。【方法】利用刺吸电位技术(electrical penetration graph, EPG)记录苜蓿斑蚜成蚜在10个苜蓿品种上的取食行为,并基于聚类分析方法筛选合适的EPG参数进行抗蚜性评价。【结果】苜蓿斑蚜成蚜在取食苜蓿过程中呈现np波、pd波、A波、B波、C波、E波、F波和G波8种波形,其中E波、F波和G波的总时间在不同品种间表现出显著差异。在5 h测试期间,E波持续时间在敖汉上最长,金皇后和中苜3号上次之,草原2号和阿尔冈金上最短;F波总时间以准格尔、阿尔冈金和金皇后上最长,敖汉、德宝和草原2号上最短,说明前者具有较强的机械抗性而后者机械抗性较弱。以第1次刺探持续时间、总刺探时间、F波总时间、C波总时间、E波总时间作为聚类分析的指标,10个苜蓿品种被聚成3类：阿尔冈金、草原2号、WL168HQ、德宝、中苜2号和新牧2号为第Ⅰ类,金皇后、中苜3号和准格尔为第Ⅱ类,敖汉为第Ⅲ类。【结论】苜蓿斑蚜成蚜在不同苜蓿品种上的取食行为存在差异,草原2号、阿尔冈金和WL168HQ在叶表皮、叶肉和韧皮部层次对苜蓿斑蚜成蚜存在抗性,金皇后在叶表皮和叶肉部位表现出抗性;苜蓿斑蚜成蚜在敖汉上能够长时间刺吸取食,因此敖汉抗性最弱。本研究为深入探讨苜蓿抗虫机理和蚜虫综合治理提供了理论参考。     

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。     

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。     

基于EPG的三种刺吸式害虫在苹果苗上的取食行为比较

【目的】苹果绵蚜Eriosoma lanigerum、绣线菊蚜Aphis citricola和梨网蝽Stephanitis nashi是苹果园的一类重要害虫,它们以刺吸式口器对苹果树造成不同程度的危害。本研究旨在明确这3种刺吸式昆虫在苹果树上取食行为差异。【方法】利用刺吸电位(electrical penetration graph, EPG)技术对苹果绵蚜和绣线菊蚜成虫在苹果苗韧皮部和非韧皮部上的EPG指标,以及苹果绵蚜、绣线菊蚜和梨网蝽成虫苹果苗上的取食行为进行了比较分析,分析了这3种害虫的成虫在苹果苗上取食8 h各种波形平均持续时间的占比。【结果】结果表明,苹果绵蚜和绣线菊蚜成蚜在苹果苗上均产生6种取食波形,即非刺探波(np)、路径波(C)、意外穿刺细胞非主动取食细胞波(pd)、木质部取食波(G)、韧皮部唾液分泌波(E1)和韧皮部取食波(E2);而梨网蝽成虫取食过程中只产生非刺探波(np)、表皮刺穿波(A)、叶肉细胞取食波(Gc)和木质部取食波(E)4种波形。从蚜虫在苹果苗非韧皮部上取食的EPG指标看,苹果绵蚜成蚜pd波平均时间显著高于绣线菊蚜的,而刺探次数、np波总时间和pd波次数均显著低于绣线菊蚜的。从蚜虫在苹果苗韧皮部上取食的EPG指标来看,除第1次出现E2波的时间外,各指标没有显著差异。从3种害虫在苹果苗上取食8 h各波形的占比来看,梨网蝽成虫的np波总时间所占比例最高(53%),其次是绣线菊蚜成虫的(24%),苹果绵蚜成虫的最低(为1%)。同时,苹果绵蚜和绣线菊蚜成虫取食波为E2波,所占总时间比例分别为35%和25%,而梨网蝽的取食波为Gc波(占总时间比例为36%)和E波(占总时间比例为11%)。【结论】本研究阐释了苹果园刺吸式口器害虫的生态位分离和取食行为学机制,为果园刺吸式口器害虫的综合治理提供了理论依据。     

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。     

Probing behavior of the tea green leafhopper on different tea plant cultivars

The probing behaviors of the tea green leafhopper, Empoasca vitis (Gothe), on 9 tea cultivars were studied using video-text Direct Current-Electrical Penetration Graph, i.e., DC-EPG. The following 7 types of waveforms produced by the leafhopper stylet probing were determined: A, stylet pathway formation; S, salivation when stylets pierce into and stay in phloems; C, active ingestion before stylets reach phloems; E and F, passive ingestion in phloems; R, the insect resting with its stylet inserted into the leaf tissue and NP without probing. The 9 tested tea cultivars were categorized into 3 groups by the cluster analysis according to the number of probes per insect, waveform durations, or duration per probe of various waveforms on different tea cultivars. Waveforms S, E and F correlated to the main feeding activity of the leafhopper and may provide valuable information on predicting the resis-tance level of the tea plants to the leafhopper. The resistance level of the 9 tea cultivars to the leafhopper was ranked based on the durations of waveforms S, E and F, as well as the duration per probe including various waveforms. The ranking order of the resis-tance was: Longjingchangye > Hangdan > Zhenghedabaicha > Qianmei 601 > Hongyafoshuo > Zhongcha 102 > Zhongcha 302 > Longjing 43 > Anjibaicha, which corresponded to the resistance level determined by the population density (infestation) of the leaf-hopper on the 9 tea cultivars in the tea fields. Our study suggests that this simple and convenient DC-EPG technique might have great potential as a reliable tool to predict the resistance of tea cultivars to the tea leafhopper.     

Probing behavior of the tea green leafhopper on different tea plant cultivars

The probing behaviors of the tea green leafhopper, Empoasca vitis (Gothe), on 9 tea cultivars were studied using video-text Direct Current-Electrical Penetration Graph, i.e., DC-EPG. The following 7 types of waveforms produced by the leafhopper stylet probing were determined: A, stylet pathway formation; S, salivation when stylets pierce into and stay in phloems; C, active ingestion before stylets reach phloems; E and F, passive ingestion in phloems; R, the insect resting with its stylet inserted into the leaf tissue and NP without probing. The 9 tested tea cultivars were categorized into 3 groups by the cluster analysis according to the number of probes per insect, waveform durations, or duration per probe of various waveforms on different tea cultivars. Waveforms S, E and F correlated to the main feeding activity of the leafhopper and may provide valuable information on predicting the resis-tance level of the tea plants to the leafhopper. The resistance level of the 9 tea cultivars to the leafhopper was ranked based on the durations of waveforms S, E and F, as well as the duration per probe including various waveforms. The ranking order of the resis-tance was: Longjingchangye > Hangdan > Zhenghedabaicha > Qianmei 601 > Hongyafoshuo > Zhongcha 102 > Zhongcha 302 > Longjing 43 > Anjibaicha, which corresponded to the resistance level determined by the population density (infestation) of the leaf-hopper on the 9 tea cultivars in the tea fields. Our study suggests that this simple and convenient DC-EPG technique might have great potential as a reliable tool to predict the resistance of tea cultivars to the tea leafhopper.     

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。     

角倍蚜干母的刺探取食行为分析

本文采用EPG技术监测角倍蚜Schlechtendalia chinensis(Bell)干母在第一寄主植物盐肤木(Rhus chinensis Mill)上的取食行为.获得干母在盐肤木小叶及叶翅上的取食波形7个,分别为np、C(A+B+C)、pd、E1、E2、F、G波.对干母在盐肤木小叶或叶翅上的取食部位组织进行切片...     

Alfalfa cyclitols in the honeydew of an aphid

The free cyclitols pinitol, ononitol and myo-inositol occur in the honeydew (excreta) of pea aphids (Acyrthosiphon pisum) which feed on pea aphid-susceptible alfalfa (Medicago sativa cv Caliverde). These cyclitols also occur in the leaves and stems of alfalfa. Aphids were incapable of de novo synthesis of these cyclitols. Honeydew production by the pea aphid results from ingesting phloem-sap, so the occurrence of cyclitols in honeydew results from their translocation in the phloem. The relatively high content of myo-inositol in honeydew indicates that it is selectively translocated. The most abundant alfalfa cyclitol, pinitol, had no effect on aphid feeding behavior at concentrations up to 1% (w/v; artificial diet).     

The role of sinigrin in host plant recognition by aphids during initial plant penetration

Plant penetration behaviour (probing) of the cabbage aphid, Brevicoryne brassicae, and the pea aphid, Acyrthosiphon pisum, was studied on excised leaves of broad beans, Vicia faba, kept in water or in a 1% aqueous solution of sinigrin. Using the DC EPG (Electrical Penetration Graph) technique it was shown that the cabbage aphid on sinigrin-untreated bean leaves showed numerous short probes into epidermis and mesophyll. None of these aphids showed either phloem salivation or ingestion waveforms on untreated leaves. In contrast, on sinigrin-treated bean leaves, 35% of the probing time was spent on phloem sap ingestion (E2) and almost all aphids reached phloem vessels and started feeding. The duration of phloem salivation before phloem ingestion and the mean duration of phloem ingestion periods were similar on a host and a sinigrin-treated non-host plant. However, the total probing time by B. brassicae was 10% longer, the total phloem sap ingestion time was twice as long, and the time to the first phloem phase within a probe was three times shorter on the host plant compared to sinigrin-treated broad beans. Acyrthosiphon pisum also responded to the addition of sinigrin to broad beans, but in this case sinigrin acted as a deterrent. On sinigrin-treated leaves, A. pisum terminated probes before ingestion from phloem vessels, and none of these aphids showed phloem salivation and ingestion on treated leaves. Glucosinolates were detected in the mesophyll cells of the brassicaceous plant, Sinapis alba. Based on this finding and in addition to the foregoing EPG analysis of aphid probing on these plants and broad beans, our hypothesis is that aphids may recognise their host plants as soon as they probe the mesophyll tissue and before they start ingestion from phloem vessels.     

基于模糊识别和刺探电位技术鉴定小麦种质资源抗蚜性

利用田间抗蚜性鉴定模糊识别技术,结合室内刺探电位(EPG)植物抗性鉴定技术,比较分析了不同遗传背景的小麦种质资源的抗蚜性,为筛选新型小麦抗蚜种质材料提供依据。2年田间抗蚜性鉴定结果表明: 小偃麦多表现为中抗或低抗,而小黑麦多为中感或低感。选取抗性性状稳定且抗性级别不同的小偃麦21(中抗)、小偃麦22(低抗)、小黑麦31(中感)和小黑麦32(低感)进行麦长管蚜取食行为分析。对非刺探波(Np)、刺探波(P)、电势落差(Pd)、水溶性唾液分泌波(E1)、韧皮部取食波(E2)、细胞机械阻碍波(F)和木质部取食波(G)等基本波形的分析显示,麦长管蚜在小偃麦上首次开始刺探的时间显著长于小黑麦,且在小偃麦上的E1波的持续时间显著大于小黑麦;麦长管蚜在小偃麦21上的F波和小偃麦22上Np波的持续时间最长,在小黑麦31上的P波和小黑麦32上的G波的持续时间最长。以E1、F和Np波的持续时间为指标,基于刺探电位的小麦种质资源抗性水平鉴定结果与田间鉴定结果基本一致。因此,使用EPG技术筛选抗蚜小麦材料时,建议采用E1、F和Np波作为评价小麦抗性水平的指标。小偃麦21、22对麦长管蚜的抗性水平较高,可作为小麦抗蚜育种的种质材料。     

不同水稻品种上白背飞虱取食行为的EPG分析

【目的】为探明不同水稻品种（丰源优272、R9810-T、华恢1号、明恢63、麻糯谷和Rathu Heenati）对白背飞虱Sogatella furcifera的抗性差异机理,利用刺吸电位技术(electronic penetration graph, EPG)记录了该虫在这6个品种三叶期稻苗上的取食行为。【方法】结合特定的EPG波形,考察了9个非韧皮部指标和22个韧皮部指标。【结果】在总记录时间8 h内,白背飞虱在品种Rathu Heenati上非刺探波np的总时间最长,其次是麻糯谷上非刺探波np的总时间,这两者间差异显著且都显著地长于其他4个品种的np总时间(P<0.05);该虫在Rathu Heenati上路径波Nc的总时间为8 523.41 s,是感虫品种明恢63 Nc总时间的2.24倍;该虫在Rathu Heenati上吸食韧皮部汁液N4-b的总时间显著地比其他品种短(P<0.05)。华恢1号和R9810-T上所有的EPG指标和感虫品种明恢63的EPG指标都没有显著的差异(P≥0.05)。白背飞虱在丰源优272上单次分泌水溶性唾液N4-a的平均时间更长,并伴随长时间的韧皮部取食。【结论】由此推测,品种Rathu Heenati可能存在不利于白背飞虱取食的忌避成分,并且在韧皮部外组织和韧皮部组织都存在抗性因子;在麻糯谷上仅存在忌避成分;然而,品种华恢1号和R9810-T可能对白背飞虱不具有明显的抗性;丰源优272可能是比明恢63更为感虫的品种。结合介体昆虫取食行为与传播持久性病毒的关系,这些结果也为利用抗白背飞虱品种控制南方水稻黑条矮缩病提供了参考。     

Characterization and correlation of DC electrical penetration graph waveforms with feeding behavior of beet leafhopper, Circulifer tenellus

Feeding behavior of beet leafhopper, Circulifer tenellus (Baker) (Homoptera: Cicadellidae), was studied with a DC electrical penetration graph. Nine different electrical penetration graph waveforms associated with feeding were identified and characterized. Waveforms were correlated with specific feeding behaviors using a number of techniques, including high magnification video recording, honeydew analysis, stylectomy, and histological processing. Waveforms were grouped into three phases based on feeding behavior: pathway phase (waveforms A, B1, B2, and C), non-phloem ingestion phase (waveform G), and phloem phase (waveforms D1, D2, D3, and D4). No ingestion was found to occur during waveforms A, B1, B2, and C. Waveform G was associated primarily with ingestion of xylem sap and occasionally with ingestion of mesophyll sap. Stylet tips were located in phloem during waveforms D1, D2, and D3, and waveforms D2 and D3 were correlated with ingestion of phloem sap. Waveform D4 probably also plays a role in phloem ingestion, because D4 is very brief and always occurs embedded in either waveform D2 or D3. In contrast to most other homopteran insects, rate of honeydew production (and hence rate of ingestion) was much lower on phloem than on xylem. More rapid rates of ingestion are expected on phloem, because its high turgor pressure drives sap into the feeding insect whereas the negative pressure of xylem sap is expected to cause a slow rate of ingestion. The very slow ingestion rate of beet leafhopper feeding on phloem suggests that it is not able to exploit the high turgor pressure of phloem to achieve the high rate of ingestion that is typical of phloem ingestion by other insects.     

Feeding behaviour of the Asiatic citrus psyllid,Diaphorina citri,on healthy and huanglongbing‐infected citrus

Diaphorina citri Kuwayama (Hemiptera: Sternorrhyncha: Psyllidae) is a vector of huanglongbing, a disease of citrus that in Asia is caused by ‘Candidatus Liberibacter asiaticus’ (α‐Proteobacteria) (Las). Acquisition of Las by D. citri appears to be variable, and this variability may be due to the suitability of the host plants and their tissues for acquisition. Therefore, this study aimed to determine the effect of symptom severity of the disease on the feeding behaviour of D. citri. Use of an electrical penetration graph showed that the pathway phase of D. citri consisted of four waveforms, A, B, C, and D; waveforms A and B have not been reported for D. citri before. The remaining waveforms, E1, E2, and G, conform to those described before for D. citri. The duration of the non‐penetration period did not differ between healthy or infected plants. However, in moderately and severely symptomatic plants, the duration of the pathway phase increased, whereas the phloem phase was shorter. In all diseased plants, the times to first and sustained salivation in the phloem were longer than those in control plants, with the times being related to symptom severity. As symptom expression increased, the percentage of time spent by psyllids salivating during the phloem phase increased; however, the percentage of time spent in phloem activities reduced gradually from ca. 74% in the control plants to ca. 8% in the severely symptomatic plants. In contrast, the percentage of time spent on xylem activities increased, as did the proportion of psyllids feeding from xylem. The differences in the durations of the E waveforms on plants showing different levels of symptom expression may account for differences in acquisition found amongst studies; therefore, future work on the acquisition and transmission of Las needs to carefully document symptom expression.     

重金属Zn2+胁迫下麦长管蚜的取食行为

为了探索重金属锌长期胁迫对麦长管蚜(Sitobion avenae)取食行为的变化影响,在模拟自然的实验室条件下,用不同浓度Zn~(2+)溶液浇灌土壤,通过土壤-小麦-蚜虫体系连续处理麦长管蚜15代,用EPG(刺探电位技术)对第1、5、10、15代成蚜的取食行为进行了监测。结果表明,第1代和第5代时,200 mg/kg的Zn~(2+)处理后np波和C波的总持续时间和数量显著低于对照,800mg/kg的Zn~(2+)使其显著增加。到第15代,高剂量的Zn~(2+)处理后np波和C波的总持续时间和数量均显著高于对照。涉及分泌唾液的E1波持续时间及涉及被动取食营养的E2波出现次数并未受到低剂量Zn~(2+)的显著影响,但高剂量的Zn~(2+)处理后单独E1波、伴随稳定E2的E1波总持续时间及E2波的数量均显著降低。麦长管蚜的取食行为会受到重金属锌的影响并且会在高剂量Zn~(2+)的胁迫条件下产生积累效应,而低剂量的Zn~(2+)则促进麦长管蚜对小麦的取食行为。针对重金属而言,此效应发生改变的关键浓度为400 mg/kg,蚜虫取食行为发生改变的关键世代为第5代和第10代。     

An analysis of the feeding behavior of three stages of Toxoptera citricida by DC electrical penetration graph waveforms

With the purpose of studying the feeding behavior of the brown citrus aphid pest, Toxoptera citricida (Kirkaldy) (Hemiptera: Aphididae), we compared stylet probing behaviors of third and fourth instars and adults on Citrus unshiu Marc (Rutaceae) seedlings using the electrical penetration graph (EPG) technique. EPG waveforms exhibited the full suite of stylet behaviors – stylet pathway, intracellular stylet puncture, phloem salivation (E1), sieve ingestion (E2), and xylem sap ingestion activities, plus the non‐penetration (Np) waveform. Before the phloem phase, the number of probes was significantly higher for third‐instar nymphs than for adults. Overall duration of Np events by adults was significantly lower than the duration of third and fourth instars. The number of short probes of the fourth instars was significantly higher than that of the adults. In the phloem phase, adults made more frequent and longer E1 events than the third and fourth instars. Third instars made more frequent but shorter E2 events, whereas adults made fewer but longer events. These results showed adults gained nutrients by increasing feeding time during phloem ingestion. Thus, the probability of phloem‐associated virus acquisition and transmission of T. citricida was higher in adults than in nymphs.     

EPG waveform characteristics of solenopsis mealybug stylet penetration on cotton

The solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae), is a polyphagous insect known to cause severe damage to cotton (especially transgenic varieties) in South Asia, and currently poses a serious threat in Asia and potentially elsewhere. Stylet penetration behavior of P. solenopsis on cotton was monitored using the electrical penetration graph (EPG) technique (DC system) and the EPG characteristics were compared with those previously published from Phenacoccus manihoti Matile‐Ferrero and Planococcus citri (Risso). We identified and further characterized typical waveforms of A, B, C, and pd (together pathway), E1 and E2 (phloem), F (derailed stylet mechanics), and G (xylem). Five novel EPG aspects were distinguished in the EPG waveforms from P. solenopsis: (1) obvious B waveforms occurred following waveform A, (2) during waveform C, some aphid‐like E1e waveforms were observed, (3) prolonged potential drops (pd) up to >1 h occurred with two continuously alternating sub‐phases pd1 and pd2, (4) the pd1 waveform always occurred as the first waveform related to phloem sieve elements, preceding the other phloem waveforms (E), the labeling of which we changed to achieve a better comparison to the aphid E waveforms, and (5) waveform F, related to derailed stylet mechanics occurred but was not reported from other mealybugs so far. This is mainly a waveform morphology study to extend existing knowledge on mealybug EPGs to investigate mealybug‐host plant interactions. Further experimental verification of waveform correlations with plant tissue positions of stylet tips and insect activities is still needed.     

B cell overexpression of FCRL5 and PD-1 is associated with low antibody titers in HCV infection

Antibodies targeting the hepatitis C virus (HCV) envelope glycoprotein E2 are associated with delayed disease progression, and these antibodies can also facilitate spontaneous clearance of infection in some individuals. However, many infected people demonstrate low titer and delayed anti-E2 antibody responses. Since a goal of HCV vaccine development is induction of high titers of anti-E2 antibodies, it is important to define the mechanisms underlying these suboptimal antibody responses. By staining lymphocytes with a cocktail of soluble E2 (sE2) glycoproteins, we detected HCV E2-specific (sE2+) B cells directly ex vivo at multiple acute infection timepoints in 29 HCV-infected subjects with a wide range of anti-E2 IgG titers, including 17 persistently infected subjects and 12 subjects with spontaneous clearance of infection. We performed multi-dimensional flow cytometric analysis of sE2+ and E2-nonspecific (sE2-) class-switched B cells (csBC). In sE2+ csBC from both persistence and clearance subjects, frequencies of resting memory B cells (rMBC) were reduced, frequencies of activated MBC (actMBC) and tissue-like MBC (tlMBC) were increased, and expression of FCRL5, an IgG receptor, was significantly upregulated. Across all subjects, plasma anti-E2 IgG levels were positively correlated with frequencies of sE2+ rMBC and sE2+ actMBC, while anti-E2 IgG levels were negatively correlated with levels of FCRL5 expression on sE2+ rMBC and PD-1 expression on sE2+ actMBC. Upregulation of FCRL5 on sE2+ rMBC and upregulation of PD-1 on sE2+ actMBC may limit anti-E2 antibody production in vivo. Strategies that limit upregulation of these molecules could potentially generate higher titers of protective antibodies against HCV or other pathogens.