Pathway phase waveform characteristics correlated with length and rate of stylet advancement and partial stylet withdrawal in AC electrical penetration graphs of adult whiteflies

A technique was developed for measuring the length of stylet insertion during adult whitefly probing. The distance that the labium shortens during a probe was shown to be equal to the length of stylets that were inserted into the plant tissue. The length of labial shortening then was measured in high-magnification video recordings of adult female silverleaf whitefly, Bemisia argentifolii, in conjunction with recording electrical penetration graphs (EPGs – AC method). Using a split-screen device, video images of the whitefly's labium during a probe and the EPG waveforms produced during the probe were recorded simultaneously on the same video tape. On playback, changes in labial length could be measured during specific EPG waveforms to determine the length of stylet insertion that occurred during the waveforms. The focus of the study was on two characteristics of the pathway phase sawtooth waveform: the frequency of voltage peaks and the increase in voltage level that occurs over time during sawtooth waveforms. The rate of stylet penetration was significantly and positively correlated with frequency of sawtooth waveform voltage peaks (r ²=0.33) and the length of stylet penetration was significantly and positively correlated (second-order polynomial) with the relative difference in voltage level between the beginning and end of the sawtooth waveform (r ²=0.43). Stylet advancement did not appear to occur during the few low-flat waveforms (unknown behavioral correlation) and high-flat waveforms (phloem phase) that were observed. Voltage drops occur sporadically during sawtooth waveforms, and these were associated with partial stylet withdrawal (indicated when the labium increased in length, but the probe was not terminated) with an accuracy of 99%.     

Stylet penetration of Cacopsylla pyri; an electrical penetration graph (EPG) study

Detailed information on plant penetration activities by pear psylla Cacopsylla pyri L. (Hemiptera Psyllidae) is essential to study phytoplasma transmission of “Candidatus Phytoplasma pyri” responsible of pear decline disease (PD) and to trace and evaluate resistant traits in new pear tree selections for advanced breeding programs. The electrical penetration graph technique or (full) EPG may relevantly contribute to this knowledge. C. pyri EPG waveforms were characterized on basis of amplitude, frequency, voltage level, and electrical origin. Additionally, stylet tracks and the putative location of stylet tips in the plant tissue were histologically related to EPG waveforms by light and transmission electron microscopy observations after stylectomy. More than one waveform occurred in the same tissue: PA, PB, PC1 and PC2 were all detected in the mesophyll, and PE1 and PE2 were both recorded in the phloem. Waveform PE1 was always preceded by transient waveform PD, as previously described in other psyllids. Interestingly, no brief intracellular punctures (potential drop waveforms) were observed during plant penetration, opposite of what is usually recorded in aphids and other Sternorrhyncha.     

Correlation of stylet activities by the glassy-winged sharpshooter, Homalodisca coagulata (Say), with electrical penetration graph (EPG) waveforms

Glassy-winged sharpshooter, Homalodisca coagulata (Say), is an efficient vector of Xylella fastidiosa (Xf), the causal bacterium of Pierce's disease, and leaf scorch in almond and oleander. Acquisition and inoculation of Xf occur sometime during the process of stylet penetration into the plant. That process is most rigorously studied via electrical penetration graph (EPG) monitoring of insect feeding. This study provides part of the crucial biological meanings that define the waveforms of each new insect species recorded by EPG. By synchronizing AC EPG waveforms with high-magnification video of H. coagulata stylet penetration in artifical diet, we correlated stylet activities with three previously described EPG pathway waveforms, A1, B1 and B2, as well as one ingestion waveform, C. Waveform A1 occured at the beginning of stylet penetration. This waveform was correlated with salivary sheath trunk formation, repetitive stylet movements involving retraction of both maxillary stylets and one mandibular stylet, extension of the stylet fascicle, and the fluttering-like movements of the maxillary stylet tips. Waveform B1 was ubitquious, interspersed throughout the other waveforms. B1 sub-type B1w was correlated with salivation followed by maxillary tip fluttering. This tip fluttering also occurred before and during B1 sub-type B1s, but was not directly correlated with either the occurrence or frequency of this waveform. Waveform B2 was correlated with sawing-like maxillary stylet movements, which usually occurred during salivary sheath branching. Waveform C was correlated with ingestion. Fluid outflow was also observed as a mechanism to clear the maxillary tips from debris during waveform C. This detailed understanding of stylet penetration behaviors of H. coagulata is an important step toward identifying the instant of bacterial inoculation which, in turn, will be applied to studies of disease epidemiology and development of host plant resistance.     

Intracellular punctures by the adult whitefly Bemisia argentifolii on DC and AC electronic feeding monitors

One of the most biologically important electrical penetration graph (EPG) waveforms recorded from aphids on DC EPG systems is the potential drop (pd), which is correlated with intracellular punctures by the stylet tips. In this study, pds of the adult female Bemisia argentifolii Bellows & Perring (Homoptera: Aleyrodidae), recorded on a DC EPG, are characterized and compared to pds of aphids. Whitefly pds consisted of 3 phases similar to those recorded from probing aphids. The major difference between aphid pds and whitefly pds was that whitefly pds lacked any observable subphases within the second phase of the pd. In addition, whitefly pds differed from aphid pds in that they: (1) did not occur frequently during stylet penetration, (2) did not occur early within probes, (3) did not occur during brief probes (<1 min). Pds produced by probing whiteflies always were preceded by a variant of waveform C which we named the pre-pd. The differences between pds of aphids and whiteflies are discussed in terms of their implications for virus transmission and host selection. Using a technique where EPG recordings can be switched back and forth between DC and AC systems, we demonstrated that the AC EPG pseudotransition waveform (Pt) was equivalent to the DC pd, and thus was correlated with intracellular punctures. Previously, intracellular punctures by whiteflies had not been detectable on AC EPG systems. The AC Pt consisted of three distinct phases (Pt1, Pt2, and Pt3) and our observations suggest that AC Pt1 correlates with the pre-pd waveform in DC EPGs and that AC Pt 2 and 3 correlate with the intracellular phase of the DC pd. AC Pts (n=47) and DC pds (n=43) were recorded on three separate plant species and were similar on all plant species.     

Electrical penetration graphs of thrips revised: combining DC- and AC-EPG signals

Within thrips feeding behaviour, sequences of four waveforms have been distinguished earlier in the DC-EPG, i.e. P, Q, R and S, representing mandibular stylet insertion, maxillary stylet insertion, ingestion, and repetitive mandibular insertion, respectively. During signal analysis it appeared that transitions from one waveform to the next were difficult to establish, making results ambiguous. In order to improve the quantitative reliability of the thrips' EPG data, the DC-EPGs were recorded concurrently with AC-EPG signals, thus providing two signals from the same activities containing different information. The additional AC information did not solve most quantification problems, however. We now propose to merge waveforms P, Q, and S, into 'puncture phase' (indicated by PQ) and waveforms R, T, and U, into 'feeding phase' (indicated by R), rather than trying to analyse all separate waveforms. This will provide a more reliable and much less laborious analysis of thrips probing behaviour. Waveforms T and U are two novel waveforms identified here by combining DC- and AC-EPG recordings with concurrent video recordings. Waveform T represents a single mandibular thrust embedded in waveform R and waveform U represents the end of a probe, presumably the retraction of the maxillary stylets.     

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。     

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.     

Stylet penetration by larvae of the greenhouse whitefly on cucumber

Probing behaviour of Trialeurodes vaporariorum (Westwood) larvae was monitored using the DC electrical penetration graph (EPG) technique on the host plant cucumber. EPGs were recorded for 16 h, simultaneously with honeydew excretion using a honeydew clock. Three waveforms were distinguished: a pathway waveform (C), and two phloem waveforms, one with a high (H), and one with a low frequency (L) signal. The C waveform mainly occurred in the crawler stage of the 1st instar larvae. EPGs recorded from larvae during and after moulting indicated that the process involves stylet withdrawal; hence the stylets of each new instar need to penetrate again from the leaf surface to the phloem.All sessile stages, from L1 to pre-pupa, spent almost their entire time in waveforms H and L. These waveforms alternated more frequently in the early instars than during the later ones, in which the H waveform became predominant. The H waveform was highly correlated with honeydew excretion and thus phloem sap ingestion. The L waveform was not related to honeydew excretion but EPGs indicated that the stylet tips remain in a sieve element during both waveforms. Periods of honeydew production demonstrated a delay of 30–40 min in relation to the onset and end of H and L waveforms. This delay is presumably related to the time needed for food passing through, or emptying of, the insect's gut. From the 1st instar to the pre-pupa, the frequency of excreted honeydew droplets decreased but their size increased, causing a net increase of the excretion rate.     

Characterisation of the feeding behaviour of western flower thrips in terms of electrical penetration graph (EPG) waveforms

Western flower thrips, Frankliniella occidentalis (Pergande) causes damage to plants when they are feeding. Also, this thrips species transmits Tomato spotted wilt virus (TSWV) during stylet penetration. We investigated the penetration behaviour (probing) of thrips on pepper leaves and on liquid diet by electrical penetration graph (EPG, DC-system) recording. In addition, we used high-magnification video observations to correlate EPG waveforms with the insect's posture, head movements, and muscle contractions. Also, EPGs were correlated with probing on liquid diets containing radio-active tracers to distinguish and quantify ingestion waveforms. The previously described waveforms P, Q, and R were distinguished and additionally, a new waveform 'S' was distinguished. Waveform P could be linked with mandibular leaf penetration, waveform Q presumably with insertion of the maxillary stylets, and waveform R with ingestion of cell contents, whereas waveform S could not be correlated with any behavioural activity. Histology of the feeding damage in pepper leaves shows that thrips ingests the contents of multiple cells per probe.     

EPG monitoring of the probing behaviour of the common brown leafhopper Orosius orientalis on artificial diet and selected host plants

The common brown leafhopper Orosius orientalis (Hemiptera: Cicadellidae) is a polyphagous vector of a range of economically important pathogens, including phytoplasmas and viruses, which infect a diverse range of crops. Studies on the plant penetration behaviour by O. orientalis were conducted using the electrical penetration graph (EPG) technique to assist in the characterisation of pathogen acquisition and transmission. EPG waveforms representing different probing activities were acquired from adult O. orientalis probing in planta, using two host species, tobacco Nicotiana tabacum and bean Phaseolus vulgaris, and in vitro using a simple sucrose-based artificial diet. Five waveforms (O1?CO5) were evident when O. orientalis fed on bean, whereas only four waveforms (O1?CO4) and three waveforms (O1?CO3) were observed when the leafhopper fed on tobacco and on the artificial diet, respectively. Both the mean duration of each waveform and waveform type differed markedly depending on the food substrate. Waveform O4 was not observed on the artificial diet and occurred relatively rarely on tobacco plants when compared with bean plants. Waveform O5 was only observed with leafhoppers probing on beans. The attributes of the waveforms and comparative analyses with previously published Hemipteran data are presented and discussed, but further characterisation studies will be needed to confirm our suggestions.     

Characterization of electrical penetration graphs of the Asian citrus psyllid, Diaphorina citri, in sweet orange seedlings

Detailed information on probing behavior of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is critical for understanding the transmission process of phloem‐limited bacteria (Candidatus Liberibacter spp.) associated with citrus ‘huanglongbing’ by this vector. In this study, we investigated stylet penetration activities of D. citri on seedlings of Citrus sinensis (L.) Osbeck cv. Pêra (Rutaceae) by using the electrical penetration graph (EPG‐DC system) technique. EPG waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration into plant tissues. The main waveforms were correlated with histological observations of salivary sheath termini in plant tissues, to determine the putative location of stylet tips. The behavioral activities were also inferred based on waveform similarities in relation to other Sternorrhyncha, particularly aphids and whiteflies. In addition, we correlated the occurrence of specific waveforms with the acquisition of the phloem‐limited bacterium Ca. Liberibacter asiaticus by D. citri. The occurrence of a G‐like xylem sap ingestion waveform in starved and unstarved psyllids was also compared. By analyzing 8‐h EPGs of adult females, five waveforms were described: (C) salivary sheath secretion and other stylet pathway activities; (D) first contact with phloem (distinct from other waveforms reported for Sternorrhyncha); (E1) putative salivation in phloem sieve tubes; (E2) phloem sap ingestion; and (G) probably xylem sap ingestion. Diaphorina citri initiates a probe with stylet pathway through epidermis and parenchyma (C). Interestingly, no potential drops were observed during the stylet pathway phase, as are usually recorded in aphids and other Sternorrhyncha. Once in C, D. citri shows a higher propensity to return to non‐probing than to start a phloem or xylem phase. Several probes are usually observed before the phloem phase; waveform D is observed upon phloem contact, always immediately followed by E1. After E1, D. citri either returns to pathway activity (C) or starts phloem sap ingestion, which was the longest activity observed.     

Compatibility of Insect Growth Regulators with Eretmocerus eremicus (Hymenoptera: Aphelinidae) for Whitefly (Homoptera: Aleyrodidae) Control on Poinsettias: I. Laboratory Assays

The compatibility of five insect growth regulators (IGRs), buprofezin, pyriproxyfen, fenoxycarb, pymetrozine, and kinoprene, were tested in the laboratory for compatibility with the whitefly parasitoid Eretmocerus eremicus Rose and Zolnerowich (Hymenoptera: Aphelinidae). The survivorship of adult parasitoids foraging on poinsettia leaves with residues 6, 24, and 96 h of age was determined. The toxicity of Bemisia argentifolii Bellows and Perring (Homoptera: Aleyrodidae) patches treated with IGRs presented to female parasitoids 24 and 96 h posttreatment was quantified. Survivorship of immature E. eremicus developing within B. argentifolii nymphs was determined by treating whitefly nymphs with IGRs 5 and 13 days postoviposition by female parasitoids. Finally, behavioral observations of female parasitoids foraging on IGR-treated and untreated B. argentifolii patches presented simultaneously were quantified to determine whether IGR residues had a repellant effect toward E. eremicus. Averaging ranks for IGRs based on their compatibility with E. eremicus and their ability to kill B. argentifolii nymphs produced the following parasitoid compatibility order: buprofezin > fenoxycarb > pymetrozine = pyriproxyfen > kinoprene. Further work in greenhouses assessing the efficacy of buprofezin with E. eremicus for B. argentifolii control on poinsettias is recommended.     

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.     

Parasrnzatlon of bemisia argentifolii (Hom.: Aleyrodidae) by encarsia pergandiella (Hym.: Apheunidae)

The biology of the arrhenotokous autoparasitoid,Encarsia pergandiella Howard, was studied in the laboratory on the silverleaf whitefly,Bemisia argentifolii Bellows & Perring. Egg to adult development of parasitoid females averaged ca. 14 days at about 25.3+0.2?C regardless of whether the whitefly host was reared on tomato, eggplant or squash. While all instars ofB. argentifolii were accepted for primary parasitization, a greater percentage of third and fourth instars were parasitized. Mortality of whitefly nymphs in the absence of parasitization did not differ among instars and averaged about 35%. Second instar to pupal parasitoid females were accepted for secondary parasitization although a greater percent of pupal females were parasitized. About 40% of immatureE. pergandiella females more than 4 days old died in the absence of secondary parasitization when exposed to adultE. pergandiella females.     

New structure in cell puncture activities by aphid stylets: a dual‐mode EPG study

Intracellular punctures by aphid stylets appear as potential drop (pd) waveforms in DC electrical penetration graph (EPG) recordings. We used a dual‐EPG device that recorded in one channel the ‘full EPG’ with R‐plus emf‐components (i.e., the usual DC EPG) and concurrently in a second channel the ‘R‐EPG’ with R‐components only. The circuit of the latter channel was an optimised amplitude modulation (AM) version derived from early (before 1990) AC systems. We also made some ‘emf‐EPG’ recordings using a separate high input resistance ‘emf‐amplifier’ sensitive to emf‐components only. The intracellular pd waveforms have previously been divided into three subphases, and we aimed to distinguish and separate these subphases more accurately by the dual‐EPG recordings than with the normal full EPG only. In this study, we temporarily distinguished five subphases (α–ε), but unequivocal distinction of only a few of these appeared possible, in spite of the information coming from the two signals. The lack of clearly separable features in R‐EPG signals often provided serious difficulties in pd recognition without the concurrent full EPG, but once located, only subphase II‐2 features were clear and supported the II‐2 data from the full EPG. Consequently, we could not distinguish subphases of complete pd waveforms better with additional R‐EPG information during cell punctures by Aphis gossypii Glover (Hemiptera: Aphididae). In Brevicoryne brassicae (L.) (Hemiptera: Aphididae), however, distinguishing II‐2 subphases in the full EPG was sometimes a problem. Our detailed dual‐EPG observations showed some waveform continuity from halfway into the II‐1 subphase (start of the newly recognised subphase β) until the end of the pd, with a strong but variable emf origin. This waveform tended to overrule other subphase waveforms in B. brassicae more than in A. gossypii and Myzus persicae (Sulzer) (Hemiptera: Aphididae). Subphase waveforms in full EPGs were especially difficult to recognise when pd periods had been interrupted in a virus inoculation experiment and additional R‐EPG information could then be useful. This inoculation experiment showed again that only the first subphase (II‐1) contributes to virus (Cucumber mosaic virus) inoculation by A. gossypii. In B. brassicae, the benefit of concurrent R‐EPG information in such virus experiments is presently under further investigation. Apart from this special application to virus experiments, we do not recommend the routine use of the dual‐EPG device. Furthermore, we do not advocate the distinction of more than the previously recognised three intracellular pd subphases as a feasible option in future studies. Analysis of EPGs with concurrent R‐EPGs requires substantially more analysis work without yielding consistently useful additional insights. This confirms earlier dual‐EPG results from thrips.     

Correlations of cibarial muscle activities of Homalodisca spp. sharpshooters (Hemiptera: Cicadellidae) with EPG ingestion waveform and excretion

Fluid flow into and out of the stylets of xylem-ingesting sharpshooters (Hemiptera: Cicadellidae: Cicadellinae) is powered by muscles of the cibarial pump. Such fluid flow is crucial for transmission of Xylella fastidiosa, the Pierce’s Disease bacterium, yet has not been rigorously studied via electrical penetration graph (EPG) technology. We correlated EPG waveforms with electromyographically (EMG) recorded muscle potentials from the cibarial dilator muscles, which power the piston-like cibarial diaphragm. There was a 1:1 correspondence of each cycle of cibarial muscle contraction/relaxation with each plateau of EPG waveform C. Results definitively showed that the C waveform represents active ingestion, i.e. fluid flow is propelled by cibarial muscle contraction. Moreover, each C waveform episode represents muscular diaphragm uplift, probably combined with a “bounce” from cuticular elasticity, to provide the suction that pulls fluid into the stylets. Fine structure of the EPG ingestion waveform represents directionality of fluid flow, supporting the primary role of streaming potentials as the electrical origin of the C waveform. Rhythmic bouts of cibarial pumping were generally correlated with sustained production of excretory droplets. However, neither the onset nor cessation of ingestion was correlated with onset or cessation of excretion, respectively. Volume of excreta is an inexact measure of ingestion. Implications for using EPG to understand the mechanism of X. fastidiosa transmission are discussed.     

Electrical recording of plant penetration by western flower thrips

Plant penetration by western flower thrips (Frankliniella occidentalis (Pergande)) was analysed with the electrical penetration graph technique (EPG, DC-system). Thrips attached to a gold wire were included in an electrical circuit to record EPGs when penetrating the plant tissues with their stylets. Three basic EPG waveforms have been distinguished, correlated with stylet penetration into cells, salivation, and ingestion, respectively. The main difference with EPGs of Homoptera is the occurrence of continued separate penetrations that are not necessarily followed by ingestion. Insertion of the stylets causes strong voltage fluctuations in the EPG. We could confirm earlier evidence that penetration of cells and subsequent ingestion of (part of) the protoplast takes less than 20 seconds. Repeated short penetrations can be followed by a continuous feeding pattern during which the stylets are not withdrawn. The same sequence of waveforms is produced on other plant parts such as fruits or pollen grains. The specific waveforms are mainly caused by electromotive force (emf). The emf component was recorded with high resolution and the correlation of waveform details with activities of the cibarial muscle system is discussed.     

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.     

Characterization of the electrical penetration graphs of the psyllid Bactericera trigonica on carrots

The psyllid Bactericera trigonica Hodkinson (Hemiptera: Triozidae) is a carrot and celery pest recently described as a vector of the plant pathogenic bacterium Candidatus Liberibacter solanacearum (Lso) on Apiaceae. Detailed information on vector stylet penetration activities is essential in the study of Lso transmission. In this study we used the electrical penetration graph (EPG) technique, characterized waveforms produced during the various stylet penetration activities in carrot leaves, and correlated them with stylet tracks and salivary sheath termini on plant tissues as well as with Lso inoculation. In addition, the effect of Lso in B. trigonica on the stylet penetration activities was tested. The EPG waveforms identified were: waveforms C1 and C2 detected in the mesophyll, waveforms D, E1, and E2 near or in the phloem sieve elements, and waveform G in the xylem vessels. A waveform pattern not previously reported for psyllids was the ‘pseudo‐potential drop’ (pseudo‐pd), characterized by sudden voltage dips similar to potential drops. However, the lowered voltage appeared to be inverted when the plant voltage is negative, indicating that it is caused by an increased resistance period and not due to a cell puncture. A direct correlation is shown between the waveform E1 and salivation into phloem sieve elements by B. trigonica as the inoculation of Lso occurred in a period as short as 30 s of E1; Lso transmission occurred in 17 of 35 plants (48%). Stylet activities during waveforms C or D had no consequences on the inoculation of Lso. In conclusion, Lso infection directly affects the probing behaviour of B. trigonica by increasing the total duration of C and D waveforms, but not variables related to phloem salivation (Lso inoculation) or ingestion (Lso acquisition). The reported information here is fundamental for identifying the psyllid vector traits of behaviour associated with transmission of Lso to Apiaceae.