Electrical penetration graphs of the nymphal stage of Bemisia argentifolii

Electrical penetration graph (EPG, DC system) waveforms were recorded from first, second, and third instar Bemisia argentifolii nymphs. Waveforms recorded were similar among the three instars. Four waveforms were recorded and were named C, J, L, and H. Waveform J is new, whereas waveforms C, L, and H of B. argentifolii nymphs were similar to those published previously from greenhouse whitefly nymphs. As in the previous study on greenhouse whitefly nymphs, there was variation in each of waveforms C, L, and H. Waveform C was recorded at an extracellular voltage level, and represents a pathway phase where the stylets penetrate the plant tissue in an intercellular pathway. At the end of waveform C, the voltage dropped to an intracellular level, indicating penetration of a living cell, and the stylet tips then remained in that cell for the rest of the EPG recording, which was sometimes as long as 16 h. Three waveforms (J, L, and H) were recorded during this intracellular phase, beginning with J, a brief (average = 31 s), low amplitude, irregular waveform. J appeared only at the beginning of the intracellular phase, and was followed by either L (five out of eight times) or H (three out of eight times). Waveforms L and H then alternated with one another for the remainder of the intracellular phase. The most conspicuous difference between L and H was the frequency of their voltage fluctuations; L had a lower frequency and H a higher frequency. Usually the shape of waveform L was dominated by voltage peaks in a positive direction, while waveform H was characterized by strong voltage peaks in a negative direction; although some variants of both L and H had distinct voltage peaks in both directions. The electrical origin of both the positive and negative voltage peaks was electromotive force (emf) fluctuation rather than resistance fluctuation. During waveform H, copious amounts of honeydew were produced, indicating that the penetrated cell was a sieve element. We conclude, therefore, that H represents phloem sap ingestion; and because J and L are produced in the same cell as H, then phloem phase is represented by waveforms J, L, and H. The biological correlations for J and L are not yet known.     

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.     

Phloem-sap feeding by animals: problems and solutions

The incidence of phloem sap feeding by animals appears paradoxical. Although phloem sap is nutrient-rich compared with many other plant products and generally lacking in toxins and feeding deterrents, it is consumed as the dominant or sole diet by a very restricted range of animals, exclusively insects of the order Hemiptera. These insects display two sets of adaptations. First, linked to the high ratio of non-essential:essential amino acids in phloem sap, these insects contain symbiotic micro-organisms which provide them with essential amino acids. For example, bacteria of the genus Buchnera contribute up to 90% of the essential amino acids required by the pea aphid Acyrthosiphon pisum feeding on Vicia faba. Second, the insect tolerance of the very high sugar content and osmotic pressure of phloem sap is promoted by their possession in the gut of sucrase-transglucosidase activity, which transforms excess ingested sugar into long-chain oligosaccharides voided via honeydew. Various other animals consume phloem sap by proxy, through feeding on the honeydew of phloem-feeding hemipterans. Honeydew is physiologically less extreme than phloem sap, with a higher essential:non-essential amino acid ratio and lower osmotic pressure. Even so, ant species strongly dependent on honeydew as food may benefit from nutrients derived from their symbiotic bacteria Blochmannia.     

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。     

Brown planthopper (N. lugens Stal) feeding behaviour on rice germplasm as an indicator of resistance

Background

The brown planthopper (BPH) Nilaparvata lugens (Stal) is a serious pest of rice in Asia. Development of novel control strategies can be facilitated by comparison of BPH feeding behaviour on varieties exhibiting natural genetic variation, and then elucidation of the underlying mechanisms of resistance.

Methodology/Principal Findings

BPH feeding behaviour was compared on 12 rice varieties over a 12 h period using the electrical penetration graph (EPG) and honeydew clocks. Seven feeding behaviours (waveforms) were identified and could be classified into two phases. The first phase involved patterns of sieve element location including non penetration (NP), pathway (N1+N2+N3), xylem (N5) [21] and two new feeding waveforms, derailed stylet mechanics (N6) and cell penetration (N7). The second feeding phase consisted of salivation into the sieve element (N4-a) and sieve element sap ingestion (N4-b). Production of honeydew drops correlated with N4-b waveform patterns providing independent confirmation of this feeding behaviour.

Conclusions/Significance

Overall variation in feeding behaviour was highly correlated with previously published field resistance or susceptibility of the different rice varieties: BPH produced lower numbers of honeydew drops and had a shorter period of phloem feeding on resistant rice varieties, but there was no significant difference in the time to the first salivation (N4-b). These qualitative differences in behaviour suggest that resistance is caused by differences in sustained phloem ingestion, not by phloem location. Cluster analysis of the feeding and honeydew data split the 12 rice varieties into three groups: susceptible, moderately resistant and highly resistant. The screening methods that we have described uncover novel aspects of the resistance mechanism (or mechanisms) of rice to BPH and will in combination with molecular approaches allow identification and development of new control strategies.     

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。     

Plant penetration activities by the flatid planthopper Metcalfa pruinosa (Hemiptera: Fulgoroidea): an electrical penetration graph‐histology analysis

The citrus flatid planthopper, Metcalfa pruinosa Say (Hemiptera: Flatidae), is a very polyphagous native insect in North America and currently a serious pest in Europe and South Korea. To understand the feeding behaviour of M. pruinosa, stylet penetration behaviour of M. pruinosa was investigated with an electrical penetration graph (EPG) system. This study reports seven EPG waveforms related to M. pruinosa feeding behaviour: np (no stylet penetration), Mp1 (initiation of stylet penetration), Mp2 (stylet movement and salivation), Mp4 (phloem feeding), Mp4‐H (honeydew excretion), Mp5 (xylem feeding) and Mp6 (unknown). To determine respective feeding behaviour related to the Mp4 and Mp5 waveforms, stylets were cut with a laser beam, and the location of the stylet tip within plant tissue was examined. We found plant sap was exuded from the severed stylets only when the Mp4 waveform was observed, suggesting phloem sap ingestion. The stylet tip was located in the xylem region, indicating xylem‐feeding activity, when the Mp5 waveform was observed. The analysis of 24 different EPG parameters suggests that M. pruinosa stylets reached the vascular bundle of a plant within ca. 5 min and spend ca. 70% of the time feeding on xylem and phloem feeding. This is the first study that reports seven distinctive EPG waveforms with respect to the feeding behaviour of M. pruinosa which could help determine host specificity and host plant susceptibility.     

Electrical penetration graphic waveforms in relation to the actual positions of the stylet tips of Nilaparvata lugens in rice tissue

The stylet penetration behavior of Nilaparvata lugens (Hemiptera: Delphacidae) in rice plants (Oryza sativa) was evaluated through the use of an electrical penetration graph (EPG). To accomplish this, we classified the EPG signals into seven different waveforms, np, N1, N2, N3, N4-a, N4-b, and N5, according to their shapes, amplitudes, and frequencies. The N4-b waveform was always preceded by N3 and N4-a, in that order. Continuous honeydew excretion only occurred during the N4-b period, and the honeydew deposited on a filter paper containing ninhydrin reagent during the N4-b period was stained violet. The tips of the stylets that were severed in the N3, N4-a, and N4-b periods were in the phloem region of rice. Moreover, the flow of plant sap after stylectomy only occurred during the N4-b period. Finally, sucrose was the only carbohydrate component identified when HPLC analysis of the plant sap was conducted. On the other hand, honeydew excretion hardly occurred during the N5 period and the tips of the stylets that were severed during the N5 period were located in the xylem region of rice. Based on the location of the stylets and honeydew excretion, the EPG waveforms for the stylet penetration behaviors of N. lugens were assigned to the following groups; np: non-penetration of stylets, N1: penetration initiation, N2: salivation and stylet movement, N3: an extracellular activity near the phloem region, N4-a: an intracellular activity in phloem region, N4-b: phloem sap ingestion, and N5: activity in the xylem region.     

Influence of pymetrozine on feeding behaviors of three rice planthoppers and a rice leafhopper using electrical penetration graphs

Pymetrozine reportedly inhibits feeding of plant sap-sucking insects, such as aphids and brown planthopper (Nilaparvata lugens (St?l)). By using electrical penetration graph (EPG), this study was conducted to investigate any differential effect of pymetrozine on the feeding behaviors of four major rice sap-sucking insect species, 1) N. lugens, 2) white-backed planthopper (Sogatella furcifera (Horváth)), 3) small brown planthopper (Laodelphax striatellus (Fallen)), and 4) green rice leafhopper (Nephotettix cincticeps Uhler). On pymetrozine-free TN1 rice plants, white-backed planthopper and small brown planthopper showed a significantly less activity in the phloem phases than brown planthopper or green rice leafhopper while green rice leafhopper engaged in relatively more xylem ingestion than brown planthopper, white-backed planthopper, and small brown planthopper. On the plants treated with 100 mg liter(-1) of pymetrozine, all four insect species showed significant increases, in total duration of nonprobing and significant decreases in the activities in phloem tissue, while all species showed similar feeding behavior during the pathway and xylem phases. This study revealed that, regardless of whether the insects on untreated plants spent more time feeding on phloem than xylem (brown planthopper) or more time on xylem than phloem (green rice leafhopper) or similar times on phloem and xylem (white-backed planthopper and small brown planthopper), their feeding behavior was disturbed by pymetrozine and exhibited similar patterns of sharp decline in activity in the phloem tissue and a significant increase the nonprobing.     

Feeding disruption of potato psyllid,Bactericera cockerelli,by imidacloprid as measured by electrical penetration graphs

The potato psyllid, Bactericera cockerelli (Sulc) (Hemiptera: Triozidae), is a serious pest of potatoes that can cause yield loss by direct feeding and by transmitting a bacterial pathogen, Candidatus Liberibacter psyllaurous (also known as Candidatus L. solanacearum), which is associated with zebra chip disease of this crop. Current pest management practices rely on the use of insecticides for control of potato psyllid to lower disease incidences and increase yields. Imidacloprid is typically applied at potato planting, and it remains unknown if imidacloprid has any effect on potato psyllid feeding behavior. Thus, our specific objectives of this study were to determine and characterize the effects of imidacloprid treatment (0.11 ml l^?1) to potato plants on adult potato psyllid feeding behavior 1, 2, and 4 weeks post‐application. Electrical penetration graph (EPG) recordings of potato psyllid feeding revealed six EPG waveforms, which include non‐probing (NP), intercellular stylet penetration (C), initial contact with phloem tissue (D), salivation into phloem sieve elements (E1), phloem sap ingestion (E2), and ingestion of xylem sap (G). The number of NP events and the duration of individual NP events significantly increased on plants treated with imidacloprid compared with untreated controls. Potato psyllids exhibited significant decreases in the number of phloem salivation events on plants treated with imidacloprid. Waveform durations and waveform durations per event for E2 and G were significantly decreased for psyllids on plants treated with imidacloprid compared with untreated controls. These data suggest that the effective use of imidacloprid to reduce transmission of Ca. Liberibacter psyllaurous is related to the negative effects of imidacloprid on psyllid feeding.     

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.     

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.     

Trade-off between foliage and tuber resistance to Phthorimaea operculella in wild potatoes

Beet leafhopper, Circulifer tenellus (Baker) (Homoptera: Cicadellidae), is the only known North American vector of beet curly top virus (Geminiviridae), which causes major economic losses in a number of crops including sugar beet, tomato, beans, and peppers. Beet curly top virus is a phloem-limited, persistently transmitted, circulative geminivirus. The strain/species of curly top virus used in this study is the CFH strain, also referred to as beet severe curly top virus (BSCTV). The direct current (DC) electrical penetration graph technique was used to determine the specific stylet penetration behavior associated with inoculation of BSCTV. Viruliferous leafhoppers were allowed to feed on healthy 3–4-week-old sugar beet plants until specific electrical penetration graph waveforms were produced, at which point feeding was artificially terminated. A series of comparisons between leafhoppers that produced different combinations of waveforms clearly implicated waveform D1 as the only waveform correlated with inoculation of BSCTV. All successful inoculations contained waveform D1, and 56 out of 64 leafhoppers that produced waveform D1 successfully inoculated test plants. Eighty-five leafhoppers did not produce waveform D1 and none of these inoculated BSCTV. While the occurrence of waveform D1 appears to be necessary for BSCTV inoculation, there was no correlation between duration of waveform D1 and inoculation success rate. The correlation of waveform D1 and BSCTV inoculation found in this study implies that waveform D1 is associated with phloem salivation.     

Waveform characterization of the soybean stem feeder Edessa meditabunda: overcoming the challenge of wiring pentatomids for EPG

Stink bugs (Hemiptera: Heteroptera: Pentatomidae) are in general robust and restless insects, which makes them difficult to wire for electropenetrograph (EPG) studies. In addition, cuticular lipids may reduce wire effectiveness, and their removal could improve success of wiring. We compared wiring effectiveness for three species of stink bugs, differing in walking behaviour and degree of cuticular waxiness, that is, Piezodorus guildinii (Westwood), Nezara viridula (L.), and Loxa deducta (Walker). Results indicated that removal of cuticular lipids by mechanical abrasion (via sanding) greatly improved attachment success with gold wire. Our hypothesis that heavier and bigger bugs would lose the wire attachment more quickly than lighter and smaller bugs was not confirmed, regardless of the sanding. In contrast, our hypothesis that greater movement of a bug would cause the wire to break more often was supported by extensive testing. Behaviour appears to be more relevant for successful wiring than body weight. We used the sanding and wiring technique to characterize and correlate direct current EPG waveforms for the large and restless stem‐feeding stink bug Edessa meditabunda (Fabricius) on soybean plants. This marks the first published example of pentatomid EPG waveforms. Edessa meditabunda recordings on soybean stems generated eight types of waveforms in three phases and two families, named as follows: non‐probing = Np and Z; pathway phase = Em1; X wave phase = X; ingestion phase, family I = Em2 and Em3; ingestion phase, family N = Em4 and Em5. These eight were described based on their frequencies, relative amplitudes, and level voltages. Histological studies of stylets within salivary sheaths correlated the Em1, Em2, and Em3 waveforms with specific penetration sites. The waveform with the longest duration when feeding was Em2, representing xylem sap ingestion; in addition, waveform Em3 (always preceded by an X wave) was correlated with phloem sap ingestion.     

Direct and indirect measurements of Phloem turgor pressure in white ash

Direct determinations and indirect calculations of phloem turgor pressure were compared in white ash (Fraxinus americana L.). Direct measurements of trunk phloem turgor were made using a modified Hammel-type phloem needle connected to a pressure transducer. Turgor at the site of the direct measurements was calculated from the osmotic potential of the phloem sap and from the water potential of the xylem. It was assumed that the water potentials of the phloem and xylem were close to equilibrium at any one trunk location, at least under certain conditions. The water potential of the xylem was determined from the osmotic potential of xylem sap and from the xylem tension of previously bagged leaves, measured with a pressure chamber. The xylem tension of bagged leaves on a branch adjacent to the site of the direct measurements was considered equivalent to the xylem tension of the trunk at that point. While both the direct and indirect measurements of phloem turgor showed clear diurnal changes, the directly measured pressures were consistently lower than the calculated values. It is not clear at present whether the discrepancy between the two values lies primarily in the calculated or in the measured pressures, and thus, the results from both methods as described here must be regarded as estimates of true phloem turgor.     

Feeding rates and carbohydrate metabolism by Bemisia tabaci (Homoptera: Aleyrodidae) on different quality phloem saps

Abstract.The effects of water stress on phloem sap quality of the melon, Cucumis melo, and how this, in turn, has an impact on the sweet potato whitefly, Bemisia tabaci were studied. Melon plants were grown under watering regimes that produced plants with or without water stress. Plants showed strong developmental responses to the treatments; water-stressed plants were shorter, with fewer, smaller leaves than those without stress. There was, however, no effect of plant water stress on the development period of whiteflies feeding on these plants, or on the weights of male or female adults. Honeydew production was used as an indirect measure to test whether the absence of insect developmental or behavioural effects was due to differential phloem sap ingestion. Feeding rates on the stressed plants were almost half those on unstressed plants, and there was also variation in the daily pattern of honeydew production. Phloem sap and honeydew were analysed to determine why the feeding behaviours differed. Amino acid composition of the phloem sap was similar in both groups of plants, but carbohydrate concentrations were greater in water-stressed plants, indicating that lower feeding rates may have been due either to the physical or nutritional quality of the phloem sap. The honeydew of insects that were feeding on water-stressed plants contained a greater concentration of carbohydrate than those on unstressed plants, and was composed of a significantly greater proportion of glucose and the disaccharide, trehalulose. This isomerization of more complex sugars from those in the diet suggests that B. tabaci uses a mechanism of osmoregulation to actively maintain its internal water status. It is concluded that transient conditions of water stress in this host plant do not affect the development of B. tabaci, due to physiological and behavioural changes in response to diets with different nutritional and physical properties. The implications of this finding for the feeding biology of B. tabaci on desert-grown crops are discussed.     

Immunological analysis of phloem sap of Bacillus thuringiensis corn and of the nontarget herbivore Rhopalosiphum padi (Homoptera: Aphididae) for the presence of Cry1Ab

Phloem sap of transgenic Bacillus thuringiensis (Bt) corn expressing a truncated form of the B. thuringiensis delta-endotoxin Cry1Ab, sap sucking aphids feeding on Bt corn and their honeydew were analysed for presence of Cry1Ab using ELISA. Phloem sap of Bt and non-Bt corn was collected using a newly developed technique with a microcapillary being directly inserted into the phloem tubes. Using this technique, no Cry1Ab was detected in the phloem sap. In contrast, measurable concentrations of Cry1Ab in the range of 1 ppb were detected when phloem sap of pooled leaf samples was extracted using EDTA buffer. This was probably because of Cry1Ab toxin released from damaged cells. When analysing apterous adults of Rhopalosiphum padi L. and their honeydew, no Cry1Ab could be detected. In contrast, Cry1Ab was clearly detected in both larvae of the leaf chewing herbivore Spodoptera littoralis (Boisduval) and their faeces, showing that Cry1Ab is detectable after ingestion and excretion by herbivores. These results suggest that R. padi ingests or contains no or only very low concentrations of Cry1Ab in the range of the detection limit. In consequence it is hypothesized that R. padi as an important prey for beneficial insects in corn is unlikely to cause any harm to its antagonists due to mediating Bt toxin.     

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.     

柑橘木虱蜜露排泄机制:进展与展望

半翅目昆虫柑橘木虱Diaphorina citri是柑橘类果树重要害虫,主要以高渗透压的植物韧皮部汁液为食,是柑橘毁灭性病害——柑橘黄龙病(HLB)的主要传播媒介。柑橘木虱取食韧皮部汁液时自身进化出一套完整的渗透调节机制调节其体内渗透压,将体内过量摄入的糖类转化成长链寡糖并以蜜露形式排出体外。本文从柑橘木虱蜜露排泄行为、蜜露组成成分以及影响蜜露排泄的多个因素进行了论述,同时综述了可能参与柑橘木虱蜜露排泄行为的渗透调节基因。研究表明,柑橘木虱雌雄成虫及若虫在蜜露排泄行为上存在显著差异,且排泄的蜜露在颜色、纹路及组成成分方面均有不同;寄主植物、杀虫剂、病原微生物及天敌化合物均会影响柑橘木虱蜜露排泄行为。分子机制探究发现,α-葡萄糖苷水解酶、水通道蛋白及糖基转移酶基因等关键渗透调节基因可能参与调控柑橘木虱蜜露排泄行为。本文可为未来有关柑橘木虱蜜露排泄行为方面的研究以及为研制柑橘木虱防治新型药剂开发新靶标提供参考。