Behavior and biology of the tomato psyllid, Bactericerca cockerelli, in response to the Mi-1.2 gene

The Mi‐1.2 gene, identified from wild varieties of tomato, Solanum peruvianum (Mill) (Solanaceae), has been incorporated into near‐isogenic commercial varieties of tomato and has been shown to confer resistance to three different species of phloem feeders: aphids, whiteflies, and nematodes. The results presented here show that plants bearing Mi‐1.2 were also resistant to the tomato psyllid, Bactericerca [Paratrioza] cockerelli (Sulc) (Homoptera: Psyllidae), a serious pest of tomato, Solanum lycopersicon (Mill), in the western half of North America. In choice studies, tomato psyllids preferred to settle on plants that did not contain the gene [Moneymaker (mi‐1.2)] compared to near‐isogenic plants with the gene [Motelle (Mi‐1.2)]. As a result, total oviposition was higher on the susceptible variety, although no‐choice studies indicated that there were no differences in numbers of eggs laid by individual females on either variety. Survival from egg to adult was higher on plants lacking the gene compared to plants containing the gene. However, there were no differences in total development time of individuals reared from either variety. The results suggest that mechanisms of resistance to the tomato psyllid observed in plants bearing the Mi‐1.2 gene are distinct from the mechanisms of resistance to the three phloem feeders examined in other studies.     

Identification and location of symbionts associated with potato psyllid (Bactericera cockerelli) lifestages

The potato psyllid (Bactericera cockerelli, Sulc) is an invasive pest of solenaceous plants including potatoes (Solanum tuberosum L.)and tomatoes (Solanum lycopersicum L.). The insect transmits the phytopathogen Candidatus Liberibacter solanacearum, which has been identified as the causal agent of Zebra Chip in potatoes. The microbiome of the potato psyllid provides knowledge of the insect's bacterial makeup which enables researchers to develop targeted biological control strategies. In this study, the microbes associated with four B. cockerelli life stages were evaluated by 16S bTEFAP pyrosequencing. The sequences were compared with a 16S-rDNA database derived from NCBI's GenBank. Some bacteria identified are initial discoveries. Species of Wolbachia, Rhizobium, Gordonia, Mycobacterium, Xanthomonas and others were also detected and an assessment of the microbiome associated with B. cockerelli was established.     

Comparative fitness of invasive and native populations of the potato psyllid (Bactericera cockerelli)

Two genetically distinct potato psyllid populations [Bactericera cockerelli (Sulc) (Homoptera: Psyllidae)] were identified in our previous study: native and invasive. The invasive population, ranging from Baja, Mexico to central California, was the result of a recent invasion, while the native population is endemic to Texas. The native (Texas) and invasive (California) populations were collected from tomato and pepper, respectively, and were examined on both hosts to test the comparative fitness of invasive populations. Our results indicated that on both plant hosts, psyllids from the native range demonstrated higher survivorship, a higher growth index, and shorter development times than the psyllids from invasive populations. The fecundity of the native-range psyllids also was significantly higher than that of invasive psyllids on tomato, but not on pepper. For the native population, host plant differences for all fitness measurements were not significant. However, within the invasive population, psyllids feeding on tomatoes showed consistently better survivorship and a higher growth index than those feeding on pepper, despite the decreased developmental time required on peppers. The LC₅₀ values (concentrations causing 50% mortality) of both populations were determined for three pesticides. Resistance to two of these pesticides was found in the invasive population. Thus, the invasive quality of the California populations may be related to increased pesticide resistance. However, it is impossible to determine if the California population was preadapted to pesticide resistance, or if the resistance developed after the range expansion and is simply a contributing factor to maintaining the expansion.     

A new Huanglongbing Species, "Candidatus Liberibacter psyllaurous," found to infect tomato and potato, is vectored by the psyllid Bactericera cockerelli (Sulc)

A new huanglongbing (HLB) "Candidatus Liberibacter" species is genetically characterized, and the bacterium is designated "Candidatus Liberibacter psyllaurous." This bacterium infects the psyllid Bactericera cockerelli and its solanaceous host plants potato and tomato, potentially resulting in "psyllid yellowing." Host plant-dependent HLB transmission and variation in psyllid infection frequencies are found.     

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.     

Ovipositional preferences, damage thresholds, and detection of the tomato-potato psyllid Bactericera cockerelli (Homoptera: Psyllidae) on selected tomato accessions

The tomato-potato psyllid Bactericera [Paratrioza] cockerelli (Sulc) has recently caused losses exceeding 50% on fresh market tomatoes in California and Baja, Mexico by injecting a toxin that results in a condition known as 'psyllid yellows'. The objectives of this study were to: (i) document oviposition preferences on a range of tomato cultivars; (ii) determine threshold levels for psyllid densities that would cause psyllid yellows on tomatoes within the first three weeks following transplanting; and (iii) identify the most important 'psyllid yellows' symptoms that might be used in surveying and monitoring for this pest. Plant lines tested included the commonly-planted commercial cultivars 'Shady Lady' and 'QualiT 21', an older, previously commercial cultivar '7718 VFN', a common cultivar planted by consumers 'Yellow Pear', and a wild type plant accession, PI 134417. When given a choice, psyllids significantly preferred 'Yellow Pear' and avoided PI 134417 for oviposition. Under no-choice conditions psyllids laid significantly fewer eggs on PI 134417, but all the other plant lines were equally good substrates for laying eggs. Thus, oviposition preference is not likely to provide a functional management strategy in large plantings. On 'Shady Lady', psyllids preferred to oviposit on plants already infested with adults. On both 'Shady Lady' and '7718 VFN' oviposition was significantly greater on plants previously infested by nymphs as compared to uninfested control plants. This suggests that, at least for some cultivars, there is a physiological change in plant attractiveness following psyllid feeding. 'Yellow Pear' and 'QualiT 21' were relatively tolerant of psyllids, requiring 18 nymphs per plant to produce the disease symptoms. Only eight nymphs per plant were needed on 'Shady Lady' and '7718 VFN'. For all cultivars, the pest density showed strong correlations with measurements such as the number of yellowing leaves and leaflets and distorted leaves, which were as good as or better than the first factor extracted from principal component analysis. Therefore, such measurements have the potential to simplify field surveys.     

Age at reproductive maturity and effect of age and time of day on sex attraction in the potato psyllid Bactericera cockerelli

Abstract The potato psyllid, Bactericera (= Paratrioza) cockerelli (?ulc) (Hemiptera: Triozidae), is a major pest of potato. Studies were conducted to determine the age at which both males and females reach reproductive maturity and the effect of age and time of day on sex attraction. Adult B. cockerelli reach reproductive maturity within 48 h post‐eclosion, with females being mature on the day of eclosion and males at 1 day post‐eclosion. Oviposition generally began 2 days after mating but was delayed when females mated within 2 days post‐eclosion. In laboratory olfactometer assays, the age of females used as odor sources and the age of males assayed to these odors did not affect sex attraction, with both younger (1–4‐day‐old) and older (8–10‐day‐old) males being attracted to females, regardless of female age (1–4‐day‐old or 8–10‐day‐old). Males assayed to live females at different times during the photophase (between 8:00 and 20:00 hours) were attracted to females between 11:00 and 17:00 hours, showing a temporal periodicity in sex attraction with B. cockerelli at least during the photophase.     

Insecticidal activity of entomopathogenic fungi (Hypocreales) for potato psyllid,Bactericera cockerelli (Hemiptera: Triozidae): Development of bioassay techniques,effect of fungal species and stage of the psyllid

The potato psyllid, Bactericera cockerelli (?ulc), is a pest of potato, tomato, and some other solanaceous vegetables and has also been incriminated in the transmission of a bacterial pathogen, Candidatus Liberibacter solanacearum, resulting in a serious disease known as ‘zebra chip’. Although there are several reports of fungal pathogens in psyllids, there are none from B. cockerelli, nor have any fungi been evaluated against it. Five isolates of fungi, one Beauveria bassiana, two Metarhizium anisopliae and two Isaria fumosorosea, were bioassayed against B. cockerelli on potato leaves under ideal conditions for the fungi. All applications were made with a Potter spray tower. With the exception of concentration-effect studies, all other applications were made using 10⁷ conidia/mL in a 2-mL aqueous suspension. All isolates except B. bassiana, produced 95–99% mortality, corrected for control mortality, in adults 2–3 days after application of conidia and 91–99% in nymphs 4 days after application. The corrected mortalities for adults and nymphs treated with B. bassiana were 53 and 78%, respectively, 4 days after application. I. fumosorosea Pfr 97 produced 95% corrected mortality in both first and late third instar nymphs. M. anisopliae (F 52) produced 96% corrected mortality in first and third instar nymphs. Pfr 97 and F 52 were evaluated for insecticidal activity against third instar B. cockerelli using 10⁵, 10⁶, and 10⁷ conidia per mL. Mortality produced by I. fumosorosea Pfr 97 ranged from 83 to 97% and that of M. anisopliae F 52 was 88 to 95% at these concentrations.     

Genetic differentiation between eastern populations and recent introductions of potato psyllid (Bactericera cockerelli) into western North America

Although tomato psyllid, Bactericera cockerelli (Sulc) (Homoptera, Psyllidae), annually causes significant losses in potato and tomato crops in eastern Mexico and the central United States, infestations in western North America have been historically rare. However, substantial populations appeared in 2001 in western North America and caused losses in tomato production exceeding 80%; losses in 2004 reached 50%. To determine if these new outbreaks were the result of a simple range expansion or the evolution of a new B. cockerelli biotype, inter simple sequence repeat (ISSR) markers, as well as mitochondrial gene cytochrome oxidase I (COI), internal transcribed spacer 2 (ITS2), and wsp sequence data were used to characterize populations of the psyllid. Western populations from Baja, Mexico, Orange County, and Ventura County were compared with populations from central USA (Colorado and Nebraska) and eastern Mexico (Coahuila). Based on ISSR markers, the psyllid populations clustered into two groups, with one group including populations from western North America and the other group including populations from central USA and eastern Mexico. For COI comparisons, there was one base‐pair difference found in the 544 bp‐long COI fragments, but the populations again segregated along the same geographic lines. Two strains of Wolbachia were identified, the maximal differences between wsp clones from all populations was 5 bp for strain Bac1 and 23 bp for strain Bac2 out of a 555‐bp fragment. The ISSR data, therefore, were consistent in indicating the development of a new psyllid biotype that has adapted to western North America rather than a simple range expansion, but the other genetic data sets were less conclusive.     

The effectiveness of double-stranded short inhibitory RNAs (siRNAs) may depend on the method of transfection

RNA interference (RNAi) is a recently described powerful experimental tool that can cause sequence-specific gene silencing, thereby facilitating functional analysis of gene function. Consequently, we became interested in using RNAi to determine the function of aberrantly expressed ErbB3 in the KAS-6/1 human myeloma cell line. Despite the wealth of information available on the use of RNAi, dsRNA target design, and the transfection of dsRNA in vitro, little information is available for transfecting dsRNA into nonadherent cells from any species. In the present study, we report that gene silencing of ErbB3 was not observed in myeloma cells when dsRNA targeting ErbB3 was introduced using conventional transfection agents and protocols that have proved successful for several adherent cell lines. Silencing of ErbB3, however, was observed in T47D cells, an adherent breast carcinoma cell line, using the same transfection methods, indicating that our target sequence was functional for gene silencing of ErbB3. Interestingly, ErbB3 was silenced in myeloma cells when the dsRNA target was introduced by electroporation. Thus, our studies illustrate the striking dependence of dsRNA-mediated gene silencing in some cells on the methods of dsRNA transfection.     

The p53-induced Wig-1 protein binds double-stranded RNAs with structural characteristics of siRNAs and miRNAs

Wig-1 is a p53-induced zinc finger protein. Here we show that human Wig-1 binds long (>or=23 bp) dsRNAs with 5'-overhangs. The first zinc finger domain is necessary but not sufficient for this dsRNA-binding in vitro. Wig-1 also binds dsRNA in living cells via zinc fingers 1 and 2. Both zinc fingers 1 and 2 are important for Wig-1-mediated growth suppression. Moreover, Wig-1 binds 21 bp dsRNAs with 3'-protruding ends. These findings demonstrate that human Wig-1 can bind different types of dsRNAs, including dsRNAs resembling small interfering RNAs (siRNAs) and microRNAs (miRNAs), and indicate that dsRNA binding has a role in Wig-1-mediated regulation of cell growth.     

Differential responses to feeding by the tomato/potato psyllid between two tomato cultivars and their implications in establishment of injury levels and potential of damaged plant recovery

An invasive new biotype of the tomato/potato psyllid （Bactericera [Paratrioza] cockerelli [Sulc.]） （Homoptera： Psyllidae） recently has caused losses exceeding 50% on fresh market tomatoes in western North America. Despite these extensive losses, little is known regarding the threshold levels at which populations must be suppressed in order to prevent economic losses. A series of experiments were therefore designed using combinations of two common tomato cultivars （QualiT 21 and Yellow Pear）, five pest-densities （0, 20, 30, 40 and 50 nymphs/plant）, and three feeding-duration （5 days, 10 days, and lifetime） treatments to test the relative importance of pest density, feeding period, and cumulative psyllid-days to establish economic threshold levels for psyllids. The cultivars differed considerably in their response to the toxin injected by the psyllid nymphs. ‘Yellow Pear＇ plants could recover from feeding by up to 40 nymphs for as long as 10 d, whereas ‘QualiT 21＇ plants were irreparably damaged by densities of 20 nymphs feeding for only 5 days. On ‘Yellow Pear＇, all plant measurements such as the number of yellow leaves and plant height were significantly better correlated with cumulative psyUid-days than with either pest density or feeding duration. On ‘QualiT 21 ＇, all plant measurements other than the number of yellow leaflets and leaves were significantly better correlated with pest density than with feeding duration or cumulative psyUid-days, and pest density was a better predictor of psyUid damage. Potential reasons for the variable responses between cultivars and the implications for psyllid sampling and integrated pest management are discussed.     

Movement of Bactericera cockerelli (Heteroptera: Psyllidae) in relation to potato canopy structure, and effects on potato tuber weights

With the threat of new plant diseases on the increase, plant disease epidemiology requires research on pathogen vector movement. Here, releases were performed in planted potato fields of different ages and canopy structures, located in the Texas Panhandle, to evaluate the range of movement of the potato psyllid Bactericera cockerelli (Sulc.). This insect is a known causative agent of psyllid yellows disease, and is a vector of the reported etiological agents of zebra chip disease of potato, 'Candidatus Liberibacter solanacearum/psyllaurous'. Based on collections of B. cockerelli immatures along transects 9 m long radiating in four cardinal directions from release points, adult females dispersed considerable distances, regardless of plant age or canopy structure. Immature abundance declined along transects and were well described by linear and nonlinear models, but abundance patterns did not differ among the different planting dates and canopy structures. However, unequal immature abundance was detected among the four cardinal directions, with more immatures generally collected along transects to the north and west of release points, opposite of prevailing winds in the area at the time of release. Plots where B. cockerelli were released had significantly lower mean potato tuber weights than control plots with no B. cockerelli. However, few plots with B. cockerelli released in them had declining trends in tuber weights with increasing distance from release points.     

Vector transmission efficiency of liberibacter by Bactericera cockerelli (Hemiptera: Triozidae) in zebra chip potato disease: effects of psyllid life stage and inoculation access period

Successful transmission of plant pathogens by insects depends on the vector inoculation efficiency and how rapidly the insect can effectively transmit the pathogen to the host plant. The potato psyllid, Bactericera cockerelli (Sulc), has recently been found to transmit "Candidatus Liberibacter solanacearum," a bacterium associated with zebra chip (ZC), an emerging and economically important disease of potato in several parts of the world. Currently, little is known about the epidemiology of ZC and its vector's inoculation capabilities. Studies were conducted in the field and laboratory to 1) assess transmission efficiency of potato psyllid nymphs and adults; 2) determine whether psyllid inoculation access period affects ZC incidence, severity, and potato yield; and 3) determine how fast the psyllid can transmit liberibacter to potato, leading to ZC development. Results showed that adult potato psyllids were highly efficient vectors of liberibacter that causes ZC and that nymphs were less efficient than adults at transmitting this bacterium. It was also determined that inoculation access period had little influence on overall ZC disease incidence, severity, and resulting yield loss. Moreover, results showed that exposure of a plant to 20 adult potato psyllids for a period as short as 1 h resulted in ZC symptom development. Furthermore, it was shown that a single adult potato psyllid was capable of inoculating liberibacter to potato within a period as short as 6 h, thereby inducing development of ZC. This information will help in developing effective management strategies for this serious potato disease.     

Oral delivery of double-stranded RNA induces prolonged and systemic gene knockdown in Metaseiulus occidentalis only after feeding on Tetranychus urticae

Metaseiulus (=Typhlodromus or Galendromus) occidentalis is an important biological control agent. Functional genomic studies on this predator have been hampered by the lack of reverse genetic tools such as RNA interference (RNAi). In the current study, we evaluated possible RNAi responses in M. occidentalis females by feeding double-stranded RNA (dsRNA) of RpL11, RpS2, RpL8, or Pros26.4 genes in 20 % sucrose solution. Females needed to subsequently feed on two-spotted spider mites (Tetranychus urticae) to elicit a nearly complete loss of egg production. The corresponding gene knockdown was robust, long-term, and was observed in the very few eggs produced (systemic or parental RNAi). Interestingly, dsRNA-mediated gene knockdown could not be induced if these predators were provided only the sucrose diet after ingesting dsRNAs; T. urticae had to be provided to elicit the RNAi response. However, the spider mite diet was not needed for sustaining the dsRNA-mediated gene knockdown once it commenced. Oral delivery of dsRNA will be a valuable tool for efficient genome-wide functional screens in this important predatory mite.     

Functional comparison of single- and double-stranded siRNAs in mammalian cells

The concept of small interfering RNA (siRNA) has been extended to include not only short double-stranded RNA of 19-25bp, but also single-stranded antisense RNA of the same length, since such single-stranded antisense siRNAs were recently found to be able to inhibit gene expression as well. We made comprehensive comparison of double- and single-stranded siRNA functions in RNA interference (RNAi), targeting multiple sites and different mRNAs, measuring RNAi effects at different time-points and in different cell lines, and examining response curves. Duplex siRNAs were found to be more potent than single-stranded antisense siRNAs. This was verified by the observation that single-stranded antisense siRNAs, which were inefficient in some cases when used alone, could be rescued from inefficiency by sequentially transfecting with the sense siRNAs. This result suggests that the structural character of siRNA molecules might be a more important determinant of siRNA efficiency than the cellular persistence of them.