Lance S. Osborne

The eubacterial endosymbionts of whiteflies (homoptera: Aleyrodoidea) constitute a lineage distinct from the endosymbionts of aphids and mealybugs

Whiteflies (superfamily Aleyrodoidea) contain eubacterial endosymbionts localized within host cells known as mycetocytes. Sequence analysis of the genes for the 16S rRNA of the endosymbionts ofBemisia tabaci, Siphoninus phillyreae, andTrialeurodes vaporariorum indicates that these organisms are closely related and constitute a distinct lineage within the γ-subdivision of theProteobacteria. The endosymbionts of whiteflies are unrelated to the endosymbionts of aphids and mealybugs, which are in two separate lineages.

The Banker Plant Method in Biological Control

In the banker plant method, long-lasting rearing units for beneficials are created in the crop by distributing plants infested with herbivores or carrying other food items, such as pollen. The method has been widely investigated over many years and used to aid establishment, development and dispersal of beneficial organisms employed in biological control. In this review, we refine the definition of the banker plant method based on previous concepts and studies and offer the term “banker plant system” to describe the unit that is purposefully added to or established in a crop for control of pests in greenhouses or open field. The three basic elements of a banker plant system (banker plant, food source, beneficials) are discussed and illustrated with examples, and the diversity of banker plant systems (classified by target pest) used or investigated is documented. The benefits of using banker plant systems, such as low cost, increased freshness of beneficials, possibility for preventive control and for integration within IPM frameworks, make the method an interesting plant protection option with potential to enhance adoption of biological control in pest management programs.

Distribution of Bemisia tabaci (Hemiptera: Aleyrodidae) Biotypes in Florida-Investigating the Q Invasion

ABSTRACT After the 2004 discovery of the Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) Q biotype in the United States, there was an urgent need to determine its distribution. As part of a coordinated country-wide effort, an extensive survey of B. tabaci biotypes was conducted in Florida, with the cooperation of growers and state and federal agencies, to monitor the introduction and distribution of both the B and Q biotypes. The biotype status of submitted B. tabaci samples was determined by polymerase chain reaction (PCR) amplification and sequencing of a 700–800-bp mitochondrial cytochrome oxidase I small subunit (mtCOI) gene fragment, PCR amplification, and size determination of two unique microsatellite markers and esterase zymogram analysis. One hundred and eighty collections were sampled from 23 counties. Of these samples, 58% were from vegetables, 37% were from ornamentals, and 5% were from peanuts, alfalfa, and weeds. Eighteen percent of all collections were found to be the Q biotype that came from greenhouse grown ornamental and herbs located in six counties. Sequence comparison of the mtCOI gene identified three separate haplotypes within Florida that were defined as Q1, Q2, and Q3. Haplotypes could be used to associate populations known to be related by grower and plant type. For example, collections from five counties were made on hibiscus linked to the same grower and all samples contained only the Q1 haplotype. Other populations contained a mix of the Q2 and Q3 haplotypes, supporting the conclusion that the Q biotype must have entered Florida through at least two separate introductions. Our data also show that two microsatellite markers are a cost-effective diagnostic alternative for biotype identification with 100% concurrence with mtCOI sequence data.

Distribution of Bemisia tabaci (Hemiptera: Aleyrodidae) Biotypes in North America After the Q Invasion

ABSTRACT After the 2004 discovery of the Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) Q biotype in the United States, there was a vital need to determine the geographical and host distribution as well as its interaction with the resident B biotype because of its innate ability to rapidly develop high-level insecticide resistance that persists in the absence of exposure. As part of a coordinated country-wide effort, an extensive survey of B. tabaci biotypes was conducted in North America, with the cooperation of growers, industry, local, state, and federal agencies, to monitor the introduction and distribution of the Q biotype. The biotype status of submitted B. tabaci samples was determined either by polymerase chain reaction amplification and sequencing of a mitochondrial cytochrome oxidase I small subunit gene fragment and characterization of two biotype discriminating nuclear microsatellite markers or esterase zymogram analysis. Two hundred and eighty collections were sampled from the United States, Bermuda, Canada, and Mexico during January 2005 through December 2011. Host plants were split between ornamental plant and culinary herb (67%) and vegetable and field crop (33%) commodities. The New World biotype was detected on field-grown tomatoes (Solanum lycopersicum L) in Mexico (two) and in commercial greenhouses in Texas (three) and represented 100% of these five collections. To our knowledge, the latter identification represents the first report of the New World biotype in the United States since its rapid displacement in the late 1980s after the introduction of biotype B. Seventy-one percent of all collections contained at least one biotype B individual, and 53% of all collections contained only biotype B whiteflies. Biotype Q was detected in 23 states in the United States, Canada (British Columbia and Ontario territories), Bermuda, and Mexico. Forty-five percent of all collections were found to contain biotype Q in samples from ornamentals, herbs and a single collection from tomato transplants located in protected commercial horticultural greenhouses, but there were no Q detections in outdoor agriculture (vegetable or field crops). Ten of the 15 collections (67%) from Canada and a single collection from Bermuda contained biotype Q, representing the first reports of biotype Q for both countries. Three distinct mitochondrial haplotypes of B. tabaci biotype Q whiteflies were detected in North America. Our data are consistent with the inference of independent invasions from at least three different locations. Of the 4,641 individuals analyzed from 517 collections that include data from our previous work, only 16 individuals contained genetic or zymogram evidence of possible hybridization of the Q and B biotypes, and there was no evidence that rare hybrid B-Q marker co-occurrences persisted in any populations.

Assessment ofBeauveria bassiana Nov. EO-1 strain, which produces a red pigment for microbial control

A new strain of the fungusBeauveria bassiana Nov. EO-1 (ATCC 74037), which produces a red pigment in solid and liquid culture, has been isolated from an infected whitefly. The red pigment was extracted and has been identified by mass spectrometry as oosporein, a potent dibenzoquinone mycotoxin.In order to assess the potential of this entomogenous fungi for microbial control purposes, a mycelium bead formulation was developed as a source for pathogenic conidial spores and oosporein production. The mycelium bead preparation was found to be a stable fungal carrier. Conidiation and germination studies have revealed the mycelium bead viability is 100% over a 1-yr period when stored at 4°C. Conidial spore production from the mycelium beads has been falling substantially per time from an initial value of 1.5 × 108 spores per bead to 3 × 105 spores per bead after a year storage at 4°C. However, the mycelium bead formulation continues to produce oosporein on agar media, at the same intensity throughout the 1 yr period.In in vitro and in vivo small scale greenhouse experimentsBeauveria bassiana Nov. EO-1 were compared with known entomogenous fungi,Beauveria sp. andPaceilomyces sp. Beauveria bassiana Nov. EO-1 was found to have a high pathogenicity against foliage insect pests (e.g., whiteflies and mealy bugs), and against soil insects (e.g., citrus root weevils). The utilization of a mycelium bead based on this strain,Beauveria bassiana Nov. EO-1, as a source of conidial spores and oosporein may have broad applications for the control of various insect pests.

Effects of the fungus Isaria fumosorosea (Hypocreales: Cordycipitaceae) on reduced feeding and mortality of the Asian citrus psyllid, Diaphorina citri (Hemiptera: Psyllidae)

Abstract The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is an important citrus pest primarily because it transmits bacteria putatively responsible for huanglongbing, a serious disease of citrus. We present a study on the effects of blastospore and conidial formulations of Isaria fumosorosea Wize on feeding rates and mortality of adult psyllids in laboratory bioassays. Information on quantities of honeydew droplets was used to make inferences on feeding rates. Psyllids treated with the blastospore formulation of I. fumosorosea produced fewer honeydew droplets compared to the conidial treatment and control beginning within the first 24 h after treatment. The highest daily mean number of droplets thereafter never exceeded 2.4 drops compared to 4 and 8 for the conidial treatment and control, respectively. The mean number (±SEM) of honeydew droplets produced per psyllid per day over 7 days was significantly higher in the control (5.5±0.5) compared to the blastospore treatment; however, there were no significant differences between the treatments. Psyllids treated with the conidial formulation of the pathogen showed no significant reduction in feeding activity until 4 days after treatment. One and 2 days post-exposure, mortality of psyllids in the blastospore treatment ranged from 8 to 25% compared to 0% in the conidial and control treatments. By 7 days post-exposure, psyllid mortality reached 100% under both fungal treatments compared to none in the controls. This study documented that adult psyllids infected by I. fumosorosea (PFR 97) produce less honeydew than healthy psyllids and suggests that they may feed less, which could potentially reduce the spread of huanglongbing.

Life History of Paracoccus marginatus (Hemiptera: Pseudococcidae) on Four Host Plant Species Under Laboratory Conditions

Abstract Life history of the mealybug, Paracoccus marginatus Williams and Granara de Willink, on three ornamental plants [Hibiscus rosa-sinensis L., Acalypha wilkesiana (Muell.-Arg.), and Plumeria rubra L.] and one weed species (Parthenium hysterophorus L.) was studied under laboratory conditions. Mealybugs were able to develop, survive, and reproduce on all four hosts; however, there were differences in the life history parameters. Adult females that developed on acalypha and parthenium emerged ≈1 d earlier than those that developed on hibiscus and plumeria. Adult males had a longer developmental time on plumeria than on the other hosts. Survival of first- and second-instar nymphs and cumulative adult survival were lowest on plumeria. Longevity was not affected by hosts for males and females and averaged 2.3 ± 0.1 and 21.2 ± 0.1 d, respectively. On plumeria, 58.9 ± 1.7% of the adults were females, which was a higher female percentage than on the other hosts. No egg production occurred in virgin females. Prereproductive and reproductive periods of the females were not affected by hosts and averaged 6.3 ± 0.1 and 11.2 ± 0.1 d, respectively. Mean fecundity of 186.3 ± 1.8 eggs on plumeria was lower than on the other three plant species. Life history parameters of P. marginatus on hibiscus, acalypha, plumeria, and parthenium show its ability to develop, survive, and reproduce on a wide variety of plant species.

OVIPOSITION AND LARVAL SURVIVAL OF DIAPREPES ABBREVIATUS (COLEOPTERA: CURCULIONIDAE) ON SELECT HOST PLANTS

Abstract In a preliminary survey in four commercial ornamental nurseries in south Florida (1998), Diaprepes abbreviatus (L.) (Coleoptera: Curculionidae) egg masses, feeding damage, or adults occurred on numerous field-grown ornamental plant species. Live oak (Quercus virginiana Mill.), silver buttonwood (Conocarpus erectus L. variety sericeus Fors. Ex DC), and black olive (Bucida buseras L.) had the highest percentage of plants with egg masses. Adult feeding damage was found on all examined plants of dahoon holly (Ilex cassine L.), cocoplum (Chrysobalanus icaco L.), black olive, live oak, Bauhinia sp., and Cassia sp. Oviposition of D. abbreviatus was evaluated in no-choice, two-choice, three-choice and multiple-choice caged tests. In no-choice tests, silver buttonwood had the highest mean number of egg masses. In two-choice tests, egg masses were laid on all plant species tested but there were significantly more egg masses on silver buttonwood than the alternate choice. The number of egg masses in the three-choice tests was low and there were no significant differences among the plant species tested. As in the no-choice and two-choice tests, significantly more egg masses were found on silver buttonwood in multiple-choice tests. Survival of larvae and their effect on plant growth was examined on several commonly grown plant species in southern Florida. Larval survival was highest on silver buttonwood and Sorghum sudanense Pers (sorghum-sudan) compared with other plant species. Root and/or total biomass was significantly reduced on green bean (Phaseolus vulgaris), silver buttonwood, Tahiti lime (Citrus aurantifolia), and sorghum-sudan.

The Scirtothrips dorsalis Species Complex: Endemism and Invasion in a Global Pest

Invasive arthropods pose unique management challenges in various environments, the first of which is correct identification. This apparently mundane task is particularly difficult if multiple species are morphologically indistinguishable but accurate identification can be determined with DNA barcoding provided an adequate reference set is available. Scirtothrips dorsalis is a highly polyphagous plant pest with a rapidly expanding global distribution and this species, as currently recognized, may be comprised of cryptic species. Here we report the development of a comprehensive DNA barcode library for S. dorsalis and seven nuclear markers via next-generation sequencing for identification use within the complex. We also report the delimitation of nine cryptic species and two morphologically distinguishable species comprising the S. dorsalis species complex using histogram analysis of DNA barcodes, Bayesian phylogenetics, and the multi-species coalescent. One member of the complex, here designated the South Asia 1 cryptic species, is highly invasive, polyphagous, and likely the species implicated in tospovirus transmission. Two other species, South Asia 2, and East Asia 1 are also highly polyphagous and appear to be at an earlier stage of global invasion. The remaining members of the complex are regionally endemic, varying in their pest status and degree of polyphagy. In addition to patterns of invasion and endemism, our results provide a framework both for identifying members of the complex based on their DNA barcode, and for future species delimiting efforts.

Population Genetics of Invasive Bemisia tabaci (Hemiptera: Aleyrodidae) Cryptic Species in the United States Based on Microsatellite Markers

ABSTRACT The Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) cryptic species complex of whitefiies contains two species, MEAM1 and MED, that are highly invasive in supportive climates the world over. In the United States, MEAM1 occurs both in the field and in the greenhouse, but MED is only found in the greenhouse. To make inferences about the population structure of both species, and the origin and recent spread of MED within the United States, 987 MEAM1 whitefiies and 340 MED whitefiies were genotyped at six and seven microsatellite loci, respectively, for population genetic analyses. Major results of the study are 1) MED exhibits more population structure and genetic differentiation than MEAM1, 2) nuclear microsatellite markers exhibit a high degree of concordance with mitochondrial markers recovering a major east-west phylogeographic break within MED, 3) both eastern and western MED are found throughout the continental United States and eastern MED is present in Hawaii, and 4) MEAM1 contains two greenhouse U.S. populations significantly differentiated from other U.S. MEAM1. The results suggest that MED was introduced into the United States on at least three occasions and rapidly spread throughout the United States, showing no discernible differentiation across 7,000 km. The results further suggest that there is an enhanced role of the protected agricultural environment in promoting genetic differentiation in both invasive B. tabaci cryptic species.