Susan E. Webb

Location of Geminiviruses in the Whitefly Bemisia tabaci (Homoptera: Aleyrodidae)

The location of tomato mottle virus (ToMoV) and cabbage leaf curl virus (CabLCV) (Geminiviridae, genus Begomovirus) in the whitefly vector Bemisia tabaci B-biotype (Homoptera: Aleyrodidae) was elucidated using a novel technique incorporating indirect immunofluorescent labeling in freshly dissected whiteflies. Begomoviruses were visualized in the whitefly by indirect-fluorescent-microscopy. Polyclonal and monoclonal primary antibodies were used to successfully detect both ToMoV and CabLCV. Both begomoviruses were located in the anterior region of the midgut and filter-chamber of adult whiteflies, with ToMoV detected in the salivary glands. CabLCV was detected at a greater frequency than ToMoV, with a positive detection of 16% (89 out of 560) for CabLCV and 3% (25 out of 840) for ToMoV. Possible sites involved in geminivirus transport from the gut lumen of whiteflies into the hemocoel were located in the filter-chamber and anterior portion of the midgut. The location of these begomoviruses suggests a possible scenario of virus movement through the whitefly, which is discussed.

Identification and Characterization of a Novel Whitefly-Transmitted Member of the Family Potyviridae Isolated from Cucurbits in Florida

ABSTRACT A novel whitefly-transmitted member of the family Potyviridae was isolated from a squash plant (Cucurbita pepo) with vein yellowing symptoms in Florida. The virus, for which the name Squash vein yellowing virus (SqVYV) is proposed, has flexuous rod-shaped particles of approximately 840 nm in length. The experimental host range was limited to species in the family Cucurbitaceae, with the most dramatic symptoms observed in squash and watermelon, but excluded all tested species in the families Amaranthaceae, Apocynaceae, Asteraceae, Chenopodiaceae, Fabaceae, Malvaceae, and Solanaceae. The virus was transmitted by whiteflies (Bemisia tabaci) but was not transmitted by aphids (Myzus persicae). Infection by SqVYV induced inclusion bodies visible by electron and light microscopy that were characteristic of members of the family Potyviridae. Comparison of the SqVYV coat protein gene and protein sequences with those of recognized members of the family Potyviridae indicate that it is a novel member of the genus Ipomovirus. A limited survey revealed that SqVYV also was present in watermelon plants suffering from a vine decline and fruit rot recently observed in Florida and was sufficient to induce these symptoms in greenhouse-grown watermelon, suggesting that SqVYV is the likely cause of this disease.

Squash vein yellowing virus Detection Using Nested Polymerase Chain Reaction Demonstrates that the Cucurbit Weed Momordica charantia Is a Reservoir Host

Squash vein yellowing virus (SqVYV) is a recently described ipomovirus from cucurbits in Florida that induces the relatively unusual symptoms in watermelon of plant death and fruit rind necrosis and discoloration, commonly known in Florida as watermelon vine decline. In this report, SqVYV infection of Momordica charantia (Balsam-apple), a common cucurbit weed, collected in 2005 and 2007 from within or adjacent to fields of declining watermelon, is demonstrated through the use of nested polymerase chain reaction (PCR). M. charantia plants located in or around fallow watermelon fields between spring and fall 2007 watermelon crops were also infected with SqVYV, indicating that this weed can serve as an oversummering host for this virus. Furthermore, whiteflies were able to acquire SqVYV from infected M. charantia and transmit it to squash and watermelon. Nested PCR was 10 to 1,000 times more sensitive than non-nested PCR for SqVYV detection in several cucurbit hosts, including M. charantia and watermelon. Melothria pendula (creeping cucumber), another common cucurbit weed, was experimentally infected with SqVYV. These results suggest that improved management of M. charantia and other cucurbit weeds needs to be incorporated into watermelon vine decline management plans to reduce sources of SqVYV and other cucurbit viruses.

Presence of P1b and absence of HC-Pro in Squash vein yellowing virus suggests a general feature of the genus Ipomovirus in the family Potyviridae

The genus Ipomovirus is one of six currently recognized genera in the family Potyviridae. The complete nucleotide sequence of Squash vein yellowing virus (SqVYV), a putative ipomovirus recently described in Florida, has been determined. The 9836 nt SqVYV genomic RNA [excluding the poly(A) tail] has one large open reading frame encoding a single polyprotein of 3172 amino acids, typical of the genome organization for most members in the family Potyviridae. The 10 mature proteins predicted to be derived from the SqVYV polyprotein include P1a and P1b but no HC-Pro, similar to Cucumber vein yellowing virus (CVYV) but different from Sweet potato mild mottle virus (SPMMV), both recognized members of the genus Ipomovirus. Phylogenetic analysis of these proteins supports classification of SqVYV as a novel species within the genus Ipomovirus. However, the similar genome organization strategy of SqVYV and CVYV, which differs from that of SPMMV, indicates that the taxonomy of the genus Ipomovirus needs to be re-examined and a new genus created within the family Potyviridae to accommodate the observed discrepancies in ipomovirus genome organization.

Precibarial and cibarial chemosensilla in the whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae)

Abstract The internal anatomy of the anterior alimentary canal of the whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) B-biotype, was examined by light, scanning, and transmission electron microscopy to elucidate the location and number of precibarial and cibarial gustatory sensilla. Elucidation of the epipharyngeal organ complex within the precibarium revealed 10 precibarial sensilla located proximal to where the paired maxillary stylets diverge on their retraction. The sensory organ complex within the cibarium consists of 8 sensilla, 6 on the epipharyngeal sclerite with 2 found within the hypopharyngeal sclerite. Fine structure investigation revealed the individual neurons to terminate at sensillar pores, which allow direct contact with passing fluids, thus supporting a chemosensory function. Ultrastructure of the neurons is similar to that of precibarial and cibarial gustatory chemosensilla found in other piercing-sucking insects. Their importance to whitefly feeding is discussed.

Trailers Transporting Oranges to Processing Plants Move Asian Citrus Psyllids

ABSTRACT Huanglongbing (citrus greening) is one of the most serious diseases of citrus. Movement of the disease occurs as a result of natural transmission by the insect vector and by movement of infected plant material. We demonstrate here that Diaphorina citri Kuwayama, the vector of citrus greening pathogens, can be transported in trailers of unprocessed fruit. Several samples of D. citri collected from trailers of oranges were positive for citrus greening pathogens, indicating that the disease can spread widely with the movement of the mature fruit as a result of moving the vectors. While movement of disease and the vector through plant materials into new areas has been recognized, our findings emphasize the need to evaluate the importance of long distance movement of psyllids on unprocessed fruit, even in the absence of vegetative tissue.

Suppression of Whiteflies, Bemisia tabaci (Hemiptera: Aleyrodidae) and Incidence of Cucurbit Leaf Crumple Virus, a Whitefly-transmitted Virus of Zucchini Squash New to Florida, with Mulches and Imidacloprid

Abstract The sweetpotato whitefly, Bemisia tabaci B biotype (Gennadius) (Homoptera: Aleyrodidae) also known as Bemisia argentifolii (Bellows and Perring) is a key pest in zucchini squash (Cucurbita pepo L.) causing both direct and indirect damage. A field experiment was conducted in the fall of 2005 and repeated in the fall of 2006 to investigate the effect of mulches alone or in combination with a reduced-risk insecticide, imidacloprid (Admire® 2F) on whiteflies and related problems in zucchini squash. UV-reflective mulch and the living mulch, buckwheat (Fagopyrum esculentum Moench), with and without imidacloprid, were evaluated in a randomized complete block design with 4 replications. White mulch (standard) was included as a control in the experiment. Mulches in combination with imidacloprid resulted in fewer whiteflies per leaf than those without imidacloprid in 2005 and 2006. More whiteflies were recorded on zucchini plants growing on white synthetic mulch than all other treatments in both years. In the 2006 growing season, these plants also showed the first symptoms of viral infection caused by Cucurbit leaf crumple virus (CuLCrV), a whitefly-transmitted geminivirus. Both living and reflective mulches were more effective than white mulch in reducing the densities of whiteflies and the incidence of CuLCrV on zucchini plants.

Effect of trap color on species composition of alate aphids (Homoptera: Aphididae) caught over watermelon plants

The species composition and abundance of alate aphids caught in water traps containing green or yellow tiles were compared. Traps were operated in a watermelon field during the spring and autumn growing seasons of 1992. Aphis spiraecola Patch, a vector of watermelon mosaic virus 2 (WMV-2), accounted for 79% of the aphids caught in the spring and 91% of those caught in the autumn in yellow traps. However, this species accounted for only 6% and 11% of aphids caught in green traps in the spring and autumn, respectively. Uroleucon pseudambrosiae (Olive), also a vector of WMV-2, was the most abundant aphid in green traps in the spring but was absent in the autumn. Aphis gossypii Glover (16% of aphids in green traps) and Aphis craccivora Koch (31%) were more common in the autumn than in the spring and may be important vectors of cucurbit potyviruses at that time.

Temperature-Dependent Development of Elasmopalpus lignosellus (Lepidoptera: Pyralidae) on Sugarcane Under Laboratory Conditions

ABSTRACT Lesser cornstalk borer, Elasmopalpus lignosellus Zeller (Lepidoptera: Pyralidae), is an important sugarcane pest in southern Florida. Development of immature stages (eggs, larvae, prepupae, and pupae) of lesser cornstalk borer was observed on sugarcane at constant temperatures (13, 15, 18, 21, 24, 27, 30, 33, and 36°C), 65–70% RH, and a photoperiod of 14:10 (L:D) h. Total development (from egg deposition to adult emergence) ranged from 22.8 ± 0.3 d at 33°C to 120.7 ± 2.8 d at 13°C. Lesser cornstalk borer required 543.48 DD to complete development. Developmental time decreased with increase in temperature from 13 to 33°C and increased markedly at 36°C in all immature stages. One linear and six nonlinear models used to model insect development (Briere-1, Briere-2, Logan-6, Lactin, Taylor, and polynomial models) were tested to describe the relationship between temperature and developmental rate (d-1). Criteria used to select the best model were the greatest r2, lowest residual sum of squares (RSS), and Akaike information criterion values. The Briere-1 model fit the data best and provided the best estimates of developmental temperature thresholds for all immature stages on sugarcane. The estimated lower and upper developmental thresholds for total development were 9.3 ± 1.8 and 37.9 ± 0.7°C, respectively. The optimal temperature estimated for the total development was 31.39 ± 0.9°C. Based on these results, we can forecast the different stages of lesser cornstalk borer at different times in sugarcane. This will enable us to choose the best time to control this pest with greater precision.

Detection of Cucurbit leaf crumple virus in Florida Cucurbits.

In October of 2006, yellow straightneck and zucchini squash plants (Cucurbita pepo L.) with crumpled, curled, thickened leaves were found in St. Johns and Marion counties in central Florida, respectively. Both locations had high populations of the whitefly, Bemisia tabaci. Incidences of symptomatic plants were greater than 95% in three squash fields (33 ha total) in St. Johns County and 35% in an experimental plot in Marion County. Twenty-three samples were collected from symptomatic plants (two from St. Johns County and 21 from Marion County). DNA was extracted for PCR and tested for the presence of begomoviruses using the following pairs of degenerate primers: AC1048/AV494, which amplifies a conserved region of the coat protein gene (2), PAR1c496/PAL1v1978, which amplifies a region of the begomovirus A component, and PBL1v2040/PCRc154, which amplifies a hypervariable region of the begomovirus B component (1). All squash samples yielded amplicons of sizes expected for a bipartite begomovirus: 1,159 nt with PAR1c496/PAL1v1978, 550 nt with AC1048/AV494, and 493 nt with PBL1v2040/PCRc154. The 1,159- and 493-nt amplicons obtained from two squash plants were cloned and sequenced. The 1,159 nt sequences from both plants shared 98% sequence identity with each other and 97% identity with equivalent regions of the A component of Cucurbit leaf crumple virus (CuLCrV) from Arizona and California (GenBank Accession Nos. AF256200 and AF224760, respectively). The 493-nt sequences amplified with PBL1v2040/PCRc154 were identical and shared a 96% identity with CuLCrV sequence (GenBank Accession No. AF327559) from Arizona and 97% identity with CuLCrV B component sequence (GenBank Accession No. AF224761) from California. Leaves were collected from eight symptomatic squash plants from Citra, FL and used for whitefly transmission assays. Approximately 100 adults of Bemisia tabaci biotype B were released onto each caged leaf and given a 24-h acquisition access period, after which a healthy squash seedling was introduced. Symptoms developed within 10 days on all test plants, and the presence of CuLCrV was confirmed by PCR assays, (primer pairs PAR1c496/PAL1v1978 and PBL1v2040/PCRc154) followed by sequencing. In 2007, similar symptoms were seen in several locations around the state. The same assays confirmed the presence of CuLCrV in watermelon (Citrullus lanatus L.) and squash in the following counties: Collier and Hendry in southwest Florida and Hillsborough, Manatee, and Sarasota in west-central Florida. To our knowledge, this is the first report of CuLCrV, and the first report of any begomovirus in cucurbits in Florida. References: (1) M. R. Rojas et al. Plant Dis. 77:340, 1993. (2) S. D. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.