Division Of Plant Industry Research Articles

A New Host Diagnosed with a Strain of Sugarcane mosaic virus in Florida: Red-Veined Prayer Plant (Maranta leuconeura erythroneura).

Prayer plants (Maranta spp.), which are indigenous to Brazil, are commercially produced in nurseries mainly to be sold as potted houseplants. However, in frost-free areas, they are also used as a ground cover. In March 2009, the Division of Plant Industry in Gainesville, FL received cuttings of red-veined prayer plant or red maranta (Maranta leuconeura erythroneura) from several nurseries in central Florida. The cuttings originated in Costa Rica. The presence of a viral infection was indicated by the mosaic pattern seen on the upper surface of the leaves and chlorotic lesions on the underside of leaves. Epidermal strips were taken and stained in Orange-Green (O-G) and Azure A (1). Microscopic examination revealed viral inclusions that stained only in O-G, indicating the presence of a potyvirus. Leaf dips were prepared for the electron microscope and flexuous rods consistent with a potyvirus were found. Indirect-ELISA using universal potyvirus antiserum (Agdia Inc., Elkhart, IN) confirmed the presence of a potyvirus infection. Total RNA was extracted from symptomatic tissue of five infected samples with Qiagen's RNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). Reverse transcription was conducted with the oligo d(T) primer M4T with AMV-RT at 37°C for 1 h. PCR was performed with primer M4 and a degenerate primer designed to amplify the 3' end of all potyviruses (2). A target amplicon of 1.7 kb was produced from all five samples. Three of these samples were cloned and sequenced. Approximately 1,250 bp were sequenced from each sample. The sequenced regions include the 3' end of the Nib gene, the complete coat protein, and the beginning of the 3' untranslated region (UTR). The nucleotide (nt) sequences were deposited into GenBank (Accession Nos. GQ853403-GQ853405). The nt sequences of the three samples were 97 to 98% identical to each other. When compared with other potyviruses in the GenBank, the samples showed closest nt identity, 92 to 93%, with several isolates of Sugarcane mosaic virus (AJ278405, AY836523, U57357, and U57356). Of the plant species mechanically inoculated (Chenopodium amaranticolor, C. quinoa, Datura stramonium, Gomphrena globosa, Nicotiana benthamiana, and Zea mays), only Z. mays (corn) showed symptoms (a mild mosaic). The same type of viral inclusions were seen in leaf strips of infected corn as in the Maranta. The corn plant reacted positively in direct-ELISA against antiserum to Sugarcane mosaic virus (Agdia Inc.). Cuttings of infected Maranta were observed in the greenhouse and indoor situations for several months. The plants infected with Sugarcane mosaic virus lacked vigor and most eventually died. ELISA tests done on a few surviving plants were positive for Sugarcane mosaic virus, but the results were inconsistent, indicating a possible low titer of virus in the plants as they were dying. To our knowledge, this is a new host for Sugarcane mosaic virus, but it does not appear that Maranta will become a significant new reservoir of this virus for sugarcane or corn growers. However, infected cuttings can greatly decrease the production of this plant as an ornamental.

Tomato spotted wilt virus Found in Five Species of the genus Tragopogon in a Florida Greenhouse.

An obviously unhealthy plant identified as Tragopogon mirus Ownbey (remarkable goatsbeard) was sent for diagnosis to the Division of Plant Industry (DPI), Gainesville, FL in May of 2008. T. mirus is a recently formed allotetraploid that has T. dubius Scop. and T. porrifolius L. (goatsbeard or salsify) as parents. The parents (family Asteraceae) are diploid and originate from Eurasia. They were introduced to the northwest United States in the early 1900s. The allotetraploid T. mirus, which does not occur in Eurasia, was discovered in 1949 and named in 1950. It has been found in the northwest states of Washington and Idaho. It has also been found in Arizona (4). The plant sent to the DPI was grown in a greenhouse for research purposes at the Botany Department of the University of Florida (Alachua County). Symptoms exhibited on the leaves included mottling, chlorotic and necrotic spots, and mild distortion. Epidermal leaf strips from a mottled leaf were stained with the Orange-Green protein stain and Azure A nucleic acid stain (1). With a light microscope, granular inclusions typical for Tomato spotted wilt virus (TSWV) (1) were seen in leaf strips from both stains. The remainder of the leaf was ground in buffer and tested serologically for TSWV by TSWV-specific ImmunoStrips (Agdia, Elkhart, IN). The ImmunoStrip was positive for the presence of TSWV. This test was confirmed by double-antibody sandwich-ELISA using antiserum and conjugate for TSWV (Agdia). Further serological testing of other Tragopogon species with similar symptoms growing in the same greenhouse revealed that T. miscellus (another recently formed allotetraploid found in the northwestern United States; parents T. dubius and T. pratensis), T. dubius, T. porrifolius, and T. pratensis were also infected with TSWV. Total RNA was extracted from symptomatic leaves of T. mirus, T. dubius, T. porrifolius, and T. miscellus. Reverse transcription-PCR was performed with universal tospovirus primers BR60 and BR65 that amplify part of the nucleocapsid protein gene (2). Target amplicons of 454 bp were produced for all four samples. The PCR product from T. porrifolius was cloned and sequenced. The resulting sequence (GenBank Accession No. FJ655913) shows high homology, 98%, to several isolates of the Tomato spotted wilt virus deposited in the GenBank (Accession Nos. AY870391, AY744477, and AF020659). T. porrifolius has been reported to be naturally infected with TSWV in Italy (3); however, to our knowledge, this is the first report of this virus in the allotetraploids T. mirus and T. miscellus and in the diploids T. dubius and T. pratensis. This report adds five new Asteraceae weeds to the list of possible reservoirs of TSWV in the United States.

Anastrepha edentataand Other Fruit Flies (Diptera: Tephritidae) Detected on Key Largo, Florida

Several Anastrepha species (Diptera: Tephritidae) are quarantine pests of commercial fruits in the New World. Florida is home to one such pest, the introduced Anastrepha suspensa (Loew) (Caribbean fruit fly), plus three other Anastrepha species that are not considered of economic importance (Foote et al. 1993; Sutton & Steck 2005). One of these latter species, Anastrepha edentata Stone, is particularly mysterious, as no host has ever been reported from anywhere in its known range of South Florida and Puerto Rico (Norrbom 2004). We attempted to document its phenology and discover its host(s) through bait trapping and rearing from fruit collections in Dagny Johnson Key Largo Hammock Botanical State Park, Key Largo, Florida, during 1994-1996. Some fruit samples were also taken from Long Key State Park, Long Key, Florida, during the same period and during earlier periods. Both areas are subtropical West Indian hardwood forests. Ten to 16 plastic McPhail traps baited with torula yeast/borax tablets dissolved in water were suspended on the branches of small trees. Traps were hung 2 m off the ground, placed at 50-100-m intervals along a narrow, unpaved trail in the park, and were checked weekly from 25 Feb 1994 to 23 Feb 1996. In total, 21 A. edentata adults were trapped on 10 different occasions (Table 1). This is the first substantial collection of the species since 1934-1937, when the fly was first discovered and trapped in relatively large numbers during an intensive fruit fly survey in South Florida (Brown 1937; Clark et al. 1996). The fly has been captured on only a handful of other occasions (see Norrbom 2007 and records of the Florida Department of Agriculture, Division of Plant Industry (FDACS/ DPI) and Florida State Collection of Arthropods (FSCA)), despite extensive, routine fruit fly trapping in nearby suburban Key Largo plus other Florida Keys and large areas of Monroe

NOTES ON THE LONGAN SCALE, THYSANOFIORINIA NEPHELII (HEMIPTERA: COCCOIDEA: DIASPIDIDAE) IN FLORIDA1

In Florida, the most common armored scales found on tropical fruits are considered exotics or not native to Florida and 3 examples of introductions in the last 10 years include the white mango scale,Aulacaspis tubercularis Newstead, the litchi scale, Andaspis punicae (Laing), and the longan scale, Thysanofiorinia nephelii (Maskell) (Hodges et al. 2005). The longan scale was first collected in the continental U.S. in Homestead, Florida in 1996. Even though the longan scale is believed to be native to the Orient, it has spread and invaded several regions worldwide prior to this detection including Autralasian, Neotropical, and Palearctic regions. Neotropical introductions detected prior to the Florida report included Brazil, Rio de Janeiro, and Cuba (Ben-Dov et al. 2003). The longan scale is now considered established in southern Florida but has not been catalogued as a significant economic pest in the United States (Miller et al. 2005). Primary hosts of concern in Florida for the longan scale include plants in the Sapindaceae, in particular longan (Dimocarpus longan) and lychee (Litchi chinensis Sonnerat). Populations may occur on leaves, stems, or fruits. Other potential host plants for the longan scale include Arecaceae (Kentia spp.) and Euphorbiaceae (Euphorbia longena) (Ben Dov et al. 1993). Increased sample submissions of pest population reports to the Florida Department of Agriculture, Division of Plant Industry have occurred particularly in Broward, Miami-Dade, and Palm Beach Counties since 2005 (Table 1). Isolated occurrences of detection have also occurred at retail stores of tropical fruit trees, but the established pest distribution will probably be restricted to climate zones suitable for tropical fruit trees. It is unknown if this species will be a significant economic pest for tropical fruits in Florida, but increased report incidences suggest that it may be an emerging pest of concern. Basic life history and control information is unknown for this species. General and taxonomic information are summarized below. In the field, the covering or armor of this scale looks similar to that of a Parlatoria scale but TABLE 1. LONGAN SCALE COLLECTION RECORDS.

Tobacco ringspot virus Found in the Cardboard Cycad (Zamia furfuracea) in Florida.

Zamia furfuracea (Zamiaceae) is native of coastal Mexico. It is a popular houseplant and easy to grow outdoors in warm climates. In November 2005, a plant of Z. furfuracea, originally from Texas, was received at the Division of Plant Industry in Gainesville, FL. The plant had numerous chlorotic spots on the leaves that eventually became necrotic. Leaves were ground in phosphate buffer (pH 7.2) with Carborundum and used to inoculate a host range that included Chenopodium amaranticolor, C. quinoa, Gomphrena globosa, Datura stramonium, and Nicotiana benthamiana. Systemic symptoms were seen in C. quinoa (necrotic lesions), G. globosa (stunting), D. stramonium (chlorotic ringspots), and N. benthamiana (wavy line patterns) 1 to 2 weeks after inoculation. C. amaranticolor showed only small necrotic local lesions. In further host range studies, systemic infections of Beta vulgaris, D. metaloides, Lactuca sativa, N. clevelandii, Pisum sativus, Petunia hybrida, Zinnia elegans (symptomless), and Cucumis sativa were observed. However, no infection of Zea mays, Verbena hybrida, Glycine max, Phaseolus vulgaris, Catharanthus roseus, Arachis hypogaea, Trifolium spp., Vigna unguiculata, Vicia faba or Digitalis spp. was detected. Inclusions observed in leaf strips of N. benthamiana and D. stramonium indicated a possible infection of this plant with a nepovirus (1). A 337-bp fragment was amplified from total RNA isolated from an inoculated D. stramonium using reverse transcription-PCR with nepovirus group primers provided by Agdia Inc. (Elkhart, IN). Sequence analysis indicated that the nucleotide (nt) and deduced amino acid (aa) sequences of the fragment were 89 to 91% and 91 to 95% identical, respectively, to sequences of the RNA-dependent RNA polymerase gene for Tobacco ringspot virus (TRSV) contained in GenBank (Accession Nos. U50869 and AJ698718). This region was only 50% (nt) and 38% (aa) identical to Cycas necrotic stunt virus (GenBank Accession No. NC_003791), a nepovirus previously reported to infect cycads (2). The original plant, symptomatic inoculated hosts, and the symptomless zinnia tested positive by double-antibody sandwich-ELISA using commercially available antiserum for TRSV (Agdia, Inc.), further confirming the presence of TRSV. Although the virus infecting Z. furfuracea has a more restricted host range than that reported for TRSV, the serology and gene sequence indicates that this virus is a unique isolate of TRSV.

Erratum to: Kenneth Baillieu Myer 1921 - 1992

Kenneth Baillieu Myer was elected to the Fellowship of the Australian Academy in April 1992, under the provision for special election of people who are not scientists but have rendered conspicuous service to the cause of science. Myer was a significant figure in Australian history by virtue of his contribution to the origins or early development of major national institutions, most notably the Howard Florey Laboratories of Experimental Physiology and Medicine, the School of Oriental Studies at the University of Melbourne, the Victorian Arts Centre and the National Library of Australia. He successfully fostered new research in organizations such as the Division of Plant Industry of the CSIRO and helped build the Oriental Collection of the Art Gallery of New South Wales.

Kenneth Baillieu Myer 1921 - 1992

Kenneth Baillieu Myer was elected to the Fellowship of the Australian Academy in April 1992, under the provision for special election of people who are not scientists but have rendered conspicuous service to the cause of science. Myer was a significant figure in Australian history by virtue of his contribution to the origins or early development of major national institutions, most notably the Howard Florey Laboratories of Experimental Physiology and Medicine, the School of Oriental Studies at the University of Melbourne, the Victorian Arts Centre and the National Library of Australia. He successfully fostered new research in organizations such as the Division of Plant Industry of the CSIRO and helped build the Oriental Collection of the Art Gallery of New South Wales.

Alternanthera mosaic virus Found in Scutellaria, Crossandra, and Portulaca spp. in Florida.

In the fall of 1998, the Division of Plant Industry (DPI) received vegetative propagations of Scutellaria longifolia (skullcap) with symptoms of foliar mosaic, chlorotic/necrotic ringspots, and wavy line patterns from a nursery in Manatee County. Flexuous particles approximately 500 nm long were found with electron microscopy. The plants tested positive for Papaya mosaic virus (PaMV) in an enzyme-linked immunosorbent assay (ELISA) test with antiserum to PaMV (Agdia, Elkhart, IN). However, in immunodiffusion tests (antiserum from D. Purcifull, University of Florida), this virus gave a reaction of partial identity indicating it was related but not identical to PaMV (1). The original infected plants were kept in a greenhouse. In January 2005, a specimen of Crossandra infundibuliformis (firecracker plant) with mosaic symptoms was submitted to the DPI from a nursery in Alachua County. Inclusions found with light microscopy and particles found with electron microscopy indicated that this plant was infected with a potexvirus. This was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) with primers designed to detect members of the virus family Potexviridae (3). These plants reacted positive to PaMV antiserum in ELISA and gave a reaction of partial identity to PaMV in immunodiffusion. A specimen of Portulaca grandiflora (moss rose) with distorted leaves found at a local retail store was also tested and gave the same results. Leaves from each of the three plant species were rubbed onto a set of indicator plants using Carborundum and potassium phosphate buffer. Total RNA was extracted from symptomatic indicator plants of Nicotiana benthamiana. RT-PCR (3) was performed, and PCR products were sequenced directly. Sequences of approximately 700 bp were obtained for all three plant species and showed 98% identity with each other. BLAST search results showed that these sequences were 93% identical to an Alternanthera mosaic virus (AltMV) sequence at the nucleotide level but only 76% identical to PaMV. The amino acid sequences were 98 and 82% identical to AltMV and PaMV, respectively. The PCR products of the virus from Scutellaria sp. were cloned, resequenced, and the sequence was entered into the GenBank (Accession No. DQ393785). The bioassay results matched those found for AltMV in Australia (2) and the northeastern United States (4), except that the Florida viruses infected Datura stramonium and Digitalis purpurea (foxglove). The virus associated with the symptoms of these three plants appears to be AltMV and not PaMV. AltMV has been found in ornamental plants in Australia, Italy, and the United States (Pennsylvania, Maryland, and now Florida). Since this virus is known to infect several plants asymptomatically and can be easily confused with PaMV serologically, it is likely that the distribution of this virus is much wider than is known at this time.

Dryocosmus kuriphilus

<i>Dryocosmus kuriphilus</i>

DISTRIBUTION OF THE ASIAN CITRUS PSYLLID, DIAPHORINA CITRI KUWAYAMA (RHYNCHOTA: PSYLLIDAE) IN THE CARIBBEAN BASIN

Asian citrus psyllid, Diaphorina citri Kuwayama 1907 (Rhynchota: Psyllidae), was described from Taiwan and is native to Asia. It has been known in the Western Hemisphere for several decades in Brazil (Hodkinson & White 1981; Lima 1942). During the past decade, its range has expanded into northern South America and the Caribbean. The focus of this paper is to document the distribution of D. citri in the Caribbean Basin. Burckhardt & Martinez (1989) reportedD. citri intercepted in France on citrus plants from Honduras and reconditione aux Etats-Unis avant d'etre importe en France. This report has been difficult to verify. There is no doubt that the intercepted insects were D. citri, but the source of the shipment is questionable. The circumstances under which these infested plants spent time being rehabilitated in the United States are not stated. Citrus plants from foreign sources are not allowed in the USA except under strict quarantine conditions (Title 7 (Agriculture), Chapter III, Part 319.19, Code of Federal Regulations). Additionally, D. citri was not present in the USA in 1989, so the infestation did not come from the USA. Visiting scientists Ronald Cave (Department of Entomology, University of Florida, pers. comm. 2003) and James Baker (Department of Entomology, North Carolina State University, pers. comm. 2003) spent many months in Honduras in the 1990s and failed to find D. citri. In 1997, specimens of D. citri were found on citrus in Corrientes, Argentina, by Sara Caiceres (Florida State Collection of Arthropods (FSCA) accession # E1997-3427). Evidently, the infestation was minor, because the Argentina Department of Agriculture was unaware of the presence of D. citri in Argentina in 1998. This low infestation level suggests that D. citri had been there prior to 1997 long enough for populations to subside to low levels controlled by local natural enemies. In 1998, D. citri was found in Guadeloupe (Etienne et al. 1998) and in south Florida (FSCA# E1998-1751) (Halbert 1998; Halbert et al. 2003). Since then, movement throughout the Caribbean has been rapid. In 1999, D. citri was found on Abaco Island and Grand Bahama Island, Bahamas, on both Citrus spp. and on Murraya paniculata (L.) Jack. (FSCA# E1999-1975, 1976, Robert C. Bullock and Robert R. Pelosi, University of Florida). Two years later, high numbers of D. citri were intercepted on citrus fruit from Abaco, Bahamas sent for processing in Ft. Pierce, FL (FSCA# E2001-747, 850, 978, 1135, and 2049, Kenneth L. Hibbard and James J. Walukiewicz, Florida Department of Agriculture and Consumer Services, Division of Plant Industry (DPI) inspectors). These repeated interceptions of D. citri indicate beyond doubt that D. citri can move on fresh, unprocessed citrus fruit. Infestations were found in West Bay, Grand Cayman, Cayman Islands in June 2000 (FSCA# E2000-2102, Joan Steer and Sasha Frederick). In 2001,D. citri was intercepted in passenger baggage on leaves of Murraya koenigii (L.) Sprengel from St. Thomas, U.S. Virgin Islands, probably indicating a population of D. citri on that island (FSCA# E2001-696). An infestation was found in Jamaica for the first time in Bodles, St. Catherine on 18 January 2003 (FSCA# E2003-259, Sharon McDonald). Diaphorina citri was first found in the Dominican Republic in September 2001, but because of its wide distribution in the country, we believe its introduction occurred at least one year before its detection. The specimens from the Dominican Republic are housed at the Museum of Natural History, Santo Domingo, Dominican Republic. All collections are by C.A. Nuiiez unless otherwise noted. They are labeled as follows: 17 adults on Murraya paniculata-Distrito Nacional, Santo Domingo, Ciudad de los Millones, 16-IX-2001; 9 adults on Citrus sinensis (L.) Osbeck, Plaza de la Cultura Juan Pablo Duarte, 20-IX-2001; 6 adults, 1 nymph on Murraya paniculata, Ensanche Miraflores, 21-IX-2001; 3 adults on Citrus limon (L.) Burm. f., 2 adults on Citrus reticulata Blanco, 6 adults on Citrus limetta Risso, 5 adults on Citrus sinensis, 7 adults on Citrus maxima (Burm.) Merr., Urbanizaci6n Las Praderas, 22IX-2001; 7 adults, 3 nymphs on Citrus aurantium L., Ensanche Quisqueya, 23-IX-2001; 11 adults on Murraya paniculata, 3 adults on Citrus limon, 3 adults on Citrus sinensis, Province of Santo Domingo, Santo Domingo Oeste, Engombe 26-VII2002; 6 adults, 9 nymphs on Murraya paniculata, Zona Industrial, Herrera, 14-XI-2002; 8 adults, 10 nymphs on Murraya paniculata, Santo Domingo Este, Ensanche Ozama, 8-VIII-2002; 7 adults on Citrus sinensis, San Cristobal, General Leger Ave., 30-IX-2001; 9 adults on Murraya paniculata, Monte Plata, Parque Central, 23-VII-

EFFECT OF MICROMITE® ON THE EGG PARASITOIDS CERATOGRAMMA ETIENNEI (HYMENOPTERA: TRICHOGRAMMATIDAE) AND QUADRASTICHUS HAITIENSIS (HYMENOPTERA: EULOPHIDAE)

The Diaprepes root weevil, Diaprepes abbreviatus (L.), was first detected near Apopka, Florida in 1964 (Woodruff 1964). Since then, it has spread throughout the citrus growing areas of the state causing growers millions of dollars in losses each year. In south Florida, D. abbreviatus is also a problem in root crops and ornamental plants (Peina & Amalin 2000). One of the components of the pest management program for this weevil is the use of Micromite? (diflubenzuron), a chitinase inhibitor that sterilizes the egg by interrupting the formation and deposition of chitin in developing embryos. Early studies indicated that diflubenzuron significantly reduces the reproductive potential of D. abbreviatus when applied to citrus foliage (Schroeder et al. 1976; Lovestrand & Beavers 1980; Schroeder et al. 1980). Later, Schroeder (1996) reported that residues of diflubenzuron significantly affected the reproductive potential of D. abbreviatus for more than one month after application to citrus foliage. Micromite has been described as a foundation product for reducing citrus root weevil populations, in part due to its compatibility with root weevil natural enemies (Anonymous 1996). However, there are no reports on the effect of micromite on egg parasitoids that have been imported and established for biological control of D. abbreviatus (Hall et al. 2001; Peina et al. 2000; Pena et al. 2003). A study was initiated to evaluate the impact of Micromite on Ceratogramma etiennei Delvare and Quadrastichus haitiensis (Gahan), two egg parasitoids of Diaprepes root weevil. Green buttonwood (Conocarpus erectus L.) seedlings were grown from cuttings in 3.7-liter pots. Adult D. abbreviatus were collected from an insecticide-free ornamental orchard in Homestead, Florida. A Guadeloupe strain of C. etiennei was obtained from J. Etienne, Institut National de la Recherche Agronomique (INRA). The culture was maintained at the Tropical Research and Education Center (TREC) insectary, Homestead, Florida. Insect cultures were maintained at 26.5 ? 1.0?C, 12:12 L:D and approximately 78% RH, on eggs of D. abbreviatus laid on strips of wax paper using the methodology of Etiennei et. al (1990). Quadrastichus haitiensis originally from Puerto Rico was obtained from Ru Nguyen, Division of Plant Industry, Gainesville, Florida, and maintained as above. The effect of Micromite ingestion by the adult Diaprepes root weevil plus absorption of residues from leaves into eggs was evaluated in experiment 1. Three green buttonwood seedlings planted separately on 3.7-liter pots were sprayed with Micromite to run-off using the simulated field rate (0.485 g/l liter of water). Another three seedlings were sprayed with water as control checks. All the seedlings were enclosed separately in screen cages (240 cm x 120 cm x 120 cm). One hundred adults of D. abbreviatus were introduced inside each cage for oviposition. After 3 days, 20 egg masses on leaves still attached to branches were collected from each seedling, arranged as bouquets in flasks of water and placed in Plexiglass cages (30 cm x 30 cm x 30 cm) separately. Six cages, 3 treated and 3 untreated, were prepared for each parasitoid species. One hundred 2to 3-dold C. etiennei and Q. haitiensis adults were introduced into each Plexiglass cage. Bouquets were removed after 3 days and portions of the leaves with individual egg masses laid between two leaf surfaces (covered with two leaf layers intact) were placed in culture tubes (12 mm x 75 mm). After 7 d, egg masses were exposed by removing one leaf surface and parasitized eggs counted. Parasitized eggs were recognized by the characteristic golden egg chorion for C. etiennei and silver transparent egg chorion for Q. haitiensis (Penia et al. 2000). Effects of ingestion of micromite by adult D. abbreviatus alone were evaluated in a separate experiment. Ten adult D. abbreviatus females were exposed to Micromite -treated seedlings and to untreated foliage for 3 days as described above. The adults were allowed to oviposit on to double strips of wax paper inside Plexiglass cages (30 cm x 30 cm x 30 cm). After 2 d, paper strips containing 20 egg masses were collected from each cage and each egg mass was placed separately inside culture tubes (12 mm x 75 mm). One 2-d-old mated female wasp was introduced into each tube (20 tubes per treatment for each parasitoid species). After 7 d, eggs were examined as described above. In a third test, 10 untreated female weevils were allowed to oviposit on wax paper for 2 days. Wax papers containing approximately 20 egg

A Virus Related to Clover Yellow Mosaic Virus Found East of the Mississippi River in Verbena canadensis in Florida.

In September 2002, several unthrifty Verbena canadensis 'Homestead purple' plants were received at the Division of Plant Industry in Gainesville, FL. Symptoms included very subtle yellowing and distortion or stunting of the younger leaves, symptoms that could be overlooked as a nutritional problem. Symptomatic leaves were ground in phosphate buffer and mechanically inoculated to a variety of plants that included Antirrhinum majus, Chenopodium amaranticolor, Datura stramonium, Gomphrena globosa, Pisum sativum, Trifolium pratense, T. repens, Vicia faba, and Vigna unguiculata. These hosts showed symptoms similar to those described for infection by Clover yellow mosaic virus (ClYMV) (3). In addition, the virus systemically infected Arachis hypogaea, Catharanthus roseus, C. quinoa, Nicotiana benthamiana, and healthy seed-grown Verbena × hybrida. No symptoms were seen in inoculated Zinnia elegans, N. glutinosa, N. clevelandii, Lycopersicon esculentum, Cucumis sativus, or Capsicum annuum. However, back inoculations of Cucumis sativus to C. amaranticolor gave typical local and systemic symptoms. Microscopic examination of leaf strips of infected V. faba revealed banded inclusions typical of the genus Potexvirus (1). Reverse-transcription polymerase chain reaction (RT-PCR) using degenerate primers for the genus Potexvirus (2) produced a 750-bp product that is the expected product for the genus Potexvirus. The RT-PCR product was cloned in pGem-T easy (Promega, Madison, WI) and sequenced. BLAST ( http://www.ncbi.nlm.nih.gov/ BLAST/ ) analysis of the sequence showed an 82% identity at the nucleotide level with the RNA polymerase gene of ClYMV. Leaf extracts of the original verbena, several inoculated hosts, and a known sample of ClYMV in N. benthamiana were tested in sodium dodecyl sulfate immunodiffusion (4) against antiserum to the degraded capsid protein of ClYMV. A reaction of identity was seen with infected samples but not with samples for comparable virus-free plants. To our knowledge, this is the first time this virus has been found in the eastern United States. It is not known how or if the presence of this noninsect transmitted virus in an ornamental will affect the agricultural, forage, or ornamental industries in the east or how widely distributed Verbena sp. infected with this virus may be at this time.

REPRODUCTIVE INCOMPATIBILITY BETWEEN TWO SUBSPECIES OF COLEOMEGILLA MACULATA (COLEOPTERA: COCCINELLIDAE)

There is interest in introducing the midwestern subspecies of Coleomegilla maculata (DeGeer), C. m. lengi Timberlake, as a biological control agent for augmentation programs in Florida. The Division of Plant Industry (DPI) of the Florida Department of Agriculture and Consumer Services has prohibited the release of C. m. lengi for fear that it could interbreed with the native Florida subspecies C. m. fuscilabris (Mulsant), causing genetic contamination or suppression of the C. m. fuscilabris populations. Two populations of these subspecies, fuscilabris from the southeastern USA and lengi from the midwestern USA, were crossed. Reciprocal single pair crosses were performed under controlled laboratory conditions (temperature, relative humidity and daylength) on two dates. Results demonstrated a nearly complete reproductive incompatibility between these two populations in the first generation (F1) and complete reproductive incompatibility in the second (F2). Further analysis is required to establish the cause of the reproductive incompatibility.

Outbreaks of Stem and Leaf Blight of Eustoma grandiflorum Caused by a Phomopsis sp. in Florida.

Between November 1997 and May 1998, numerous lisianthus (Eustoma grandiflorum) cultivars exhibited severe stem and leaf blight at two pot-flower production sites in Hillsborough and Dade counties, FL. Blight occurred in mature plants and ranged in incidence from 3 to 5% in Dade County and from 40 to 80% in Hillsborough County. Initial stem necrosis was rapidly followed by leaf blight and production of numerous dark pycnidia in diseased tissue. As stem blight progressed, infected plants collapsed and died. Pycnidia contained biguttulate, 7.2 × 2.2-μm spores typical of Phomopsis alpha conidia; beta conidia were not observed. A Phomopsis sp. was isolated consistently when pycnidia from symptomatic stems were placed on acidified 25% potato dextrose agar after surface-disinfestation in 0.5% NaOCl; only alpha conidia were observed in culture. Pathogenicity was confirmed using a suspension of hyphae, pycnidia, and conidia made by comminuting one 95-mm-diameter acidified carnation leaf agar plate containing a 4-week-old colony of the Phomopsis sp. in 100 ml of deionized water. Six plants of lisianthus 'Maurine Blue' (three wounded at the crown, three nonwounded) each were inoculated with 10 ml of the fungal suspension. An equal number of noninoculated lisianthus (three wounded, three nonwounded) served as controls. After inoculation, plants were maintained in a greenhouse with average high and low temperatures of 38 and 25°C, respectively. Stem and leaf blight symptoms were observed in two of three wounded plants and in all nonwounded plants within 11 and 15 days after inoculation, respectively. Infection by a Phomopsis sp. was confirmed by reisolation from symptomatic tissue. Although this Phomopsis sp. has been detected previously in lisianthus exhibiting leaf and stem lesions (1), this report establishes the ability of the of the fungus to act as a primary pathogen and to cause extensive losses in this crop. Reference: (1) Alfieri et al. 1994. Diseases and Disorders of Plants in Florida. Bull. No. 14. Division of Plant Industry, Gainesville, FL.

Seasonal Flight of Plecia nearctica (Diptera: Bibionidae) in Southern Florida

The lovebug, Plecia nearctica Hardy, is a serious nuisance to motorists traveling in southern states. The insects are smashed against windshields obscuring vision and cars may overheat when radiators become clogged. The smashed insects also damage car paint if the body fluids are not removed soon after contact (Callahan and Denmark 1973). The insect was first described by Hardy (1940) from Galveston, Texas, who reported it to be widely spread, but more common in Texas and Louisiana than other Gulf Coast states. It has now progressed to all states bordering on the Gulf of Mexico, as well as Georgia, South Carolina, and parts of Central America. It was first collected in Florida in 1949 and today is found throughout Florida (Denmark and Mead 1992). Adult flights of lovebugs have been reported to occur primarily during May and September in different areas of the insect's range (Hetrick 1970; Callahan and Denmark 1973; Buschman 1976; Thornhill 1976; Callahan 1985). However, in spite of these previous reports, no data have ever been presented actually showing seasonal flights of the lovebugs. Our objective was to determine seasonal flight patterns of P nearctica in southern Florida. Additional data on temporal sychronony of flights at different locations are also presented. Yellow sticky traps (Pherocon AM, no bait) made by Trece Inc., Salinas, California and baited with anethole (Cherry 1998) were used to monitor adult flights of lovebugs. Sampling was conducted by putting out 15 new sticky traps each month at approximately mid-month. These 15 traps were located at three different locations (5 traps/ location) to provide data on temporal synchrony between locations. One location was the Everglades Research and Education Center (location one), University of Florida, located near Belle Glade, Florida. The other two locations were located approximately 24 km southwest of the research center (location two) and 31 km east of the research center (location three). All three locations were located within Palm Beach County, Florida, and surrounded by agricultural lands (sugarcane, rice, vegetables). Traps at each location were placed 250 m apart in a straight line (i.e. a one km transect) adjacent roads. Each trap was hung one m above the ground on a metal rod and had a sponge (3 x 3 x 3 cm) containing 5 ml of anethole wedged into it. All 15 traps were hung and baited on the same day and collected seven days later. The seven day exposure period was used to avoid short term inclement weather factors such as rain, extreme winds, etc. that might greatly reduce adult catches within a shorter trap exposure time. Traps were collected by wrapping each trap in clear cellophane in the field. Lovebugs on each trap were counted by microscopic identification in a laboratory. A sample of lovebug adults collected on the traps during different major flight periods (May, September, December) was sent to Dr. Gary Steck, Division of Plant Industry, Gainesville, Florida who corroborated that the insects were P nearctica. Statistical analysis was conducted by SAS (1996). Temporal synchrony of adult flights between locations was determined by multiple correlation using the total number of adults caught each month at each location. Differences in mean numbers of adults caught at each location during different months were determined by using Tukey's test. A figure to visually show the overall seasonal flight pattern of the lovebugs is also presented. Although different numbers of lovebugs were observed between locations (Table 1), there was temporal synchrony in flight between locations. Multiple correlation of

Field Assessment of a Fiber Optic Spectral Reflectance System

Spectral reflectance measurements were taken from immature and mature leaves of `Rio Red' grapefruit `McCarty' grapefruit `Minneola' tangelo, `Satsuma' mandarin, `Dancy' tangerine, `Nagami' oval kumquat, and `Valencia' orange from 1330 to 1530 hr 1 day at the Florida Citrus Arboretum, Division of Plant Industry, Winter Haven, Fla. A PS-1000 spectrometer was used with fiber optic cables and a lens source (visible range of the spectrum 400-800 sun) coupled with a tungsten halogen light source. A data acquisition card was connected to a notebook computer with a SpectraScope computer program for processing and data storage. Immature and mature leaves of `Minneola' tangelo had greater percentage reflectance in the 500-800 sun range than the other cultivars and leaf ages measured. More detailed information was obtained with the PS-1000 than with conventional spectrometers. The slope of the citrus spectral curves in the 800 nm range was not as sharp as conventional spectrometers, but had a much higher reflectance value than those obtained with a different spectrometer. The system used here was convenient to transport and use in the field and produced clear, interpretable data.

Invertebrate Pets and the Florida Department of Agriculture and Consumer Services

The Division of Plant Industry (DPI) of the Florida Department of Agriculture and Consumer Services now regulates importation into Florida of all arthropods except Crustacea, no longer just those of actual or potential agricultural importance. The operating law is Chapter 581.083 of the Florida Statutes, and the operating procedure is Title 5B-57.004 of the Florida Administrative Code. The current law was proposed because of importation by the pet trade of species that did not already occur in Florida and were potentially harmful to the environment. The Division requires specimens (for confirmation of identification) to accompany applications for permits.

Velvetbean Caterpillar (Lepidoptera: Noctuidae): Surviving Freezing Weather in Louisiana

The efforts of R. C. Bullock (University of Florida, IFAS Agricultural Experiment Station, Ft. Pierce, Florida), and W. N. Dixon and R. Nguyen (FDACS, DPI, Gainesville, Florida), in obtaining fruit samples with miners are appreciated. The many plant protection specialists of the Florida Dept. of Agriculture and Consumer Services, Division of Plant Industry, are thanked for their extensive search for fruit miners. Contribution No. 795, Section of Entomology, DPI, FDACS.

Validation of the CASPR Aerial Spray Efficiency Model

This article describes the implementation of the Baltin-Amsden formula, a method for estimating the cost of an aerial spray operation, onto personal computers through the CASPR (Computer Assisted Spray Productivity Routine) program. The CASPR predictions are compared to observed data taken during the 1991 Gypsy Moth Eradication Program run by the Utah Department of Agriculture, Division of Plant Industry. The model is able to predict the total times of the aerial spray operation to within 23%, on average, and therefore, provides a means of estimating quickly the cost of any aerial spray operation scenario.

Manipulation of Chromosomal Segments From Wild Wheat for the Improvement of Bread Wheat

Specific alien chromatin segments have been used for many years to introduce novel disease resistance genes into bread wheat. The 1RS chromosome segment being used in Australian wheats will be discussed in detail to present the molecular biological studies carried out on this chromosome segment as well as its agronomic contributions to wheat. Studies are also being carried out on the manipulation of the D genome of wheat. Within Australia we have initiated the screening of a collection of 420 accessions of Triticum tauschii (the donor of the D genome) involving groups in the Departments of Agriculture in New South Wales, Victoria, Western Australia and the University of Sydney as well in the Division of Plant Industry. Disease resistance has been sought and found for cereal cyst nematode, Septoria nodorum, Septoria tritici, Puccinia striiformis, P. recondita and P. graminis. At the DNA level the collection has been screened for variation at the 5Sdna and Nor loci, as well as generating Pst (a restriction endonuclease) genomic clones, in preparation for providing RFLP markers for a genetic linkage map. Lines of T. tauschii showing high levels of disease resistance have been crossed directly to Australian wheat cultivars or are being used to form synthetic hexaploids by crossing to tetraploid wheats.