David G. Riley

Thrips Vectors of Tospoviruses

Tospoviruses belong to the sole phytovirus genus, Tospovirus , in the family Bunyaviridae . Tospoviruses are known to be exclusively transmitted by thrips belonging to the family Thripidae and subfamily Thripinae. Of the known 1,710 species of Thripidae only 14 thrips species are currently reported to transmit tospoviruses. Thrips-transmitted tospoviruses cause severe yield losses to several economically important crops in the United States and worldwide. For instance, a single Tospovirus ( Tomato spotted wilt virus ) alone caused an estimated

Tactics for Management of Thrips (Thysanoptera: Thripidae) and Tomato Spotted Wilt Virus in Tomato

1.4 billion in losses in the U.S. over 10 years. Global trade and associated movement of plant materials across borders have introduced tospoviruses and their vectors into newer areas. Advances in serological and molecular techniques have also led to identification of new tospoviruses. This scenario has also initiated new vector-pathogen interactions between introduced and native thrips species and tospoviruses. The goal of this manuscript is to provide a comprehensive and updated list of thrips species that serve as vectors of tospoviruses along with information pertaining to common names, key diagnostic characters, distribution, important crops economically affected, and thrips and Tospovirus -induced symptoms. The manuscript is prepared with special emphasis to the U.S., but information pertaining to other countries is also included.

Temperature and Host Plant Effects on Development, Survival, and Fecundity of Bemisia argentifolii (Homoptera: Aleyrodidae)

Abstract Four studies were conducted in Georgia during spring 1999, 2000, 2001, and 2002 to evaluate various management tactics for reducing thrips and thrips-vectored tomato spotted wilt virus (TSWV) in tomato and their interactions relative to fruit yield. Populations of thrips vectors of TSWV, Frankliniella occidentalis (Pergande) and Frankliniella fusca (Hinds), were determined using flower and sticky trap samples. The management practices evaluated were host plant resistance, insecticide treatments, and silver or metallic reflective mulch. Averaged over all tests, the TSWV-resistant tomato ‘BHN444′ on silver mulch treatment had the largest effect in terms of reducing thrips and spotted wilt and increasing marketable yield. Of the insecticide treatments tested, the imidacloprid soil treatment followed by early applications of a thrips-effective foliar insecticide treatment provided significant increase in yield over other treatments. Tomato yield was negatively correlated with the number of F. fusca and percentage of TSWV incidence. F. occidentalis per blossom was positively correlated with percentage of TSWV incidence, but not with yield. No significant interactions were observed between cultivar reflective mulch main plot treatments and insecticide subplot treatments; thus, treatment seemed to be additive in reducing the economic impact of thrips-vectored TSWV. Control tactics that manage thrips early in the growing season significantly increased tomato yield in years when the incidence of TSWV was high (>17%).

Genetic parameters for FAMACHA© score and related traits for host resistance/resilience and production at differing severities of worm challenge in a Merino flock in South Africa.

Abstract Development time and percent survival of the silverleaf whitefly, Bemisia argentifolii Bellows & Perring (formerly referred to as B strain of sweetpotato whitefly), were determined at five constant temperatures ranging from 20 to 35°C on two cultivars of cotton, cantaloupe, and pepper (Capsicum sp.). Fecundity was recorded at 30°C during the first 10 d of adulthood. Developmental, survival, and fecundity rates were not significantly different between cultivars within crops of either cotton or cantaloupe, but varied between crops. Development time from egg to adult at 20–32°C ranged from 14.6 ± 0.8 (mean ± SE) to 36.0 ± 1.0 d on cantaloupe and from 16.3 ± 0.7 to 37.9 ± 2.1 d on cotton, respectively. Whiteflies did not develop at 35°C or on pepper at any temperature. Minimum developmental thresholds and degree-days requirements from egg to adult were 11.1°C and 312.5 DD on cotton and 13.2°C and 250.0 DD on cantaloupe, respectively. Immature survival was high (76.5 ± 11.5–100%) on cantaloupe, intermediate (37.3 ± 13.3–64.4 ± 10.2%) on cotton, and very low (0–8.3 ± 8.3%) on pepper from 20–32°C. Fecundity ranged from 153.3 ± 10.8–158.3 ± 9.3 eggs per female on cantaloupe, from 117.0 ± 6.0–117.5 ± 22.1 eggs per female on cotton, and from 2.1 ± 0.7–40.5 ± 5.8 eggs per female on pepper, at 30°C.

Aggregation pheromone for the pepper weevil,Anthonomus eugenii cano (Coleoptera: Curculionidae): Identification and field activity

The objectives of this study were to estimate genetic parameters for FAMACHA score and related traits at different levels of worm challenge in sheep and to assess the effect of different methods for modelling records from treated lambs on estimates of genetic parameters. Data were collected over five consecutive Haemonchus seasons from a total of l671 Merino lambs using the FAMACHA clinical evaluation system, and anaemic individuals were treated as needed, until flock health necessitated mass treatment at the peak of the worm season. Records of each sampling occasion were classified into low, moderate, or peak levels of worm challenge. Animal models were run separately for traits within each data set. Alternative analyses were conducted in which records of treated lambs were (1) included without adjustment, (2) included along with a fixed effect representing treatment status of the lamb for each record, and (3) included after application of a penalty to offset any phenotypic improvement or advantage due to that treatment. Estimates of heritability for individual FAMACHA data sets ranged from 0.06+/-0.04 to 0.24+/-0.05, the highest estimates being obtained for peak worm challenge data. Estimates of genetic correlation for FAMACHA with other traits varied, but were always near negative unity for FAMACHA score with haematocrit value. When data of treated lambs were penalised, higher estimates of heritability were obtained than when not penalised, hence this may be an effective method for allowing for early treatment of overly susceptible animals before the level of worm challenge is at an optimum level for BLUP (Best Linear Unbiased Prediction) evaluation. The estimate of genetic correlation for FAMACHA score in moderate worm challenge with that in peak worm challenge was almost unity. This suggests that estimation of breeding values for this trait using data from moderate worm challenge may be as effective as that from peak challenge.

Transmission of Iris Yellow Spot Virus by Frankliniella fusca and Thrips tabaci (Thysanoptera: Thripidae)

This study describes the identification of an aggregation pheromone for the pepper weevil,Anthonomus eugenii and field trials of a synthetic pheromone blend. Volatile collections and gas chromatography revealed the presence of six male-specific compounds. These compounds were identified using chromatographic and spectral techniques as: (Z)-2-(3,3-dimethylcyclohexylidene)ethanol, (E)-2-(3,3-dimethylcyclohexylidene)ethanol, (Z)-(3,3-dimethylcyclohexylidene)acetaldehyde, (E)-(3,3-dimethylcyclohexylidene)acetaldehyde, (E)-3,7-dimethyl-2,6-octadienoic acid (geranic acid), and (E)-3,7-dimethyl-2,6-octadien-1-ol (geraniol). The emission rates of these compounds from feeding males were determined to be about: 7.2, 4.8, 0.45, 0.30, 2.0, and 0.30µg/male/day, respectively. Sticky traps baited with a synthetic blend of these compounds captured more pepper weevils (both sexes) than did unbaited control traps or pheromone-baited boll weevil traps. Commercial and laboratory formulations of the synthetic pheromone were both attractive. However, the commercial formulation did not release geranic acid properly, and geranic acid is necessary for full activity. The pheromones of the pepper weevil and the boll weevil are compared. Improvements for increasing trap efficiency and possible uses for the pepper weevil pheromone are discussed. A convenient method for purifying geranic acid is also described.

Intramuscular fat and fatty acid composition of longissimus muscle from divergent pure breeds of cattle

ABSTRACT Thrips-transmitted Iris yellow spot virus (IYSV) (Family Bunyaviridae, Genus Tospovirus) affects onion production in the United States and worldwide. The presence of IYSV in Georgia was confirmed in 2003. Two important thrips species that transmit tospoviruses, the onion thrips (Thrips tabaci (Lindeman)) and the tobacco thrips (Frankliniella fusca (Hinds)) are known to infest onion in Georgia. However, T. tabaci is the only confirmed vector of IYSV. Experiments were conducted to test the vector status of F. fusca in comparison with T. tabaci. F. fusca and T. tabaci larvae and adults reared on IYSV-infected hosts were tested with antiserum specific to the nonstructural protein of IYSV through an antigen coated plate ELISA. The detection rates for F. fusca larvae and adults were 4.5 and 5.1%, respectively, and for T. tabaci larvae and adults they were 20.0 and 24.0%, respectively, indicating that both F. fusca and T. tabaci can transmit IYSV. Further, transmission efficiencies of F. fusca and T. tabaci were evaluated by using an indicator host, lisianthus (Eustoma russellianum (Salisbury)). Both F. fusca and T. tabaci transmitted IYSV at 18.3 and 76.6%, respectively. Results confirmed that F. fusca also can transmit IYSV but at a lower efficiency than T. tabaci. To attest if low vector competency of our laboratory-reared F. fusca population affected its IYSV transmission capability, a Tomato spotted wilt virus (Family Bunyaviridae, Genus Tospovirus) transmission experiment was conducted. F. fusca transmitted Tomato spotted wilt virus at a competent rate (90%) suggesting that the transmission efficiency of a competent thrips vector can widely vary between two closely related viruses.

Direct and indirect effects of a thrips-transmitted Tospovirus on the preference and fitness of its vector, Frankliniella fusca

The objective of this study was to compare the fatty acid (FA) composition of intramuscular fat from the LM of 3 divergent breeds of cattle: Angus (AN, n = 9), Brahman (BR, n = 7), and Romosinuano (RM, n = 11). Cattle were blocked by breed and finished 129 d before slaughter in one year and 157 d in the next year. Longissimus muscle samples were collected from each carcass between the 10th and 13th ribs, trimmed of external fat, frozen in liquid nitrogen, homogenized, and used for fat extraction, using a modified Folch procedure. Extracted fat was analyzed for FA by using a GLC system with an HP-88 capillary column. Fatty acid composition was expressed using both a normalized percentage (%) and gravimetric calculation (mg/g of fresh muscle tissue) in relation to degree of saturation, which was determined using a saturation index (ratio of total SFA to total unsaturated FA). Crude fat determination revealed that LM from AN purebred cattle had the greatest amount of intramuscular fat (7.08%; P = 0.001). Although intramuscular fat of LM from RM contained a reduced percentage of total SFA (P = 0.002) compared with AN, it had the greatest percentage of total PUFA (P < 0.001 and P = 0.020). The percentages of total MUFA were similar among the 3 breeds (P = 0.675). The gravimetric calculation, a measure of actual FA concentration, showed significantly greater concentrations of SFA (26.67 mg/g), MUFA (26.50 mg/g), and PUFA (2.37 mg/g) in LM from AN cattle, as compared with LM from BR and RM cattle (P < 0.001). Interestingly, BR purebreds had the least PUFA concentration (1.49 mg/g; P <or= 0.001) in the LM, although their intramuscular fat content was similar to that of RM (P = 0.924). Regardless of breed, the MUFA proportion was always the greatest (47.58%; P <or= 0.005), whereas PUFA was the least contributor to FA composition (1.49 to 2.37 mg/g and 4.36 to 8.78%; P < 0.001). Beef LM fatty acid composition was characterized by palmitic and oleic acids being the most abundant FA (P < 0.001). These results suggested a genetic variation in FA synthesis and deposition among breeds that influenced both marbling and its composition.

First report of Vidalia onion (Allium cepa) naturally infected with tomato spotted wilt virus and Iris yellow spot virus (family Bunyaviridae, genus Tospovirus) in Georgia.

Phytoviruses including tospoviruses are known to affect the behavior and fitness of their vectors both positively and negatively. In this study, we investigated the effects of Tomato spotted wilt virus (TSWV) (family Bunyaviridae, genus Tospovirus) infection on the fitness and feeding ability of tobacco thrips, Frankliniella fusca (Hinds) (Thysanoptera: Thripidae) using peanut, Arachis hypogaea L. (Fabaceae), as a host. Potentially viruliferous F. fusca laid more eggs than non‐viruliferous F. fusca. In contrast, fewer potentially viruliferous F. fusca developed into adults and required a longer developmental time than non‐viruliferous F. fusca, indicating a direct negative effect of the virus on thrips fitness. In addition, no‐choice feeding tests indicated that non‐viruliferous F. fusca fed more rapidly than potentially viruliferous F. fusca. Typically, phytovirus infections are known to enhance the availability of vital nutrients such as free amino acids in infected host plants and to affect other important physiological processes negatively. Free amino acids are known to play a vital role in egg production and development. Further investigations in this study revealed that leaflets of infected plants had ca. 15 times more free amino acids than non‐infected leaflets. TSWV‐infected leaflets were used to rear potentially viruliferous thrips. Higher amino acid levels in TSWV‐infected leaflets than in non‐infected leaflets could have contributed to increased oviposition by potentially viruliferous F. fusca compared to non‐viruliferous F. fusca. Taken together, these results suggest that increased concentrations of free amino acids in TSWV‐infected plants might serve as an incentive for thrips feeding on otherwise unsuitable hosts, thereby facilitating TSWV acquisition and transmission.

Host Plant Resistance Against Tomato spotted wilt virus in Peanut (Arachis hypogaea) and Its Impact on Susceptibility to the Virus, Virus Population Genetics, and Vector Feeding Behavior and Survival

Vidalia onion is an important crop in Georgias agriculture with worldwide recognition as a specialty vegetable. Vidalia onions are shortday, Granex-type sweet onions grown within a specific area of southeastern Georgia. Tomato spotted wilt virus (TSWV) has been endemic to Georgia crops for the past decade, but has gone undetected in Vidalia onions. Tobacco thrips (Frankliniella fusca) and Western flower thrips (Frankliniella occidentalis) are the primary vectors for TSWV in this region, and a number of plant species serve as reproductive reservoirs for the vector or virus. Iris yellow spot virus (IYSV), an emerging tospovirus that is potentially a devastating pathogen of onion, has been reported in many locations in the western United States (2,4). Thrips tabaci is the known vector for IYSV, but it is unknown if noncrop plants play a role in its epidemiology in Georgia. During October 2003, a small (n = 12) sampling of onions with chlorosis and dieback of unknown etiology from the Vidalia region was screened for a variety of viruses, and TSWV and IYSV infections were serologically detected. Since that time, leaf and bulb tissues from 4,424 onion samples were screened for TSWV and IYSV using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) with commercial kits (Agdia Inc., Elkhart, IN). Samples were collected from 53 locations in the Vidalia region during the growing season between November 2003 and March 2004. Plants exhibiting stress, such as tip dieback, necrotic lesions, chlorosis or environmental damage were selected. Of these, 306 were positive for TSWV and 396 were positive for IYSV using positive threshold absorbance of three times the average plus two standard deviations of healthy negative onion controls. Positive serological findings of the onion tissues were verified by immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) for TSWV (3) and RT-PCR for IYSV (1). In both instances, a region of the viral nucleocapsid (N) gene was amplified. The PCR products were analyzed with gel electrophoresis with an ethidium bromide stain in 0.8% agarose. Eighty-six percent (n = 263) of the TSWV ELISA-positive samples exhibited the expected 774-bp product and 55 percent (n = 217) of the IYSV ELISA-positive samples exhibited the expected 962-bp product. The reduced success of the IYSV verification could be attributed to the age and deteriorated condition of the samples at the time of amplification. Thrips tabaci were obtained from onion seedbeds and cull piles within the early sampling (n = 84) and screened for TSWV by the use of an indirect-ELISA to the nonstructural (NSs) protein of TSWV. Of the thrips sampled, 25 were positive in ELISA. While the incidence of IYSV and TSWV in the Vidalia onion crop has been documented, more research is needed to illuminate their potential danger to Vidalia onions. References: (1) I. Cortês et al. Phytopathology 88:1276, 1998. (2) L. J. du Toit et al. Plant Dis. 88:222, 2004. (3) R. K. Jain et al. Plant Dis. 82:900, 1998. (4) J. W. Moyer et al. (Abstr.) Phytopathology 93(suppl.):S115, 2003.