Acquisition Feeding Research Articles

A neurotropic route for Maize mosaic virus (Rhabdoviridae) in its planthopper vector Peregrinus maidis

To investigate the dissemination route of Maize mosaic virus (MMV, Rhabdoviridae) in its planthopper vector Peregrinus maidis (Delphacidae, Hemiptera), temporal and spatial distribution of MMV was studied by immunofluorescence confocal laser scanning microscopy following 1-week acquisition feeding of planthoppers on infected plants. MMV was detected 1-week post first access to diseased plants (padp) in the midgut and anterior diverticulum, 2-week padp in the esophagus, nerves, nerve ganglia and visceral muscles, and 3-week padp in hemocytes, tracheae, salivary glands and other tissues. MMV is neurotropic in P. maidis; infection was more extensive in the nervous system compared to other tissues. A significantly higher proportion of planthoppers had infected midguts (28.1%) compared to those with infected salivary glands (20.4%) or to those that transmitted MMV (15.7%), suggesting the occurrence of midgut and salivary gland barriers to MMV transmission in P. maidis. In this planthopper, the esophagus and anterior diverticulum are located between the compound ganglionic mass and the salivary glands. We postulate that MMV may overcome transmission barriers in P. maidis by proceeding from the midgut to the anterior diverticulum and esophagus, and from these to the salivary glands via the nervous system: a neurotropic route similar to that of some vertebrate-infecting rhabdoviruses.

Persistently Infected Calves as Reservoirs for Acquisition and Transovarial Transmission of Babesia bovis by Rhipicephalus ( Boophilus ) microplus

Babesia bovis is a deadly disease of cattle resulting in severe economic losses in the vast regions of the world where it is endemic. If reintroduced into the United States, babesiosis would cause significant mortality in the naïve cattle population. In order to address the risk to U.S. cattle, it is essential to quantify the transovarial transmission efficiency in adult female Boophilus microplus ticks following acquisition feeding on persistently infected cattle. This study tested the hypothesis that infection rates are the same for larval progeny derived from females fed to repletion during persistent or acute infection. Increasing parasite levels during acute infection correlated with an increasing number of females harboring kinetes detectable in hemolymph (r = 0.9). The percent infected larvae ranged from 0 to 20% when derived from females fed to repletion on persistently infected calves and from 4 to 6% when derived from females fed to repletion during acute parasitemia. There was no significant difference in infection rates of larval progeny, implying that the risk associated with the introduction of either persistently infected or acutely infected cattle is equal. Parasite levels ranged from 2.4 x 10(2) to 1.9 x 10(5) in 3-day-fed larvae derived from females fed to repletion on persistently infected cattle. One group of larvae failed to transmit the parasite, suggesting that a threshold level of parasites must be obtained by larval progeny via transovarial transmission in order for larvae to deliver sufficient parasites to infect a naïve host.

Xiphinema rivesi from Slovania Transmit Tobacco ringspot virus and Tomato ringspot virus to Cucumber Bait Plants.

The dagger nematode, Xiphinema rivesi Dalmasso, a member of the X. americanum group, was detected in 2002 for the first time in Slovenia and for the fourth time in Europe (4). X. rivesi is a vector of at least four North American nepoviruses including Cherry rasp leaf virus (CRLV), Tobacco ringspot virus (TRSV), Tomato ringspot virus (ToRSV), and Peach rosette mosaic virus (PRMV) (1,2). All of these viruses are included on the EPPO and EU lists of quarantine organisms, but none of the Xiphinema species found in Europe have been reported to transmit these nepoviruses. Three virus isolates, including TRSV (from Lobelia spp.; virus collection of the Plant Protection Service, Wageningen, The Netherlands), ToRSV (grapevine isolate PV-0381; DSMZ, Braunschweig, Germany), and Arabis mosaic virus (ArMV) (from Vinca spp.; virus collection of the Plant Protection Service), were used in transmission tests with a population of X. rivesi found in Slovenia. X. rivesi is not known to transmit ArMV and this virus was included as a check. The nematodes were extracted from peach orchard soil collected near the village of Dornberk, and transmission tests fulfilled the set of criteria proposed by Trudgill et al. (3). Cucumis sativus cv. Eva, grown in a growth chamber at 25°C, was used as acquisition hosts and transmission bait plants. The acquisition hosts were mechanically inoculated and showing systemic symptoms before the introduction of nematodes. Noninoculated acquisition plants were included as controls. After a 10-day acquisition feeding period, the nematodes were transferred to healthy bait plants and allowed a 14-day inoculation feeding period. X. rivesi transmitted TRSV and ToRSV but not ArMV. TRSV and ToRSV bait plants developed systemic symptoms 4 to 6 weeks after the nematodes were transferred. Transmission of TRSV and ToRSV was confirmed by testing leaf and root sap of bait plants in a double antibody sandwich (DAS)-ELISA. High virus concentrations were detected in the roots and leaves of TRSV and ToRSV symptomatic plants. DAS-ELISA on bait plants from nematodes that had been allowed to feed on ArMV-infected or the virus-free control acquisition plants gave negative results. No symptoms appeared on bait plants used for ArMV transmission or the control bait plants. To our knowledge, this is the first report of transmission of TRSV and ToRSV with a Xiphinema population from Europe.

Conservation of Transmission Phenotype of <I>Anaplasma marginale</I> (Rickettsiales: Anaplasmataceae) Strains Among <I>Dermacentor</I> and <I>Rhipicephalus</I> Ticks (Acari: Ixodidae)

Before the eradication of Boophilus ticks from the United States, Rhipicephalus (Boophilus) microplus (Canestrini) and Rhipicephalus (Boophilus) annulatus (Say) were important biological vectors of the cattle pathogen Anaplasma marginale Theiler. In the absence of Boophilus ticks, A. marginale continues to be transmitted by Dermacentor ticks. However, a few U.S. strains are not transmissible by Dermacentor andersoni Stiles, Dermacentor variabilis (Say), or both, raising the question of how these strains evolved and how they are maintained. We hypothesize that the U.S. non-Dermacentor-transmissible strains of A. marginale were formerly Boophilus-transmitted strains that have been maintained by a combination of persistent infection and mechanical transmission since the eradication of their biological vector from the United States. To test this hypothesis, we attempted to transmit a well-documented non-Dermacentor-transmissible A. marginale strain (Florida), by using D. andersoni and the two Boophilus species that formerly occurred in the United States. For comparison, we examined tick-borne transmission of a strain of A. marginale (Puerto Rico), which has previously been shown to be transmissible by both D. andersoni and B. microplus. All three species of tick transmitted the Puerto Rico strain, and immunohistochemical (IHC) analysis confirmed the presence ofA. marginale colonies in their salivary glands. All three tick species failed to transmit the Florida strain. Although both D. andersoni and B. microplus acquired transient midgut and salivary gland infections after acquisition feeding, we were unable to detect colonies of the Florida strain in the salivary glands with IHC. This demonstrates that the transmission phenotype ofA. marginale strains is conserved among tick species, and it suggests that the failure of the Florida strain to be transmitted by ticks is related to a general inability to efficiently invade or replicate in tick cells, rather than to a failure to invade or replicate in cells of a specific tick species.

Identification of midgut and salivary glands as specific and distinct barriers to efficient tick-borne transmission of Anaplasma marginale.

Understanding the determinants of efficient tick-borne microbial transmission is needed to better predict the emergence of highly transmissible pathogen strains and disease outbreaks. Although the basic developmental cycle of Anaplasma and Ehrlichia spp. within the tick has been delineated, there are marked differences in the ability of specific strains to be efficiently tick transmitted. Using the highly transmissible St. Maries strain of Anaplasma marginale in Dermacentor andersoni as a positive control and two unrelated nontransmissible strains, we identified distinct barriers to efficient transmission within the tick. The Mississippi strain was unable to establish infection at the level of the midgut epithelium despite successful ingestion of infected blood following acquisition feeding on a bacteremic animal host. This inability to colonize the midgut epithelium prevented subsequent development within the salivary glands and transmission. In contrast, A. marginale subsp. centrale colonized the midgut and then the salivary glands, replicating to a titer indistinguishable from that of the highly transmissible St. Maries strain and at least 100 times greater than that previously associated with successful transmission. Nonetheless, A. marginale subsp. centrale was not transmitted, even when a large number of infected ticks was used for transmission feeding. These results establish that there are at least two specific barriers to efficient tick-borne transmission, the midgut and salivary glands, and highlight the complexity of the pathogen-tick interaction.

Transovarial Transmission Efficiency of Babesia bovis Tick Stages Acquired by Rhipicephalus ( Boophilus ) microplus during Acute Infection

The protozoan parasite Babesia bovis, a reemerging threat to U.S. cattle, is acquired by adult female ticks of the subgenus Boophilus and is transovarially transmitted as the kinete stage to developing larval offspring. Sporozoites develop within larvae and are transmitted during larval feeding on a bovine host. This study evaluated the efficiency of B. bovis infection within Rhipicephalus (Boophilus) microplus following acquisition feeding on acutely parasitemic cattle. Parasite levels were quantified in blood from experimentally infected cattle and within hemolymph and larvae derived from acquisition-fed female B. microplus. There was a positive correlation between blood parasite levels in acutely parasitemic cattle and kinete levels in the hemolymph of adult female Boophilus ticks following acquisition feeding; however, there was no relationship between kinete levels in females and infection rates of larval progeny. Boophilus microplus females that acquisition fed produced larval progeny with infection rates of 12% to 48%. Importantly, larvae derived from replete females with very low levels of kinete infection, as demonstrated by microscopy and PCR, had infection rates of 22% to 30% and transmitted B. bovis during transmission feeding. These data demonstrate that although hemolymph infection may be undetectable, transmission to larval progeny occurs at a level which ensures transmission to the bovine host.

Aphis glycines as a Vector of Persistently and Nonpersistently Transmitted Viruses and Potential Risks for Soybean and Other Crops.

The recently introduced soybean aphid (Aphis glycines), which is widespread in the soybean-growing regions in the United States, is the only aphid able to develop large colonies on soybean. Although its potential as a vector of plant viruses is recognized, reports on virus transmission efficiency by this aphid species are limited. In the present study, we examined the ability of A. glycines to transmit several economically important viruses. The results showed that A. glycines transmitted the potyviruses Bean yellow mosaic virus (BYMV) and Soybean mosaic virus from soybean to soybean more efficiently than Myzus persicae. However, M. persicae transmitted the alfamovirus Alfalfa mosaic virus and the potyviruses Tobacco etch virus (TEV) and Tobacco vein mottling virus (TVMV) from tobacco to tobacco more efficiently than A. glycines. This is the first report to demonstrate that the soybean aphid can vector TEV and TVMV, two economically important tobacco viruses. This also is the first report to document successful transmission of BYMV by A. glycines. All attempts to transmit the nepovirus Tobacco ringspot virus by A. glycines were unsuccessful, regardless of the length of the acquisition and inoculation feeding periods. Although the luteovirus Soybean dwarf virus (SbDV) was widely distributed in red and white clover in Kentucky, it was not detected in soybean. All transmission experiments of SbDV by A. glycines were unsuccessful. A reverse-transcription polymerase chain reaction (RT-PCR) assay was developed to detect SbDV in single aphids using a pair of primers designed to amplify a 372-bp PCR fragment in the coding region of SbDV coat protein. Although A. glycines was not a vector of SbDV, the virus was detected in 100% of tested aphids by RT-PCR after a 24- to 48-h virus acquisition access feeding. The practical applications of RT-PCR in detecting persistently transmitted viruses are discussed.

Quantifying the Feeding Periods Required by Corn Flea Beetles to Acquire and Transmit Pantoea stewartii.

The feeding periods required by corn flea beetles to acquire and transmit Pantoea stewartii were investigated in the Stewart's disease of corn pathosystem. To quantify the effect of acquisition feeding period on percentage of acquisition, field-collected corn beetles were allowed to feed for 6, 12, 24 36, 48, and 72 h on corn seedlings previously inoculated with a rifampicin- and nalidixic acid-restraint strain of P. stewartii. Acquisition of P. stewartii by corn flea beetles was considered positive if the rifampicin- and nalidixic acid-marked strain was recovered on selective media. To quantity the effect of transmission feeding period on percent transmission of P. stewartii by corn flea beetles, P. stewartii- infested corn flea beetles were allowed to feed on healthy corn seedlings for periods of 3, 6, 12, 24, 36, 48, and 72 h. After the appropriate transmission feeding period, leaf tissues surrounding the sites of feeding scars were cultured for the presence of the P. stewartii-marked strain. Transmission of P. stewartii was considered positive if the marked strain was recovered on selective media. Acquisition of P. stewartii occurred within 6 h and the percentage of corn flea beetles that had acquired P. stewartii after 72 h ranged from 68 to 94%. The change in P. stewartii acquisition by corn flea beetles (Y) with respect to acquisition feeding period (X) was best described by the Gompertz model, with R2 values ranging from 91 to 99%. The mean time for acquisition by 50% of the corn flea beetles was 36.5 ± 11.6 h. The minimum transmission feeding time required for corn flea beetles to transmit P. stewartii following a 48-h acquisition feeding period was less than 3 h. The percent transmission of P. stewartii by corn flea beetles was nearly 100% after a 48-h transmission feeding period and was 100% by 72 h. Among population growth models evaluated, the monomolecular model best described the relationship between percent transmission (Y) and transmission feeding periods (X), with R 2 values of up to 84%. However, a nonlinear form of the monomolecular model better quantified the relationship between percent transmission and transmission feeding period, because pseudo-R2 values ranged between 98.1 and 99.5%. The predicted transmission feeding time required for 50% of P. stewartii-infested corn flea beetles to transmit the pathogen was 7.6 ± 0.87 h. These results suggest that the corn flea beetle is a highly efficient vector that can quickly acquire and transmit P. stewartii, thereby requiring insecticide seed treatments and foliar insecticides that act quickly to prevent corn flea beetles from acquiring and transmitting P. stewartii to corn plants.

Citrus tristeza virus transmission by the Toxoptera citricida vector: In vitro acquisition and transmission and infectivity immunoneutralization experiments

Citrus tristeza virus (CTV) is transmitted by several aphid species in a semi-persistent manner with Toxoptera citricida, the brown citrus aphid (BrCA), being the most efficient. As yet, the molecular interactions between the virus and its aphid vectors have not been determined. This is the first report of aphids acquiring CTV from preparations through an artificial membrane and then transmitting it to receptor plants. The BrCA fed across artificial membranes on crude tissue preparations made from CTV-infected bark tissue were able to transmit CTV to virus-free receptor plants at low rates. CTV p20, p27 and p25 proteins, detected by Western blots, were present in all crude tissue preparations from CTV-infected plants. Partially purified CTV preparations were not transmitted by the BrCA in this manner. Infectivity immunoneutralization experiments were conducted where aphids were forced to feed in vitro on three CTV-specific antibodies (p25, p27 and p20) before being placed on receptor plants following a 48 h acquisition feed on CTV-infected source plants. There were no differences in transmission rates among the majority of treatments and the control treatments. However, in one infectivity immunoneutralization experiment, the CTV p20 antibodies significantly enhanced CTV transmission compared to buffer only, pre-immune antiserum or no antibody control treatments. This suggests the inactivity of CTV p20 aids BrCA transmission of virions.

Transmission studies on an indian isolate of sugarcane mosaic potyvirus

An Indian isolate of sugarcane mosaic virus was found easily transmitted by mechanical sap inoculation to sugarcane and sorghum plants. Out of different methods tried for mechanical inoculation, Bain’s and Matz’s method gave maximum per cent of infection in inoculated sugarcane and three varieties ofSorghum vulgare (cv. M.P.cherry, CSH-5 and CSH-9). The virus was also found to be transmitted by four aphids, viz.Aphis gossypii, Longiunguis sacchari, Myzus persicae andRophalosiphum maidis. Since the virus could be transmitted most efficiently byR. maidis the detailed study was made with it to establish the virus-vector relationship.R. maidis could acquire virus without any pre-acquisition fasting, but fasting has beneficial effect up to 4h on the acquisition power. In case, fasting is increased beyond 4 h, the efficiency of transmission decreased. The aphid acquire the virus within 30 sec. However, optimum acquisition feeding was obtained in 2 min. but the maximum infection was obtained after 30 min infection feeding of test plants. Even a single viruliferous aphid was enough to initiate the infection. The transmission per cent decreased when the aphids were fasted after acquisition feeding of 2h from 60 to 13 per cent. In serial transfers, the aphids cease to be infective very soon while feeding on the test plants. Thus the virus under study was transmitted apparently by typically non-persistent manner and was stylet borne.

Virus-Vector Relationship of Bittergourd yellow mosaic virus and the Whitefly Bemisia tabaci Genn.

Bittergourd yellow mosaic virus (BGYMV) is one of the virus affecting the bittergourd (Momordica charantia). It is transmitted by whitefly Bemisia tabaci Genn. Minimum of five whiteflies were required to transmit the virus. However, hundred per cent transmission of BGYMV disease was obtained when 45 whiteflies were released per plant. Twelve hours of acquisition access feeding period (AAFP) and inoculation access feeding period (IAFP) were required for the whiteflies to transmit the disease. The percentage of transmission increased with increase in both acquisition and inoculation feeding period. A pre-acquisition starvation period of 3.5 h ensured hundred per cent transmission, while post-acquisition starvation period affected the transmission.

Transmisión del Mal de Río Cuarto virus por ninfas de primer y tercer estadio de Delphacodes kuscheli

Se comparó la capacidad de ninfas de primer y tercer estadío de Delphacodes kuscheli Fennah (Hemiptera: Delphacidae) para adquirir y posteriormente transmitir el Mal de Río Cuarto virus (MRCV), bajo condiciones controladas. Ninfas I y III avirulíferas se alimentaron separadamente de plantas de trigo infectadas durante 48 horas, para luego ser colocadas en subgrupos de tres individuos sobre plantas de trigo sanas. Se realizaron transmisiones seriales utilizando períodos de inoculación de 24 horas. Ambos estadií os lograron adquirir y transmitir el MRCV, pero se evidenció una mayor cantidad de subgrupos infectivos cuando la adquisición se efectuó como ninfas I, así como una disminución significativa en la duración del período de latencia del virus respecto de los insectos que adquirieron el MRCV durante el tercer estadio ninfal.

Evaluation of Tomato (Lycopersicon Esculentum Mill.) Genotypes for resistance to the Tomato Yellow Leaf Curl Tanzania Virus

Field experiments were conducted for two seasons (2001 and 2002) to determine the reaction of tomato genotypes to Tomato yellow leaf curl Tanzania virus. Tomato plants at the first leaf stage were inoculated with the virus in insect-proof cages in the screenhouse following 48-h and 72-h acquisition and inoculation feeding times by Bemisia tabaci, respectively. After inoculation, the whiteflies were killed with Karate 5EC (a.i 50 g/l λ-cylohalothrin). Test plants were transplanted to the field 4 weeks after inoculation in paired inoculated and non-inoculated plots replicated three times. The plants were maintained insect-free for the duration of the experiment by spraying weekly with Karate 5EC. Disease severity was scored weekly on a scale of 0 – 4 (0 = no symptoms and 4 = very severe symptoms). Average fresh plant weight after harvest (without roots), fruit number per plant, fruit weight and average total yield were recorded. There were significant differences in the effect of disease on yield components between genotypes compared with uninfected controls, except for genotype TY172, which produced insignificant differences (P = 0.5) in all the parameters assessed between the inoculated and uninoculated control plants. The susceptible cultivar Moneymaker had the most severe symptoms of TYLCTZV of all the cultivars tested, while no symptoms were observed on the resistant tomato line TY172. The line TY172 consistently performed better than the rest of the tomato genotypes, followed by Tengeru 97. Tengeru 97 produced the highest mean plant fresh weight and yielded more than the rest of the tomato genotypes tested. It is therefore, suggested that the resistance found in TY172 should be introgressed into Tengeru 97, Cal-J and Moneymaker, which are local tomato varieties preferred by growers in Tanzania. Such efforts will increase and improve the yield and quality of these varieties.

Whitefly transmission of sweet potato viruses

Three genera of plant viruses, Begomovirus (Geminiviridae), Crinivirus (Closteroviridae) and Ipomovirus (Potyviridae), contain members that infect sweet potato ( Ipomoea batatas) and are transmitted by whiteflies. The begomoviruses, Sweet potato leaf curl virus (SPLCV) and Ipomoea leaf curl virus (ILCV), and the ipomovirus Sweet potato mild mottle virus are transmitted by Bemisia tabaci, the sweet potato whitefly. The crinivirus, Sweet potato chlorotic stunt virus (SPCSV), is transmitted by B. tabaci and Trialeurodes abutilonea, the bandedwinged whitefly. Transmission experiments were done with three of these viruses using laboratory-reared whiteflies and 2-day acquisition and transmission feeding periods. SPLCV and ILCV were transmitted from single and double infections by B. tabaci at rates of 5–10%. Transmission rates for SPLCV by B. tabaci were 15–20%. T. abutilonea transmitted SPCSV at a rate of ca. 3% but did not transmit ILCV or SPLCV.

ミツバてんぐ巣病及びタマネギ萎黄病のヒシモンモドキによる伝搬

Cryptotaenia japonica witches' broom phytoplasma (CJWP) and onion yellows phytoplasma (OYP) were transmitted by the leafhopper H. sellatiformis. CJWP and OYP were also detected by PCR in H. sellatiformis fed on infected plants. Latent periods in H. sellatiformis of CJWP and OYP were 13-17 days. The phytoplasmas were detected by PCR in leafhoppers at about 7 days after the start of acquisition feeding. This is the first report of the transmission of CJWP and OYP by H. sellatiformis.

The effect of feeding time on potato virus S transmission byMyzus persicae (Sulz.) andAphis nasturtii Kalt, aphids

The relationship between the feeding time and potato virus S (PVS) transmission byMyzus persicae andAphis nasturtii was studied. The optimal acquisition feeding time forM. persicae was at 30 sec to 2 min (2.9% infected plants). For transmission byA. nasturtii the highest level of infection was after acquisition feeding at 15 sec (11%), 30 sec and 2 min (9.6% each) and there was a significant linear decrease of plant infection as feeding time extended from 7 sec to 64 min. A different relationship has been obtained for inoculation feeding. In PVS transmission byM. persicae the plant infection increased from 0% at 7 sec feeding to 2.9% at 8 and 32 min. In virus transmission byA. nasturtii the infection was 4% after 7 sec feeding and increased significantly to 12.2% at 64 min feeding time.

Biomolecular relationships among isolates ofTomato Yellow Leaf Curl Tanzania Virus

An investigation of the biological properties of the virus causing tomato yellow leaf curl disease in Tanzania was initiated to compare it with other known tomato yellow leaf curl viruses. Properties relating to acquisition and inoculation feeding time, persistence, mechanical inoculation, seed transmission and host range were studied. Results obtained indicate that the virus was transmitted persistently byBemisia tabaci Genn., but it was not mechanically, sap- or seed-transmissible. Minimum acquisition and inoculation feeding time was 30 min.Capsicum annuum, Datura stramonium, Nicotiana glutinosa, N. tabacum andLycopersicon esculentum were found to be hosts of the virus among the plant species tested, whereasPhaseolus vulgaris was not. It is concluded that the properties of the agent causing yellow leaf curl symptoms in tomato plants from different regions in Tanzania are similar to those ofTomato yellow leaf curl Sardinia virus species studied elsewhere.

Effect of Aphid Behavior on Efficiency of Transmission of Soybean mosaic virus by the Soybean-Colonizing Aphid, Aphis glycines.

The soybean aphid (Aphis glycines) was a poor vector (0.83% transmission) when the aphids were allowed overnight acquisition feed on Soybean mosaic virus (SMV)-infected soybean leaves. However, A. glycines was shown to be a very efficient vector (34.72% transmission) when individual aphids were allowed a 1-min acquisition probe on the same infected leaves used for the feeding treatment. Similar results were obtained with Myzus persicae and tobacco in transmission experiments of the potyviruses Tobacco etch virus (feeding: 1.36%; probing: 45.5%) and Tobacco vein mottling virus (feeding: 2.0%; probing: 47.5%). A reverse transcription-polymerase chain reaction (RT-PCR) assay was developed to detect SMV in single soybean aphids using a pair of primers designed to amplify a 469-bp PCR fragment in the coding region of SMV coat protein. In contrast to the low transmission rate obtained with the soybean aphids that acquired virus through overnight feeding, RT-PCR detected SMV in 100% of these aphids. Interestingly, the rate of SMV detection by RT-PCR in aphids that were allowed a 1-min acquisition probe (31.67%) coincided with percent transmission (34.72%). The practical application of RT-PCR in detecting nonpersistently transmitted viruses and its implications for virus epidemiology are discussed.

First Report of Tobacco Bushy Top Disease in China.

In 1993, a severe epidemic of a new disease of flue-cured tobacco (Nicotiana tabacum) occurred in western Yunnan Province, People's Republic of China. Since then, over 40,000 ha of tobacco have been affected, with an average incidence of ≈15%. Infected plants were stunted, and leaves showed symptoms of vein distortion, vein clearing, mottling, and rounding. Axillary buds sprouted from the main stem of infected plants early and formed lateral shoots, on which other shoots were produced. As a result, the infected plants presented the characteristic "bushy" appearance. Aphid- (Myzus persicae (Sulzer)) and sap-transmission experiments were conducted. In aphid-transmission tests, after an acquisition feeding period of 24 h on diseased tobacco, the shortest test feeding that resulted in infection was 5 min. The causal agent(s) was readily sap-transmissible, but it could not be transmitted from sap-inoculated plants to healthy plants by aphids. Thirty-two species of plants in eleven families were tested by sap-inoculation for infectivity to alternative hosts by the casual agent(s). All the species of Nicotiana tested were infected. All the hosts were restricted to the solanaceae. The symptoms, transmission, and host range of this disease were identical to those of tobacco bushy top disease in Zimbabwe (1). Using umbravirus-specific primers (3), a 550-bp DNA fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from diseased tobacco, whose sequence (GenBank Accession No. AF402620) indicated the occurrence of an umbravirus. The results coincided with the taxonomic status of Tobacco bushy top virus, one of the causal agents of tobacco bushy top disease (1), a tentative species of the genus Umbravirus (2). To our knowledge, this is the first report of tobacco bushy top disease in China and the first report of a large outbreak of the disease outside sub-Saharan Africa.

Variation in Transmission Efficiency Among Barley yellow dwarf virus-RMV Isolates and Clones of the Normally Inefficient Aphid Vector, Rhopalosiphum padi

The RMV strain of Barley yellow dwarf virus (BYDV-RMV) is an unassigned member of the Luteoviridae that causes barley yellow dwarf in various cereal crops. The virus is most efficiently vectored by the aphid Rhopalosiphum maidis, but can also be vectored with varying efficiency by R. padi and Schizaphis graminum. Field collections of alate aphids migrating into the emerging winter wheat crop in the fall of 1994 in central New York identified a high proportion of R. padi transmitting BYDV-RMV. This prompted a comparison of the BYDV-RMV isolates and the R. padi populations found in the field with type virus and aphid species maintained in the laboratory. A majority of the field isolates of BYDV-RMV were similar to each other and to the type BYDV-RMV isolate in disease severity on oat and in transmission by the laboratory-maintained population of R. maidis and a field-collected population of R. maidis. However, several field populations of R. padi differed in their ability to transmit the various BYDV-RMV isolates. The transmission efficiency of the R. padi clones was increased if acquisition and inoculation feeding periods were allowed at higher temperatures. In addition, the transmission efficiency of BYDV-RMV was significantly influenced by the aphid that inoculated the virus source tissue. In general, BYDV-RMV transmission by R. padi was higher when R. padi was the aphid that inoculated the source tissue than when R. maidis was the inoculating aphid. The magnitude of the change varied among virus isolates and R. padi clones. These results indicate that, under certain environmental conditions, R. padi can play a significant role in the epidemiology of BYDV-RMV. This may be especially significant in regions where corn is a major source of virus and of aphids that can carry virus into a fall-planted wheat crop.