FACTORS AFFECTING THE SPREAD OF APHID-BORNE VIRUSES IN POTATO IN EASTERN SCOTLAND

Experiments at Invergowrie, south-east Perthshire, showed that the extent of spread of potato leaf-roll and Y viruses varied from year to year and that virus Y consistently spread more than leaf roll. Most spread of Virus Y occurred before the end of June and of leaf-roll virus before the end of July. Both viruses spread slightly more in late- than in early-planted crops. When plants with leaf roll and already colonized by Myzus persicae were placed in a healthy crop of Majestic potatoes at intervals during the season, the amount of virus spread decreased rapidly with increasing age of the crop. Spread of leaf roll occurred in all of twenty-five 'seed' crops in different districts of eastern Scotland in 1955 but in only twenty out of thirty-six similar crops in 1956. Annual and regional differences in virus spread appear to reflect differences in the time at which migrant aphids reach potato crops in early summer and the rate at which infestation builds up in the crops.     

STUDIES ON THE APHID TRANSMISSION OF A STRAIN OF HENBANE MOSAIC VIRUS

A virus causing a wilt of Datura stramonium was identified as a strain of henbane mosaic virus. It causes necrotic local lesions in Nicotiana rustica , and local lesions are demonstrable in tobacco by staining with iodine. Some of the factors affecting its transmission by Myzus persicae (Sulz.) were studied quantitatively using these lesions.
Infective aphids differed little in their ability to cause infection, and usually produced two or three lesions. The duration of the feeding puncture did not affect the number of infections and had little effect on the percentage of aphids becoming infective. Transmissible virus did not seem to be continually imbibed while aphids fed on infected plants, and there were indications that it was acquired immediately before aphids withdrew their stylets from the leaf. Aphids became infective when allowed to make feeding punctures into epidermis stripped from infected leaves.
M. persicae transmitted during feeding punctures as brief as 5–10 sec; the probability of single feeding punctures resulting in infection reached a maximum with those lasting from 20 to 30 sec, during which the stylets did not penetrate as far as the centre of the epidermal cell and little or no saliva appeared to be ejected. M. persicae did not transmit the virus when its stylets were artificially wetted with infective sap.
Periods of darkness before inoculation with datura wilt virus increased the susceptibility of Nicotiana rustica to infection by rubbing, but not to infection by aphids.     

STUDIES ON TWO APHID-TRANSMITTED VIRUSES OF LEGUMINOUS CROPS

Pea mosaic virus was transmitted by Myzus persicae Sulz., Macrosiphum pisi Kalt., M. solanifolii Ash. and Aphis fabae Scop., but not by Hyperomyzus staphyleae Koch. It is a 'non-persistent' virus (Watson & Roberts, 1939), and is most readily transmitted when vectors are fasted and then given a short infection feeding. Vector efficiency was not increased by increases in preliminary fasting beyond 15 min. or with increasing infection feeding beyond 1 hr. Most aphids became non-infective within 15 min. when feeding, but fasting aphids remained infective for 3 hr. Species that fed readily on the infected plants were less efficient vectors than those which did not. Seed set by infected plants produced healthy seedlings.
Pea enation mosaic virus persisted in Myzus persicae and Macrosiphum pisi for more than 140 hr.; its transmission was unaffected by preliminary treatments of aphids. No transmission was obtained until at least 4 hr. after aphids had left infected plants; usually the 'latent' period exceeded 1 day and its duration was apparently unaffected by the length of the infection feeding.     

THE CORRELATION OF APHID NUMBERS WITH THE SPREAD OF LEAF ROLL AND RUGOSE MOSAIC IN POTATO CROPS

An analysis of the results of experiments in different parts of England and Wales from 1941 to 1947 on the spread of potato leaf roll and rugose mosaic showed that leaf roll spread was correlated with the number of alate Myzus persicae (Sulzer) caught on sticky traps throughout the potato-growing season; there was some correlation with the maximum count of M. persicae per 100 leaves, but this possibly results from the correlation between trapped aphids and the number per 100 leaves. Spread of rugose mosaic (potato virus Y) was correlated to a lesser degree with number of M, persicae , perhaps because other aphid species are often vectors. With both diseases higher correlations were obtained when the infected plants were dispersed among the healthy crop than when they were placed together in a row. It is concluded that it is possible to predict the average health of potato stocks in the following year from average trap data; further work may enable the health of individual stocks to be predicted.     

Experiments on the spread of rugose mosaic and leaf roll in potato crops in 1946

A series of experiments on the spread of potato rugose mosaic (virus Y ), and leaf roll, which has been in progress on a uniform plan since 1943, was ended in 1946. Mean values for thirteen centres in England and Wales showed that in 1946 69% of the infections with virus Y and 48 % of those with leaf-roll virus reached the tubers of Majestic potatoes by the beginning of August. There was usually little subsequent increase of rugose mosaic, but a late increase of leaf roll was associated with a relatively high initial spread. Three-quarters of the virus Y and over half the leaf-roll infections occurred within five plants distance of the source. There was no close correlation between the spread of either virus and the maximum number of Myzus persicae , either apterous forms on the plants or alate forms caught on adhesive traps, but centres with high trap catches in July and August showed pronounced late season spread of leaf roll. There were marked differences at different centres in the relative spread of the two viruses. The amount of spread and the gradients from source of infection of the two viruses are compared over the period 1943–6.     

THE SPREAD OF VIRUS DISEASES TO SINGLE POTATO PLANTS BY WINGED APHIDS

Young potato plants in pots exposed in the open near plots of potatoes for limited periods at intervals during the summer, became infested with large numbers of winged aphids only during warm, calm and dry weather. Although visited by aphids during May and June, when much of the spread of viruses occurred in nearby potato crops, few of the potted plants became infected. Most potted plants became infected in July when alate aphids were leaving neighbouring potato crops. Widely different proportions of the exposed plants became infected in different years; in two of the three years, many more plants were infected with virus Y than with leaf roll virus.     

EXPERIMENTS ON THE COLONIZATION OF POTATO PLANTS BY APTEROUS AND BY ALATE APHIDS IN RELATION TO THE SPREAD OF VIRUS DISEASES

Batches of potato plants in pots were placed in the field for limited periods among plants infected with potato virus Y and leaf roll virus. Some of the potted plants were surrounded by sticky boards which prevented apterous aphids from reaching them. Almost as many plants within the boards as without became infected, indicating that most of the spread of virus was by winged aphids.
Apterae were probably responsible for spreading the viruses throughout a hill after one or more stems were infected. They may carry infection to neighbouring plants, but most of these will have been infected already by alatae.
The number of plants contracting infection was unaffected by watering.     

QUANTITATIVE STUDIES ON THE TRANSMISSION OF CABBAGE BLACK RING SPOT VIRUS BY MYZUS PERSICAE (SULZ.)

Factors affecting the transmission of cabbage black ring spot virus by Mysus persicae (Sulz.) were studied quantitatively using the local lesions produced on tobacco leaves. Aphids prevented from feeding for 15 min. or more, before feeding for a few minutes on an infected plant, caused more infections than unfasted aphids. Fasted aphids acquired virus from infected plants in feeding times as short as 10 sec., and infected healthy plants in test-feeding times of 5 sec. Increasing test-feeding times to 30 min. increased the numbers of infections. Increasing infection- feeding times from 10 sec. to 5 min. had little effect, but increasing to more than 5 min. greatly reduced the number of transmissions. This reduction was partly offset if the aphids were prevented from feeding continuously while on the infected plants. With undisturbed infection-feeding periods of 15 min. or longer, previously fasted aphids caused no more infections than unfasted aphids.
Infective aphids lost their ability to produce lesions more rapidly when feeding than when fasting.
Winged and wingless aphids were equally efficient vectors.     

THE TRANSMISSION OF PLANT VIRUSES BY BITING INSECTS, WITH PARTICULAR REFERENCE TO COWPEA MOSAIC

Experiments on the virus-vector relationship of the Trinidad cowpea mosaic virus, transmitted by Ceratoma ruficornis , gave the following results: ability to infect decreased with increasing time after ceasing to feed on infected plants, but vectors remained infective for 14 days (much longer than the longevity in vitro of the virus at glasshouse shade temperatures of 23–31°C.); the beetles transmitted more consistently after longer feeding on infected plants, though feeds of under 5 min. made them efficient vectors; the proportion of plants infected increased with the amount of feeding damage on them; fasting the vectors before feeding on infected plants increased voracity but had no effect on their ability to transmit; beetles became infective immediately after feeding on infected plants. Cowpeas were infected by inoculation with macerated infective vectors or with juice regurgitated by vectors. There is no evidence that aphids or other sucking insects can transmit the virus. It seems similar to squash mosaic and turnip yellow mosaic, for vectors of all three viruses probably transmit by regurgitating infective juice during feeding.     

Infection of potato plants with potato leafroll virus changes attraction and feeding behaviour of Myzus persicae

Potato leafroll virus (PLRV; genus Polerovirus, family Luteoviridae) is a persistently transmitted circulative virus that depends on aphids for spreading. The primary vector of PLRV is the aphid Myzus persicae (Sulzer) (Homoptera: Aphididae). Solanum tuberosum L. potato cv. Kardal (Solanaceae) has a certain degree of resistance to M. persicae: young leaves seem to be resistant, whereas senescent leaves are susceptible. In this study, we investigated whether PLRV‐infection of potato plants affected aphid behaviour. We found that M. persicae's ability to differentiate headspace volatiles emitted from PLRV‐infected and non‐infected potato plants depends on the age of the leaf. In young apical leaves, no difference in aphid attraction was found between PLRV‐infected and non‐infected leaves. In fact, hardly any aphids were attracted. On the contrary, in mature leaves, headspace volatiles from virus infected leaves attracted the aphids. We also studied the effect of PLRV‐infection on probing and feeding behaviour (plant penetration) of M. persicae using the electrical penetration graph technique (DC system). Several differences were observed between plant penetration in PLRV‐infected and non‐infected plants, but only after infected plants showed visual symptoms of PLRV infection. The effects of PLRV‐infection in plants on the behaviour of M. persicae, the vector of the virus, and the implications of these effects on the transmission of the virus are thoroughly discussed.     

Transmission of potato leafroll virus in relation to the honeydew excretion of Myzus persicae

Honeydew excretion of single Myzus persicae nymphs on potato leafroll virus (PLVR)-infected Physalis floridana was studied during the acquisition access period (AAP) in relation to the efficiency of virus transmission.
With increasing length of the AAP, the percentage of nymphs that transmitted the virus increased. These nymphs produced significantly more honeydew droplets during the AAP on PLRV-infected P. floridana plants than nymphs which failed to transmit the virus. However, the number of honeydew droplets excreted during the AAP by transmitting nymphs did not affect the length of the latency period. Nymphs which infected the first test plant after a short latency period produced a similar amount of honeydew during the AAP to those with a longer latency period.
Honeydew excretion recorded on plants of varied age, showed that nymphs feeding on bottom leaves of infected plants produced more honeydew droplets than on comparable leaves of healthy plants. On infected plants, nymphs produced more honeydew droplets on bottom leaves with pronounced symptoms than on top leaves that hardly showed any symptom of PLRV infection.
The concentration of viral antigen measured by ELISA was lower in top leaves than in bottom leaves of infected plants. Nevertheless, nymphs feeding on top leaves transmitted the virus more efficiently than those which used bottom leaves as virus source. When bottom leaves were used as a virus source, the percentage of viruliferous nymphs decreased with plant age. These results indicate that the availability of virus for acquisition by aphids declines with increasing plant age and symptom severity.     

Volatiles from potato plants infected with potato leafroll virus attract and arrest the virus vector, Myzus persicae (Homoptera: Aphididae)

The influence of viral disease symptoms on the behaviour of virus vectors has implications for disease epidemiology. Here we show that previously reported preferential colonization of potatoes infected by potato leafroll virus (genus Polerovirus) (luteovirus) (PLRV) by alatae of Myzus persicae, the principal aphid vector of PLRV, is influenced by volatile emissions from PLRV-infected plants. First, in our bioassays both differential immigration and emigration were involved in preferential colonization by aphids of PLRV-infected plants. Second, M. persicae apterae aggregated preferentially, on screening above leaflets of PLRV-infected potatoes as compared with leaflets from uninfected plants, or from plants infected with potato virus X (PVX) or potato virus Y (PVY). Third, the aphids aggregated preferentially on screening over leaflet models treated with volatiles collected from PLRV-infected plants as compared with those collected from uninfected plants. The specific cues eliciting the aphid responses were not determined, but differences between headspace volatiles of infected and uninfected plants suggest possible ones.     

INFECTIVITY OF APHIDS BRED ON VIRUS-INFECTED CAULIFLOWER PLANTS

Caged cauliflower plants infected with either cabbage black ring spot virus (CBRSV) or cauliflower mosaic virus (CIMV) were colonized with Myzus persicae or Brevicoryne brassicae. Winged and wingless aphids that voluntarily flew or walked from these plants were transferred singly to healthy cauliflower or other brassica seedlings to compare their feeding behaviour and ability to transmit the viruses. Wingless aphids settled to probe more readily than winged, and B. brassicae was initially more restless than M. persicae. CIMV was more readily transmitted than CBRSV by both species, and B. brassicae rarely transmitted CBRSV. Wingless aphids transmitted less often than winged ones, and no wingless B. brassicae transmitted CBRSV, although they did CIMV. Fewer aphids transmitted CBRSV from old plants than from young ones, but plant age had little effect on CIMV transmission.     

Experiments on the Spread of Potato Virus X Between Plants in Contact

Experiments on the spread of five strains of potato virus X were made with seven potato varieties and with tomato plants both under glass and in the field. Spread by leaf contact between healthy and infected plants was confirmed, and it was also found that spread could occur between plants whose only contact was below ground.
The rate of spread was much greater in tomato than in potato plants, and virulent strains of the virus, which achieve a high concentration in infected plants, spread more rapidly than avirulent strains. In only one experiment with potatoes did more than 10% of the healthy potato plants exposed to infection become infected during one season.
Datura stramonium and tomato plants became infected when growing in soil containing sap or residues from X -infected plants.
It was common in the field for potato plants whose foliage gave no reaction for virus X at the end of the season to yield a mixed progeny of healthy and infected tubers. Such infections are thought to result from underground spread.
Attempts to transmit virus X from infected to healthy potatoes by means of Rhizoctonia solani failed. No examples of infection were found except when healthy plants came into direct contact with sources of the virus.     

THE USE OF SOIL INSECTICIDES TO CONTROL POTATO APHIDS AND VIRUS DISEASES

A replicated trial was done to find whether the insecticides Thimet and Rogor applied in the soil affected the spread of aphid-transmitted viruses from infected to healthy plants with potato crops. The insecticides were applied at planting as activated carbon formulations at rates equal in cost to three sprays with DDT emulsion at 2 lb. DDT per acre. The infected plants were removed (rogued) in late June.
Thimet applied along the furrows with the fertilizer, and Rogor applied in individual doses beneath each tuber, kept the plants free from aphids from a week after the plants emerged until early August. Thimet in individual doses was less effective but greatly decreased the aphid infestation. All treatments prevented or greatly decreased the spread of leaf-roll virus, but they only slightly decreased the spread of virus Y. No treatment damaged the plants or depressed yields significantly.
Tubers harvested from the plots treated with insecticides contained only very small quantities of the insecticides, but shoots from them, when infested with adult Myzus persicae (Sulz.), carried fewer aphids a week after infestation than did shoots from control tubers. Shoots of tubers from treated plots also grew more slowly than those from the controls.
The aphicidal efficiency of Thimet applied as individual doses separated from the tubers by distances of up to 6 in., decreased as the distance increased, but the effect of distance became less as time passed.
Reasons for the differences in the behaviour of the insecticides are discussed, and the possibilities that the method offers to control virus diseases. The application of insecticides to soil promises to be a useful way of controlling the spread of viruses, provided the harvested crop is free from toxic residues.     

Transmission characteristics and some other properties of bean yellow vein-banding virus, and its association with pea enation

Bean yellow vein-banding virus (BYVBV) has been found occasionally in mixed infection with pea enation mosaic virus (PEMV) in spring-sown field beans (Vicia faba minor) in southern England. Glasshouse tests confirmed that, like PEMV, BYVBV is transmissible by manual inoculation and by aphids in the persistent manner. However, BYVBV can be transmitted by aphids only from plants that are also infected with a helper virus, usually PEMV. Thus after separation from PEMV by passage through Phaseolus vulgaris it was no longer aphid-transmissible. It became aphid-transmissible again only after re-mixing in plants with PEMV or with a substitute helper, bean leaf roll virus (BLRV). It was not transmitted by aphids that fed sequentially on plants singly infected with PEMV and BYVBV. Thus the interaction between BYVBV and PEMV (or BLRV) that enables BYVBV to be transmitted by aphids seems to occur only in doubly infected plants. However, it was not transmitted by aphids from plants doubly infected with BYVBV and broad bean wilt virus (BBWV). BYVBV and PEMV were transmitted more readily by Acyrthosiphon pisum than by Myzus persicae; neither virus was transmitted by Aphis fabae. Phenol extracts of BYVBV-infected leaves were more infective than phosphate buffer or bentonite-clarified extracts and were sometimes infective when diluted to 1/1000. The infectivity of BYVBV in phosphate buffer extracts of leaves singly infected with BYVBV, unlike that in extracts of leaves doubly infected with BYVBV and PEMV (or BLRV), was destroyed by treatment with organic solvents. BYVBV infected 11 of 28 plant species that were inoculated with phenol extracts; seven of the infected species were legumes. No transmission of BYVBV was detected through seed harvested from infected field bean plants. Isometric particles c. 30 nm in diameter were seen in extracts of plants doubly infected with BYVBV and PEMV but not in extracts of plants infected with BYVBV alone. Leaves of plants infected with BYVBV, alone or with PEMV, contained membrane-bound structures c. 50–90 nm in diameter associated with the tonoplast in cell vacuoles. These structures were not found in healthy leaves. BYVBV has several properties in common with other known aphid-borne viruses that are helper-dependent and transmitted in a persistent manner. Possibly, as suggested for some of them, aphid transmission of BYVBV depends on the coating of its nucleic acid with helper virus coat protein.     

The adaptation of tomato mosaic virus to resistant tomato plants

Tomato mosaic virus derived from susceptible tomato plants (the standard virus) was cultured in resistant plants. Sap from non-inoculated leaves of resistant tomato plants infected with virus from the resistant host was more infective and contained more virus particles than leaf sap of resistant plants infected with the standard virus. Leaves of resistant tomatoes infected with virus from the resistant host also showed more obvious symptoms. Susceptible plants infected with virus from resistant plants not only showed fewer symptoms than when infected with standard virus, but samples were less infective and contained less virus up to 26 weeks, when values for infectivity were similar. This modification in activity was not reversible and was obtained with two lines of tomato having different types of resistance. Passage of virus from resistant plants through susceptible plants did not impair its ability to infect resistant plants.     

STUDIES IN RUBUS VIRUS DISEASES

Raspberry leaf mottle virus is acquired by Amphorophora rubi in feeding periods of 1/2-2 hr. Longer feeding periods cause no significant fluctuations in infectivity. During continuous feeding on healthy plants, infectivity declines rapidly after 2 hr., but persists for at least 5 hr. if aphids are transferred frequently to fresh healthy plants. The estimated probability of infection by single aphids is 1 in 20. Rarely more than 50% of test plants, Rubus occidentalis , seedlings became infected, and this is thought to be due mainly to variation in plant susceptibility. Young leaves of infected Norfolk Giant are, however, better sources of virus than older leaves.     

New approaches for restricting spread of potato leafroll virus by different methods of eradicating infected plants from potato crops

Experiments were made at Invergowrie in 1984 and 1985 to compare the spread of potato leafroll virus (PLRV) after removing infected plants by three different methods; conventional roguing, desiccation with diquat, or incineration for 45–60s using a propane gas flame. Potato leaf roll 'infector' plants, grown in plots of virus-free Maris Piper seed potatoes, were artificially infested in June with aphids (Myzus persicae) from a laboratory culture, and removed from the plots after 2 or 3 wk. In both years, natural infestations of potato aphids were scarce during this period. There was no significant difference in the proportion of tubers infected with PLRV in adjacent plants after the neighbouring infector plants had been rogued by hand or desiccated with diquat, but the proportion was considerably reduced following incineration of the infector plants. In 1984, the spread of PLRV in conventionally rogued plots was also significantly reduced by a mixture of deltamethrin plus heptenophos, applied four times from 80% crop emergence, and was almost eliminated by a treatment with aldicarb granules, either at planting, or as a side-dressing 5 wk later. In 1985, delaying infector removal by 8 days in early July significantly increased the spread of PLRV to neighbouring plants from 2.3% (1 July) to 8.3% (9 July). A single application of deltamethrin plus heptenophos to infectors 1 wk before removal did not significantly decrease spread. Although incineration was quick and effective, the value of this method of eradicating infector plants in seed potato crops is limited because it failed to destroy infected tubers.     

SOME PROPERTIES OF FOUR STRAINS OF CUCUMBER MOSAIC VIRUS

Different strains of cucumber mosaic virus differ in their host range, symptoms caused, virulence towards different plants, transmissibility by aphids, dilution end-point and thermal inactivation point.
There are seasonal variations in the susceptibility of some host species; French bean is apparently immune during the summer but during the winter produces countable local lesions suitable for quantitative assays.
Different host species differ in the ease with which cucumber mosaic virus is transmitted to and from them; systemic infection in beet rarely occurs unless the virus is introduced into young tissues. Inhibitors of infectivity in sap of sugar beet and Phytolacca sp. make mechanical transmission from these to other hosts difficult; the inhibitors interfere less with the infection of hosts in which they occur than with the infection of tobacco.
Cucumber mosaic virus has a low temperature coefficient of thermal inactivation and much infectivity is destroyed by heating at temperatures below the thermal inactivation point.
Myzus persicae (Sulz.) is a more efficient vector than M. ornatus Laing which is more efficient than Macrosiphum euphorbiae (Thomas); although individual aphids can cause more than one infection, most cease to be infective in feeding periods of from one to five minutes.