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.     

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.     

SOME FACTORS AFFECTING THE TRANSMISSION OF LEAF-ROLL VIRUS BY APHIDS

Datura tatula is a more suitable host than potato for studying the factors influencing the transmission of potato leaf-roll virus by Myzus persicae ; it is more easily infected, provides a better source of virus for feeding aphids, produces symptoms more quickly and over a longer period of the year.
Loughnane's (1943) claim that leaf-roll virus is transmitted by starved aphids that feed for only 5 min. on infected potato plants was not confirmed. The shortest infection-feeding time in which M. persicae aphids became infective was 2 hr.; such aphids did not infect healthy plants in the first 2 days and, when transferred to a series of healthy plants at intervals, infected only few. The ability to cause infections was increased by increasing the length of infection feeding. Aphids fed for many days on infected plants could infect healthy plants in the first 15 min. of test feeding, and they continued to cause infections for long periods.
Aphids became infective more readily when feeding on recently infected Datura tatula , showing only slight symptoms, than on older plants with pronounced chlorosis; similarly, young potato sprouts showing no symptoms were better sources of virus for aphids than older plants showing severe leaf roll.
The differences in severity of symptoms shown by potato plants with leaf roll in the field mainly occur because of differences in virulence of accompanying strains of potato virus X , but isolates of leaf-roll virus were found that also varied in virulence.     

THE DISTRIBUTION OF VIRUSES IN DIFFERENT LEAF TISSUES AND ITS INFLUENCE ON VIRUS TRANSMISSION BY APHIDS

Exposing both surfaces of leaves systemically infected with cabbage black ring spot virus (CBRSV) or henbane mosaic virus to ultra-violet radiation decreases the infectivity of expressed sap to about one-fifth. As irradiation probably inactivates virus mainly in the epidermis, which occupies about one-quarter the volume of the leaves, these viruses seem to occur at much higher concentrations in sap from the epidermis than in sap from other cells. By contrast, tobacco mosaic virus seems not to occur predominantly in the epidermis.
CBRSV and henbane mosaic virus are normally transmitted most frequently by previously fasted aphids that feed for only short periods on infected leaves, but aphids treated like this transmit rarely from leaves that have been exposed to ultraviolet radiation. Irradiation has relatively little effect on the proportion of aphids that transmit after long infection feedings. Fasting seems to increase transmission by increasing the probability that aphids will imbibe sap from the epidermis of leaves they newly colonize. With longer periods on infected leaves, the ability of fasted aphids to transmit probably decreases because they then feed from deeper cells and their stylets contain sap with less virus. Only virus contained in the stylets seems to be transmitted, not virus taken into the stomach. About half the transmissions of henbane mosaic virus by aphids that have colonized tobacco leaves for hours may be caused by insects that temporarily cease feeding on the phloem and newly penetrate the epidermis.
Irradiating infected leaves affected the transmission of sugar-beet mosaic virus in the same way as that of henbane mosaic virus, but had little effect on the transmission of beet yellows virus, whose vectors become more likely to transmit the longer they feed on infected plants.     

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.     

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.     

STUDIES ON DANDELION YELLOW MOSAIC AND OTHER VIRUS DISEASES OF LETTUCE

The symptoms caused by dandelion yellow mosaic virus on cultivated lettuce, Lactuca serriola and L. virosa , are described and compared with those caused by lettuce mosaic virus. Lettuce is much more susceptible than dandelion to the yellow mosaic virus; no infections of dandelion were obtained by mechanical inoculation and only three by aphides, whereas infection of lettuce is regularly obtained by aphides and by inoculation provided an abrasive is used. Myzus ornatus, M. ascalonicus and Aulacorthum solani transmitted dandelion yellow mosaic virus but not lettuce mosaic virus, whereas Myzus persicae transmitted the latter but not the former. Nasonovia ribicola , the common lettuce aphis, transmitted neither. Aphides became infective only after feeding periods of some hours on the diseased plants and ceased to be infective within an hour of the infective feeding. Their efficiency as vectors was not increased by a preliminary starving period, as happens with Myzus persicae and lettuce mosaic virus. Lettuce mosaic virus was found in most samples of commercial seed, which explains its prevalence; no evidence was found for the seed-transmission of dandelion mosaic virus and it is doubtful if it occurs, for infected lettuce are so severely affected that they rarely set seed.
Cucumber mosaic virus was isolated from naturally infected lettuce.     

SOME PROPERTIES OF THREE VIRUSES ISOLATED FROM A DISEASED PLANT OF SOLANUM JASMINOIDES PAXT. FROM INDIA

Three mechanically transmissible viruses were isolated from a diseased Solanum jasminoides plant obtained from India. One is a strain of potato virus Y , which in some potato varieties produces symptoms resembling those caused by potato virus C , but unlike potato virus C it is readily transmitted by Myzus persicae. The second, named tobacco wilt virus, is also transmitted by M. persicae but much less readily, whereas the third, named datura necrosis virus, is not. All three have a wide host range, but neither tobacco wilt nor datura necrosis viruses infects potato plants. All three have long flexuous particles and similar general properties.
Simultaneous infection with datura necrosis virus usually decreases the concentration reached by potato virus Y in tobacco plants but not in Nicotiana glutinosa.     

THE EFFECT OF DIFFERENT HOST PLANTS OF POTATO VIRUS C IN DETERMINING ITS TRANSMISSION BY APHIDS

A stock of potato virus C derived from Edgecote Purple potatoes in 1945 was not then transmitted by aphids, although more than 2000 aphids were used in conditions optimal for transmitting the serologically related potato virus Y. This stock of virus C has been propagated continuously since, by manual inoculation in a series of Nicotiana glutinosa and N. tabacum , and in 1955 it was transmitted by the aphid Myzus persicae (Sulz.): about one in twenty of the aphids transmitted it compared with one in two for potato virus Y.
Virus C derived from the Edgecote Purple potatoes in 1955 was not transmitted by aphids; both stocks of virus C produced only local lesions in Majestic potato leaves, and gave similar symptoms in tobacco.
When inoculated to Majestic potatoes and then returned to tobacco plants, potato virus C usually ceased to be aphid transmitted and did not recover this property in any of the subsequent subcultures.
Transmission from stock by aphids did not isolate a strain of virus C which was any more readily transmitted by aphids, indeed, for the first two or three subcultures, aphids usually transmitted more readily from plants inoculated manually. But the few isolates which remained aphid transmissible, after a second passage through potato, were rather readily transmitted.
These results suggest that the ability of a virus to be aphid transmitted is, at least in part, determined by the host plant in which it is multiplying, but the nature of the changes which determine this ability are unknown.     

RESOLUTION OF STRAWBERRY VIRUS COMPLEXES IV. THE LATENT PERIOD OF VIRUS 3 IN THE VECTOR

From the time of first feeding on plants infected with strawberry virus 3, 10–19 days elapsed before Capitophorus fragariae became infective, a longer 'latent period' than any previously recorded for an aphid-transmitted virus. The time taken for aphids to develop infectivity after leaving infected plants decreased with increasing duration of the infection feed. Aphids which had fed for 16 days on an infected plant caused infection in the first day of test feeding.     

SOME EFFECTS OF HOST NUTRITION ON THE SUSCEPTIBILITY OF PLANTS TO INFECTION BY CERTAIN VIRUSES

Fertilizer treatments that greatly influenced the growth of tobacco and potato plants in pots had little effect on the number that became infected with potato virus Y when the plants were colonized by equal numbers of infective aphids, though the number was slightly decreased by nitrogen and increased by phosphorus.
The number of local lesions produced on leaves of tobacco and Nicotiana glutinosa by tomato aucuba mosaic and tobacco mosaic viruses was increased by additions of both nitrogen and phosphorus, provided that these also increased growth. The predominant effect of both nutrients in increasing susceptibility was indirect by increasing plant size, but over certain critical ranges both elements also increased the numbers of lesions produced per unit leaf area. Conditions of maximum susceptibility approximated closely to those producing optimal growth, and susceptibility, whether measured by lesions per half-leaf or per unit area, was decreased by a deficiency or excess of either element. Sometimes the addition of nitrogen reduced susceptibility when still increasing plant growth.     

The role of the helper virus, anthriscus yellows, in the transmission of parsnip yellow fleck virus by the aphid Cavariella aegopodii

Transmission of parsnip yellow fleck virus (PYFV) by the aphid Cavariella aegopodii occurs only when the aphids are also carrying the helper virus, anthriscus yellows (AYV). None of five other viruses tested was able to act as helper. In experiments in which aphids were allowed to feed through membranes on crude or treated extracts from infected plants, aphids already carrying AYV acquired PYFV, but virus-free aphids failed to acquire either AYV or PYFV. PYFV was not transmitted by insects injected with haemolymph from aphids carrying both viruses, or with purified preparations of PYFV. PYFV was transmitted when AYV-carrying aphids, except those whose stylets had been removed, were contaminated externally with PYFV preparations. Ultraviolet irradiation of infected leaves did not prevent aphids from acquiring AYV, presumably because it is confined to deeply-lying tissues. AYV-carrying aphids could acquire PYFV from u.v.-irradiated leaves after acquisition access times of 2 h but not after feeds of only 2 or 15 min (which are adequate on unirradiated leaves), suggesting that PYFV is present in all parts of the leaf. No ‘helper agent’ distinct from AYV itself was detected in these experiments or in experiments on minimum acquisition feeding time or maximum period of persistence in the aphid. U.v.-inactivated PYFV competed with infective PYFV for retention sites in AYV-carrying aphids, whereas AYV apparently did not. It is suggested that there is no helper agent for PYFV, other than AYV particles. The possibility that there is one for AYV is not excluded.     

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.     

CHANGES IN THE TRANSMISSIBILITY BY APHIDS OF A STRAIN OF CUCUMBER MOSAIC VIRUS

A strain of cucumber mosaic virus isolated from a spinach plant in 1946 was readily transmitted by Myzus persicae until 1955 when it lost this property, although it was still being propagated in conditions in which other strains remained transmissible. M. circumflexus also transmitted other strains but not this one. It was transmitted as readily as other strains by Aphis gossypii and Myzus ascalonicus. M. ascalonicus transmitted less frequently than Aphis gossypii. Transmission of the spinach strain by other aphids did not make it transmissible by Myzus persicae ; nor did propagation in different plant species or several passages through spinach. In 1955 the spinach strain was occasionally transmitted by M. persicae , but the cultures isolated in this way were no more readily transmissible by the aphid than was the bulk culture maintained by manual inoculation of sap, and after a few weeks all cultures ceased to be transmitted by M. persicae.     

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.     

The influence of size and duration of aphid infestation on host plant quality, and its effect on sugar beet yellowing virus epidemiology

This paper studies the influence of previous infestation on the host quality of sugar beet (Beta vulgaris L.) for aphids and the influence of previous infestation on sugar beet yellowing virus epidemiology. Sugar beet previously infested with Myzus persicae (Sulzer) or Aphis fabae Scopoli (Homoptera: Aphididae) had an improved host quality for subsequently infesting aphids of the same species. There was a significant negative relationship between the number of M. persicae infesting a plant and the proportion of those that died with a dark deposit in their stomachs, and a significant positive relationship between the number that settled on a plant and the number that infested it previously. Nymphs feeding on previously infested plants grew more rapidly than those on control plants. The beneficial effect of previous infestation persisted for at least 2 weeks and prolongation of the infestation beyond 2 weeks was of no further benefit to the aphids. Field grown sugar beet, previously colonised by M. persicae, was more susceptible to natural infestation by M. persicae up to 5 days after exposure. Previously infested plants were also more susceptible to infection with beet mild yellowing virus (BMYV) but not beet yellows virus (BYV), suggesting that the aphids on the previously infested sugar beet settled more readily and were more inclined to feed (and thus transmit BMYV) than aphids on the previously uninfested plants. The consequences for the control of sugar beet yellowing virus vectors are discussed.     

THE DETECTION OF CUCUMBER MOSAIC VIRUS IN SINGLE APHIDS

Abstract  Cucumber mosaic virus (CMV) was detected in single aphids Myzus persicae and Aphis gossy pii by dot immunobinding assay (DIBA). The DIB A procedure with the lowest detectable concentration endpoint of 0.16ng/ml of purified CMV, or 0. 32pg per sample dot of 2μI antigens, satisfied the sensitivity required for the detection of CMV in single virus-carrying aphids. The aphid extracting method of dipping the stylets dissected from aphid heads in the droplets of small volume (2–5p1) of suitable buffer not only ensured the highly concentrated Firuses fully released from the aphid stylets, but decreased the interference of non-virus materials from aphids to minimum as well, and thus overcame the two major difficulties in the detection of plant viruses in vectors. The virus-carrying rates tested by DIBA had good coincidence with the transmission rates by bioassay. Undoubtedly, such breakthroughs in the technique of detecting nonpersistent viruses in aphid vectors are of great epidemiological importance.     

THE SPREAD OF BEET YELLOWS AND BEET MOSAIC VIRUSES IN THE SUGAR-BEET ROOT CROP I. FIELD OBSERVATIONS ON THE VIRUS DISEASES OF SUGAR BEET AND THEIR VECTORS MYZUS PERSICAE SULZ. AND APHIS FABAE KOCH

A survey of aphids and virus diseases of sugar-beet root crops in eastern England was made between 1940 and 1948. Prior to 1943 the observations were made on fertilizer experiments; from 1943 onwards they were made on commercial fields selected for position in relation to beet and mangold seed crops. The incidence of beet yellows increased with increasing numbers of Myzus persicae , but not of Aphis fabae. The relation with M. persicae was sufficiently close to suggest that it is the most important, possibly the only important, vector of beet yellows virus. Beet mosaic virus also increased with increasing numbers of M. persicae , but the relation was not close enough to exclude the possibility of other vectors.
Numbers of A. fabae on sugar beet were slightly, but consistently, depressed by the use of salt as a fertilizer. Other fertilizers had variable effects. Neither aphids nor virus are likely to be greatly affected by fertilizers.
Beet yellows is most prevalent in areas where seed crops are grown, but within these areas nearness to individual seed crops did not appear to increase its incidence. M. persicae were more numerous on sugar beet in seed-crop areas than elsewhere, and this alone might account for the prevalence of yellows. Beet mosaic virus is more closely associated with seed crops than is beet yellows. It is most prevalent near to seed crops within the seed-crop areas.     

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.