Peanut mottle virus in East Africa

A very common and widespread virus pathogen of groundnut and soybean in East Africa was identified as peanut mottle virus (PnMV)* on the basis of particle morphology, serology, host range and reaction, transmission and physical properties. Virus concentration adequate for serological tests was obtained from cowpea (Vigna unguiculata) cultured at 27 °C but not at 23 °C. Purified preparations from this source gave a single, specific light-scattering zone in sucrose density gradients. PnMV was purified using 0.5 M sodium citrate buffer containing 1% mercapto-ethanol; an antiserum made against such preparations had a homologous titre of 1/8192. Groundnut and soya isolates from N., N.E., N.W. and S. districts of Uganda, N.W. Tanzania, and W. and E. (coastal) districts of Kenya were serologically similar and varied, within narrow limits, in symptoms induced in certain groundnut and soya varieties. A serologically related but distinct virus was isolated from Voandzeia subterranea. PnMV was not related serologically to any of ten viruses of the PVY group. Glasshouse experiments simulating groundkeeper conditions in the field indicated 20% seed transmission in groundnut; PnMV was transmitted by Aphis craccivora in the non-persistent manner. All twenty-one varieties and breeding lines of soybean tested were highly susceptible. The prevalence of PnMV in East Africa and the reduction in yield caused in groundnut indicates the virus to be economically important, and groundnut and soybean improvement programmes should include routine PnMV susceptibility tests.     

Peanut stripe virus - a new seed-borne potyvirus from China infecting groundnut (Arachis hypogaea)1

A new virus, peanut stripe (PStV), isolated from groundnut (Arachis hypogaea) in the USA, induced characteristic striping, discontinuous vein banding along the lateral veins, and oakleaf mosaic in groundnut. The virus was also isolated from germplasm lines introduced from the People's Republic of China. PStV was transmitted by inoculation of sap to nine species of the Chenopodiaceae, Leguminosae, and Solanaceae; Chenopodium amaranticolor was a good local lesion host. PStV was also transmitted by Aphis craccivora in a non-persistent manner and through seed of groundnut up to 37%. The virus remained infective in buffered plant extracts after diluting to 10^-3, storage for 3 days at 20°C, and heating for 10 min at 60°C but not 65°C. Purified virus preparations contained flexuous filamentous particles c. 752 nm long, which contained a major polypeptide of 33 500 daltons and one nucleic acid species of 3·1 × 10⁶ daltons. In ELISA, PStV was serologically related to blackeye cowpea mosaic, soybean mosaic, clover yellow vein, and pepper veinal mottle viruses but not to peanut mottle, potato Y, tobacco etch, and peanut green mosaic viruses. On the basis of these properties PStV is identified as a new potyvirus in groundnut.     

Groundnut eyespot virus, a new member of the potyvirus group

A virus causing ‘eyespot’ leaf symptoms in groundnut plants was transmitted by sap-inoculation and by Aphis craccivora in the non-persistent manner. It infected 16 of 72 species from five of 12 families and was easily propagated in Arachis hypogaea and Physalis floridana. The virus has particles c. 13 × 755 nm and is serologically closely related to soybean mosaic and pepper veinal mottle viruses, and more distantly to four other potyviruses. The virus differs in host range, in vitro properties and serological properties from previously described strains of soybean mosaic and pepper veinal mottle viruses. It seems to be a distinct member of the potyvirus group and we propose the name groundnut eyespot virus.     

Detection of a Luteovirus in Groundnut Rosette Diseased Groundnuts (Arachis hypogaea) by Enzyme-linked Immunosorbent Assay and Immunoelectron Microscopy

In groundnut rosette diseased groundnut plants collected near Zaria, Nigeria, a luteovirus was detected by ELISA and ISEM. In ELISA only beet western yellows virus antiserum reacted, while in ISEM luteovirus particles were trapped by antisera beet western yellows virus, potato leafroll virus, pea leafroll virus and barley yellow dwarf virus. The data are in agreement with the interpretation that the assistor of groundnut rosette virus is possibly a member of the luteovirus group.     

A Simple Procedure to Detect a dsRNA Associated with Groundnut Rosette

A low-molecular weight double-stranded (ds) RNA [900 base pairs (bp)] associated with groundnut rosette disease can be used as a diagnostic tool. A simple procedure has been developed that is rapid, reliable, and requires only standard electrophoresis equipment and ultraviolet light for detection of nucleic acid bands. Using this procedure, the dsRNA was detected only in groundnut plants with green rosette of chlorotic rosette symptoms. It was not found in uninoculated groundnut plants, in symptomless groundnut plants with groundnut rosette assistor virus, or in groundnut plants infected with several other known groundnut viruses. In studies with northern blots of extracts from rosette-diseased and healthy plants, 5′-endlabeled dsRNA only hybridized to a 900 bp dsRNA from diseased plants. The 900 bp dsRNA was not infectious and its origin remains obscure.     

The nature of the resistance in groundnut to rosette disease

Groundnut rosette disease is caused by a complex of three agents, groundnut rosette virus (GRV) and its satellite RNA, and groundnut rosette assistor virus (GRAV); the satellite RNA is mainly responsible for the disease symptoms. Groundnut genotypes possessing resistance to rosette disease were shown to be highly resistant (though not immune) to GRV and therefore to its satellite RNA, but were fully susceptible to GRAV.     

Dissemination and detection of peanut clump virus in groundnut seed

Experiments were done to assess the role of seed-transmission in the dissemination of peanut clump virus (PCV) in groundnut (Arachis hypogea L.), and the usefulness of enzyme-linked immunosorbent assay (ELISA) for detecting the virus in infected groundnut seed. The virus was present in 7.5% of seedling progeny from infected plants and could be detected in 16.5% of the seeds by ELISA. When groundnut seeds were grown in a field contaminated by the virus, it was shown that by roguing the infected plants, only 0.1% of the seeds from the remaining plants contained the virus. It was also established that the level of contamination of seeds by the virus was inversely proportional to the seed size.     

Chrysanthemum Stem Necrosis Caused by a Possible Novel Tospovirus

Necrotic lesions surrounded by yellow areas on leaves followed by stem necrosis were observed on chrysanthemum plants cultivated in Atibaia, Sào Paulo State, Brazil. The host range, in vitro properties and particle morphology of the causal virus were typical of a Tospovirus. Serological studies demonstrated that the virus differed from tomato spotted wilt. tomato chlorotic spot, groundnut ringspot and inpatiens necrotic spot Tospoviruses. The virus isolate is thus possibly a representativeof a new serogroup of a new virus.     

A Serious Disease of Groundnut Caused by Cowpea Mild Mottle Virus in the Sudan

A serious disease of groundnut (Arachis hypogaea L.) characterized by stunting of plants, downward rolling, mottling, general chlorosis and reduced size of the leaflets was observed in the Sudan. Surveys conducted from 1992 to 1994 showed that this disease was restricted to irrigated groundnut crops grown between the two Niles. The virus had slightly flexuous filamentous particles (626 nm long) and was transmitted by whiteflies. It was identified serologically as cowpea mild mottle virus (CPMMV). This appears to be the first record of natural occurrence of CPMMV on groundnut in the Sudan and the first evidence that it causes a disease of major economic importance.     

Epidemiology of groundnut rosette virus disease: current status and future research needs@

Rosette is the most destructive virus disease of groundnut in sub-Saharan Africa. It is caused by a complex of three agents, namely groundnut rosette assistor virus, groundnut rosette virus and its satellite RNA. The disease appears to be indigenous to Africa as it has not been recorded elsewhere. Thus rosette represents a new-encounter situation as the disease is thought to have spread to the introduced groundnut from indigenous host plants. Rosette has been known since 1907 and much information has been obtained on the main features of the disease, viz. its biology, transmission, viral aetiology and diagnosis, and the impact of chemical control of the aphid vector, cultural practices and virus-resistant varieties on disease management. However, there are still many gaps in the available knowledge, especially the reasons for the large and unpredictable fluctuations in the incidence and severity of rosette disease throughout sub-Saharan Africa. Three unresolved issues of particular importance concern the nature of the primary source(s) of inoculum, the means of survival of virus and vector during unfavourable periods, and the distances over which the aphid vector can disperse and disseminate virus. Now that the aetiology of the disease is understood and diagnostic tools have been developed, the time is opportune for new initiatives in understanding the ecology and epidemiology of rosette. Substantial progress can be made by developing a co-ordinated multi-disciplinary research programme and making full use of the latest techniques, approaches and experience gained elsewhere with other insect-borne viruses. This information would help to explain the sporadic disease epidemics that cause serious crop losses and sometimes total crop failure, and would also facilitate the development of disease forecasting methods and sustainable integrated disease management strategies.     

Effect of Fungicides on the Viability of Phylophthora cactorum Propagules m the Soil

Vein-clearing followed by downward rolling and necrosis of leaves and severe stunting of groundnut (Arachis hypogaea) plants were caused by cowpea mild mottle virus (CMMV). The virus was readily transmitted by mechanical sap inoculations to groundnut and to 10 plant species belonging to Leguminosae, Chenopodiaceae and Solanaceae. Chenopodium quinoa and Beta vulgaris were good diagnostic hosts. Diseased sap remained infective at 10^–3 but not 10^–4, when stored 8 to 9 days at 25 °C; for 10min at 75 °C but not 80°C. In limited tests, virus was not seed-transmitted m groundnut or soybean. Virus was transmitted by Bemisia tabaci but not by Aphis craccivora or Myzus persicae. An antiserum for CMMV was produced and virus was serologically related to CMMV reported on cowpea and groundnut crinkle virus (GCV) from West Africa. Employing carbon diffraction grating replica as a standard the modal length of virus particles to be 610 nm. Infected cells contained large number of virus particles associated with endoplasmic reticulum.     

Peanut green mosaic virus — a member of the potato virus Y group infecting groundnut (Arachis hypogaea) in India

A virus, now named peanut green mosaic virus (PGMV), was isolated from groundnut (Arachis hypogaea) in India and identified as a member of the potato virus Y group by electron microscopy, aphid transmission, and its chemical properties. It was sap transmissible to 16 species of the Leguminosae, Solanaceae, Chenopodiaceae, Aizoaceae and Pedaliaceae; Phaseolus vulgaris was a good local lesion host. PGMV remained infective in buffered groundnut leaf sap at dilutions of 10^-3 after 3 to 4 days at 25 °C, or heating for 10 min to 55 °C but not 60 °C. PGMV was transmitted in the non-persistent manner by Aphis gossypii and Myzus persicae but was not seed-borne. Purified virus preparations contained flexuous filamentous particles c. 750 nm long which sedimented as a single component with a sedimentation coefficient (S°_20w) of 171S, and contained a single polypeptide (mol. wt 34 500 daltons) and one nucleic acid species (mol. wt 3.25 × 10⁶ daltons). PGMV is serologically unrelated to peanut mottle virus (PMV) and other viruses infecting leguminous crops. Infected leaves contained cylindrical, cytoplasmic inclusions.     

Viruses associated with chlorotic rosette and green rosette diseases of groundnut in Nigeria*

Groundnut (Arachis hypogaea) plants from Nigeria with chlorotic rosette disease contained a manually transmissible virus, considered to be a strain of groundnut rosette virus (GRV(C)). GRV(C) infected nine out of 32 species in three out of nine families. It caused local lesions without systemic infection in Chenopodium amaranticolor, C. murale and C. quinoa, and systemic symptoms in Glycine max, Nicotiana benthamiana, N. clevelandii and Phaseolus vulgaris as well as in groundnut. Some ‘rosette-resistant’ groundnut lines were also infected. GRV(C) was transmitted by Aphis craccivora, but only from groundnut plants that were also infected with an aphid-transmissible second virus, which was not manually transmissible and was considered to be groundnut rosette assistor virus (GRAV). Plants infected with GRAV contained isometric particles c. 25 nm in diameter which were detectable by immunosorbent electron microscopy on grids coated with antisera to several luteoviruses, especially with antisera to bean leaf roll, potato leafroll and beet western yellows viruses. No virus-like particles were observed in extracts from plants infected with GRV(C) alone. A single groundnut plant obtained from Nigeria with symptoms of green rosette contained luteovirus particles, presumed to be of GRAV, and yielded a manually transmissible virus that induced symptoms similar to those of GRV(C) in C. amaranticolor but gave only mild or symptomless infection of N. benthamiana and N. clevelandii. It was considered to be a strain of GRV and designated GRV(G).     

Identification of Peanut Green Mosaic Virus Strains in India*

During field surveys, three peanut green mosaic virus isolates differing in symptomatology on groundnut and a few other hosts were collected. Ultrathin sections of infected groundnut leaflets showed cytoplasmic inclusions with pin wheels and scrolls. In enzyme-linked immunosorbent assay they reacted strongly with antisera to peanut green mosaic and soybean mosaic virus antisera, and moderately with adzuki bean mosaic and peanut stripe virus antisera. All isolates also reacted positively with antisera to peanut eye spot, blackeye cowpea mosaic, pea seed-borne mosaic, potato virus Y and tobacco etch viruses, and did not react with antisera to peanut mottle, bean yellow mosaic, bean common mosaic, clover yellow vein and sugarcane mosaic viruses. SDS-PAGE analysis of purified virus preparations of the three isolates showed a single polypeptide with mol. wt. of 34,500 daltons. Based on these results, the three isolates are identified as biologically distinct strains of peanut green mosaic virus.     

Studies on the transmission of groundnut rosette virus by Aphis craccivora Koch

Four strains of groundnut rosette virus were transmitted by a race of Aphis craccivora (Koch) from groundnut in Nigeria. Two of these strains, both from East Africa, were transmitted only by A. craccivora from Kenya. A fifth isolate, from Nigeria, was not transmissible by either race. The two races of aphids have been shown elsewhere to be distinct biotypes. Most A. craccivora needed longer than 24 h feeding on infected groundnuts to acquire virus, and many needed 2–3 days of feeding on healthy plants to cause infection, even after several days on infected plants. The delays partly reflect the slow uptake of virus and possibly a period needed for virus multiplication in aphid tissue but some is lost through resistance of the test plants to infection. In consecutive feeding experiments Natal Common variety could be infected soon after aphids had left the source of virus, but a more resistant Nigerian variety sometimes needed several more days. The frequency of inoculation by aphids, or the concentration of virus in the inocula or both, increased with time, but the times at which aphids were able to infect plants was also dependent on variety.     

Different variants of the satellite RNA of groundnut rosette virus are responsible for the chlorotic and green forms of groundnut rosette disease*

Groundnut plants with symptoms of rosette disease contain groundnut rosette virus (GRV), but GRV is transmitted by Aphis craccivora only from plants that also contain groundnut rosette assistor virus (GRAV). Two main forms of rosette disease are recognised, ‘chlorotic rosette’ and ‘green rosette’. GRV cultures invariably possess a satellite RNA and this is the major cause of rosette symptoms: satellite-free isolates derived from GRV cultures from Nigerian plants with chlorotic or green rosette, or from Malawian plants with chlorotic rosette, induced no symptoms, or only transient mild mottle or interveinal yellowing, in groundnut. When the satellite RNA species from GRV cultures from Nigerian green or Malawian chlorotic rosette were reintroduced into the three satellite-free isolates in homologous and heterologous combinations, the ability to induce rosette symptoms was restored and the type of rosette induced was that of the cultures from which the satellite RNA was derived. Thus different forms of the satellite are responsible for the different forms of rosette disease. Other forms of the satellite induce only mild chlorosis or mottle symptoms in groundnut. Individual plants may contain more than one form of the satellite, and variations in their relative predominance are suggested to account for the variable symptoms (ranging from overall yellowing to mosaic) seen in some plants graft-inoculated with chlorotic rosette.     

Comparison of the coat protein of groundnut rosette assistor virus with those of other luteoviruses

The coat protein gene of groundnut rosette assistor virus (GRAV) was cloned and sequenced. The deduced amino acid sequences of the coat protein and of another protein encoded in a different, overlapping, reading frame resemble those of other luteoviruses. Four monoclonal antibodies against GRAV, prepared using denatured coat protein as immunogen, also reacted with some other luteoviruses in ELISA. Nevertheless, they will be useful as reagents for the identification of GRAV infections in groundnut.     

Additional sources of resistance to groundnut rosette disease in groundnut germplasm and breeding lines

Groundnut rosette, a virus disease of groundnut (Arachis hypogaea) transmitted by the aphid, Aphis craccivora Koch, reduces yield in susceptible cultivars by 30–100%. Additional sources were sought in germplasm accessions involving 2301 lines from different sources and from 252 advanced breeding lines derived from crosses involving earlier identified sources of resistance to rosette. The lines were evaluated in field screening trials using an infector row technique during 1996 and 1997 growing seasons. Among the germplasm lines, 65 accessions showed high levels of resistance while 134 breeding lines were resistant. All rosette disease resistant lines were susceptible to groundnut rosette assistor virus. This work identified germplasm and breeding lines that will contribute to an integrated management of groundnut rosette disease. These new sources also provide an opportunity to eliminate yield losses due to the rosette disease.     

TRANSMISSION OF GROUNDNUT ROSETTE VIRUS

The rosette virus was transmitted to groundnut plants, if previously etiolated, bymechanical inoculation of juice with 'Celite' addition; but only a small proportion of the inoculations succeeded.
Aphis craccivora (Koch), the known vector, transmitted the virus by feeding on germinating groundnut seeds; and from this fact we developed an experimental technique that is convenient and flexible.
Different races of this species appeared to vary in inherent efficiency in transmission, and one failed ever to transmit. Within races that could transmit, all larval forms and alate and apterous adults might transmit; but alatae were sometimes significantly more efficient than apterae, and at other times the converse held. A field experiment showed that wingless forms, moving over the soil surface, might play a predominant part in secondary spread around a rosetted plant.
Comparative tests with groups of I, 2, 3 and 4 infective aphids supported the hypothesis that infections by this vector are individual and independent.
Single aphids, tested for 24 hr. on 10 successive days without access to an outside source of virus, might infect on any day up to the tenth. Similar results were obtained in a succession of I hr. tests on a single day. In the infected seed the virus rapidly became available to feeding aphids; previously non-infective aphids acquired the virus by feeding on a seed during the third day from the first exposure of this seed to infective aphids.     

Involvement of Volatile Substances in Systemic Resistance of Barley Against Erysiphe graminis f. sp. hordei Induced by Pruning of Leaves

Horsegram yellow mosaic disease was shown to be caused by a geminivirus; horsegram yellow mosaic virus (HYMV). The virus could not be transmitted by mechanical sap inoculation. Leaf dip and purified virus preparations showed geminate virus particles, measuring 15-18 * 30 nm. An antiserum for HYMV was produced and in enzyme-linked immunosorbent assay (ELISA) and immunosorbent electron microscopy (ISEM) tests HYMV was detected in leaf extracts of fieldinfected bambara groundnut, french bean, groundnut, limabean, mungbean, pigeonpea and soybean showing yellow mosaic symptoms. Bemisia tabaci fed on purified HYMV through a parafilm membrane transmitted the virus to all the hosts listed above but not to Ageratum conyzoides, okra, cassava, cowpea, Croton bonplandianus, Lab-lab purpureus, Malvastrum coromandalianum and tomato. No reaction was obtained in ELISA and ISEM tests between HYMV antibodies and extracts of plants diseased by whitefly-transmitted agents in India such as A. conyzoides yellow mosaic, okra yellow vein mosaic, C. bonplandianus, yellow vein mosaic, M. coromandalianum yellow vein mosaic, tomato leaf curl and cassava mosaic. HYMV was also not found to be related serologically to bean golden mosaic, virus.