Sweet potato virus disease (SPVD) is the name used to describe a range of severe symptoms in different cultivars of sweet potato, comprising overall plant stunting combined with leaf narrowing and distortion, and chlorosis, mosaic or vein-clearing. Affected plants of various cultivars were collected from several regions of Uganda. All samples contained the aphid-borne sweet potato feathery mottle potyvirus (SPFMV) and almost all contained the whiteflyborne sweet potato chlorotic stunt closterovirus (SPCSV). SPCSV was detected by a mix of monoclonal antibodies (MAb) previously shown to react only to a Kenyan isolate of SPCSV, but not by a mixture of MAb that detected SPCSV isolates from Nigeria and other countries. Sweet potato chlorotic fleck virus (SPCFV) and sweet potato mild mottle ipomovirus (SPMMV) were seldom detected in SPVD-affected plants, while sweet potato latent virus (SPLV) was never detected. Isolates of SPFMV and SPCSV obtained by insect transmissions together induced typical symptoms of SPVD when graft-inoculated to virus-free sweet potato. SPCSV alone caused stunting and either purpling or yellowing of middle and lower leaves when graft-inoculated to virus-free plants of two cultivars. Similarly diseased naturally inoculated field plants were shown consistently to contain SPCSV. Both this disease and SPVD spread rapidly in a sweet potato crop.
The movement function of poa semilatent hordeivirus (PSLV) is mediated by the triple gene block (TGB) proteins, of which two, TGBp2 and TGBp3, are membrane proteins. TGBp3 is localized to peripheral bodies in the vicinity of the plasma membrane and is able to re-direct TGBp2 from the endoplasmic reticulum (ER) to the peripheral bodies.
Four hundred and forty-eight symptomatic and 638 asymptomatic samples were collected from sweet potato fields throughout Kenya and analysed serologically using antibodies to Sweet potato feathery mottle virus (SPFMV), Sweet potato chlorotic stunt virus (SPCSV), Sweet potato mild mottle virus (SPMMV), Cucumber mosaic virus (CMV), Sweet potato chlorotic fleck virus (SPCFV), Sweet potato latent virus (SwPLV), Sweet potato caulimo-like virus (SPCaLV), Sweet potato mild speckling virus (SPMSV) and C-6 virus in enzyme-linked immunosorbent assays (ELISA). Only SPFMV, SPMMV, SPCSV, and SPCFV were detected. Ninety-two percent and 25% of the symptomatic and asymptomatic plants respectively tested positive for at least one of these viruses. Virus-infected plants were collected from 89% of the fields. SPFMV was the most common and the most widespread, detected in 74% of the symptomatic plants and 86% of fields surveyed. SPCSV was also very common, being detected in 38% of the symptomatic plants and in 50% of the fields surveyed. SPMMV and SPCFV were detected in only 11% and 3% of the symptomatic plant samples respectively. Eight different combinations of these four viruses were found in individual plants. The combination SPFMV and SPCSV was the most common, observed in 22% of symptomatic plants. Virus combinations were rare in the asymptomatic plants tested. Incidence of virus infection was highest (18%) in Kisii district of Nyanza province and lowest (1%) in Kilifi and Malindi districts of Coast province.
In a survey of most sweetpotato-growing areas of Uganda, virus-like diseases were observed in all districts surveyed. Out of 338 fields sampled in 35 of the then 42 districts, 219 (65%) had some plants with symptoms. The most common symptoms included vein clearing, mottling, leaf distortion, yellowing, stunting and leaf strapping. Particularly high viruslike disease incidences (means of 34-86%) were encountered in districts around Lake Victoria and in the Rift Valley in southern and western parts of Uganda; particularly low incidences were encountered in the east and north of Uganda. Using four formats of enzyme-linked immunosorbent assay in combination with immunoelectron microscopy and polymerase chain reaction assays, five viruses were identified. Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) were most commonly detected, being found in about 90% of samples. Sweet potato mild mottle virus at 10%, Sweet potato chlorotic fleck virus (SPCFV) at 8% and Sweet potato caulimo-like virus (SPCaLV) at 0·07% were more rarely detected. Most infections were multiple, SPCSV + SPFMV constituting > 90% of all double infections. Triple infections, involving mainly SPFMV, SPCSV and either SPMMV or SPCFV, and quadruple infections of SPFMV + SPCSV + SPMMV + SPCFV were observed in < 10% of the diseased samples. The identification of SPCaLV is the first evidence of its occurrence in Africa.
An immunodominant membrane protein (IMP) of the apple proliferation (AP) phytoplasma was detected in preparations from AP-diseased periwinkle plants using monoclonal and polyclonal antibodies to the AP agent. Following isolation from Western blots and partial sequencing, degenerate oligonucleotides derived from the IMP sequence were used as probes to identify a DNA fragment containing the ORF encoding the IMP. Complete sequencing and subsequent analysis of the cloned DNA fragment revealed the presence of two ORFs, predicted to encode proteins with molecular masses of 25 kDa (P-318A) and 19 kDa (P-318B). Whilst database searches failed to assign a possible function to P-318A, analysis of P-318B predicted an amphiphilic membrane protein with a positively charged N-terminal portion, followed by a hydrophobic segment forming an a-helix, and a hydrophilic C-terminal part located outside of the cell. The amphiphilic nature of P-318B was confirmed by its solubility in T r i t o n X-114. The gene encoding P-318B was expressed in Escherichia coli and the resulting protein was used to immunize rabbits. The antiserum obtained reacted specifically with P-318B. The same protein was also detected by an antiserum raised against antigen preparations from APdiseased plants. The P-318B antiserum did not react with antigen preparations from plants infected with the closely related pear decline phytoplasma. However, in Southern hybridization studies, the gene encoding the IMP hybridized to genomic fragments of the pear decline and European stone fruit yellows phytoplasmas. It also showed significant sequence similarity to a gene encoding an antigenic membrane protein of the sweet potato witches' broom phytoplasma, but not to a gene encoding a major immunogenic membrane protein of an aster yellows group phytoplasma. Since it appears that most phytoplasmas possess a major immunogenic membrane protein which may have a function in pathogenesis, this work may be a basis for further studies on fundamental aspects of host-pathogen interactions. It also describes a new approach to obtain suitable immunogens to produce specific antibodies for detection and characterization of the non-culturable phytoplasmas.
SummaryA virus causing sunken veins on ‘Georgia Jet’ sweet potato, and yellow brittle leaves and stunting on Ipomoea setosa, was purified and a specific antiserum was prepared. Flexuous particles with a normal length of 850 nm and a diameter of 12 nm with an open helical structure typical of closteroviruses were observed. The virus particle protein has an apparent mol. wt of c. 34 kD. Double‐stranded RNA isolated from SPSVV‐infected I. setosa and subjected to electrophoresis in agarose consisted of one major band with an estimated Mr of 10.5 kbp and two minor bands with Mr of 9.0 and 5.0 kbp. Fibril‐containing vesicles in phloem cells were observed in ultrathin sections of infected leaf tissues. The virus was transmitted by the whitefly Bemisia tabaci in a semi‐persistent manner and by grafting, but not mechanically. The virus could be transmitted to various Ipomoea species, to Nicotiana clevelandii, N. benthamiana and Amaranthus palmeri. The virus did not react with an antiserum to lettuce infectious yellows virus. Based on particle morphology, serology and symptom expression, the virus appears unique and different from all other reported whitefly‐transmitted closteroviruses. We propose it be named “sweet potato sunken vein virus” (SPSVV).
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