Molecular and biological characterization of Macroptilium yellow mosaic virus from Jamaica

The molecular and biological characterization of a begomovirus infecting the common weed Macroptilium lathyroides from Jamaica are reported. The virus showed 92% sequence identity to an isolate of Macroptilium yellow mosaic virus (MaYMV) from Cuba, but was distinct from the two other begomoviruses isolated from M. lathyroides , namely Macroptilium yellow mosaic Florida virus (80% identity) and Macroptilium mosaic Puerto Rico virus (68% identity). Hence, the Jamaican begomovirus was considered an isolate of MaYMV and called Macroptilium yellow mosaic virus -[Jamaica] (MaYMV-[JM]). In infectivity studies using cloned DNA-A and DNA-B genomic components, MaYMV-[JM] infected red kidney bean ( Phaseolus vulgaris ) and produced mild symptoms in Scotch Bonnet pepper ( Capsicum chinense ), but did not infect cabbage ( Brassica oleracea ). This information has implications for the development of strategies to control begomovirus diseases in Jamaica and elsewhere.     

Bean golden yellow mosaic virus from Chiapas, Mexico: Characterization, Pseudorecombination with Other Bean-Infecting Geminiviruses and Germ Plasm Screening

ABSTRACT The complete nucleotide (nt) sequences of the cloned DNA-A (2644 nts) and DNA-B (2609 nts) components of Bean golden yellow mosaic virus (BGYMV-MX) from Chiapas, Mexico were determined. The genome organization of BGYMV-MX is similar to that of other Western Hemisphere bipartite geminiviruses (genus Begomovirus). Infectivity of the cloned BGYMV-MX DNA components in common bean (Phaseolus vulgaris) plants was demonstrated by particle bombardment and agroinoculation. BGYMV-MX was identified as a BGYMV (previously type II BGMV) isolate based on sequence analyses, sap-transmissibility, and pseudorecombination experiments with other bean-infecting begomoviruses. On the basis of differences in the DNA-B hypervariable region, symptom phenotype, and properties of infectious pseudorecombinants, BGYMV-MX may represent a distinct strain of BGYMV. Pseudorecombination experiments further established that BGYMV symptom determinants mapped to DNA-B, and that BGYMV-MX was most closely related to BGYMV from Guatemala. A Tomato leaf crumple virus (TLCrV) DNA-A/BGYMV-MX DNA-B pseudorecombinant was infectious in bean, establishing that a viable reassortant can be formed between begomovirus species from different phylogenetic clusters. Bean germ plasm representing the two major gene pools (Andean and Mesoamerican) was screened for response to BGYMV-MX with three methods of inoculation: sap-inoculation, particle bombardment, and agroinoculation. Andean germ plasm was very susceptible and similar results were obtained with all three methods, whereas Mesoamerican germ plasm showed resistance to BGYMV-MX, particularly with agroinoculation.     

Identifying resistance gene analogs associated with resistances to different pathogens in common bean

ABSTRACT A polymerase chain reaction approach using degenerate primers that targeted the conserved domains of cloned plant disease resistance genes (R genes) was used to isolate a set of 15 resistance gene analogs (RGAs) from common bean (Phaseolus vulgaris). Eight different classes of RGAs were obtained from nucleotide binding site (NBS)-based primers and seven from not previously described Toll/Interleukin-1 receptor-like (TIR)-based primers. Putative amino acid sequences of RGAs were significantly similar to R genes and contained additional conserved motifs. The NBS-type RGAs were classified in two subgroups according to the expected final residue in the kinase-2 motif. Eleven RGAs were mapped at 19 loci on eight linkage groups of the common bean genetic map constructed at Centro Internacional de Agricultura Tropical. Genetic linkage was shown for eight RGAs with partial resistance to anthracnose, angular leaf spot (ALS) and Bean golden yellow mosaic virus (BGYMV). RGA1 and RGA2 were associated with resistance loci to anthracnose and BGYMV and were part of two clusters of R genes previously described. A new major cluster was detected by RGA7 and explained up to 63.9% of resistance to ALS and has a putative contribution to anthracnose resistance. These results show the usefulness of RGAs as candidate genes to detect and eventually isolate numerous R genes in common bean.     

Isolation and characterization of a potyvirus associated with bushy dwarf symptom in chickpea, Cicer arietinum, in India

A potyvirus that induced stunting and a characteristic bushy appearance at the apical region, due to proliferation of terminal branches with narrowed, reduced and deformed leaflets, was isolated from chickpea in India. The virus was sap-transmissible to 14 species of Chenopodiaceae, Leguminosae, Solanaceae and Malvaceae; Chenopodium amaranticolor was a good local lesion host. Virus particles, trapped by immunosorbent electron microscopy and stained with uranyl acetate, were 710 ×10 nm long. Purified virus preparations contained a single polypeptide species of 32,500 Da and one nucleic acid species of 3.1 · 10⁶ Da. The virus was serologically related to soybean mosaic, azuki bean mosaic and peanut mottle viruses but not to clover yellow vein, pea seed-borne mosaic and bean yellow mosaic viruses.
On the basis of these properties, the virus was identified as a previously undescribed potyvirus in chickpea, for which the name chickpea bushy dwarf virus is proposed.     

Serological relationships between five fungally transmitted cereal viruses and other elongated viruses

F(ab')₂ and protein A ELISA tests were used to investigate the serological affinities of five fungally transmitted cereal viruses: barley yellow mosaic (BaYMV), barley mild mosaic (BaMMV), oat mosaic (OMV), wheat yellow mosaic (WYMV) and oat golden stripe (OGSV). Within this group only BaYMV and WYMV were related. Chinese and UK isolates of BaYMV appeared to be similar. In tests using antisera to 29 other elongated viruses, BaYMV was related to one isolate of bean yellow mosaic poty virus (BYMV-G) and OGSV had affinities with BYMV-G, potato virus M, red clover vein mosaic (both carlaviruses) and perhaps Hordeum mosaic virus. The results were confirmed in immunoelectron microscopic tests. No affinities were found for BaMMV, OMV or WYMV.     

Genetic and Phenotypic Variation of the Pepper golden mosaic virus Complex

ABSTRACT Three isolates of the bipartite begomovirus Pepper golden mosaic virus (PepGMV) were characterized for genomic and biological properties. The complete nucleotide sequences of the DNA-A and DNA-B components were determined from infectious clones of PepGMV-Serrano (PepGMV-Ser), PepGMV-Mosaic (PepGMV-Mo), and PepGMV-Distortion (PepGMV-D). Nucleotide sequence identity among PepGMV components ranged from 91 to 96% for DNA-A and from 84 to 99% for DNA-B, with each PepGMV component most closely related to the corresponding component of Cabbage leaf curl virus (CaLCV). However, phylogenetic relationships among begomovirus components were incongruent because DNA-A of PepGMV and CaLCV share an inferred evolutionary history distinct from that of DNA-B. The cloned components of PepGMV-Ser, -Mo, and -D were infectious by biolistic inoculation to pepper but differed in symptom expression: PepGMV-Ser exhibited a bright golden mosaic, PepGMV-Mo produced a yellow-green mosaic, and PepGMV-D caused only a mild mosaic and foliar distortion followed by a "recovery" phenotype in which leaves developing after initial symptom expression appeared normal. Differences in symptoms also were observed on tomato, tobacco, and Datura stramonium. Progeny virus derived from clones of PepGMV-Ser and -Mo were transmitted from pepper to pepper by the B biotype of Bemisia tabaci; progeny virus derived from PepGMV-D clones was not transmissible by the B biotype. Reassortant genomes derived from heterologous DNA components of the three isolates were infectious in all possible pairwise combinations, with symptom phenotype in pepper determined by the DNA-B component. Collectively, these results indicate that the three virus isolates examined may be considered distinct strains of PepGMV that have the capacity to exchange genetic material.     

Emergence of a New Cucurbit-Infecting Begomovirus Species Capable of Forming Viable Reassortants with Related Viruses in the Squash leaf curl virus Cluster

ABSTRACT Cucurbit leaf curl virus (CuLCV), a whitefly-transmitted geminivirus previously partially characterized from the southwestern United States and northern Mexico, was identified as a distinct bipartite begomovirus species. This virus has near sequence identity with the previously partially characterized Cucurbit leaf crumple virus from California. Experimental and natural host range studies indicated that CuLCV has a relatively broad host range within the family Cucurbitaceae and also infects bean and tobacco. The genome of an Arizona isolate, designated CuLCV-AZ, was cloned and completely sequenced. Cloned CuLCV-AZ DNA A and B components were infectious by biolistic inoculation to pumpkin and progeny virus was transmissible by the whitefly vector, Bemisia tabaci, thereby completing Koch's postulates. CuLCV-AZ DNA A shared highest nucleotide sequence identity with Squash leaf curl virus-R (SLCV-R), SLCV-E, and Bean calico mosaic virus (BCaMV) at 84, 83, and 80%, respectively. The CuLCV DNA B component shared highest nucleotide sequence identity with BCaMV, SLCV-R, and SLCV-E at 71, 70, and 68%, respectively. The cis-acting begomovirus replication specificity element, GGTGTCCTGGTG, in the CuLCV-AZ origin of replication is identical to that of SLCV-R, SLCV-E, and BCaMV, suggesting that reassortants among components of CuLCV-AZ and these begomoviruses may be possible. Reassortment experiments in pumpkin demonstrated that both reassortants of CuLCV-AZ and SLCV-E A and B components were viable. However, for CuLCV-AZ and SLCV-R, only one reassortant (SLCV-R DNA A/CuLCV-AZ DNA B) was viable on pumpkin, even though the cognate component pairs of both viruses infect pumpkin. These results demonstrate that reassortment among sympatric begomovirus species infecting cucurbits are possible, and that, if generated in nature, could result in begomoviruses bearing distinct biological properties.     

Variability in Geminivirus Isolates Associated with Phaseolus spp. in Brazil

ABSTRACT Bean golden mosaic geminivirus (BGMV) is the single most devastating virus of common beans in the tropical and subtropical Americas and the Caribbean Basin. The BGMV from Brazil, named BGMV-BZ, is considered distinct from BGMV-PR isolates from Puerto Rico, Guatemala, and the Dominican Republic because of DNA sequence data, the ability to form pseudorecombinants, and mechanical transmissibility properties. In bean-growing areas of Brazil, samples were collected from beans, lima beans, and the weed Leonurus sibiricus displaying typical symptoms of infection by geminiviruses. Viral DNA fragments comprising part of the rep gene, the common region, and part of the cp gene were amplified by polymerase chain reaction, cloned, and sequenced. The bean samples had geminivirus with sequences nearly identical to that of BGMV-BZ collected in Goiania, state of Goiás, in 1986. The sample from lima bean contained a new species of geminivirus that induces symptoms similar to those induced by BGMV-BZ and was named lima bean golden mosaic virus (LBGMV-BR). While all sequences from bean samples clustered with BGMV-BZ, the sequence from the lima bean isolate stood alone. A mixed infection with abutilon mosaic geminivirus was also found in a single sample from the state of S?o Paulo. DNA sequence comparisons indicate that the virus isolate from L. sibiricus represents a new geminivirus species, designated here as leonurus mosaic virus.     

Biological and Molecular Properties of a Begomovirus from Dicliptera sexangularis

ABSTRACT Sixangle foldwing, Dicliptera sexangularis (Acanthaceae), showing severe yellow mottle and leaf distortion symptoms was collected from the shoreline of Calusa Island (Lee County, FL). The putative virus was transmitted from infected D. sexangularis to healthy seedlings by mechanical, whitefly (Bemisia tabaci biotype B), and graft-inoculations. Different forms of geminivirus-like DNAs were detected in total DNA extracted from infected plants by Southern blot hybridization analyses using DNA-A and -B of Bean golden mosaic virus (BGMV) from Guatemala as probes. Preliminary polymerase chain reaction experiments and sequence comparisons indicated that the virus was a distinct bipartite begomovirus. The virus was designated Dicliptera yellow mottle virus (DiYMV). Replicative dsDNAs of DiYMV were extracted, digested with selected restriction enzymes, and cloned into a plasmid vector. Both DNA-A and -B were sequenced and compared with those of other begomoviruses. Phylogenetic analyses using AV1, AC1, and BV1 nucleotide sequences indicated that DiYMV has a close relationship with the New World begomoviruses, especially those distributed in the nearby geographic areas of the Florida coast and the Caribbean Basin. However, different percent nucleotide sequence identities and phylogenetic relationships were detected when different open reading frames (ORFs) of DiYMV were compared with their counterparts from begomoviruses from the Caribbean Basin. Based on phylogenetic analyses of the AC1 and BV1 ORFs, DiYMV was closely related to BGMV type II isolates, whereas sequence comparisons of the common region and the AC4-derived amino acid sequences indicated its close relationship with Potato yellow mosaic virus from Venezuela.     

北京菜豆(Phaseolus Vulgaris)上的一种新病毒—和性黄色花叶病毒

 从北京郊区患有类似花叶病害的菜豆株上分离到一种线条状病毒(长约700至750nm)并于1983至1986年间加以研究。在温室内接种可以侵染菜豆、蚕豆、豌豆、大豆、决明、苋色藜及昆诺藜而不能侵染被接种的任何茄科植物.同标准的菜豆普遍花叶病毒(BCMV)相比,它不侵染茄科植物例如黄花烟(Nicotiana rustica)和矮牵牛(Petunia hybrida)在菜豆叶上也不产生坏死枯斑.此外它能侵染蚕豆、大豆、豌豆及决明而BCMV则不能.BCMV能侵染豇豆而这一分离物则不能.此分离物的物理性状为:钝化温度=56-580℃(十分钟),硫释限点=10^-3至10^-4,180℃下存治期为3天,A260/280=1.12.菜豆受侵叶组织易用光学显微镜观察到内含体包括一种片层叠合体.在电镜下超薄切片中可以看到风轮状体.极易用汁液摩接及用桃蚜(Myzus persic(接种.此病毒的衣壳蛋白亚基的分子量经测定为32,000道尔顿.此病毒与下列病毒即菜豆普通花叶病毒,菜豆黄色花叶病毒,黑眼豇豆花叶病毒,豌豆种传花叶病毒,三叶草黄脉病毒,莴苣花叶病毒及甜菜花叶病毒的抗血清均无反应.由于它在菜豆的"一窝猴"品种的叶片上产生沿脉黄色小点,因此认为这是一种新的独立的病毒,称之为菜豆和性黄色花叶病毒.     

Avoiding Secondary Leaf Fall Disease of Rubber by Chemical Defoliation

Abstract

An intensive survey of cultivated plants throughout the island of Montserrat resulted in the identification of a number of viruses. Squash mosaic virus was found in cantaloupe, pumpkin and squash. There were cases of multiple infection of pumpkin and squash by cucumber and squash mosaic viruses. Potato virus Y was recovered from pepper and tomato. Bean yellow mosaic and cowpea mosaic viruses were found in red kidney bean and string bean while cowpea mosaic virus was found in cowpea and broad bean (Jamaica bean). Other viruses that were previously identified on the basis of field symptomatology were confirmed by host range and serology. Viruses were not recovered from sea island cotton or from sweet potato, although field symptoms on the sweet potato are similar to those caused by sweet potato mosaic virus. The leaf tissues of crop plants were inoculated to a wide range of indicator plants. Some of those that reacted systemically were examined by electron microscopy for virus particles and the sap extracted from them was used in serodiagnostic assays for virus identification. This is the first attempt to identify positively the viruses of crop plants on the island.     

Persistence of infectivity of three viruses in plant material dried over CaCl2 and stored under different conditions

The infectivity of three viruses in leaf material dried over CaCl₂ and stored under different conditions was tested over a period of seven years. Alfalfa mosaic virus and cucumber mosaic virus kept very well for the entire period both at 4°C and at room temperature, provided they were stored over CaCl₂ in well-closed tubes. The infectivity of bean yellow mosaic virus, however, gradually declined, especially at room temperature. In open or loosely closed containers without CaCl₂ none of the viruses survived more than three months of storage and the infectivity of bean yellow mosaic virus declined after only two weeks.     

First report of fig mosaic virus infecting common fig (Ficus carica) in Japan

For the first time, fig mosaic virus (FMV) was detected in common fig (Ficus carica) trees in Shimane, Japan. These trees exhibited mosaic, ringspots, or distortion, accompanied by chlorosis on leaves and yellow spots on fruits. Some of the symptomatic trees were infested with the eriophyid mite Aceria ficus. The virus was detected based on RT-PCR, followed by sequencing. The amplified 300 base-pair fragments shared 83.5–91.5% identity with the corresponding region of RNA-dependent RNA polymerase gene of FMV isolates previously reported in Turkey, Iran, and Italy.     

Characterization of an isolate of beet mosaic virus from South Kazakhstan

A filamentous virus isolated from a sugar-beet plant showing systemic mosaic collected in South Kazakhstan was identified as an isolate of beet mosaic virus (BMV-K). BMV-K was transmitted by the green peach aphid Myzus persicae in a non-persistent manner, and by sap inoculation to 11 out of 19 species from seven families tested. The virus could not be transmitted to Nicotiana tabacum, N. debneyi, N. glutinosa and N. clevelandii, cither mechanically or with M. persicae. The thermal inactivation point of BMV-K in sugar-beet sap was 55-60 C, dilution end point 1:1000 and longevity in vitro 2 days at 20 C. A purification procedure produced 1-5-3 mg of purified virus from 100 g of infected Stellaria media plants. Purified virus contained a single protein species of molecular weight 34 700 Da. In ELISA tests, BMV-K reacted positively with BMV-specifc antisera obtained from Japan. Germany and Portugal. By competitive DAS- ELISA, the virus isolate was shown to be closely serologically related to all the three isolates of BMV, and very distantly related to bean yellow mosaic and soy bean mosaic viruses.     

Virus-induced changes in the response of faba bean to infection by Botrytis

Prior infection of faba bean with the viruses bean yellow mosaic and bean leaf roll increased host susceptibility to subsequent infection by Botrytis fabae and B. cinerea. Cell necrosis beneath inoculum droplets, rate and extent of lesion spread and sporulation of B. fabae were all increased on detached leaves from virus-infected compared with healthy plants. Changes were most marked in young leaves showing conspicuous symptoms of systemic virus infection and in plants virus-infected for at least 2-4 weeks. Localized lesions produced by B. cinerea or a low concentration of B. fabae conidia (10³ spores/ ml) showed increased cell necrosis but were not transformed into aggressive, spreading lesions on virus-infected leaves.     

豌豆病毒病病原研究

 1986年至1990年,从豌豆田中采集了150余份病毒病样本,鉴定出蚕豆萎蔫病毒(BB-WV)、芜菁花叶病毒(TuMV)、马铃薯Y病毒组分离物、黄瓜花叶病毒(CMV)、莴苣花叶病毒(LMV)、大豆花叶病毒(SMV)、豌豆花叶病毒(PMV)、菜豆黄花叶病毒(BYMV)和苜蓿花叶病毒(AMV)等9种病毒。样本中,BBWV所占的比例最高,达59.2%,其次为CMV,占15.5%。BBWV常与CMV复合侵染豌豆,LMV发生也较普遍。田间调查表明,豌豆病毒病发病率因种植地区及品种不同而有差异,平均发病率为12.4%。     

Species diversity,phylogeny and genetic variability of begomovirus populations infecting leguminous weeds in northeastern Brazil

A survey of begomoviruses infecting leguminous weeds (family Fabaceae) was carried out in four states of northeastern Brazil. A total of 26 full‐length begomovirus components (19 DNA‐A and seven DNA‐B, with three pairs of cognate A and B components) were amplified using rolling‐circle amplification, then cloned and sequenced. Sequence analysis indicated the presence of six species, four of them novel. In phylogenetic analysis five of the viruses clustered with other Brazilian begomoviruses, but one of them (Euphorbia yellow mosaic virus, EuYMV) clustered with viruses from other countries in Central and South America. Evidence of recombination was found among isolates of Macroptilium yellow spot virus (MaYSV). The MaYSV population had a high degree of genetic variability. Macroptilium lathyroides was revealed as a common host for several of these viruses, and could act as a mixing vessel from which recombinant viruses could emerge. The results indicate that leguminous weeds are reservoirs of several begomoviruses in Brazil, and could play a significant role in begomovirus epidemics, both as inoculum sources and as sources of emerging novel viruses.