scholarly journals Diversity and Mealybug Transmissibility of Ampeloviruses in Pineapple

Plant Disease ◽  
2005 ◽  
Vol 89 (5) ◽  
pp. 450-456 ◽  
Author(s):  
D. M. Sether ◽  
M. J. Melzer ◽  
J. Busto ◽  
F. Zee ◽  
J. S. Hu
Keyword(s):  
Deletion Mutant ◽  
Open Reading Frames ◽  
Ananas Comosus ◽  
Sequence Information ◽  
Rt Pcr ◽  
Degenerate Primers ◽  
Pcr Assays ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug

Mealybug wilt of pineapple (MWP) is one of the most destructive diseases of pineapple (Ananas comosus) worldwide. At least one Ampelovirus species, Pineapple mealybug wilt associated virus-2 (PMWaV-2), and mealybug feeding are involved in the etiology of MWP. A previously undescribed Ampelovirus sharing highest homology with PMWaV-1 and a putative deletion mutant sharing highest homology with PMWaV-2 were detected with reverse transcription-polymerase chain reaction (RT-PCR) assays using degenerate primers. Results were verified with additional sequence information and by immunosorbent electron microscopy. Sequence homology between the virus tentatively designated PMWaV-3, and PMWaV-1 and PMWaV-2, decreases toward the N-terminal across the HSP70 homolog, small hydrophobic protein, and RNA-dependent RNA polymerase open reading frames (ORF). Putative PMWaV-3 could not be detected with four different monoclonal antibodies specific for PMWaV-1 and PMWaV-2. The potential deletion mutant spanning the N-terminal of the HSP70 region was obtained from a pineapple accession from Zaire maintained at the USDA-ARS National Clonal Germplasm Repository in Hawaii. Putative PMWaV-3, like PMWaV-1 and PMWaV-2, is transmissible separately or in combination with other PMWaVs by Dysmicoccus brevipes and D. neobrevipes mealybugs. Plants infected with PMWaV-3 that were continuously exposed to mealybugs did not develop symptoms of MWP in the absence of PMWaV-2. Specific RT-PCR assays were developed for detection of putative PMWaV-3 and the deletion mutant.

scholarly journals Differentiation, Distribution, and Elimination of Two Different Pineapple mealybug wilt-associated viruses Found in Pineapple

Plant Disease ◽  
2001 ◽  
Vol 85 (8) ◽  
pp. 856-864 ◽  
Author(s):  
D. M. Sether ◽  
A. V. Karasev ◽  
C. Okumura ◽  
C. Arakawa ◽  
F. Zee ◽  
...  
Keyword(s):  
Agricultural Research ◽  
The United States ◽  
United States Department ◽  
Rt Pcr ◽  
Apical Bud ◽  
The World ◽  
Pcr Assays ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug

Surveys for Pineapple mealybug wilt-associated virus-1 (PMWaV-1) and PMWaV-2 were conducted on pineapple samples from Hawaii and around the world. Tissue blot immunoassays (TBIAs) with two different monoclonal antibodies (MAb) specific to either PMWaV-1 or PMWaV-2 indicated that both closteroviruses are widely distributed throughout the pineapple-growing areas of the world. In the worldwide survey, PMWaV-1 was found in 80% of the mea-lybug wilt of pineapple (MWP)-symptomatic and 78% of the asymptomatic pineapple plants tested. A subset of plants was tested for PMWaV-2; 100% of the symptomatic plants and 12% of the asymptomatic plants were positive for this virus. A reverse transcription-polymerase chain reaction (RT-PCR) assay was developed to differentiate between PMWaV-1 and PMWaV-2. Oligonucleotide primers were designed using distinct regions of the HSP 70 homolog genes of the two viruses. PMWaV-specific RT-PCR assays and TBIAs were used to screen the pineapple accessions maintained at the United States Department of Agriculture-Agricultural Research Service National Clonal Germplasm Repository for PMWaV infection; 73% of the accessions were found infected with at least one PMWaV. Pineapple accessions found PMWaV-free were challenged with viruliferous mealybugs to test for immunity to PMWaV-1. No immune germ plasm was identified. Potential alternative virus hosts were screened for infection with virus-specific RT-PCR assays and TBIAs and were also challenged with viruliferous mealybugs. No alternate hosts of PMWaV-1 or PMWaV-2 were identified. PMWaV-1 infection was eliminated through axillary and apical bud propagation from infected crowns. Strategies to manage MWP are discussed.

scholarly journals Spatial and Temporal Incidences of Pineapple mealybug wilt-associated viruses in Pineapple Planting Blocks

Plant Disease ◽  
2010 ◽  
Vol 94 (2) ◽  
pp. 196-200 ◽  
Author(s):  
D. M. Sether ◽  
W. B. Borth ◽  
M. J. Melzer ◽  
J. Hu
Keyword(s):  
Ananas Comosus ◽  
Ratoon Crop ◽  
Virus Incidence ◽  
Central Regions ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug ◽  
Fruit Harvest ◽  
Propagation Material

A complex of Pineapple mealybug wilt-associated viruses (PMWaVs) that can infect pineapple (Ananas comosus) is correlated with reduced yields and mealybug wilt of pineapple. The incidences of PMWaV-1 and PMWaV-2 at planting, fruit harvest, the beginning of the ratoon crop, and ratoon fruit harvest were determined for end, side, and central regions of planting blocks in eight commercial fields. Differences in virus incidence for the three regions at ratoon harvest were highly significant (P = 0.0018). Central regions of planting blocks had lower virus incidences at the time of ratoon fruit harvest. Collection of propagation material from the central regions of planting blocks will help to minimize PMWaV incidence in fields planted with this material.

scholarly journals Interactions Between Rotylenchus reniformis and Pineapple mealybug wilt associated virus-1 in Pineapple

Plant Disease ◽  
2002 ◽  
Vol 86 (9) ◽  
pp. 933-938 ◽  
Author(s):  
B. S. Sipes ◽  
D. M. Sether ◽  
J. S. Hu
Keyword(s):  
Vegetative Growth ◽  
Fruit Weight ◽  
Field Conditions ◽  
Ananas Comosus ◽  
Limiting Factor ◽  
Ratoon Crop ◽  
Nematode Reproduction ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug

The individual and combined effects of Pineapple mealybug wilt associated virus-1 (PMWaV-1) infection in pineapple, Ananas comosus, and Rotylenchus reniformis on pineapple growth were evaluated under greenhouse and field conditions. Under greenhouse conditions, no effect of PMWaV-1 infection on pineapple growth or nematode reproduction was observed. Under field conditions, the interaction of PMWaV-1 and nematodes was evaluated in plant and ratoon crops. In the plant crop, pineapple in plots treated with the nematicide 1,3-dichloropropane showed increased vegetative growth, whereas virus infection had no effect on vegetative growth. Nematodes reduced the average fruit weight (P = 0.01), whereas PMWaV-1 infection did not (P > 0.14). However, more fruit in the largest size classes (sizes 7 and 8) were produced in PMWaV-1-free plots than in PMWaV-1-infected plots (P = 0.03). The average fruit weight decreased in the presence of virus or nematodes. The smallest fruits were from PMWaV-1-infected plants infested with R. reniformis. More early-ripening fruit (30%) were produced by plants infected with PMWaV-1 than by PMWaV-free plants (P < 0.05). PMWaV-1 infection may be one of the reasons for asynchronous fruit ripening, which is a top limiting factor for pineapple production in Hawaii. In the ratoon crop, PMWaV-1 infection reduced fruit weight by 9% (P < 0.01), whereas nematode effects were similar across treatments (P > 0.10). More fruit in the three largest size categories were produced in PMWaV-1-free plots than in PMWaV-1-infected plots (P < 0.01). PMWaV-1 infection reduced the number of fruit produced in the ratoon crop (P < 0.02). An interaction (P < 0.03) between R. reniformis and PMWaV-1 infection status was detected in the ratoon crop. The fewest fruit were produced in plots with PMWaV-1-infected plants that were nematode infected.

scholarly journals Prevalence De La Maladie Du Deperissement De L’ananas (Wilt) Et Perception Des Producteurs Au Benin

2016 ◽  
Vol 12 (33) ◽  
pp. 350
Author(s):  
Houndédji C. D. ◽  
Zandjanakou-Tachin M. ◽  
Affokpon A. ◽  
Bokonon-Ganta A. H.
Keyword(s):  
Linear Models ◽  
Multinomial Distribution ◽  
Viral Disease ◽  
Quantitative Information ◽  
Ananas Comosus ◽  
Mealybug Wilt ◽  
Distribution Family ◽  
Severity Of The Disease ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug

Pineapple [Ananas comosus (L.) Merril], cultivated in Benin for it economical importance is affected by Pineapple mealybug wilt, a viral disease. The objectives of this study were to evaluate the determining factors of its prevalence and farmers knowledge on the disease. To achieve these, a total number of 180 farmers, distributed in five communes of southern Benin have been interviewed in 2014 and 2015. In order to identify factors that influence the disease distribution, generalized linear models with the binomial and multinomial distribution family were used to explain the effect of the variables (field and plant levels) and disease severity, respectively. Only factors ‘commune’ 'and' year ' had a significant effect on the incidence at field level (69% in 2014 and 78% in 2015) while three predictors (township, year and variety) explained the severity of the disease and the interaction year -township was significant. Farmers perceive the disease and attribute five causes responsible for its expression. They use height methods to manage the disease. These findings constitute a qualitative as well as quantitative information to build on in designing methods of controlling the disease in order to boost the productivity of pineapple in Benin.

scholarly journals Nucleotide sequence, genome organization and phylogenetic analysis of pineapple mealybug wilt-associated virus-2

2001 ◽  
Vol 82 (1) ◽  
pp. 1-7 ◽  
Author(s):  
M. J. Melzer ◽  
A. V. Karasev ◽  
D. M. Sether ◽  
J. S. Hu
Keyword(s):  
Phylogenetic Analysis ◽  
Coat Protein ◽  
Genome Organization ◽  
Open Reading Frames ◽  
Double Stranded Rna ◽  
The Family ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug ◽  
Reading Frames

The genome of pineapple mealybug wilt-associated closterovirus-2 (PMWaV-2) was cloned from double-stranded RNA isolated from diseased pineapple and its sequence determined. The 3′-terminal 14861 nt of the single-stranded RNA genome contains ten open reading frames (ORFs) which, from 5′ to 3′, potentially encode a >204 kDa polyprotein containing papain-like protease, methyltransferase and helicase domains (ORF1a), a 65 kDa RNA-dependent RNA polymerase (ORF1b), a 5 kDa hydrophobic protein (ORF2), a 59 kDa heat shock protein 70 homologue (ORF3), a 46 kDa protein (ORF4), a 34 kDa coat protein (ORF5), a 56 kDa diverged coat protein (ORF6), a 20 kDa protein (ORF7), a 22 kDa protein (ORF8) and a 6 kDa protein (ORF9). A 132 nt untranslated region was present at the 3′ terminus of the genome. This genome organization is typical of the monopartite closteroviruses, including the putative +1 ribosomal frameshift allowing expression of ORF1b. Phylogenetic analysis revealed that within the family Closteroviridae the mealybug-transmitted PMWaV-2 is more closely related to other mealybug-transmitted members than to those which are transmitted by aphids or whiteflies. Within this group, PMWaV-2 shares the greatest sequence identity with grapevine leafroll-associated virus-3, another mealybug-transmitted closterovirus.

scholarly journals Occurrence and Molecular Characterization of Three Pineapple Mealybug Wilt-Associated Viruses in Pineapple in Taiwan

Plant Disease ◽  
2009 ◽  
Vol 93 (2) ◽  
pp. 196-196 ◽  
Author(s):  
B. N. Shen ◽  
Y. X. Zheng ◽  
W. H. Chen ◽  
T. Y. Chang ◽  
H.-M. Ku ◽  
...  
Keyword(s):  
Amino Acid ◽  
Molecular Characterization ◽  
Amino Acid Identity ◽  
Rt Pcr ◽  
Acid Identity ◽  
Cp Gene ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug

Pineapple (Ananas comosus) is one of the major fruit crops in Taiwan, accounting for 275 million U.S. dollars in 2006, following betel nut and citrus production in crop value. Tainung No. 17 is the most important cultivar, accounting for more than 70% of pineapples planted. Mealybug wilt of pineapple (MWP) is one of the most destructive diseases of pineapple. Pineapple mealybug wilt-associated virus-1 (PMWaV-1), PMWaV-2, and PMWaV-3 were identified as three distinct species in Ampelovirus from diseased Hawaiian pineapple (1,2). In November of 2007, pineapples (cv. Tainung No. 17) planted in Pingtung County of southern Taiwan showed symptoms similar to MWP. Mealybugs (Dysmicoccus brevipes) were also found. Three primer pairs, 225/226, 223/224, and 263/264 described previously specific for the HSP70h genes of PMWaV-1 (1), -2, and -3 (2), respectively, were used to detect the presence of these three viruses by reverse transcription (RT)-PCR. Expected DNA fragments of 590, 610, and 499 nt were obtained from the total RNA isolated from the leaves of diseased pineapples with primer pairs 225/226, 223/224, and 263/264, respectively. The RT-PCR amplified fragments were cloned, sequenced, and analyzed. The 590-nt fragment (Accession No. EU769113) shared 91.6 to 99.5% nucleotide and 96.8 to 99.5% amino acid identity to those of five isolates of PMWaV-1 available in the GenBank; one each from Hawaii (Accession No. AF414119) and Thailand (Accession No. EF620774) and three from Australia (Accession Nos. EF488752, EF467923, and EF467925). The 610-nt fragment (Accession No. EU769115) showed 98.7 and 99.7% nucleotide and 98% and 100% amino acid identity to those of PMWaV-2 from Hawaii (Accession No. AF283103) and Thailand (Accession No. EU016675), respectively. The 499-nt fragment (Accession No. FJ209047) shared 86.8 to 99.0% nucleotide and 94.0 to 100.0% amino acid identity to those of five PMWaV-3 isolates available in the GenBank; one from Hawaii (Accession No. DQ399259) and four from Australia (Accession Nos. EF467918, EF467919, EF488754, and EF488755). Using primer pairs FJ08-1 (5′-ATGGCTGATTCGAGC)/FJ08-2 (5′-TTATTTGCGTCCACC), FJ08-7 (5′-AGTGAGATTGATCGT)/FJ08-8 (5′-TGCAGGTATCCGCTG), and FJ08-35 (5′-AACGACCGAACTCGC)/FJ08-36 (5′-ATACTACAGATATTG) specific to the coat protein (CP) genes of PMWaV-1, -2, and -3, respectively, expected DNA fragments of 774, 909, and 789 nt were amplified by RT-PCR. The 774-nt CP gene of PMWaV-1 (Accession No. EU769114) shared 99% nucleotide and 98.4% amino acid identity to those of Hawaiian isolate (Accession No. AF414119). The 909-nt CP gene of PMWaV-2 (Accession No. EU769116) shared 99.0 and 99.1% nucleotide identity with isolates from Hawaii (Accession No. AF283103) and Cuba (Accession No. DQ225114), respectively, and 99.3% amino acid identity with both. The 789-nt CP gene of PMWaV-3 (Accession No. FJ209048) shared 99.1% nucleotide and 98.1% amino acid identity to those of the Hawaiian isolate (Accession No. DQ399259). One to two viruses among PMWaV-1, -2, and -3 were detected in all 40 samples collected from diseased pineapples. To our knowledge, this is the first report to identify three PMWaVs in the most important and widely planted pineapple cultivar in Taiwan, Tainung No. 17, by molecular characterization of the HSP70h and CP genes. References: (1) D. M. Sether et al. Plant Dis. 85:856, 2001. (2) D. M. Sether et al. Plant Dis. 89:450, 2005.

scholarly journals Detection of Pineapple mealybug wilt-associated virus 1 and 3 in Mexico

2016 ◽  
Author(s):  
Daniel Leobardo Ochoa-Martínez ◽  
Daniel Emigdio Uriza-Ávila ◽  
Reyna Isabel Rojas-Martínez
Keyword(s):  
Australian Isolate ◽  
Hsp70 Gene ◽  
Rt Pcr ◽  
Specific Primers ◽  
The World ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug ◽  
Wilt Virus ◽  
Mexican Isolate

<p>In El Bajo Papaloapan, the main producing area of pineapple of Mexico, leaves with typical symptoms of viral infection consisting in chlorosis, flaccidity, reduced growth and reddening were collected. By RT-PCR with specific primers for the hsp70 gene and subsequent sequencing were detected Pineapple mealybug wilt virus associated-virus 1 (PMWaV-1) and Pineapple mealybug wilt virus associated-3 (PMWaV-3). From the sequences obtained a tree was done with sequences from different regions of the world available in GenBank in order to know their similarity. The sequence obtained from the Mexican isolate PMWaV-1 was genetically related to the sequences of isolates from Cuba, Taiwan, Thailand and Hawaii and more distant from the Australian isolate. The sequence obtained for the Mexican isolate PMWaV-3 was more related to isolates from Hawaii, Cuba, Australia and Taiwan and more distant from the Thailand isolate. This is the first report of the presence of these two viruses in Mexico.</p>

scholarly journals First Report of a Closterovirus-Like Particle Associated with Pineapple Plants (Ananas comosus cv. Smooth Cayenne) Affected with Pineapple Mealybug Wilt in Cuba

Plant Disease ◽  
1998 ◽  
Vol 82 (2) ◽  
pp. 263-263 ◽  
Author(s):  
Eduviges G. Borroto ◽  
Mayra Cintra ◽  
Justo González ◽  
Carlos Borroto ◽  
Pedro Oramas
Keyword(s):  
Electron Microscope ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
System Size ◽  
Ananas Comosus ◽  
Meristem Culture ◽  
First Report ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug

Pineapple mealybug wilt (PMW) was first described in Hawaii in 1910 and has since spread wherever pineapple is cultivated. PMW is currently one of the most important field diseases of pineapple in Cuba, causing up to 40% crop loss. Leaves of plants affected with PMW turn bronze-red and lose turgidity, and the tips curve down. Root system size is decreased and fruits are unmarketable because the flesh is fibrous and sour. Leaves from wilt-affected pineapple plants were selected from the Experimental Station “Tomás Roig” at Ciego de Avila, Cuba. Asymptomatic plants derived from meristem tissue and cultivated under greenhouse conditions were also collected. Both samples were analyzed by several virus purification procedures and finally purified with a protocol modified from Gunashinge and German (1). Partially purified samples from symptomatic and asymptomatic plants were negatively stained and examined by a JEOL (JEM-2000 EX) transmission electron microscope at a magnification of ×30,000. Sixteen, long, flexuous, rod-shaped, viruslike particles (estimated length 1,200 to 1,450 nm; width about 12 nm) were observed in symptomatic leaves, using electron microscope micrographs. These particles were similar to those of the pineapple clostero-like virus found in Hawaii (1) and Australia (2). Particles were observed from symptomatic plants but not from plants propagated by primary meristem culture. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of viral proteins indicated that the coat protein has a molecular mass of approximately 23 kDa. This is the first report of closterovirus-like particles associated with pineapple plants (Ananas comosus cv. Smooth Cayenne) affected with PMW in Cuba. References: (1) U. B. Gunasinge and T. L. German. Phytopathology 79:1337, 1989. (2) W. Wakman et al. Aust. J. Agric. Res. 46:947, 1995.

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