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 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 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 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 Mealybug Wilt of Pineapple and Associated Viruses

Horticulturae ◽  
2018 ◽  
Vol 4 (4) ◽  
pp. 52 ◽  
Author(s):  
Kishore Dey ◽  
James Green ◽  
Michael Melzer ◽  
Wayne Borth ◽  
John Hu
Keyword(s):  
Symptom Development ◽  
Tropical Regions ◽  
Virus Particles ◽  
Subgroup Ii ◽  
The World ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug ◽  
Shed Light ◽  
Complex Genome

Mealybug wilt of pineapple (MWP) is a disease of pineapple that has a long history in Hawaii, but is present throughout the world where pineapples are grown in tropical regions. The disease has an interesting etiology that is poorly understood but involves an association with virus particles, mealybug vectors, and ants which spread the mealybug vectors. Several distinct pineapple mealybug wilt-associated virus (PMWaV) species have been identified thus far with potential further member species yet to be characterized. Pineapple mealybug wilt-associated viruses are member species of the Ampelovirus genus of the Closteroviridae family. Ampeloviruses are split into two subgroups, subgroup I and subgroup II. PMWaV-2 is a subgroup II member, and these have a longer and more complex genome with additional genes on the 3’ terminus of the RNA genome compared to subgroup I ampeloviruses. PMWaV-2, along with the presence of mealybug vectors, have been shown to be necessary factors in symptom development in Hawaii. Some of these extra genes in the 3’ of PMWaV-2 have recently been shown to function as silencing suppressors, and may play a role in the virulence of PMWaV-2 and symptom development. In other regions of the world, reports of symptomatic plants without PMWaV-2 infection, but with PMWaV-1, -3 or some combination, contradict the requirement of PMWaV-2 for symptom development in MWP. It is possible that further, uncharacterized PMWaVs may be present in symptomatic pineapple plants that test negative for PMWaV-2, explaining the inconsistency in symptom development. More research is necessary to explore the confusing etiology of the MWP disease, and to perhaps shed light upon the symptom development.

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 Probing behavior of Dysmicoccus brevipes mealybug in pineapple plants1

2016 ◽  
Vol 46 (4) ◽  
pp. 458-463
Author(s):  
Lenira Viana Costa Santa-Cecília ◽  
◽  
Ernesto Prado ◽  
Brígida Souza ◽  
Keyword(s):  
Insect Vector ◽  
Recording Time ◽  
Defense Strategies ◽  
Phenolic Contents ◽  
Probing Behavior ◽  
Epg Technique ◽  
Dysmicoccus Brevipes ◽  
Mealybug Wilt ◽  
Pineapple Mealybug Wilt ◽  
Pineapple Mealybug

ABSTRACT Differences in susceptibility to viruses in plants may be partially explained by the insect vector probing behavior and by the presence of phenolic compounds, which are often associated with defense strategies. This study aimed at detecting barriers that may difficult the probing activity of the Dysmicoccus brevipes (Pseudococcidae) pineapple mealybug, a vector of the pineapple mealybug wilt-associated virus, as well as evaluating the phenolic content of plants, in order to verify any possible relationship with the probing behavior, by using the electrical penetration graphs (EPG) technique. Seedlings of 'Smooth Cayenne' and 'Pérola' pineapple cultivars were used in the experiments. Only 28 % and 21 % of the mealybugs reached the phloem of the 'Smooth Cayenne' and 'Pérola' cultivars, respectively, over 16 h of recording, with an average of 9 h to reach the phloem vessels. The xylem phase was extended in both cultivars and represented approximately 31 % ('Smooth Cayenne') and 44 % ('Pérola') of the recording time. The phenolic contents of both cultivars were similar.

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