Molecular Evidence of Cacao Swollen Shoot Virus Acquisition and Retention by Planococcus Citri (Risso) and Pseudococcus Longispinus (Targioni-Tozzetti) and Pseudococcus Viburni (Signoret) Mealybugs (Hemiptera: Pseudococcidae)

Mealybugs (Hemiptera: Coccomorpha: Pseudococcidae) are important pests in fruit production in Uruguay; however, very little is known about the species involved. A survey of mealybugs associated especially with fruit crops (apple, citrus, figs, grapes, pears, quince and strawberry), and other crops like vegetables and sugar cane, ornamentals and weeds was performed between 2017 and 2019 in Uruguay, using integrated taxonomy (morphology and DNA analyses) for their identification. A total of 19 mealybug species were identified. The most common species were Planococcus ficus (Signoret), Pseudococcus scatoterrae Granara de Willink and Pseudococcus viburni (Signoret) on fruits, and Phenacoccus madeirensis Green, Phenacoccus peruvianus Granara de Willink and Planococcus citri (Risso) on ornamental plants, all of them causing damage to their hosts. This study presents nine new species records for Uruguay, besides the description of two new species. An identification key to the mealybugs in Uruguay is provided.

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Planococcus citri (citrus mealybug).

10.1079/cpc.45082.20210102911 ◽

2021 ◽

Author(s):

Gillian Watson

Keyword(s):

Plant Material ◽

Plant Virus ◽

Host Plants ◽

Planococcus Citri ◽

Virus Diseases ◽

Citrus Mealybug ◽

Fresh Plant Material ◽

The World ◽

Cacao Swollen Shoot Virus ◽

Human Transport

Abstract Planococcus citri is a highly polyphagous, adaptable mealybug that can feed on many host plants in a variety of conditions, and can reproduce rapidly. It has been reported on over 200 host-plant species belonging to 191 genera and 82 families, and can seriously damage many crops, particularly citrus and glasshouse tomatoes. It is known to transmit some plant virus diseases like Cacao swollen shoot virus. The mealybug is of Old World origin, but its polyphagy has facilitated its spread about the world by human transport of infested plants over many years, and it is now established in in all the temperate and tropical zoogeographic regions, and lives under glass in higher latitudes. Its small size and cryptic habits makes it difficult to detect and identify at plant quarantine inspection. The increase in international trade in fresh plant material in recent years is facilitating its continued spread.

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Preparation of Mealybugs (Hemiptera: Pseudococcidae) for Genetic Characterization and Morphological Examination

Journal of Insect Science ◽

10.1093/jisesa/iev086 ◽

2015 ◽

Vol 15 (1) ◽

Cited By ~ 3

Author(s):

B. W. Bahder ◽

M. L. Bollinger ◽

M. R. Sudarshana ◽

F. G. Zalom

Keyword(s):

Taxonomic Status ◽

Planococcus Citri ◽

Morphological Identification ◽

Agricultural Pests ◽

Current Standard ◽

Physical Damage ◽

Morphological Examination ◽

High Quality ◽

Voucher Specimen ◽

Pseudococcus Viburni

Abstract Mealybugs (Hemiptera: Pseudococcidae) are economically significant agricultural pests on many different crops. Because of their small size and lack of easily visible characters for identification, determination of their taxonomic status is difficult and requires technical competency to prepare a slide-mounted specimen. The standard mounting technique does not allow for analysis of the genome of the specimen. Conversely, preparatory techniques for genetic analysis of mealybugs cause either loss of the entire individual or physical damage that can make morphology-based identification difficult. This study describes a simple protocol that does not impact physical integrity of the specimen for fixation and microscopic examination yet enables simultaneous DNA extraction for DNA-based identification of four mealybug species. All species prepared yielded high quality slide mounts, identified as Planococcus citri Risso, Pseudococcus viburni Signoret, Rhizoecus kondonis Kuwana, or Rhizoecus californicus Ferris. DNA extracted in this manner had higher purity and yield in the final eluate than in samples extracted using standard methods. DNA extracted was successfully amplified by polymerase chain reaction using primers for the cytochrome oxidase I gene and subsequently sequenced for all specimens. This protocol is likely to be applicable to other Hemiptera taxa that are preserved by slide mounting, allowing for both the preparation of a high-quality voucher specimen for morphological identification and simultaneous analysis of DNA for the same specimen. The methods used are technically less challenging than current standard procedures.

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Molecular identification of three of the most important mealybug species (Hemiptera: Sternorrhyncha: Coccoidea: Pseudococcidae) on ornamental plants in Guilan province, Iran

Zootaxa ◽

10.11646/zootaxa.3009.1.2 ◽

2011 ◽

Vol 3009 (1) ◽

pp. 46 ◽

Cited By ~ 4

Author(s):

REZA HOSSEINI ◽

JALIL HAJIZADEH

Keyword(s):

Ornamental Plants ◽

Accurate Method ◽

Planococcus Citri ◽

Morphological Identification ◽

Specific Primers ◽

Pseudococcus Viburni ◽

Species Specific ◽

Pseudococcus Comstocki ◽

Guilan Province ◽

Study Species

Mealybugs (Coccoidea: Pseudococcidae) are serious pests, particularly as invasive species on many agricultural products. Morphological identification of mealybugs is based on adult female characters that, in the absence of adult females or with damaged specimens, can be problematic, especially when identification is required urgently, such as that involving the exportation/importation market. In this study, species-specific primers were designed to identify three of the most abundant mealybug species found on ornamental plants in Guilan province, Iran: Planococcus citri (Risso), Pseudococcus viburni (Signoret) and Pseudococcus comstocki (Kuwana). By generating amplification products of different sizes, the three species-specific primers, along with universal COI primers, were successfully used in multiplex PCR tests to identify all three mealybug species in a single reaction. Analysis of a large array of specimens from different geographic locations on different host plants showed that this was a reliable and accurate method.

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Molecular and morphological characterization of mealybugs (Hemiptera: Pseudococcidae) from Chilean vineyards

Bulletin of Entomological Research ◽

10.1017/s0007485312000053 ◽

2012 ◽

Vol 102 (5) ◽

pp. 524-530 ◽

Cited By ~ 23

Author(s):

M.C.G. Correa ◽

J-F. Germain ◽

T. Malausa ◽

T. Zaviezo

Keyword(s):

Management Strategies ◽

Common Species ◽

Morphological Characterization ◽

Economic Losses ◽

Morphological Examination ◽

Pseudococcus Longispinus ◽

Main Production ◽

Pseudococcus Viburni ◽

Production Areas

AbstractMealybugs are major pests of grapevines worldwide. They cause economic losses by lowering the cosmetic value of fruits, reducing yields, transmitting viruses and resulting in the quarantine or rejection of produce in international trade. Knowledge of the species present in a vineyard is important for the adjustment of management strategies. We surveyed and accurately characterized the mealybugs infesting vineyards in one of the main production areas of Chile; 164 mealybugs were sampled from 26 vineyards in four regions of Chile and identified by DNA sequencing for two markers (cytochrome oxidase I and internal transcribed spacer 2) and morphological examination.Pseudococcus viburni(Signoret) was the most common species, followed byPseudococcus meridionalisPrado andPseudococcus cribataGonzález. Molecular variability at the COI and ITS2 loci was observed in bothP. viburniandP. cribata. A comparison of haplotypes ofP. viburniworldwide provides support for a recent hypothesis that this species is native to South America, a finding with direct consequences for management. NeitherPseudococcus longispinus(Targioni & Tozzetti) norPlanococcus ficusSignoret were found.

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Transmission of Activated-Episomal Banana streak OL (badna)virus (BSOLV) to cv. Williams Banana (Musa sp.) by Three Mealybug Species

Plant Disease ◽

10.1094/pdis-92-8-1158 ◽

2008 ◽

Vol 92 (8) ◽

pp. 1158-1163 ◽

Cited By ~ 29

Author(s):

J. B. Meyer ◽

G. G. F. Kasdorf ◽

L. H. Nel ◽

G. Pietersen

Keyword(s):

Tissue Culture ◽

Enzyme Linked Immunosorbent Assay ◽

Planococcus Citri ◽

Pseudococcus Longispinus ◽

Musa Sp ◽

Transmission Study ◽

Mother Plants ◽

History Of ◽

Tissue Culture Propagation ◽

Transmission Studies

Four different mealybug species (Dysmicoccus brevipes, Planococcus citri, P. ficus, and Pseudococcus longispinus) were evaluated for their ability to transmit putative activated-episomal Banana streak OL (badna)virus (BSOLV) to banana cv. Williams (Cavendish subgroup, AAA). Expressible endogenous sequences of banana streak viruses (BSVs) have been reported to be present in the DNA of various Musa hybrids, including FHIA-21 (AAAB). To obtain activated episomal BSOLV for this experimental transmission study, intentional stress by tissue culture propagation was applied to indexed FHIA-21 which, while free of other viruses, can contain activated episomal BSOLV. Immunocapture polymerase chain reaction and triple-antibody sandwich enzyme-linked immunosorbent assay results revealed that 13.4% of the derived progeny of the mother plants were infected with episomal BSOLV. Four of these BSOLV-infected progeny were used as sources of episomal virus for transmission studies. D. brevipes, Planococcus citri, and P. ficus mealybugs were able to transmit the putative activated episomal BSOLV. Control plants for the transmission experiments included FHIA-21 corms with no background history of tissue culture, as well as virus-free Williams plants. Episomal Banana streak GF (badna)virus (BSGFV) was transmitted from asymptomatic corm-derived FHIA-21 plants by P. citri and P. ficus. This is the first report of P. ficus as a vector of BSVs.

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A multiplex PCR assay for the simultaneous identification of three mealybug species (Hemiptera: Pseudococcidae)

Bulletin of Entomological Research ◽

10.1017/s000748530700538x ◽

2007 ◽

Vol 98 (1) ◽

pp. 27-33 ◽

Cited By ~ 37

Author(s):

D.L. Saccaggi ◽

K. Krüger ◽

G. Pietersen

Keyword(s):

Multiplex Pcr ◽

Dna Extraction ◽

Single Step ◽

Planococcus Citri ◽

Morphological Identification ◽

Citrus Mealybug ◽

Pseudococcus Longispinus ◽

Multiplex Pcr Assay ◽

High Level ◽

Simultaneous Identification

AbstractMolecular species identification is becoming more wide-spread in diagnostics and ecological studies, particularly with regard to insects for which morphological identification is difficult or time-consuming. In this study, we describe the development and application of a single-step multiplex PCR for the identification of three mealybug species (Hemiptera: Pseudococcidae) associated with grapevine in South Africa: Planococcus ficus (vine mealybug), Planococcus citri (citrus mealybug) and Pseudococcus longispinus (longtailed mealybug). Mealybugs are pests on many commercial crops, including grapevine, in which they transmit viral diseases. Morphological identification of mealybug species is usually time-consuming, requires a high level of taxonomic expertise and usually only adult females can be identified. The single-step multiplex PCR developed here, based on the mitochondrial cytochrome c oxidase subunit 1 (CO I) gene, is rapid, reliable, sensitive, accurate and simple. The entire identification protocol (including DNA extraction, PCR and electrophoresis) can be completed in approximately four hours. Successful DNA extraction from laboratory and unparasitized field-collected individuals stored in absolute ethanol was 97%. Specimens from which DNA could be extracted were always correctly identified (100% accuracy). The technique developed is simple enough to be implemented in any molecular laboratory. The principles described here can be extended to any organism for which rapid, reliable identification is needed.

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Natural enemies of three mealybug species (Hemiptera: Pseudococcidae) found on citrus and effects of some insecticides on the mealybug parasitoid Coccidoxenoides peregrinus (Hymenoptera: Encyrtidae) in South Africa

Bulletin of Entomological Research ◽

10.1079/ber2003235 ◽

2003 ◽

Vol 93 (3) ◽

pp. 243-254 ◽

Cited By ~ 12

Author(s):

W.M. Wakgari ◽

J.H. Giliomee

Keyword(s):

South African ◽

Common Species ◽

Planococcus Citri ◽

Citrus Limon ◽

Nymphal Stage ◽

Predator Species ◽

Pseudococcus Longispinus ◽

Host Trees ◽

Citrus Orchards ◽

Intensive Sampling

AbstractThe population density of mealybug species in some South African citrus orchards has increased to pest status in recent years. The characterization of the natural enemy complex and quantification of their contribution to the control of Planococcus citri (Risso), Pseudococcus longispinus (Targioni-Tozzetti) and Pseudococcus calceolariae (Maskell) on Citrus limon (L.) and Citrus reticulata (Blanco) was investigated through intensive sampling. Eight primary and four secondary parasitoids, and two predator species were identified from P. citri and P. calceolariae. Anagyrus pseudococci (Girault) and Coccidoxenoides peregrinus (Timberlake) were the most common species, accounting for 44% and 21% of the total. Of the five primary parasitoids reared from P. longispinus, A. pseudococci and Anagyrus sp. were predominant, comprising 41% and 30%. Nymphal and adult parasitism (range = 0–26% vs. 0–66%) and predation (range = 0–5.6% vs. 0–4.1%) varied significantly between host trees and mealybug species (P < 0.001). The numbers of nymphal instars and adult stages of P. calceolariae and P. longispinus and the nymphal stage of P. citri that were parasitized and killed by predators correlated significantly with the total number of hosts on which they acted (P < 0.01), suggesting a density-dependent association. Laboratory bioassay of nine contact insecticides (methidathion, methomyl, methyl-parathion, parathion, profenofos and prothiofos) against C. peregrinus indicated that all were highly toxic, causing 98–100% mortality in < 6 h of treatment. The IGRs fenoxycarb and triflumuron did not cause significant parasitoid mortality (P > 0.05). However, a mixture of pyriproxyfen and mineral oil caused a marginally significant mortality (P < 0.05).

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Identification of mealybug pest species (Hemiptera: Pseudococcidae) in Egypt and France, using a DNA barcoding approach

Bulletin of Entomological Research ◽

10.1017/s0007485312000041 ◽

2012 ◽

Vol 102 (5) ◽

pp. 515-523 ◽

Cited By ~ 27

Author(s):

S. Abd-Rabou ◽

H. Shalaby ◽

J.-F. Germain ◽

N. Ris ◽

P. Kreiter ◽

...

Keyword(s):

Management Strategies ◽

Ornamental Plants ◽

Planococcus Citri ◽

Pest Species ◽

Morphological Examination ◽

Phenacoccus Solenopsis ◽

Major Barrier ◽

Maconellicoccus Hirsutus ◽

Pseudococcus Longispinus ◽

Cryptic Taxa

AbstractPseudococcidae (mealybugs) is a large taxonomic group, including a number of agronomic pests. Taxonomic identification of mealybug species is a recurrent problem and represents a major barrier to the establishment of adequate pest management strategies. We combined molecular analysis of three DNA markers (28S-D2, cytochrome oxidase I and internal transcribed spacer 2) with morphological examination, for the identification of 176 specimens collected from 40 mealybug populations infesting various crops and ornamental plants in Egypt and France. This combination of DNA and morphological analyses led to the identification of 17 species: seven in Egypt (Planococcus citri (Risso), Planococcus ficus (Signoret), Maconellicoccus hirsutus (Green), Ferrisia virgata (Cockerell), Phenacoccus solenopsis Tinsley, Phenacoccus parvus Morrison and Saccharicoccus sacchari (Cockerell)) and 11 in France (Planococcus citri, Pseudococcus viburni Signoret, Pseudococcus longispinus (Targioni-Tozzetti), Pseudococcus comstocki (Kuwana), Rhizoecus amorphophalli Betrem, Trionymus bambusae (Green), Balanococcus diminutus (Leonardi), Phenacoccus madeirensis Green, Planococcus vovae (Nasonov), Dysmicoccus brevipes (Cockerell) and Phenacoccus aceris Signoret), Pl. citri being found in both countries. We also found genetic variation between populations considered to belong to the same species, justifying further investigation of the possible occurrence of complexes of cryptic taxa.

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