Development of silverleaf assay, protein and nucleic acid-based diagnostic techniques for the quick and reliable detection and monitoring of biotype B of the whitefly, Bemisia tabaci (Gennadius)

AbstractThe aim of this study was to develop and optimize silverleaf bioassay, esterase analysis and PCR-based techniques to distinguish quickly and reliably biotype B of the whitefly, Bemisia tabaci (Gennadius), from Indian indigenous biotypes. Zucchini and squash readily develop silverleaf symptoms upon feeding by the B biotype, but they are not readily available in Indian markets. A local pumpkin variety ‘Big’ was, therefore, used in silverleaf assay, which developed symptoms similar to those on zucchini and squash and can be used reliably to detect B biotype. Analysis of non-specific esterases of B and the indigenous biotypes indicated both quantitative and qualitative differences in esterase patterns. Two high molecular weight bands were unique to B biotype and they occurred in abundance. These esterases were used to develop quick and field-based novel detection methods for differentiating B from the indigenous biotypes. Development of these simple and cost-effective protocols has wider application as they can be potentially used to identify other agricultural pests. Mitochondrial cytochrome oxidase I gene sequences and randomly amplified polymorphic DNA (RAPD) polymorphisms, generated using the primer OpB11, were also found useful for detecting B. tabaci biotypes. A B biotype-specific RAPD band of 800 bp was sequenced, which was used to a develop sequence characterized amplified region (SCAR) marker. The SCAR marker involved the development of B biotype-specific primers that amplified 550 bp PCR products only from B biotype genomic DNA. Silverleaf assay, esterases, RAPDs or a SCAR marker were used in combination to analyse whitefly samples collected from selected locations in India, and it was found that any of these techniques can be used singly or in combination to detect B biotype reliably. The B biotype was found in southern parts of India but not in the north in 2004–06.

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Identification of the B, Q, and native Brazilian biotypes of the Bemisia tabaci species complex using Scar markers

Pesquisa Agropecuária Brasileira ◽

10.1590/s0100-204x2016000500016 ◽

2016 ◽

Vol 51 (5) ◽

pp. 555-562 ◽

Cited By ~ 3

Author(s):

Paulo Roberto Queiroz ◽

Erica Soares Martins ◽

Nazaré Klautau ◽

Luzia Lima ◽

Lilian Praça ◽

...

Keyword(s):

Bemisia Tabaci ◽

Species Complex ◽

Scar Marker ◽

Random Amplified Polymorphic Dna ◽

Scar Markers ◽

Sequence Characterized Amplified Region ◽

Field Samples ◽

Scar Primer ◽

Q Biotype ◽

B Biotype

Abstract: The objective of this work was to develop sequence-characterized amplified region (Scar) markers to identify the B, Q, and native Brazilian biotypes of the sweet potato whitefly [Bemisia tabaci (Hemiptera: Aleyrodidae)]. Random amplified polymorphic DNA (RAPD) amplification products, exclusive to the B and Brazilian biotypes, were selected after the analysis of 12,000 samples, in order to design a specific Scar primer set. The BT-B1 and BT-B3 Scar markers, used to detect the B biotype, produced PCR fragments of 850 and 582 bp, respectively. The BT-BR1 Scar marker, used to identify the Brazilian biotype, produced a PCR fragment of 700 bp. The Scar markers were tested against the Q biotype, and a flowchart was proposed to indicate the decision steps to use these primers, in order to correctly discriminate the biotypes. This procedure allowed to identify the biotypes that occur in field samples, such as the B biotype. The used set of primers allowed to discriminate the B, Q, and native Brazilian biotypes of B. tabaci. These primers can be successfully used to identify the B biotype of B. tabaci from field samples, showing only one specific biotype present in all cultures.

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Diagnosis of sexually transmitted infections (STI) using self-collected non-invasive specimens

Sexual Health ◽

10.1071/sh03014 ◽

2004 ◽

Vol 1 (2) ◽

pp. 121 ◽

Cited By ~ 26

Author(s):

Suzanne M. Garland ◽

Sepehr N. Tabrizi

Keyword(s):

Sexually Transmitted Infections ◽

Control Strategies ◽

Cost Effective ◽

Diagnostic Techniques ◽

Detection Methods ◽

Molecular Diagnostic ◽

Molecular Diagnostic Techniques ◽

Sexually Transmitted ◽

Diagnostic Assays ◽

Remote Areas

Paramount in control of transmission of sexually transmitted infections (STIs) is their prompt recognition and appropriate treatment. In countries where definitive diagnoses are difficult, a ‘syndromic approach’ to management of STIs is recommended and practiced, yet many STIs have common symptoms or are asymptomatic and therefore go undetected and untreated. This is of particular concern with the recognition that HIV transmission is increased with co-existent STIs: the attributable risk for each STI varying with the prevalence within a particular population. Hence, HIV public health prevention approaches must include STI preventative strategies to be effective. Even then, microbiological screening is incorporated into STI control strategies; lack of access to appropriate services (especially in rural and remote areas), reluctance of at-risk populations to attend for treatment, fear of invasive genital examinations, and lower sensitivities of conventional diagnostic assays reduces the effectiveness of such programmes. Therefore, accurate, cost-effective, reliable diagnostic assays (preferably those which can be used in the field) are needed to impact on the incidence of the various STIs, as well as HIV. With the advent of molecular technologies, including target and signal amplification methods, diagnoses of STIs have been revolutionised and allow the use of non or minimally invasive sampling techniques, some of which are self-collected by the patient, e.g. first-void urine, cervico-vaginal lavage, low vaginal swabs, and tampons. Most studies evaluating such self-sampling with molecular diagnostic techniques have demonstrated an equivalent or superior detection of STIs as compared to conventional sampling and detection methods. These sampling methods can also be used to determine prevalence of STIs in various populations, but particularly those with difficult access to medical care. In this article, the utility of self-sampling collection devices for detection of various STIs, particularly in women, is reviewed as one step towards formulating appropriate strategies in control of STIs, and which are especially suited for remote areas.

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Post-release evaluation of Eretmocerus hayati Zolnerowich and Rose in Australia

Bulletin of Entomological Research ◽

10.1017/s0007485308006445 ◽

2008 ◽

Vol 99 (2) ◽

pp. 193-206 ◽

Cited By ~ 32

Author(s):

P.J. De Barro ◽

M.T. Coombs

Keyword(s):

Bemisia Tabaci ◽

Host Plants ◽

World Wide ◽

Order Of Magnitude ◽

Biotype B ◽

Magnitude Increase ◽

B Biotype

AbstractBemisia tabaci biotype B is a significant pest of agriculture world-wide. It was first detected in Australia in 1994. Assessments of the potential of parasitoids already present in Australia to control this pest indicated that two species of Eretmocerus and 11 species of Encarsia were present, but they did not exert sufficient control with a combined average of 5.0±0.3% apparent parasitism of 4th instars. Further, only 25% of samples containing biotype B had parasitised individuals present. The surveys also identified that fewer B biotype were being parasitised compared with the Australian indigenous biotype. Overall, Er. mundus was the most abundant parasitoid prior to the introduction. Previous research indicated that Er. hayati offered the best prospects for Australia and, in October 2004, the first releases were made. Since then, levels of apparent parasitism have averaged 29.3±0.1% of 4th instars with only 24% of collections having no parasitism present. Eretmocerus hayati contributed 85% of the overall apparent parasitism. In addition, host plants of the whitefly with low or no parasitism prior to the release have had an order of magnitude increase in levels of parasitism. This study covers the establishment of the case to introduce Er. hayati and the post-release establishment period November 2004–March 2008.

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SCAR molecular markers of the B biotype and two non-B populations of the whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae)

Chinese Journal of Agricultural Biotechnology ◽

10.1079/cjb2006108 ◽

2006 ◽

Vol 3 (3) ◽

pp. 189-194 ◽

Cited By ~ 13

Author(s):

Zang Lian-Sheng ◽

Jiang Tong ◽

Xu Jing ◽

Liu Shu-Sheng ◽

Zhang You-Jun

Keyword(s):

Bemisia Tabaci ◽

Random Amplified Polymorphic Dna ◽

Trialeurodes Vaporariorum ◽

Chain Reaction ◽

Specific Sequence ◽

Sequence Characterized Amplified Region ◽

Scar Primer ◽

Polymerase Chain ◽

Scar Primers ◽

B Biotype

AbstractRandom amplified polymorphic DNA (RAPD) analyses were performed with random primer H16 for the B biotype and two non-B populations of Bemisia tabaci collected from Zhejiang (China). The specific sequence fragments containing 446, 390 and 1317 nucleotides were amplified for the B biotype, ZHJ-1, ZHJ-2 populations, respectively. The three specific fragments were cloned and sequenced, and three pairs of SCAR primers were designed according to the sequences determined. With improvement of the conditions of the polymerase chain reaction (PCR), the specific fragments of B biotype, ZHJ-1 and ZHJ-2 populations, namely 439, 366 and 1238 nucleotides, respectively, were amplified with the sequence characterized amplified region (SCAR) primer of the corresponding population, while specific fragments of the other populations of B. tabaci or Trialeurodes vaporariorum could not be amplified.

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Developing a non-destructive metabarcoding protocol for detection of pest insects in bulk trap catches

Scientific Reports ◽

10.1038/s41598-021-85855-6 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Jana Batovska ◽

Alexander M. Piper ◽

Isabel Valenzuela ◽

John Paul Cunningham ◽

Mark J. Blacket

Keyword(s):

Cost Effective ◽

Diagnostic Techniques ◽

Morphological Identification ◽

Bactericera Cockerelli ◽

Agricultural Pests ◽

Community Size ◽

Field Samples ◽

Trap Catches ◽

Non Destructive ◽

Mock Communities

AbstractMetabarcoding has the potential to revolutionise insect surveillance by providing high-throughput and cost-effective species identification of all specimens within mixed trap catches. Nevertheless, incorporation of metabarcoding into insect diagnostic laboratories will first require the development and evaluation of protocols that adhere to the specialised regulatory requirements of invasive species surveillance. In this study, we develop a multi-locus non-destructive metabarcoding protocol that allows sensitive detection of agricultural pests, and subsequent confirmation using traditional diagnostic techniques. We validate this protocol for the detection of tomato potato psyllid (Bactericera cockerelli) and Russian wheat aphid (Diuraphis noxia) within mock communities and field survey traps. We find that metabarcoding can reliably detect target insects within mixed community samples, including specimens that morphological identification did not initially detect, but sensitivity appears inversely related to community size and is impacted by primer biases, target loci, and sample indexing strategy. While our multi-locus approach allowed independent validation of target detection, lack of reference sequences for 18S and 12S restricted its usefulness for estimating diversity in field samples. The non-destructive DNA extraction proved invaluable for resolving inconsistencies between morphological and metabarcoding identification results, and post-extraction specimens were suitable for both morphological re-examination and DNA re-extraction for confirmatory barcoding.

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Identification, geographical distribution and host plants of Bemisia tabaci (Genn.) biotypes (Homoptera: Aleyrodidae) in the State of Paraná, Brazil

Anais da Sociedade Entomológica do Brasil ◽

10.1590/s0301-80592000000300023 ◽

2000 ◽

Vol 29 (3) ◽

pp. 597-603 ◽

Cited By ~ 3

Author(s):

Sueli S. Martinez ◽

Alfredo O. R. de Carvalho ◽

Luiz G. Vieira ◽

Liliane M. Nunes ◽

Anésio Bianchini

Keyword(s):

Geographical Distribution ◽

Bemisia Tabaci ◽

Banding Pattern ◽

Host Plants ◽

Ornamental Plants ◽

The State ◽

The North ◽

Rapd Pcr ◽

Biotype B

This work was carried out in order to identify Bemisia tabaci (Genn.) biotypes present in the state of Paraná and to determine their geographical distribution and host plants. About 50 adults were collected in several host crops, weeds and ornamental plants in North, Northwest, Northeast, West and Central areas of the state, from January to May, 1998 and 1999. The species were identified by means of RAPD-PCR, using the primer Operon H-16. Whitefly populations were detected mainly from February on, in both years, seldom achieving more than one adult per leaf. The insect was found in only 66% of the sampled areas. Bean golden mosaic was almost never observed during the period. Both biotype A and biotype B of B. tabaci were found in the State of Paraná, the last one being more restricted to the north region. Although biotype B was found colonising a wider range of host plants, it has not spread out in all the state and most of the whitefly specimens found are still the biotype A. A banding pattern distinct of those obtained for A and biotype B of B. tabaci was obtained exclusively with the populations collected from cassava, thus indicating the possible presence of a biotype specific for this crop.

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Biology of Bemisia tabaci (Genn.) B-biotype and parasitism by Encarsia formosa (Gahan) on collard, soybean and tomato plants

Scientia Agricola ◽

10.1590/s0103-90162008000600011 ◽

2008 ◽

Vol 65 (6) ◽

pp. 639-642 ◽

Cited By ~ 4

Author(s):

Karina Manami Takahashi ◽

Evoneo Berti Filho ◽

André Luiz Lourenção

Keyword(s):

Plant Species ◽

Bemisia Tabaci ◽

Comparison Study ◽

Encarsia Formosa ◽

Tomato Plants ◽

Glycine Max L ◽

Biotype B ◽

Polyphagous Insect ◽

Hatching Eggs ◽

B Biotype

The silverleaf whitefly Bemisia tabaci (Genn.) B-biotype (= B. argentifolii) (Hemiptera: Aleyrodidae) is a polyphagous insect attacking many plant species of economic importance. A comparison study was conducted on the duration of the egg-to-adult period, and the percentage of hatching eggs of Bemisia tabaci (Genn.) B-biotype on collard (Brassica oleracea L. var. acephala D.C.), soybean(Glycine max (L.) Merr.) and tomato (Lycopersicon esculentum Mill.) plants, as well as the egg-to-adult period of Encarsia formosa (Gahan) on the 1st, 2nd, 3rd and 4th whitefly nymphal instars on these three plant species. The experiments were conducted in a laboratory (25ºC, 70 ± 10% RH, 14-hour photophase). The duration of the egg-to-adult period of B. tabaci was 19.8 days on collard, 21.2 days on soybean and 22.0 days on tomato. The number of hatched eggs was higher on collard when compared to soybean and tomato plants. Concerning E. formosa regardless of plant species, the duration for the egg-to-adult period was shorter for the 3rd and 4th instar nymphs as compared with the other instars.

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Siklus hidup dan statistik demografi kutukebul Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) biotipe B dan non-B pada tanaman cabai (Capsicum annuum L.)

Jurnal Entomologi Indonesia ◽

10.5994/jei.14.3.87 ◽

2018 ◽

Vol 14 (3) ◽

pp. 143

Author(s):

Purnama Hidayat ◽

Hazen Arrazie Kurniawan ◽

Lutfi Afifah ◽

Hermanu Triwidodo

Keyword(s):

Life Cycle ◽

Bemisia Tabaci ◽

Chili Pepper ◽

Reproduction Rate ◽

Increase Rate ◽

Biological Aspects ◽

Biotype B ◽

Length Of Life ◽

Net Reproduction ◽

B Biotype

The whitefly Bemisia tabaci (Gennadius) biotype B also known as Bemisia argentifolii (Gennadius) is a more malignant whitefly biotype in damaging plants compared to non-B biotype. Currently the whitefly B. tabaci biotype B has been reported its existence in Indonesia. Basic information such as life cycle, length of life, fecundity, and breeding ability of a whitefly is important information as a basis in preparing the whitefly control strategy. The aim of this research was to study the life cycle and demographic statistic of the B. tabaci biotype B and the non-B biotype on chili pepper. The study was conducted by observing the development of the whiteflies from eggs to adult in a grow chamber that the temperature and lighting were controlled. Observations were made on several aspects of biology and some parameters of demographic statistics. The results showed that the biotype B of B. tabaci has several different biological aspects with the non-B whitefly in chili pepper. The life cycle of the biotype B of B. tabaci and the non-B were different, 33.27 and 30.86 days respectively. The biotype B of B. tabaci had a net reproduction rate (R0) which was similar to that of the non-B biotype as well as the average of its generation. However, the intrinsic increase rate (r) of the biotype B B. tabaci was 2.5 times shorter than the non-B biotype. The biotype B of B. tabaci doubled its population (DT) 2 times faster than the non-B biotype. It is clear that the biotype B of B. tabaci potentially more dangerous than the non-B.

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Developing a Non-destructive Metabarcoding Protocol for Detection of Pest Insects in Bulk Trap Catches

10.21203/rs.3.rs-125070/v1 ◽

2020 ◽

Author(s):

Jana Batovska ◽

Alexander Piper ◽

Isabel Valenzuela ◽

John Cunningham ◽

Mark Blacket

Keyword(s):

Cost Effective ◽

Diagnostic Techniques ◽

Morphological Identification ◽

Bactericera Cockerelli ◽

Agricultural Pests ◽

Community Size ◽

Field Samples ◽

Trap Catches ◽

Non Destructive ◽

Mock Communities

Abstract Metabarcoding has the potential to revolutionise insect surveillance by providing high-throughput and cost-effective species identification of all specimens within mixed trap catches. Nevertheless, incorporation of metabarcoding into insect diagnostic laboratories will first require the development and evaluation of protocols that adhere to the specialised regulatory requirements of invasive species surveillance. In this study, we develop a multi-locus non-destructive metabarcoding protocol that allows sensitive detection of agricultural pests, and subsequent confirmation using traditional diagnostic techniques. We validate this protocol for the detection of tomato potato psyllid (Bactericera cockerelli) and Russian wheat aphid (Diuraphis noxia) within mock communities and field survey traps. We find that metabarcoding can reliably detect target insects within mixed community samples, including specimens that morphological identification did not initially detect, but sensitivity appears inversely related to community size and is impacted by primer biases, target loci, and sample indexing strategy. While our multi-locus approach allowed independent validation of target detection, lack of reference sequences for 18S and 12S restricted its usefulness for estimating diversity in field samples. The non-destructive DNA extraction proved invaluable for resolving inconsistencies between morphological and metabarcoding identification results, and post-extraction specimens were suitable for both morphological re-examination and DNA re-extraction for confirmatory barcoding.

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