Banana Tree Infected with Banana Bunchy Top Virus Attracts Pentalonia nigronervosa Aphids Through Increased Volatile Organic Compounds Emission.

Banana plants are affected by various viral diseases, among which the most devastating is the "bunchy top", caused by the Banana bunchy top virus (BBTV) and transmitted by the aphid Pentalonia nigronervosa Coquerel. The effect of BBTV on attraction mechanisms of dessert and plantain banana plants on the vector remains far from elucidated. For that, attractiveness tests were carried out using a two columns olfactometer for apterous aphids, and a flight cage experiment for alate aphids. Volatile Organic Compounds (VOCs) emitted by either healthy or BBTV-infected banana plants were identified using a dynamic extraction system and gas-chromatography mass-spectrometry (GC-MS) analysis. Behavioral results revealed a stronger attraction of aphids towards infected banana plants (independently from the variety), and towards the plantain variety (independently from the infection status). GC-MS results revealed that infected banana plants produced VOCs of the same mixture as healthy banana plants but in much higher quantities. In addition, VOCs produced by dessert and plantain banana plants were different in nature, and plantains produced higher quantities than dessert banana trees. This work opens interesting opportunities for biological control of P. nigronervosa, for example by luring away the aphid from banana plants through manipulation of olfactory cues.

ICTV Virus Taxonomy Profile: Nanoviridae.

Nanoviridae is a family of plant viruses (nanovirids) whose members have small isometric virions and multipartite, circular, single-stranded (css) DNA genomes. Each of the six (genus Babuvirus) or eight (genus Nanovirus) genomic DNAs is 0.9-1.1 kb and is separately encapsidated. Many isolates are associated with satellite-like cssDNAs (alphasatellites) of 1.0-1.1 kb. Hosts are eudicots, predominantly legumes (genus Nanovirus), and monocotyledons, predominantly in the order Zingiberales (genus Babuvirus). Nanovirids require a virus-encoded helper factor for transmission by aphids in a circulative, non-propagative manner. This is a summary of the ICTV Report on the family Nanoviridae, which is available at ictv.global/report/nanoviridae.

Estimating a novel stochastic model for within-field disease dynamics of banana bunchy top virus via approximate Bayesian computation.

The Banana Bunchy Top Virus (BBTV) is one of the most economically important vector-borne banana diseases throughout the Asia-Pacific Basin and presents a significant challenge to the agricultural sector. Current models of BBTV are largely deterministic, limited by an incomplete understanding of interactions in complex natural systems, and the appropriate identification of parameters. A stochastic network-based Susceptible-Infected-Susceptible model has been created which simulates the spread of BBTV across the subsections of a banana plantation, parameterising nodal recovery, neighbouring and distant infectivity across summer and winter. Findings from posterior results achieved through Markov Chain Monte Carlo approach to approximate Bayesian computation suggest seasonality in all parameters, which are influenced by correlated changes in inspection accuracy, temperatures and aphid activity. This paper demonstrates how the model may be used for monitoring and forecasting of various disease management strategies to support policy-level decision making.

Cardamom Bushy Dwarf Virus Infection in Large Cardamom Alters Plant Selection Preference, Life Stages, and Fecundity of Aphid Vector, Micromyzus kalimpongensis (Hemiptera: Aphididae).

Cardamom bushy dwarf virus (CBDV) causes foorkey disease of large cardamom (Ammomum subulatum Roxburgh) in the eastern sub-Himalayan mountains. Although the aphid Micromyzus kalimpongensis Basu (Hemiptera: Aphididae) is known as a vector of CBDV, its behavior in dissemination of CBDV has not been investigated. In the present study, M. kalimpongensis was observed to colonize in higher number on CBDV-infected large cardamom plants compared with the healthy plants in the several plantations in Sikkim and Darjeeling hills. The affinity of M. kalimpongensis to the diseased large cardamom plants was further confirmed in a contained field experiment with intact plant as well as in a laboratory bioassay with the plant extract, where significantly higher number of aphids settled on the diseased plants or extracts compared with the healthy counterparts. Aphids grown on CBDV-infected large cardamom plants had shortened nymphal period and increased longevity and fecundity compared with those grown on the healthy plants. In the contained field experiment, M. kalimpongensis migrated to the CBDV-infected plants, colonized there, acquired CBDV, and once the diseased plants withered, migrated to healthy plants, which eventually became diseased. Our results suggest a general pattern of spread of CBDV by M. kalimpongensis where CBDV-infected plants attract or arrest and stimulate emergence and migration of viruliferous aphids that otherwise are sedentary in the underground plant parts of large cardamom. To our knowledge, this is the first study that shows the influence of a plant virus from the family Nanoviridae in altering behavior of its insect vector that favors its dissemination.

Localization, concentration, and transmission efficiency of Banana bunchy top virus in four asexual lineages of Pentalonia aphids.

Banana bunchy top virus (BBTV) is the most destructive pathogenic virus of banana plants worldwide. The virus is transmitted in a circulative non-propagative manner by the banana aphid, Pentalonia nigronervosa Coquerel. In this work, we examined the localization, accumulation, and transmission efficiency of BBTV in four laboratory-established lineages of Pentalonia aphids derived from four different host plants: taro (Colocasia esculenta), heliconia (Heliconia spp.), red ginger (Alpinia purpurata), and banana (Musa sp.). Mitochondrial sequencing identified three and one lineages as Pentalonia caladii van der Goot, a recently proposed species, and P. nigronervosa, respectively. Microsatellite analysis separated the aphid lineages into four distinct genotypes. The transmission of BBTV was tested using leaf disk and whole-plant assays, both of which showed that all four lineages are competent vectors of BBTV, although the P. caladii from heliconia transmitted BBTV to the leaf disks at a significantly lower rate than did P. nigronervosa. The concentration of BBTV in dissected guts, haemolymph, and salivary glands was quantified by real-time PCR. The BBTV titer reached similar concentrations in the guts, haemolymph, and salivary glands of aphids from all four lineages tested. Furthermore, immunofluorescence assays showed that BBTV antigens localized to the anterior midguts and the principal salivary glands, demonstrating a similar pattern of translocations across the four lineages. The results reported in this study showed for the first time that P. caladii is a competent vector of BBTV.

Tropism, compartmentalization and retention of banana bunchy top virus (Nanoviridae) in the aphid vector Pentalonia nigronervosa.

Plant viruses of the families Luteoviridae and Geminiviridae rely on hemipteran vectors for the infection of their hosts. Several lines of evidence have revealed that these viruses are transmitted by competent vectors in a circulative manner, involving entry into the vector's body and the crossing of epithelial tissues forming the alimentary tract and the salivary glands. Similar to luteovirids and geminiviruses, a third family of plant viruses, the family Nanoviridae, have also been reported to be transmitted by aphids in a circulative manner. However, there is limited direct evidence of a possible path of translocation through the aphid vectors. Here, we used time-course experiments and transmission assays coupled with real-time PCR and immunofluorescence assays on dissected tissues to examine the translocation, compartmentalization and retention of banana bunchy top virus (BBTV) into the aphid vector Pentalonia nigronervosa. Our results indicate that BBTV translocates rapidly through the aphid vector; it is internalized into the anterior midgut in which it accumulates and is retained at concentrations higher than either the haemolymph or the principal salivary glands. Despite the large increase in viral concentration, we have failed to detect BBTV transcripts with RT-PCR. When tissues were not permeabilized, BBTV localized as distinct puncta in the proximity of the basal surface of the cells forming the anterior midgut and principal salivary glands, suggesting an on-going process of virion escape and internalization, respectively. Interestingly, we document that those organs can have direct contact within the aphid body, suggesting a possible haemolymph-independent translocation path.

Predicting the benefits of banana bunchy top virus exclusion from commercial plantations in Australia.

Benefit cost analysis is a tried and tested analytical framework that can clearly communicate likely net changes in producer welfare from investment decisions to diverse stakeholder audiences. However, in a plant biosecurity context, it is often difficult to predict policy benefits over time due to complex biophysical interactions between invasive species, their hosts, and the environment. In this paper, we demonstrate how a break-even style benefit cost analysis remains highly relevant to biosecurity decision-makers using the example of banana bunchy top virus, a plant pathogen targeted for eradication from banana growing regions of Australia. We develop an analytical approach using a stratified diffusion spread model to simulate the likely benefits of exclusion of this virus from commercial banana plantations over time relative to a nil management scenario in which no surveillance or containment activities take place. Using Monte Carlo simulation to generate a range of possible future incursion scenarios, we predict the exclusion benefits of the disease will avoid Aus$15.9-27.0 million in annual losses for the banana industry. For these exclusion benefits to be reduced to zero would require a bunchy top re-establishment event in commercial banana plantations three years in every four. Sensitivity analysis indicates that exclusion benefits can be greatly enhanced through improvements in disease surveillance and incursion response.

Population distribution and density of Pentalonia nigronervosa (Hemiptera: Aphididae) within banana mats: influence of plant age and height on sampling and management.

The banana aphid, Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae), is the most economically important pest of banana (Musa spp.) fields in Hawaii. Recently, there has been a concerted effort in Hawaii to learn more about the biology and ecology of this pest. However, limited work has been directed at determining the distribution of P. nigronervosa in banana fields and developing an integrated pest management plan. Therefore, a survey was conducted in banana fields throughout the Hawaiian Islands to determine the distribution and density of P. nigronervosa within banana mats from plants of different stages. Another aim was to determine whether the presence of ants on banana plants could be used as a reliable indicator of aphid infestations. Results of the survey showed that plants < or = 1.5 m (small sucker) in height contain the highest aphid populations per meter in plant height and that mother plants (> or = 2.5 m) had the lowest aphid counts and rate of infestation compared with small and intermediate suckers (> 1.5 < 2.5 m). More specifically, aphid population was reduced by approximately 12 aphids for every meter increase in plant height and that aphids are rarely found > or = 2.5 m within the plant canopy. Although there was an increase likelihood of finding ants on banana plants with higher aphid densities, results suggest that ants would be present on plants in the absence of aphids. Implications of these and other findings with respect to sampling and managing P. nigronervosa and associated Banana bunchy top virus are discussed.

Immunofluorescence localisation of Banana bunchy top virus (family Nanoviridae) within the aphid vector, Pentalonia nigronervosa, suggests a virus tropism distinct from aphid-transmitted luteoviruses.

We have applied immunocapture PCR and developed an immunofluorescence assay to specifically detect Banana bunchy top virus (BBTV; family Nanoviridae, genus Babuvirus) within its aphid vector, Pentalonia nigronervosa (Hemiptera, Aphididae). BBTV was localised using either monoclonal or polyclonal antibodies into the anterior midgut (stomach) and into specific cells forming the principal salivary glands. These results suggest a distinct path of virus translocation that likely differs from the one described for aphid-transmitted luteovirus, which enter hemocoels through the hindguts and posterior midguts and that penetrate the accessory salivary glands of their competent vectors. To our understanding, this is the first work analysing the localisation of a virus member of the family Nanoviridae within an aphid vector.

Identification of two RNA silencing suppressors from banana bunchy top virus.

In order to suppress RNA silencing, many plant and some animal viruses encode RNA silencing suppressors to achieve infection. In this study, we report that B3 and B4, encoded by DNA3 and DNA4 of banana bunchy top virus (BBTV), exhibit RNA silencing suppression activity. B3 and B4 were able to increase the transient expression of green fluorescent protein (GFP) and dramatically enhanced the pathogenicity of potato virus X (PVX) in Nicotiana benthamiana. B4 was able to reverse established gene silencing on an inoculated leaf or on an upper leaf. B3, however, was only active during infection of an inoculated leaf. Furthermore, B4, but not B3, was able to enhance GFP expression in the transgenic N. benthamiana line 16c. In conclusion, B3 and B4 are the RNA silencing suppressors of BBTV, and they may act at different steps in the RNA silencing pathways.

Comparative susceptibility of two banana cultivars to Banana bunchy top virus under laboratory and field environments.

Field and laboratory experiments were carried out on the island of Oahu, HI, to compare the susceptibility of the two most commonly grown banana (Musa sp.) cultivars in the state ('Dwarf Brazilian' or Santa Catarina [locally known as dwarf apple] and 'Williams') to the aphid-borne Banana bunchy top virus (genus Babuvirus, family Nanoviridae, BBTV). Several morphological and physiological features of the two cultivars were monitored to determine whether the banana aphid, Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae), transmits BBTV to both cultivars at a similar rate; and whether after successful inoculation, does each cultivar respond similarly to viral infection. Results from the laboratory experiment showed that a similar percentage of both cultivars were infected with BBTV by aphid vectors (> 90% for both cultivars). However, field results showed a significantly lower percentage of dwarf apple (39%) infected with BBTV compared with Williams (79%). We also found that all physiological and morphological features measured (i.e., plant height, leaf area, canopy, chlorophyll level, and moisture content) for both cultivars were impacted similarly by BBTV. The incubation period, or the time between plant infection and initial appearance of disease symptoms, was similar for both cultivars. Results also showed that BBTV transmission efficiency was lower in the field than in the laboratory, despite that more aphids per plant were used for field than laboratory inoculation tests. The results highlight the potential use of less susceptible cultivars to help manage BBTV and the importance of screening banana varieties in the field to determine their response to vectors and associated diseases.

Unusual events involved in Banana bunchy top virus strain evolution.

Banana bunchy top virus (BBTV) can be transmitted by aphids and consists of at least six integral components (DNA-R, -U3, -S, -M, -C, and -N). Several additional replication-competent components (additional Reps) are associated with some BBTV isolates. A collected BBTV strain (TW3) that causes mild symptoms was selected to study the processes in BBTV evolution. Southern blot hybridization, polymerase chain reaction (PCR), and real-time PCR did not detect DNA-N in TW3. Real-time PCR quantification of BBTV components revealed that, except for the copy number of TW3 DNA-U3, each detected integral component of BBTV TW3 was at least two orders lower than that of the severe strains. No infection was observed in plants inoculated with aphids, which were first given acquisition access to the TW3-infected banana leaves. Recombination analysis revealed recombination between the integral component TW3 DNA-U3 and the additional Rep DNA-Y. All BBTV integral components contain a replication initiation region (stem-loop common region) that share high sequence identity. Sequence alignment revealed that TW3 DNA-R, -S, -M, and -C all have a stem-loop common region containing a characteristic 9-nucleotide deletion found only in all reported DNA-N. Our data suggest that the additional Rep DNAs can serve as sources of additional genetic diversity for integral BBTV components.

Aphid transmission of Banana bunchy top virus to bananas after treatment with a bananacide.

Field and laboratory studies were conducted to determine the impact of using a herbicide as a bananacide on aphid transmission of Banana bunchy top virus (family Nanoviridae, genus Babuvirus, BBTV) to healthy banana (Musa spp.) plants. BBTV-infected banana plants in a commercial orchard were treated with Roundup Weathermax herbicide. Using polymerase chain reaction, the time after herbicide treatment that BBTV could no longer be detected in the infected plants was determined. The impact of the herbicide treatment on Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae) virus acquisition and ability to inoculate healthy banana plants with BBTV also were determined. Generally, banana plants were dead beyond 42 d after herbicide injection (DAI), and BBTV was detected in a similar high percentage of treated plants from 0 up to 21 DAI. During two field trials, 0 and 32% of P. nigronervosa acquired the virus from treated plants at 42 DAI, respectively, but none successfully inoculated a healthy banana plant beyond 35 DAI. Finally, 22% of P. nigronervosa colonies collected directly from the pseudostem of injected plants at the final sample date (42 DAI) tested positive for BBTV and infected 9.5% of the healthy banana plants. The findings indicate that banana plants may remain a potential source of virus inoculum 6 wk after injection with a bananacide. The implications of these findings with respect to BBTV management are discussed.