The New Zealand perspective of an ecosystem biology response to grapevine leafroll disease.

Grapevine leafroll-associated virus 3 (GLRaV-3) is a major pathogen of grapevines worldwide resulting in grapevine leafroll disease (GLD), reduced fruit yield, berry quality and vineyard profitability. Being graft transmissible, GLRaV-3 is also transmitted between grapevines by multiple hemipteran insects (mealybugs and soft scale insects). Over the past 20 years, New Zealand has developed and utilized integrated pest management (IPM) solutions that have slowly transitioned to an ecosystem-based biological response to GLD. These IPM solutions and combinations are based on a wealth of research within the temperate climates of New Zealand's nation-wide grape production. To provide context, the grapevine viruses present in the national vineyard estate and how these have been identified are described; the most pathogenic and destructive of these is GLRaV-3. We provide an overview of research on GLRaV-3 genotypes and biology within grapevines and describe the progressive development of GLRaV-3/GLD diagnostics based on molecular, serological, visual, and sensor-based technologies. Research on the ecology and control of the mealybugs Pseudococcus calceolariae and P. longispinus, the main insect vectors of GLRaV-3 in New Zealand, is described together with the implications of mealybug biological control agents and prospects to enhance their abundance and/or fitness in the vineyard. Virus transmission by mealybugs is described, with emphasis on understanding the interactions between GLRaV-3, vectors, and plants (grapevines, alternative hosts, or non-hosts of the virus). Disease management through grapevine removal and the economic influence of different removal strategies is detailed. Overall, the review summarizes research by an interdisciplinary team working in close association with the national industry body, New Zealand Winegrowers. Teamwork and communication across the whole industry has enabled implementation of research for the management of GLD.

Grapevine Leafroll-Associated Virus 3 Genotype Influences Foliar Symptom Development in New Zealand Vineyards.

Grapevine leafroll disease (GLD) constrains wine production worldwide. In New Zealand, the main causal agent of GLD is grapevine leafroll-associated virus 3 (GLRaV-3). To control GLD, an integrated management program is used and includes removing (roguing) GLRaV-3-infected vines from the vineyard. The classical foliar symptoms from virus-infected red-berry cultivars are leaves with dark red intervein, green veins, and downward rolling of margins. Growers use these phenotypic cues to undertake visual symptom identification (VSI) for GLD. However, the influence of the known large genetic variation among GLRaV-3 isolates on the foliar symptoms from different grapevine cultivars remains undescribed, especially in cool-climate growing environments, such as New Zealand. Over three vintages (2015, 2016, and 2017), VSI for GLD was undertaken at three field sites in New Zealand (Auckland, Hawke's Bay, and Marlborough), each including four cultivars (Merlot, Pinot noir, Sauvignon blanc, and Pinot gris) infected with three GLRaV-3 genotypes (Groups I, VI, and X) or GLRaV-3-uninfected control plants. Throughout this study, no visual symptoms were observed on white-berry cultivars infected with GLRaV-3. For red-berry cultivars, the greatest variability in observed foliar symptoms among regional study sites, cultivars, and GLRaV-3 genotypes was observed early in the growing season. In particular, Group X had significantly delayed symptom expression across all three sites compared with Groups I and VI. As the newly infected, young vines matured in years 2 and 3, the GLRaV-3 genotype, cultivar, region, and environmental conditions had minimal influence on the accuracy of VSI, with consistently high (>95%) within-vintage identification by the end of each vintage. The results from this study strongly support the use of VSI for the GLD management of red-berry cultivar grapevines, Merlot and Pinot noir, as a reliable and cost-effective tool against GLD.

Retention and Transmission of Grapevine Leafroll-Associated Virus 3 by Pseudococcus calceolariae.

Grapevine leafroll-associated virus 3 (GLRaV-3), an economically significant pathogen of grapevines, is transmitted by Pseudococcus calceolariae, a mealybug commonly found in New Zealand vineyards. To help inform alternative GLRaV-3 control strategies, this study evaluated the three-way interaction between the mealybug, its plant host and the virus. The retention and transmission of GLRaV-3 by P. calceolariae after access to non-Vitis host plants (and a non-GLRaV-3 host) White clover (Trifolium repens L. cv. "Grasslands Huia white clover"), Crimson clover (T. incarnatum), and Nicotiana benthamiana (an alternative GLRaV-3 host) was investigated. For all experiments, P. calceolariae first instars with a 4 or 6 days acquisition access period on GLRaV-3-positive grapevine leaves were used. GLRaV-3 was detected in mealybugs up to 16 days on non-Vitis plant hosts but not after 20 days. GLRaV-3 was retained by second instars (n = 8/45) and exuviae (molted skin, n = 6/6) following a 4 days acquisition period on infected grapevines leaves and an 11 days feeding on non-Vitis plant hosts. Furthermore, GLRaV-3 was transmitted to grapevine (40-60%) by P. calceolariae second instars after access to white clover for up to 11 days; 90% transmission to grapevine was achieved when no alternative host feeding was provided. The 16 days retention period is the longest observed in mealybug vectoring of GLRaV-3. The results suggest that an alternative strategy of using ground-cover plants as a disrupter of virus transmission may be effective if mealybugs settle and continue to feed on them for 20 or more days.

The Long-Term Effects of Reduced Competitive Ability on Foraging Success of an Invasive Pest Species.

Ant species like Pheidole megacephala (F.), Solenopsis invicta (Buren), and the Argentine ant, Linepithema humile (Mayr), have repeatedly been reported to be strongly associated with honeydew-producing arthropods like aphids, scale insects, and mealybugs, effectively protecting them from biological control agents like parasitoids. Here we report the results of a successful trial using pheromone dispensers to suppress Argentine ant activity over large sections in a commercial vineyard over a period of two months and preventing ant access into and foraging within the vine canopy. We found Argentine ant activity to be significantly reduced in pheromone-treated plots for the duration of the trial period compared with control plots. Our results showed a significant reduction in the numbers of Argentine ant workers recruited to randomly placed food resources within treated plots compared with untreated plots. Furthermore, spatial distribution of Argentine ants alongside transects in untreated plots remained relatively continuous, while increasing sharply beyond the borders of treated plots. Lastly, we measured the body fat content of workers and found a significant reduction in fat among workers from treated plots compared with untreated plots, suggesting an adverse effects on nest fitness. Additionally, we provide an initial assessment of the feasibility of the presented approach. Our results showed that it is possible to control Argentine ant, preventing them access to and foraging within the vine canopy, thereby reducing Argentine ants' access to honeydew.

Synthetic pheromones as a management technique - dispensers reduce Linepithema humile activity in a commercial vineyard.

BACKGROUND: Invasive ants, such as the Argentine ant, have often been reported to facilitate honeydew-producing hemipteran pests such as mealybugs, which can be vectors of plant pathogens. Synthetic pheromones may offer a target-specific method to control such ants and consequently lower the abundance of honeydew-producing pests. Here we report the results of a trial to suppress Argentine ants in grapevines using ant pheromone dispensers. RESULTS: Compared with untreated controls, we observed a significant drop in Argentine ant activity on the ground, irrespective of whether pheromone dispensers were placed at ground level, within the canopy or in both locations. Ant counts in the canopy confirmed that Argentine ant abundance was reduced under the influence of the pheromone dispenser placed at ground level compared with untreated controls. However, placing dispensers only in the canopy did not reduce the numbers of ants within the canopy compared with untreated controls. CONCLUSION: Our results showed that pheromone dispensers can significantly reduce Argentine ant foraging in grapevines if they are positioned appropriately. This technique could potentially reduce the abundance of associated mealybugs and potentially attendant virus vectoring areawide.

Enantiospecific Synthesis of Both Enantiomers of the Longtailed Mealybug Pheromone and Their Evaluation in a New Zealand Vineyard.

The irregular monoterpenoid sex pheromone of Pseudococcus longispinus and its enantiomer were prepared from the corresponding bornyl acetates. The use of readily accessible chiral starting materials and lactone-lactone rearrangement are the highlights of the present synthesis. The biological activities of the two enantiomers and racemic mixture were tested in a New Zealand vineyard. The (S)-(+)-enantiomer was significantly more attractive to P. longispinus males than the racemic mixture or the (R)-(-)-enantiomer.

Spatial analysis of mass trapping: how close is close enough?

BACKGROUND: The identification of new attractants can present opportunities for developing mass trapping, but standard screening methods are needed to expedite this. We have developed a simple approach based on quantifying trap interference in 4 × 4 trap arrays with different spacings. We discuss results from sex pheromones in Lepidoptera (light brown apple moth, Epiphyas postvittana), Diptera (apple leaf curling midge, Dasineura mali) and Homoptera (citrophilous mealybug, Pseudococcus calceolariae), compared with a kairomone for New Zealand flower thrips (Thrips obscuratus). RESULTS: The ratio of catch in corner traps to catch in centre traps was 25:1 at 750 D. mali traps ha(-1) , and was still ∼5:1 at 16 traps ha(-1) , suggesting trap interference even at such low trap densities. Trap competition for sex pheromone lures at close spacing (<5 m) was evident in 16-trap arrays of P. calceolariae, but less so for E. postvittana. No trap competition was observed at 4 m spacings with the kairomone for T. obscuratus. CONCLUSIONS: The ratio of catch in traps in the corner and centre of a 16-trap array at different spacings offers a rapid preliminary assessment method for determining the potential for mass trapping. Additional knowledge of vital rates and dispersal is needed for predicting population suppression. Our approach should have value in mass trapping development. © 2014 Society of Chemical Industry.

Ecology and management of grapevine leafroll disease.

Grapevine leafroll disease (GLD) is caused by a complex of vector-borne virus species in the family Closteroviridae. GLD is present in all grape-growing regions of the world, primarily affecting wine grape varieties. The disease has emerged in the last two decades as one of the major factors affecting grape fruit quality, leading to research efforts aimed at reducing its economic impact. Most research has focused on the pathogens themselves, such as improved detection protocols, with limited work directed toward disease ecology and the development of management practices. Here we discuss the ecology and management of GLD, focusing primarily on Grapevine leafroll-associated virus 3, the most important virus species within the complex. We contextualize research done on this system within an ecological framework that forms the backbone of the discussion regarding current and potential GLD management strategies. To reach this goal, we introduce various aspects of GLD biology and ecology, followed by disease management case studies from four different countries and continents (South Africa, New Zealand, California-USA, and France). We review ongoing regional efforts that serve as models for improved strategies to control this economically important and worldwide disease, highlighting scientific gaps that must be filled for the development of knowledge-based sustainable GLD management practices.

The absolute configuration of the sex pheromone of the citrophilous mealybug, Pseudococcus calceolariae.

The absolute configuration of the sex pheromone of the citrophilous mealybug, Pseudococcus calceolariae, was determined to be (1R,3R)-[2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropyl]methyl (R)-2-acetoxy-3-methylbutanoate. NMR, derivatization reactions, chiral gas chromatography-mass spectrometry, and comparison with synthetic chiral reference compounds, were used to determine the absolute configuration of this compound. This activity of this compound was further confirmed by testing synthetic stereoisomers of the compound as lures in traps for adult male mealybugs. Traps baited with 1,000 µg of the pheromone compound caught 36 times more males than traps baited with virgin females. A mixture of stereoisomers of the pheromone compound can be used for field trapping without adverse effects on trap catch. A comparison with the structures of other sex pheromones of mealybugs is presented.

Trapping Dasinuera mali (Diptera: Cecidomyiidae) in apples.

The midge Dasineura mali (Kieffer) (Diptera: Cecidomyiidae) is a significant pest of apples (Malus spp.), and the recent identification of the female sex pheromone is enabling new direct control tactics to be considered. Direct control using male suppression will require knowledge of the frequency of multiple mating, dispersal and colonization rates, and the efficiency of male removal. Males were able to mate up to five times, with a mean of 2.7 times when presented in a 10 female-to-1 male group, designed to simulate male suppression. Male catch in response to the pheromone loading was curvilinear over 4 orders of magnitude from 3 microg to 30 mg on rubber septa. Trapping using a high-dose pheromone lure was combined with oil-based traps similar to the inexpensive New Zealand "Lynfield trap" used for tephritid surveillance, to test male suppression in young orchard blocks at 500 traps per ha. Monitoring traps indicated 96% lower catch in the treated plots compared with control plots, over 137 d. However, a lack of shoot tip infestation in both treated and untreated plots indicated limited colonization and prevented an assessment of potential population suppression. Furthermore, a contribution to these results from communication disruption cannot be ruled out. Replicated transects of frequency of infested shoots from a mature orchard across the adjacent young block confirmed that colonization by ovipositing females was essentially limited to the first 30 m.