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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.

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