Cryptophlebia leucotreta granulovirus-SA (CrleGV-SA) is used as a commercial biopesticide for the false codling moth, Thaumatotibia leucotreta, in citrus and other crops. The virus is sensitive to UV irradiation from sunlight, which reduces its efficacy as a biopesticide in the field. We selected a UV-resistant CrleGV-SA isolate, with more than a thousand-fold improved virulence compared to the wild-type isolate, measured by comparing LC50 values. CrleGV-SA purified from infected T. leucotreta larvae was exposed to UV irradiation under controlled laboratory conditions in a climate chamber mimicking field conditions. Five cycles of UV exposure, followed by propagating the virus that retained infectivity in vivo with re-exposure to UV, were conducted to isolate and select for UV-resistant virus. Serial dilution bioassays were conducted against neonates after each UV exposure cycle. The concentration-responses of the infectious UV-exposed virus populations were compared by probit analysis with those from previous cycles and from the original CrleGV-SA virus population. NGS sequences of CrleGV-SA samples from UV exposure cycle 1 and cycle 5 were compared with the GenBank CrleGV-SA sequence. Changes in the genomes of infective virus from cycles 1 and 5 generated SNPs thought to be responsible for establishing UV tolerance. Additional SNPs, detected only in the cycle 5 sequence, may enhance UV tolerance and improve the virulence of the UV-tolerant population.

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Cryptophlebia leucotreta granulovirus (CrleGV), a double-stranded DNA virus (genus Betabaculovirus, family Baculoviridae), is highly infective to the citrus insect pest Thaumatotibia leucotreta. The South African isolate CrleGV-SA is formulated into a commercial biopesticide and registered for use in several countries. In South Africa, it is used as a biopesticide in a multi-faceted integrated pest management approach for citrus crops involving chemical and biological control methods. The virus nucleocapsid is surrounded and protected by an occlusion body (OB) composed of granulin protein in a crystalline matrix. Like all other baculoviruses, CrleGV is susceptible to ultraviolet (UV) radiation from sunlight. This reduces its efficacy as a biopesticide in the field and necessitates frequent respraying. UV damage to baculovirus biopesticides is detected by means of functional bioassays. However, bioassays do not give an indication of whether any structural damage has occurred that may contribute to functional loss. In this study, transmission electron microscopy (TEM) was used to observe damage to the OB and nucleocapsid (NC) of CrleGV-SA, following controlled UV irradiation in the laboratory to mimic field conditions. The resultant images were compared with images of non-irradiated CrleGV-SA virus. TEM images of irradiated CrleGV-SA samples revealed changes to the OB crystalline faceting, a reduction in the size of the OBs, and damage to the NC following UV exposure for 72 h.