Characterization and Distribution of a Potyvirus Associated with Passion Fruit Woodiness Disease in Uganda

This article describes the incidence and etiology of a viral disease of passion fruit in Uganda. Symptoms, including those characteristic of passion fruit woodiness disease (PWD), were observed on 32% of plants in producing areas. Electron microscopic observations of infected tissues revealed flexuous filaments of ca. 780 nm. Enzymelinked immunosorbent assays indicated a serological relationship with Cowpea aphid-borne mosaic virus (CABMV) and Passion fruit ringspot virus (PFRSV). In host range studies, only species in the families Solanaceae and Chenopodiaceae were susceptible, and neither Vigna unguiculata nor Phaseolus vulgaris became infected. Coat protein (CP) gene sequences of eight isolates exhibited features typical of potyviruses and were highly similar (88 to 100% identity). However, the sequences had limited sequence identity with CP genes of two of the three potyviruses reported to cause PWD: East Asian Passiflora virus and Passion fruit woodiness virus (PWV). Deduced amino acid sequences for the CP of isolates from Uganda had highest identity with Bean common mosaic necrosis virus (BCMNV) (72 to 79%, with evolutionary divergence values between 0.17 and 0.19) and CABMV (73 to 76%, with divergence values between 0.21 and 0.25). Based on these results and in accordance with International Committee for Taxonomy of Viruses criteria for species demarcation in the family Potyviridae, we conclude that a previously unreported virus causes viral diseases on passion fruit in Uganda. The name “Ugandan Passiflora virus” is proposed for this virus.

Resistance to Passion fruit woodiness virus in Transgenic Passionflower Expressing the Virus Coat Protein Gene

We report the use of the coat protein (CP) gene from Passion fruit woodiness virus (PWV) to produce resistant transgenic plants of yellow passion fruit. A full-length CP gene from a severe PWV isolate from the state of São Paulo, Brazil (PWV-SP) was cloned into pCAMBIA 2300 binary vector, which was further introduced into Agrobacterium tumefaciens strain EHA 105. Leaf disks were used as explants for transformation assays, e.g., 2,700 and 2,730 disks excised from plants from the Brazilian cultivars IAC-275 and IAC-277, respectively. In vitro selection was performed in kanamycin. After transferring to the elongation medium, 119 and 109 plantlets of IAC-275 and IAC-277, respectively, were recovered. Integration of the PWV CP gene was confirmed in seven of eight plants evaluated by Southern blot analysis, showing different numbers of insertional events for the CP gene. Three transgenic plants (T3, T4, and T7) expressed the expected transcript, but the 32 kDa PWV CP was detected by Western blot in only two plants (T3 and T4). The results of three successive mechanical inoculations against the transgenic plants using three PWV isolates showed that the primary transformant T2 of IAC-277 was immune to all isolates.

Protection between strains of Passion fruit woodiness virus in sunnhemp

The main objective of the present study was to evaluate the effect of the sunhemp (Crotalaria juncea) host species on the protective ability of two mild strains of Passion fruit woodiness virus (PWV), named F-101 and F-144, which had failed to protect passion flowers (Passiflora edulis f. flavicarpa) in previous experiments. The nucleotide sequences of the capsid protein (CP) gene and the 3'-non-translated region (3'-NTR) of these mild strains and the severe strain of PWV-SP were compared to confirm their relationship. The results of two protective tests with sunhemp plants in the greenhouse and one test under field conditions showed that all plants infected with either mild strain were protected against infection and/or symptom expression of the severe strain of PWV-SP. Evaluation of the relative concentration of the mild strains in sun hemp leaves showed an apparent uniformity in virus distribution in the leaf tissues, different than that which was previously reported for these mild strains in passion flower leaves. These results agree with previous studies that showed the effect of the concentration of the protective strains and the host species in the protection process.