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Passiflora mottle virus (PaMoV), an aphid-borne potyvirus, is the primary causal virus of the devastating passionfruit woodiness disease in Vietnam. Here we generated a non-pathogenic, attenuated PaMoV strain for disease control by cross protection. A full-length genomic cDNA of PaMoV strain DN4 from Vietnam was constructed to generate an infectious clone. The green fluorescent protein was tagged at the N-terminal region of the coat protein gene to monitor in planta the severe PaMoV-DN4. Two amino acids within the conserved motifs of HC-Pro of PaMoV-DN4 were mutated individually or in combination as K53E or/and R181I. Mutants PaMoV-E53 and PaMoV-I181 induced local lesions in Chenopodium quinoa plants, while PaMoV-E53I181 caused infection without apparent symptoms. In passionfruit plants, PaMoV-E53 elicited severe leaf mosaic, PaMoV-I181 induced leaf mottling, while PaMoV-E53I181 caused transient mottling followed by symptomless recovery. PaMoV- E53I181 was stable after six serial passages in yellow passionfruit plants. Its temporal accumulation levels were lower than those of the wild type, with a zigzag accumulation pattern, typical of a beneficial protective virus. An RNA silencing suppression (RSS) assay revealed that all three mutated HC-Pros are defective in RSS. Triplicated cross-protection experiments with a total of 45 plants showed that the attenuated mutant PaMoV-E53I181 provided a high protection rate (91%) against the homologous wild-type virus in passionfruit plants. This work revealed that PaMoV-E53I181 can be used as a protective virus to control PaMoV by cross protection.

Passionfruit plantings in Vietnam increased to 10,000 ha in 2019. However, outbreaks of passionfruit woodiness disease (PWD) have become a serious threat to production. In this study, five virus isolates (DN1, DN4, NA1, GL1, and GL2) were collected from different areas of Vietnam. Their causal roles in PWD were verified by back-inoculation to passionfruit. Analyses of coat protein (CP) and genomic sequences revealed that the GL1 isolate is closely related to East Asia Passiflora virus (EAPV) AO strain of Japan (polyprotein nt and aa identities of 98.1 and 98.2%, respectively), and the GL2 isolate is related to Telosma mosaic virus (TelMV) isolate PasFru, China (polyprotein nt and aa identities of 87.1 and 90.9%, respectively). CP comparison, host range, and cytological characterization indicated that DN1, DN4, and NA1 are potyviruses but are different from EAPV and TelMV. Phylogenic analyses of their CP and genome sequences indicated that these three isolates and the passionfruit severe mottle-associated virus Fujian isolate of China belong to a distinct clade, which does not meet the threshold (76% nt identity of polyprotein) to be regarded as any of potyviral species. Thus, a new species name, Passiflora mottle virus, (PaMoV), has been proposed by the International Committee on Taxonomy of Viruses. A rabbit antiserum was produced against the CP of DN1, and it can distinguish PaMoV from TelMV and EAPV in western blotting and enzyme-linked immunosorbent assay (ELISA) without cross-reactions. Field surveys of 240 samples by ELISA and reverse transcription PCR found that PWD in Vietnam is caused mainly by PaMoV, followed by EAPV, mixed infection of PaMoV and EAPV, and rare cases of TelMV.

A survey conducted on newly cultivated lowland rice fields by direct seeding method in Hai Duong Province, Viet Nam, in March 2017 revealed high devastation of the field. In these fields, farmers used an annual crop rotation cycle of rice-scallion-rice. Investigations on the devastated fields revealed that the chemical and physical soil properties were appropriate for rice cultivation. On the other hand, observations done on the root systems showed that the dead plants have symptomatic root galls suggesting the presence of plant parasitic nematodes. Sequencing of the internal transcribed spacer (ITS) region of the rDNA genes of the nematodes showed that the root nematodes extracted from the infested fields belonged to Meloidogyne graminicola. The reproductive factor of the isolated M. graminicola population on the IR64 rice variety (Oryza sativa indica) was normal, suggesting that the impact of this plant pest was not due to the emergence of an unusual virulent population. The combination of the three factors (wrong cropping choice for rotation, using rice variety susceptible to M. graminicola and direct seeding) were obviously promoting the nematode infection and its high proliferation in the surveyed fields. Meloidogyne graminicola could parasitize and propagate in scallions of Vietnam. Since this plant is annually cultivated on a paddy field for crop rotation, preventive measures or alternative plant for crop rottion is necessary.