Abstract
Watermelon (Citrullus lanatus) ranks among the top ten fruits cultivated globally, being prized for its fresh, nutritious fruit. In Uganda, watermelon is a significant fruit and cash crop, however, watermelon production is constrained by both abiotic and biotic factors, including viruses (Masika et al., 2022a). Fifteen leaf samples (four diseased and one non-symptomatic) were collected in 2020 from three commercial watermelon farms in Kween, Masaka and Mukono districts, Uganda. Two sets of leaf tissue were collected, each containing 12 diseased and three non-symptomatic leaf samples. One set was analysed using high-throughput sequencing (HTS) and the other was kept at -80°C to confirm the HTS results by RT-PCR and Sanger sequencing. Total RNA was extracted according to Lodhi et al. (1994); the HTS libraries were constructed from one RNA sample containing a pool of the 15 leaves using the TruSeq Stranded Total RNA with a Ribo-Zero Plant kit (Illumina Inc., USA), and sequenced by HTS using the HiSeq 2500 platform (Illumina Inc., USA) (150 bp paired ends) in South Africa. De novo assembly of HTS data was performed using the Genome Detective virus tool (Vilsker et al., 2019), generating a report containing the viral consensus sequences. A total of 8,682,658 raw reads were obtained (http://www.ncbi.nlm.nih.gov/bioproject/1191351). After cleaning and removal of non-viral reads, 7,038,818 and 865,648 reads were obtained. From the clean reads and assembled contigs, six viruses: Citrullus lanatus cryptic virus (CiLCV), Cucumis melo cryptic virus (CmCV), Moroccan watermelon mosaic virus (MWMV), pepo aphid-borne yellows virus (PABYV), Watermelon crinkle leaf-associated virus (WCLaV), and one novel virus, Escherichia coli virus, known to infect prokaryotes were identified. Such novel viruses were also reported by Togoobat et al. (2023). The complete genome of MWMV (GenBank Accession No. OM718862) was 9,672 nt without the poly(A) tail, and the PABYV genome (OM802496) was 5,798 nt. Since sequencing was conducted on pooled samples, we recognise the potential risk of chimera formation during the generation of consensus sequences. As a result, we confirmed the presence of viruses in the second set of samples which had been frozen. RT-PCR was performed with specific primers (MWMV-F: 5'-CATTCTCAACTTGGTGGGACGC-3' and R: 5'-CTGATGCTCTCCTCTGTGACAAG-3') for the cylindrical inclusion gene of MWMV (582 bp), and PABYV primers according to Masika et al. (2022b). Of the15 individual samples, 13 and 11 tested positive for MWMV and PABYV, respectively. One MWMV-positive amplicon was Sanger sequenced (Masaka, PQ397541) and showed 97% sequence identity with the HTS genome isolates. Nine of the 15 samples were infected with both viruses. One non-symptomatic sample from Kween tested positive for both viruses, while the two others were singly infected by MWMV. In addition, two diseased samples were singly infected with PABYV and MWMV, respectively. The singly-infected samples generally exhibited mild leaf symptoms, while mixed infections involving both viruses resulted in more severe symptoms, including mosaic patterns, distortion and pronounced growth retardation. The other identified viruses in our HTS analysis are likely latent or asymptomatic in watermelon under the conditions we surveyed. While detailed quantitative data on disease incidence was not undertaken, observational estimates indicated that approximately 30–40% of plants in the affected fields exhibited virus-like symptoms, including mosaic patterns, leaf mottling, vein clearing, and stunting (Fig. 1). This is the first report of MWMV in watermelon plants in Uganda; the reports of CiLCV, CmCV and WCLaV require further confirmation. The authors would like to acknowledge the financial support provided by the International Foundation for Science, Grant/Award Number: 1-1-C-6140-2
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Claims 12
Claim · model-written summary
MWMV is reported for the first time infecting watermelon plants in Uganda.
- Taxon
- watermelon
Citrullus lanatust0 - Interactor
- MWMVe1pathogen
- Outcome
- first report of MWMV in watermelon plantse1
- Setting
- Ugandae1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e1“This is the first report of MWMV in watermelon plants in Uganda”
Claim · model-written summary
Six viruses were identified by high-throughput sequencing in watermelon samples, including one novel virus known to infect prokaryotes.
- Taxon
- watermelonCitrullus lanatust0
- Interactor
- Citrullus lanatus cryptic virusCitrullus lanatus cryptic viruse2other
- Outcome
- six virusese2
- Qualifier
- Cucumis melo cryptic virus (CmCV), Moroccan watermelon mosaic virus (MWMV), pepo aphid-borne yellows virus (PABYV), Watermelon crinkle leaf-associated virus (WCLaV)e2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e2“From the clean reads and assembled contigs, six viruses: Citrullus lanatus cryptic virus (CiLCV), Cucumis melo cryptic virus (CmCV), Moroccan watermelon mosaic virus (MWMV), pepo aphid-borne yellows virus (PABYV), Watermelon crinkle leaf-associated virus (WCLaV), and one novel virus, Escherichia coli virus, known to infect prokaryotes were identified”
Claim · model-written summary
The MWMV complete genome is 9,672 nt without the poly(A) tail.
- Taxon
- watermelonCitrullus lanatust0
- Interactor
- MWMVe3pathogen
- Outcome
- complete genomee3
- Value
- 9,672 nte3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e3“The complete genome of MWMV (GenBank Accession No. OM718862) was 9,672 nt without the poly(A) tail”
Claim · model-written summary
The PABYV genome is 5,798 nt.
- Taxon
- watermelonCitrullus lanatust0
- Interactor
- PABYVe4pathogen
- Outcome
- PABYV genomee4
- Value
- 5,798 nte4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e4“the PABYV genome (OM802496) was 5,798 nt”
Claim · model-written summary
MWMV tested positive in 13 of the 15 individual samples.
- Taxon
- watermelonCitrullus lanatust0
- Interactor
- MWMVe5pathogen
- Outcome
- tested positivee5
- Value
- 13e5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e5“Of the15 individual samples, 13 and 11 tested positive for MWMV and PABYV”
Claim · model-written summary
PABYV tested positive in 11 of the 15 individual samples.
- Taxon
- watermelonCitrullus lanatust0
- Interactor
- PABYVe5pathogen
- Outcome
- tested positivee5
- Value
- 11e5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e5“Of the15 individual samples, 13 and 11 tested positive for MWMV and PABYV”
Claim · model-written summary
Nine of the 15 samples were infected with both viruses.
- Taxon
- watermelonCitrullus lanatust0
- Outcome
- infected with both virusese6
- Value
- Nine of the 15 samplese6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e6“Nine of the 15 samples were infected with both viruses”
Claim · model-written summary
Mixed infections with both viruses resulted in more severe symptoms than single infections.
- Taxon
- watermelonCitrullus lanatust0
- Treatment
- mixed infections involving both virusese7
- Outcome
- symptomse7
- Direction
- more severee7
- Comparator
- singly-infected samplese7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e7“The singly-infected samples generally exhibited mild leaf symptoms, while mixed infections involving both viruses resulted in more severe symptoms”
Claim · model-written summary
Singly infected watermelon samples generally exhibited mild leaf symptoms.
- Taxon
- watermelonCitrullus lanatust0
- Outcome
- leaf symptomse7
- Qualifier
- milde7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e7“The singly-infected samples generally exhibited mild leaf symptoms, while mixed infections involving both viruses resulted in more severe symptoms”
Claim · model-written summary
The other identified viruses are likely latent or asymptomatic in watermelon under the surveyed conditions.
- Taxon
- watermelonCitrullus lanatust0
- Outcome
- latent or asymptomatice0
- Qualifier
- likelye0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e0“The other identified viruses in our HTS analysis are likely latent or asymptomatic in watermelon under the conditions we surveyed”
Claim · model-written summary
Approximately 30–40% of plants in the affected fields exhibited virus-like symptoms.
- Taxon
- watermelonCitrullus lanatust0
- Outcome
- exhibited virus-like symptomse0
- Value
- 30–40%e0
- Qualifier
- approximatelye0
- Setting
- affected fieldse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e0“approximately 30–40% of plants in the affected fields exhibited virus-like symptoms”
Claim · model-written summary
One MWMV amplicon showed 97% sequence identity with the HTS genome isolates.
- Taxon
- watermelonCitrullus lanatust0
- Interactor
- MWMVe1pathogen
- Outcome
- sequence identity with the HTS genome isolatese1
- Value
- 97%e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“infecting watermelon (Citrullus lanatus) in Uganda”
e1“One MWMV-positive amplicon was Sanger sequenced (Masaka, PQ397541) and showed 97% sequence identity with the HTS genome isolates”
The same questions in other papers
infection · disease · Citrullus lanatus (watermelon)Positive and negative results
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