Abstract
Thirty-one clones of sweetpotatoes collected from some parts of Zimbabwe were used as inoculum sources to mechanically inoculate 13 experimental hosts: Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Datura stramoniumitalic, Gomphrena globosa, Ipomoea purpurea, I. quamoclit, I. rubrocorulea, Nicotiana benthamiana, N. clevelandii, N. glutinosa, N. rustica, and N. tabacum. Systemic vein clearing was observed in N. benthamiana inoculated with buffered sap from nine clones. Purification of the vein clearing inducing agent from one of the sweetpotato clones gave yields ranging from 2 to 17 mg/kg and the A260 nm /A280 nm was around 1.2. Electron microscopy revealed flexuous filamentous particles with a modal length of 830 nm. Protein analysis of purified virus preparations by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a major protein band of 40 kDa, and this was assumed to be the viral coat protein. Minor protein bands of 27, 37, and 46 kDa were also observed. The viral protein degraded upon storage at 4°C over time to yield a protein band of 27 kDa. Polyclonal antiserum was produced against the purified virus. Protein A gold labeling of the purified virus incubated with available antisera; sweetpotato chlorotic stunt virus (SPCSV), sweetpotato feathery mottle virus strain russet crack (SPFMV-RC), sweetpotato feathery mottle virus, sweetpotato mild mottle virus, sweetpotato latent virus, sweetpotato chlorotic fleck virus, and sweetpotato caulimo-like virus resulted in a higher labeling density with the antiserum of SPFMV-RC than with the antiserum of SPCSV, while the other sera did not react. Further characterization of the vein clearing inducing agent was attempted by reverse transcription-polymerase chain reaction amplification of total RNA with degenerate primers for potyviruses and an oligo dT primer and PCR products of correct size were obtained. The nucleotide sequence was determined and the amino acid of the polyprotein deduced. Comparison with other strains of SPFMV showed strong similarity except for an insertion of 22 amino acids at the N-terminus of the coat protein. The coat protein size of 335 amino acids is the biggest SPFMV so far determined.
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Claims 10
Claim · model-written summary
Systemic vein clearing appeared in N. benthamiana inoculated with buffered sap from nine sweetpotato clones.
- Taxon
- N. benthamiana
Nicotiana benthamianae0 - Outcome
- Systemic vein clearinge0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Systemic vein clearing was observed in N. benthamiana inoculated with buffered sap from nine clones”
Claim · model-written summary
Purification of the vein clearing agent from a sweetpotato clone gave yields of 2 to 17 mg/kg.
- Taxon
- sweetpotato
_Flower.jpg?width=60)
Ipomoea batatase0 - Outcome
- yieldse0
- Value
- 2 to 17 mg/kge0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
_Flower.jpg?width=500)
Verbatim
e0“Purification of the vein clearing inducing agent from one of the sweetpotato clones gave yields ranging from 2 to 17 mg/kg”
Claim · model-written summary
Electron microscopy showed flexuous filamentous particles with a modal length of 830 nm.
Not reportedTaxon
Verbatim
e0“Electron microscopy revealed flexuous filamentous particles with a modal length of 830 nm”
Claim · model-written summary
SDS-PAGE of purified virus showed a major protein band of 40 kDa, assumed to be the coat protein.
Not reportedTaxon
Verbatim
e0“revealed a major protein band of 40 kDa”
Claim · model-written summary
Minor protein bands of 27, 37 and 46 kDa were also observed.
Not reportedTaxon
Verbatim
e0“Minor protein bands of 27, 37, and 46 kDa were also observed”
Claim · model-written summary
The viral protein degraded on storage at 4 C to a 27 kDa band.
Not reportedTaxon
Verbatim
e0“The viral protein degraded upon storage at 4°C over time to yield a protein band of 27 kDa”
Claim · model-written summary
Labeling density of the purified virus was higher with SPFMV-RC antiserum than with SPCSV antiserum; the other antisera did not react.
Verbatim
e0“resulted in a higher labeling density with the antiserum of SPFMV-RC than with the antiserum of SPCSV, while the other sera did not react”
Claim · model-written summary
RT-PCR with degenerate potyvirus primers and an oligo dT primer gave PCR products of correct size.
- Outcome
- PCR products of correct sizee0
Not reportedTaxon
Verbatim
e0“PCR products of correct size were obtained”
Claim · model-written summary
Compared with other SPFMV strains, the sequence was strongly similar except for a 22 amino acid insertion at the N-terminus of the coat protein.
- Taxon
- SPFMVSweet potato feathery mottle viruse0
- Outcome
- strong similarity except for an insertione0
- Value
- 22 amino acidse0
Verbatim
e0“Comparison with other strains of SPFMV showed strong similarity except for an insertion of 22 amino acids at the N-terminus of the coat protein”
Claim · model-written summary
The coat protein of 335 amino acids is the biggest SPFMV coat protein determined so far.
- Taxon
- SPFMVSweet potato feathery mottle viruse0
- Outcome
- coat protein sizee0
- Value
- 335 amino acidse0
Verbatim
e0“The coat protein size of 335 amino acids is the biggest SPFMV so far determined”