Abstract
During surveys of tomato (Solanum lycopersicum) fields in Niono, Mali, conducted in March 2011, unusual disease symptoms, including stunted growth, epinasty, and chlorosis of leaves and necrosis of leaf veins and stems were observed in multiple fields. The incidence of these symptoms was low (~1 to 5%), but they were distinct from those associated with known diseases in the region. A representative leaf sample with these symptoms was applied to filter paper (FTA cards, Whatman), and DNA and RNA extracts were prepared according to manufacturer instructions. RT-PCR tests for Tomato spotted wilt virus, Tobacco streak virus, Tomato necrotic spot virus, Tobacco/tomato mosaic viruses, Cucumber mosaic virus, Alfalfa mosaic virus, torradoviruses, and potyviruses, and PCR tests for begomoviruses, phytoplasmas, and 'Candidatus Liberibacter' infection were also negative. However, virus-like symptoms developed in all 16 tomato seedlings (cv. Early Pak 7) 7 to 10 days after mechanical (sap) inoculation with inoculum prepared from the FTA sample. No symptoms developed in mock-inoculated control plants (n = 3). Symptoms induced included stunted growth and severe epinasty of leaves, followed by necrosis of leaf veins, petioles, and stems. These symptoms were similar to those observed in plants in Mali. When RNA extracts prepared from leaves of these symptomatic plants were mechanically inoculated onto 24 tomato seedlings, similar symptoms developed in all plants, suggesting the causal agent might be a viroid. RT-PCR tests with RNA from symptomatic tomato leaves and universal (3) and various specific Pospiviroid primer pairs were negative. However, equivalent RT-PCR tests conducted with the pCLV4/pCLVR4 primer pair specific for Columnea latent viroid (CLVd) (2) generated a DNA fragment of the expected size (~370 bp). The sequence of this DNA fragment (GenBank Accession No. JQ362419) was 99% identical with those of CLVd isolates from the Netherlands (AY373446 and AY372396). In host range studies, the CLVd isolate from Mali induced symptoms in all 48 mechanically-inoculated tomato plants, whereas no symptoms developed (up to 90 days after inoculation) in inoculated Chenopodium quinoa, C. amaranticolor, Nicotiana benthamiana, N. tabacum (cvs. Havana, Glurk and Turkish), N. glutinosa, Datura stramonium, common bean (cvs. Topcrop and Pinto bean), pumpkin (cv. Small Sugar), pepper (Capsicum annuum, cv. Yolo Wonder) and cucumber (cvs. Emparator and Poinsett 76) plants (results of three independent experiments with six plants per experiment). Symptomless infections were detected in pepper (24 of 30), N. benthamiana (25 of 25), and N. tabacum cv. Turkish (11 of 24) plants by RT-PCR with the pCLV4/pCLVR4 primer pair. To our knowledge, this is the first report of CLVd infecting tomato in Mali. RT-PCR tests of seeds collected from CLVd-infected tomato, pepper, and N. benthamiana plants also detected CLVd (1). Thus, it is possible that CLVd was introduced into Mali in association with seed. References: (1) O. Batuman and R. L. Gilbertson. Phytopathology 102:S4.9, 2012. (2) R. L. Spieker. Arch. Virol. 141:1823, 1996. (3) J. T. J. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004.
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Claims 12
Claim · model-written summary
Unusual disease symptoms were observed in multiple tomato fields in Niono, Mali, at low incidence (~1 to 5%).
- Taxon
- tomato (Solanum lycopersicum)
Solanum lycopersicume1 - Outcome
- unusual disease symptomse1
- Value
- ~1 to 5%e2
- Setting
- Niono, Malie1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“During surveys of tomato (Solanum lycopersicum) fields in Niono, Mali, conducted in March 2011, unusual disease symptoms, including stunted growth, epinasty, and chlorosis of leaves and necrosis of leaf veins and stems were observed in multiple fields”
e2“The incidence of these symptoms was low (~1 to 5%)”
Claim · model-written summary
RT-PCR and PCR tests for known viruses, phytoplasmas, begomoviruses and Candidatus Liberibacter were negative in the symptomatic tomato sample.
- Taxon
- tomato (Solanum lycopersicum)Solanum lycopersicume1
- Outcome
- RT-PCR testse3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“tomato (Solanum lycopersicum) fields in Niono, Mali”
e3“RT-PCR tests for Tomato spotted wilt virus, Tobacco streak virus, Tomato necrotic spot virus, Tobacco/tomato mosaic viruses, Cucumber mosaic virus, Alfalfa mosaic virus, torradoviruses, and potyviruses, and PCR tests for begomoviruses, phytoplasmas, and 'Candidatus Liberibacter' infection were also negative”
Claim · model-written summary
Mechanical sap inoculation with the FTA-sample inoculum produced virus-like symptoms in all 16 tomato seedlings (cv. Early Pak 7), while mock-inoculated controls showed none.
- Taxon
- tomatoSolanum lycopersicume4
- Material
- cv. Early Pak 7e4
- Treatment
- mechanical (sap) inoculation with inoculum prepared from the FTA samplee4
- Outcome
- virus-like symptomse4
- Comparator
- mock-inoculated control plantse5
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e4“virus-like symptoms developed in all 16 tomato seedlings (cv. Early Pak 7) 7 to 10 days after mechanical (sap) inoculation with inoculum prepared from the FTA sample”
e5“No symptoms developed in mock-inoculated control plants (n = 3)”
Claim · model-written summary
Mechanically inoculating RNA extracts from symptomatic plants onto 24 tomato seedlings produced similar symptoms in all plants.
- Taxon
- tomatoSolanum lycopersicume6
- Treatment
- RNA extracts prepared from leaves of these symptomatic plantse6
- Outcome
- similar symptomse6
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e6“RNA extracts prepared from leaves of these symptomatic plants were mechanically inoculated onto 24 tomato seedlings, similar symptoms developed in all plants”
Claim · model-written summary
RT-PCR with the pCLV4/pCLVR4 primer pair specific for Columnea latent viroid gave a DNA fragment of the expected size (~370 bp) in tomato.
- Taxon
- tomato (Solanum lycopersicum)Solanum lycopersicume1
- Interactor
- Columnea latent viroid
Columnea latent viroide7pathogen - Outcome
- DNA fragment of the expected sizee7
- Value
- ~370 bpe7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“tomato (Solanum lycopersicum) fields in Niono, Mali”
e7“equivalent RT-PCR tests conducted with the pCLV4/pCLVR4 primer pair specific for Columnea latent viroid (CLVd) (2) generated a DNA fragment of the expected size (~370 bp)”
Claim · model-written summary
The sequenced RT-PCR fragment was 99% identical to CLVd isolates from the Netherlands.
- Taxon
- tomato (Solanum lycopersicum)Solanum lycopersicume1
- Interactor
- CLVde2pathogen
- Outcome
- sequence of this DNA fragmente2
- Value
- 99%e2
- Comparator
- CLVd isolates from the Netherlandse2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e1“tomato (Solanum lycopersicum) fields in Niono, Mali”
e2“The sequence of this DNA fragment (GenBank Accession No. JQ362419) was 99% identical with those of CLVd isolates from the Netherlands (AY373446 and AY372396)”
Claim · model-written summary
The CLVd isolate from Mali induced symptoms in all 48 mechanically inoculated tomato plants.
- Taxon
- tomatoSolanum lycopersicume2
- Treatment
- CLVd isolate from Malie2
- Interactor
- CLVde2pathogen
- Outcome
- symptomse2
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“the CLVd isolate from Mali induced symptoms in all 48 mechanically-inoculated tomato plants”
Claim · model-written summary
No symptoms developed up to 90 days after inoculation in Chenopodium quinoa and the other listed non-tomato species (C. amaranticolor, Nicotiana, Datura, bean, pumpkin, pepper, cucumber).
- Taxon
- Chenopodium quinoa
Chenopodium quinoae0 - Outcome
- symptomse0
- Qualifier
- up to 90 days after inoculatione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“no symptoms developed (up to 90 days after inoculation) in inoculated Chenopodium quinoa, C. amaranticolor, Nicotiana benthamiana, N. tabacum (cvs. Havana, Glurk and Turkish), N. glutinosa, Datura stramonium, common bean (cvs. Topcrop and Pinto bean), pumpkin (cv. Small Sugar), pepper (Capsicum annuum, cv. Yolo Wonder) and cucumber (cvs. Emparator and Poinsett 76) plants”
Claim · model-written summary
Symptomless CLVd infection was detected by RT-PCR in 24 of 30 pepper plants.
Verbatim
e0“Symptomless infections were detected in pepper (24 of 30), N. benthamiana (25 of 25), and N. tabacum cv. Turkish (11 of 24) plants by RT-PCR with the pCLV4/pCLVR4 primer pair”
Claim · model-written summary
Symptomless CLVd infection was detected by RT-PCR in 25 of 25 Nicotiana benthamiana plants.
- Taxon
- N. benthamiana
Nicotiana benthamianae0 - Outcome
- symptomless infectionse0
- Value
- 25 of 25e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Symptomless infections were detected in pepper (24 of 30), N. benthamiana (25 of 25), and N. tabacum cv. Turkish (11 of 24) plants by RT-PCR with the pCLV4/pCLVR4 primer pair”
Claim · model-written summary
Symptomless CLVd infection was detected by RT-PCR in 11 of 24 Nicotiana tabacum cv. Turkish plants.
- Taxon
- N. tabacum
Nicotiana tabacume0 - Material
- cv. Turkishe0
- Outcome
- symptomless infectionse0
- Value
- 11 of 24e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Symptomless infections were detected in pepper (24 of 30), N. benthamiana (25 of 25), and N. tabacum cv. Turkish (11 of 24) plants by RT-PCR with the pCLV4/pCLVR4 primer pair”
Claim · model-written summary
This is the first report of CLVd infecting tomato in Mali.
- Taxon
- tomatoSolanum lycopersicume0
- Interactor
- CLVde0pathogen
- Outcome
- CLVd infecting tomatoe0
- Setting
- Malie0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“To our knowledge, this is the first report of CLVd infecting tomato in Mali”
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