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Teruo Sano - One of the best experts on this subject based on the ideXlab platform.
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First report of Coleus blumei viroid 5 infection in vegetatively propagated clonal coleus cv. 'Aurora black cherry' in Japan
New Disease Reports, 2015Co-Authors: Taro Tsushima, Teruo SanoAbstract:Plectranthus scutellarioides (synonym: Coleus blumei ) is a popular garden plant, cultivated worldwide and is known to be infected with several viroids of the genus Coleviroid , family Pospiviroidae . Coleus blumei viroid 1 (CbVd-1) is the…
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First Report of Coleus blumei viroid Infecting Coleus in India.
Plant disease, 2013Co-Authors: Charith Raj Adkar-purushothama, H. Nagaraja, M. Y. Sreenivasa, Teruo SanoAbstract:Members of the genus Coleviroid (Coleus blumei viroid [CbVd]), family Pospiviroidae have been reported to infect Coleus (Solenostemon sp.). CbVd-1 was first reported from Brazil, CbVd-2, -3, and -4 were first reported from Germany, whereas CbVd-5 and -6 were recently identified in China (2). In India, Coleus is extensively cultivated as an ornamental plant in home gardens. In March to June 2012, Coleus leaf samples with irregular chlorotic spots/patches were collected from home gardens of two different districts of Karnataka (Kodagu and Mysore districts), India, suspecting the presence of Coleus blumei viroids (CbVd 1 to 6). Low molecular weight RNAs were extracted using 2% CTAB buffer containing 1.4 M NaCl, 20 mM EDTA, pH 8.0 and 100 mM Tris-Cl, pH 8.0 (1). Viroid-like RNA was enriched by fractionation 2M LiCl soluble nucleic acids (4). A DNA fragment with the expected size of CbVd-1 was detected in 10 (including both districts) of 14 analyzed (incident rate of 71%) from reverse transcription (RT)-PCR assay using Coleviroid specific primers (forward 5'-TGGATCCAGCGCTGCAACGGAATCCA-3' and reverse 5'-TTGGATCCGCCAGGGAACCCAGGTAAG-3'). RT was performed at 37°C for 60 min in 25 μl reaction mix containing 5 μl RNA extracts, 1 μl reverse primer, 1× first strand buffer, 10 mM dNTPs, and 200U M-MuLV-RT (Invitrogen, USA). For PCR, 2 μl RT was mixed in 25 μl PCR mix containing 0.2 μM each forward and reverse primers and 2U LA Taq (Takara-Bio, Japan) according to the manufacturer's instruction. PCR parameter was one cycle at 94°C for 2 min, 35 cycles at 94°C for 45 s, 53°C for 30 s, and 72°C for 60 s, followed by final extension at 72°C for 10 min (4). Sequence analysis of cloned amplicons detected CbVd-1 in India. To confirm the sequence of the primer regions, an additional set of tail-tail primers were designed, CbVd1-136F (5'-CTTCGTGGAACGGCTCCGCG-3') and CbVd1-136Rev (5'-GAAGAAGCCGAAGCAACTCTC-3') and were used for RT-PCR. Amplified products were cloned, sequenced and compared with previously obtained data. Further, the presences of CbVd-1 in Coleus samples was confirmed by a RNA gel blot assay using digoxigenin-labeled CbVd-1 cRNA probe (3). Alignment of 19 sequences obtained from four representative Coleus samples found the presence of two sequence variants of CbVd-1, namely Ind-1 (GenBank Accession No. AB740017) and Ind-2 (AB740018). Ind-1 was found to differ from Ind-2 by two nucleotide substitutions at position 40 (C to T) and 211 (T to C). BLAST analysis of Ind-1 showed 100% sequence similarity with CbVd-1 isolates from China (DQ178399) and South Korea (EU 410620), whereas Ind-2 was 99% identical to these two Chinese and Koreans isolates. Furthermore, 97% and 96% sequence identity with CbVd 1-RL RNA (Accession no. X95366) was observed for Ind-1 and Ind-2, respectively. Isolates from India were 88% similar with Coleus blumei viroid 1-RG (X95291). To the best of our knowledge, this is the first molecular evidence for the presence of CbVd-1 infecting Coleus in India. Coleus harbors various viroid species and CbVd-1, reported widely, can transmit efficiently through seed and also could infect the other herbaceous plants (3). This report from India will contribute further understanding of a potential risk of Coleus viroids in ornamental species. References: (1) J. J. Doyle and J. L. Doyle. Phytochem. Bull. 19:11, 1987. (2) F. H. Fu et al. Plant Dis. 95:494, 2011. (3) Ishiguro et al. Ann. Phytopathol. Soc. Jpn. 62:84, 1996. (4) S.-F. Li et al., Plant Pathol. 55:565, 2006.
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Rapid detection and identification of viroids in the genus Coleviroid using a universal probe
Journal of virological methods, 2012Co-Authors: Dongmei Jiang, Teruo Sano, Wanying Hou, Ni Kang, Lü Qin, Lianhui XieAbstract:A simple, low-cost hybridization assay using a universal DIG-labeled riboprobe for the rapid detection and identification of coleus viroids is presented. An octamer of 32-nucleotide sequence derived from the central conserved region (CCR) of viroids in the genus Coleviroid was used to develop a universal cRNA probe (8-central-conserved-region probe, 8CCR probe) for coleus viroids. Dot-blot hybridization assays demonstrated that the sensitivity of this probe was similar to specific probes for each CbVd, and Northern hybridization results revealed that at least four coleus viroids could be distinguished readily and simultaneously using the 8CCR probe. Batch detection assay showed that hybridization using the 8CCR probe can identify coleus viroids rapidly and effectively. This rapid and low-cost molecular hybridization technique is an effective way to survey the occurrence of coleus viroids, and has reference for the detection of other viroids and possibly viruses.
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sap direct rt pcr for the rapid detection of coleus blumei viroids of the genus Coleviroid from natural host plants
Journal of Virological Methods, 2011Co-Authors: Dongmei Jiang, Lianhui Xie, Teruo SanoAbstract:A simple and fast sap-direct RT-PCR (reverse transcription-polymerase chain reaction) for the rapid detection of 3 viroids of the genus Coleviroid is presented. The templates for cDNA synthesis were obtained directly from the sap of coleus using a pipettor, a common tool in molecular biology laboratories, and 3 coleus blumei viroids (CbVds) were detected simultaneously using a pair of universal primers designed according to sequences in the central conserved region (CCR) of CbVds. RT-PCR results demonstrated that CbVd-1, CbVd-5, and CbVd-6 can be detected accurately in viroid-infected plants but not in viroid-free plants. The results of RT-PCR, dot-blot, sequencing, and batch-detection revealed that this method can be used to identify CbVds rapidly. The method also reduces cross-contamination among different samples to a minimum. It is considered that this rapid and simple technique is an effective method for the identification and cloning of CbVds.
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Coleus blumei viroid 6: a new tentative member of the genus Coleviroid derived from natural genome shuffling.
Archives of virology, 2009Co-Authors: Wanying Hou, Dongmei Jiang, Teruo SanoAbstract:Coleus blumei can be infected by several viroids of the genus Coleviroid. One year after detecting a mixed infection of coleus blumei viroid 1 (CbVd-1) and 5 (CbVd-5) in coleus seedlings inoculated with these two viroids, we found an additional viroid-like RNA. Sequence analysis revealed a viroid of 342 nucleotides that contains the central conserved region of Coleviroids and is a chimera of the left half of CbVd-3 and the right half of CbVd-5. This new viroid, tentatively referred to as coleus blumei viroid 6 (CbVd-6), appears to have arisen from a natural recombination event or genome shuffling.
Dongmei Jiang - One of the best experts on this subject based on the ideXlab platform.
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High-Throughput Sequencing Analysis of Small RNAs Derived from Coleus Blumei Viroids.
Viruses, 2019Co-Authors: Dongmei Jiang, Meng Wang, Zhixiang ZhangAbstract:Characterization of viroid-derived small RNAs (vd-sRNAs) is important to understand viroid–host interactions; however, vd-sRNAs belonging to the genus Coleviroid are yet to be identified and characterized. Herein, we used coleus plants singly infected with coleus blumei viroid (CbVd)-1, -5, or -6 and doubly infected with CbVd-1 and -5 to identify and analyze their vd-sRNAs. We found sense and antisense vd-sRNAs for CbVd-1, -5 and -6, and 22-nt vd-sRNAs were the most abundant; moreover, the 5′-terminal nucleotides (nts) of CbVd-1, -5, and -6 were biased toward U and C, and sRNAs derived from these three viroids were unevenly distributed along their genomes. We also noted that CbVd-5 and -6 share a fragment that forms the right half of the rod-like secondary structure of these viroids, which implied that they generated almost the same type of vd-sRNAs. This finding indicated that vd-sRNA biogenesis is mainly determined by the primary sequence of their substrates. More importantly, we found two complementary vd-sRNAs (22 nt) that were generated from the central conserved region (CCR) of these three viroids, suggesting an important role of CCR in vd-sRNA biogenesis. In conclusion, our results provide novel insight into the biogenesis of vd-sRNAs and the biological roles of CCR.
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Probe design and microarray detection of Coleviroid.
2013Co-Authors: Yongjiang Zhang, Jun Yin, Dongmei Jiang, Yanyan Xin, Fang Ding, Ziniu Deng, Guoping WangAbstract:(A) Two oligonucleotide probes Cole1 and Cole6 were designed matching conserved regions of Coleus blumei viroid 1. Black bars are high-scoring segment pairs (HSP) from the BLASTN alignments of the other Coleviroids to Coleus blumei viroid 1 (X95291). Red bars are the alignments of Cole1 and Cole6 to Coleus blumei viroid 1. (B) Pseudo-color image of Coleus blumei viroid 1 hybridization on the microarray. All probes with feature signal intensity minus background signal intensity more than 1500 are marked on the images. HEX, positive control (PC) and negative control (NC) are marked on the image. (C) Feature and background signal intensities of all the viroid probes on the microarray. Probes targeting Coleviroid genera are depicted as red triangles. Other probes are depicted as black circles. (D) Sequence alignment of Coleviroid probes Cole1, Cole4 and Cole6 with the Coleus blumei viroid 1 (KC581915) sequenced by CAAS-IPP, and Coleus blumei viroid 1 (CbVd 1, X95291) and Coleus blumei viroid 6 (CbVd 6, NC_012805) genomic sequences from NCBI.
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First report of Coleus blumei viroid 5 from Coleus blumei in India and Indonesia.
Plant disease, 2013Co-Authors: Dongmei Jiang, L. H. XieAbstract:Coleus blumei, which was found originally in Indonesia, is an ornamental plant grown worldwide. It can be infected by several viroids of the genus Coleviroid, family Pospiviroidae. Six main viroids that infect coleus have been reported: Coleus blumei viroid 1 through 6 (CbVd-1 ~ CbVd-6). Although CbVd-1 was first reported in a commercial coleus in Brazil in 1989 (1), and then in Germany, Japan, Canada, Korea, China, and India, CbVd-5 was reported only in China in 2009 (2). Symptoms caused by CbVd-5 varied depending on different cultivars, and in case of an unknown cultivar of "Red with dark green edge," are very clear albino symptoms. From 2010 to 2011, 60 and 3 leaf samples of coleus were collected from Hyderabad, India, and Java, Indonesia, respectively, and subjected to low molecular weight RNA extraction according to Li et al. (3). The results of dot-blot hybridization using CbVd-5 cRNA probes and RT-PCR using CbVd-5 specific primers (CbVd-5-PF: 5'-TGACTAGAACAGTAGTAAAG-3' / CbVd-5-PR: 5'-AATTGAGGTCAAACCTCTTT-3') demonstrated that 28 out of the 60 samples from India and all three samples from Indonesia were positive for CbVd-5. The resulting RT-PCR fragments from one sample selected randomly from each country were cloned into the pMD18-T vector (Takara) and transformed into E. coli DH5α competent cells. Five positive clones of each sample were sequenced. The result of sequence analysis revealed that the similarities of CbVd-5 between the sequences we obtained and the reference sequence (GenBank Accession No. NC003683) were 97.8 to 100%. Bioassay using nine viroid-free coleus plants from three cultivars (three from each cultivar), inoculated with CbVd-5 infectious clones by stem slashing, demonstrated that CbVd-5 could induce albino symptom on the leaves of the unknown cultivar "Red with dark green edge" 2 months after inoculation. To our knowledge, this is the first report of CbVd-5 from India and Indonesia, and the second report of CbVd-5 in the world. Considering the effect of CbVd-5 on the appearance of coleus and its recombination ability, a certification program may be needed to control the spread of this viroid. References: (1) M. E. N. Fonseca et al. Fitopatol. Bras. 14:94, 1989. (2) W. Y. Hou et al. Arch. Virol. 154:315, 2009. (3) S. F. Li et al. Ann. Phytopathol. Soc. Jpn. 61:381, 1995.
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Rapid detection and identification of viroids in the genus Coleviroid using a universal probe
Journal of virological methods, 2012Co-Authors: Dongmei Jiang, Teruo Sano, Wanying Hou, Ni Kang, Lü Qin, Lianhui XieAbstract:A simple, low-cost hybridization assay using a universal DIG-labeled riboprobe for the rapid detection and identification of coleus viroids is presented. An octamer of 32-nucleotide sequence derived from the central conserved region (CCR) of viroids in the genus Coleviroid was used to develop a universal cRNA probe (8-central-conserved-region probe, 8CCR probe) for coleus viroids. Dot-blot hybridization assays demonstrated that the sensitivity of this probe was similar to specific probes for each CbVd, and Northern hybridization results revealed that at least four coleus viroids could be distinguished readily and simultaneously using the 8CCR probe. Batch detection assay showed that hybridization using the 8CCR probe can identify coleus viroids rapidly and effectively. This rapid and low-cost molecular hybridization technique is an effective way to survey the occurrence of coleus viroids, and has reference for the detection of other viroids and possibly viruses.
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sap direct rt pcr for the rapid detection of coleus blumei viroids of the genus Coleviroid from natural host plants
Journal of Virological Methods, 2011Co-Authors: Dongmei Jiang, Lianhui Xie, Teruo SanoAbstract:A simple and fast sap-direct RT-PCR (reverse transcription-polymerase chain reaction) for the rapid detection of 3 viroids of the genus Coleviroid is presented. The templates for cDNA synthesis were obtained directly from the sap of coleus using a pipettor, a common tool in molecular biology laboratories, and 3 coleus blumei viroids (CbVds) were detected simultaneously using a pair of universal primers designed according to sequences in the central conserved region (CCR) of CbVds. RT-PCR results demonstrated that CbVd-1, CbVd-5, and CbVd-6 can be detected accurately in viroid-infected plants but not in viroid-free plants. The results of RT-PCR, dot-blot, sequencing, and batch-detection revealed that this method can be used to identify CbVds rapidly. The method also reduces cross-contamination among different samples to a minimum. It is considered that this rapid and simple technique is an effective method for the identification and cloning of CbVds.
Wanying Hou - One of the best experts on this subject based on the ideXlab platform.
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Rapid detection and identification of viroids in the genus Coleviroid using a universal probe
Journal of virological methods, 2012Co-Authors: Dongmei Jiang, Teruo Sano, Wanying Hou, Ni Kang, Lü Qin, Lianhui XieAbstract:A simple, low-cost hybridization assay using a universal DIG-labeled riboprobe for the rapid detection and identification of coleus viroids is presented. An octamer of 32-nucleotide sequence derived from the central conserved region (CCR) of viroids in the genus Coleviroid was used to develop a universal cRNA probe (8-central-conserved-region probe, 8CCR probe) for coleus viroids. Dot-blot hybridization assays demonstrated that the sensitivity of this probe was similar to specific probes for each CbVd, and Northern hybridization results revealed that at least four coleus viroids could be distinguished readily and simultaneously using the 8CCR probe. Batch detection assay showed that hybridization using the 8CCR probe can identify coleus viroids rapidly and effectively. This rapid and low-cost molecular hybridization technique is an effective way to survey the occurrence of coleus viroids, and has reference for the detection of other viroids and possibly viruses.
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Coleus blumei viroid 6: a new tentative member of the genus Coleviroid derived from natural genome shuffling.
Archives of virology, 2009Co-Authors: Wanying Hou, Dongmei Jiang, Teruo SanoAbstract:Coleus blumei can be infected by several viroids of the genus Coleviroid. One year after detecting a mixed infection of coleus blumei viroid 1 (CbVd-1) and 5 (CbVd-5) in coleus seedlings inoculated with these two viroids, we found an additional viroid-like RNA. Sequence analysis revealed a viroid of 342 nucleotides that contains the central conserved region of Coleviroids and is a chimera of the left half of CbVd-3 and the right half of CbVd-5. This new viroid, tentatively referred to as coleus blumei viroid 6 (CbVd-6), appears to have arisen from a natural recombination event or genome shuffling.
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Identification and characterization of a new Coleviroid (CbVd-5)
Archives of virology, 2008Co-Authors: Wanying Hou, Teruo SanoAbstract:A viroid-like RNA was detected from coleus (Coleus blumei) in China. It consisted of 274 nucleotides and had 66% sequence identity with a member of the closest known viroid species. The predicted secondary structure is rod-shaped with extensive base pairing, and it has the conserved region characteristic of the genus Coleviroid. Two terminal sequences that are highly conserved among some members of the genus were also identified. The viroid-like RNA was successfully transmitted to coleus by slash-inoculation. This viroid was identified as a new member of the genus Coleviroid, and we tentatively propose the name Coleus blumei viroid 5 (CbVd-5).
L. H. Xie - One of the best experts on this subject based on the ideXlab platform.
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First report of Coleus blumei viroid 5 from Coleus blumei in India and Indonesia.
Plant disease, 2013Co-Authors: Dongmei Jiang, L. H. XieAbstract:Coleus blumei, which was found originally in Indonesia, is an ornamental plant grown worldwide. It can be infected by several viroids of the genus Coleviroid, family Pospiviroidae. Six main viroids that infect coleus have been reported: Coleus blumei viroid 1 through 6 (CbVd-1 ~ CbVd-6). Although CbVd-1 was first reported in a commercial coleus in Brazil in 1989 (1), and then in Germany, Japan, Canada, Korea, China, and India, CbVd-5 was reported only in China in 2009 (2). Symptoms caused by CbVd-5 varied depending on different cultivars, and in case of an unknown cultivar of "Red with dark green edge," are very clear albino symptoms. From 2010 to 2011, 60 and 3 leaf samples of coleus were collected from Hyderabad, India, and Java, Indonesia, respectively, and subjected to low molecular weight RNA extraction according to Li et al. (3). The results of dot-blot hybridization using CbVd-5 cRNA probes and RT-PCR using CbVd-5 specific primers (CbVd-5-PF: 5'-TGACTAGAACAGTAGTAAAG-3' / CbVd-5-PR: 5'-AATTGAGGTCAAACCTCTTT-3') demonstrated that 28 out of the 60 samples from India and all three samples from Indonesia were positive for CbVd-5. The resulting RT-PCR fragments from one sample selected randomly from each country were cloned into the pMD18-T vector (Takara) and transformed into E. coli DH5α competent cells. Five positive clones of each sample were sequenced. The result of sequence analysis revealed that the similarities of CbVd-5 between the sequences we obtained and the reference sequence (GenBank Accession No. NC003683) were 97.8 to 100%. Bioassay using nine viroid-free coleus plants from three cultivars (three from each cultivar), inoculated with CbVd-5 infectious clones by stem slashing, demonstrated that CbVd-5 could induce albino symptom on the leaves of the unknown cultivar "Red with dark green edge" 2 months after inoculation. To our knowledge, this is the first report of CbVd-5 from India and Indonesia, and the second report of CbVd-5 in the world. Considering the effect of CbVd-5 on the appearance of coleus and its recombination ability, a certification program may be needed to control the spread of this viroid. References: (1) M. E. N. Fonseca et al. Fitopatol. Bras. 14:94, 1989. (2) W. Y. Hou et al. Arch. Virol. 154:315, 2009. (3) S. F. Li et al. Ann. Phytopathol. Soc. Jpn. 61:381, 1995.
Butkovic Anamarija - One of the best experts on this subject based on the ideXlab platform.
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Molecular characterization of Coleus blumei viroids 1 and 3 in Plectranthus scutellarioides in Croatia
'Springer Science and Business Media LLC', 2019Co-Authors: Škorić Dijana, Černi Silvija, Jezernik Karlo, Butkovic AnamarijaAbstract:Asymptomatic Plectranthus scutellarioides (syn. Coleus blumei) plants of four different cultivars were investigated for the presence of Coleus blumei viroid (CbVd) species (genus Coleviroid, family Pospiviroidae). Cloned and sequenced amplicons obtained by generic Coleviroid primers were analyzed for their primary and secondary structure. CbVd-1 was found in one of the coleus cultivars in a single infection and in another cultivar in a mixed infection with CbVd-3. Additionally, CbVd-3 was found in a third coleus cultivar in a single infection. In singly and doubly infected cultivars, both CbVd-1 and CbVd-3 variants displayed higher variability regarding both length and nucleotide composition in a doubly infected cultivar. Sequences from different cultivars showed different phylogenetic grouping suggesting distinct infection sources, probably introduced infected seeds. For all phylogenetic subgroups, characteristic nucleotide changes were identified and their influence on the secondary structure was predicted. The most noticeable structural changes were observed in nucleotide positions 25–26 in CbVd-1 and in the position 138 in CbVd-3 variants. This work includes the first report of Coleviroids in Croatia, the second reported Coleviroids sequences from Europe and the third report of CbVd-3 in the world enlarging the pool of knowledge on the variability and distribution of viroids in horticultural crops.This study was funded by the University of Zagreb grants to D. Škorić and S. Černi numbers: 20281214, 20282308 and 20283119.Peer reviewe
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Biological and molecular properties of Coleus blumei viroid 3
University of Zagreb. Faculty of Science. Department of Biology., 2017Co-Authors: Butkovic AnamarijaAbstract:Viroidi su gole, prstenaste molekule RNA duge 246 – 401 nukleotida. Najmanje su autonomne infektivne nukleinske kiseline dosada poznate, ne kodiraju proteine, a isključivo su biljni patogeni. Plectranthus scutellarioides (L.) R. Br. (syn. Coleus blumei) ili šarena kopriva je domaćin nekoliko Coleus blumei viroida (CbVd). Šarena kopriva je široko rasprostranjena ukrasna biljka i domaćin za istraživanje roda Coleviroid (Pospiviroidae). Najbolje istraženi Coleviroid je Coleus blumei viroid 1 (CbVd-1) dok se za CbVd-3 to ne može reći. Cilj ovog diplomskog rada je bio potvrditi inficiranost prethodno istraživanih kultivara šarene koprive viroidom CbVd-3 te istražiti njegove glavne molekularne i biološke značajke (krug domaćina, prijenos sjemenom, prijenos cijepljenjem). Iz ukupne RNA potencijalnih domaćina metodom reverzne transkripcije i lančane reakcije polimeraze (RT-PCR) detektirana je prisutnost viroida uz primjenu generičkih početnica za rod Coleviroid. Od istraženih potencijalnih alternativnih domaćina iz porodice Lamiaceae, jedino se Salvia hispanica L. pokazala kao mogući novi domaćin Coleviroida. Transgeni korjenčići šarene koprive također su domaćini Coleviroida koji u njima perzistira. Analiza parcijalnih sekvenci viroida iz S. hispanica i transgenih korjenčića šarene koprive ukazuje na CbVd-1 i potvrđuje njegov širi krug domaćina u odnosu na CbVd-3. Dokazana je prenosivost CbVd-3 sjemenom šarene koprive od 40%, no prijenos cijepljenjem nije bio uspješan. Zbog nedostatnih tehničkih uvjeta pokus cijepljenja bi trebalo ponoviti. Istraživanjem domaćina metodom sekvencijske poliakrilamidne gel-elektroforeze (sPAGE) za laboratorijski uzorak šarene koprive inficirana s CbVd-3 u smjesi s CbVd-1 infekcija je potvrđena.Viroids are naked, circular RNA molecules with length between 246 and 401 nucleotides. They are the smallest autonomous infectious noncoding nucleic acids known so far. They infect only plants. Plectranthus scutellarioides (L.) R. Br. (syn. Coleus blumei) is a host to many Coleus blumei viroid species (CbVd). Coleus blumei is grown worldwide and it is a host for studying Coleviroid members (Pospiviroidae). Coleus blumei viroid 1 (CbVd-1) is the best studied Coleviroid so far while the knowledge on CbVd-3 biology is scarce. The goals of my thesis were to confirm infections with CbVd-3 in previously analyzed coleus cultivars and to study the main molecular and biological properties (hosts, seed transmission, transmission by graft-inoculation) of CbVd-3. Total plant RNA samples from potential hosts were tested by reverse transcription and polymerase chain reaction (RT-PCR) with generic Coleviroid primers targeting the central conserved region of the viroid molecule. Several potential new hosts from Lamiaceae family were tested but only Salvia hispanica L. was found positive for Coleviroid presence. Transgenic hairy roots of coleus were also found as persistent hosts of a Coleviroid. The partial sequence analysis from S. hispanica and coleus hairy roots suggest the presence of CbVd-1 confirming its wide host range as opposed to CbVd-3. In one of the coleus cultivars tested, the CbVd-3 seed transmission was 40%. Graft-transmission experiments were unsuccessful as yet but due to technical difficulties, they should be repeated to draw relevant conclusions. Sequential polyacrylamide gel electrophoresis (sPAGE) analyses confirmed the infection of one of the coleus plants with CbVd-3 in mixed infection with CbVd-1