Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group
A real time PCR assay conjugated with the fluorescent SYBR(R) Green I dye has been developed for rapid, sensitive and quantitative ;detection of 'Ca. Phytoplasma pyri', 'Co. P. prunorum' and 'Ca. P. mali', quarantine members of apple proliferation (16SrX) group. The selected...
| Autores principales: | , , , , |
|---|---|
| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
|
| Acceso en línea: | http://hdl.handle.net/20.500.11939/4609 |
| _version_ | 1855491784951463936 |
|---|---|
| author | Torres, E. Bertolini, Edson Cambra, Mariano Monton, C. Martin, M. P. |
| author_browse | Bertolini, Edson Cambra, Mariano Martin, M. P. Monton, C. Torres, E. |
| author_facet | Torres, E. Bertolini, Edson Cambra, Mariano Monton, C. Martin, M. P. |
| author_sort | Torres, E. |
| collection | ReDivia |
| description | A real time PCR assay conjugated with the fluorescent SYBR(R) Green I dye has been developed for rapid, sensitive and quantitative ;detection of 'Ca. Phytoplasma pyri', 'Co. P. prunorum' and 'Ca. P. mali', quarantine members of apple proliferation (16SrX) group. The selected primers amplify specifically a target of 217-bp fragment from the 16Sr gene region of the 16SrX group and not from any other tested phytoplasma groups. An artificial template consisting in a plasmid clone of a 1785-bp DNA fragment of the 16S rRNA gene, 16S/23S rDNA spacer region, tRNA-Ile and partial 23S rRNA gene of a 'Ca. P. prunorum' isolate, was used to establish a calibration curve to evaluate the number of amplified targets per sample. The sensitivity of the technique was similar to nested-PCR (10 copies of the amplified target per mu l). The estimated concentration of phytoplasmas in infected pear, plum and apricot trees ranged from 9.7 X 10(3) to 3.0 X 10(5) phytoplasmas per gram of tissue. The method offers the possibility to detect simultaneously, in a single reaction, all quarantine phytoplasmas affecting fruit trees hosts in Europe. (C) 2005 Elsevier Ltd. All rights reserved. |
| format | Artículo |
| id | ReDivia4609 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia46092025-04-25T14:43:59Z Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group Torres, E. Bertolini, Edson Cambra, Mariano Monton, C. Martin, M. P. A real time PCR assay conjugated with the fluorescent SYBR(R) Green I dye has been developed for rapid, sensitive and quantitative ;detection of 'Ca. Phytoplasma pyri', 'Co. P. prunorum' and 'Ca. P. mali', quarantine members of apple proliferation (16SrX) group. The selected primers amplify specifically a target of 217-bp fragment from the 16Sr gene region of the 16SrX group and not from any other tested phytoplasma groups. An artificial template consisting in a plasmid clone of a 1785-bp DNA fragment of the 16S rRNA gene, 16S/23S rDNA spacer region, tRNA-Ile and partial 23S rRNA gene of a 'Ca. P. prunorum' isolate, was used to establish a calibration curve to evaluate the number of amplified targets per sample. The sensitivity of the technique was similar to nested-PCR (10 copies of the amplified target per mu l). The estimated concentration of phytoplasmas in infected pear, plum and apricot trees ranged from 9.7 X 10(3) to 3.0 X 10(5) phytoplasmas per gram of tissue. The method offers the possibility to detect simultaneously, in a single reaction, all quarantine phytoplasmas affecting fruit trees hosts in Europe. (C) 2005 Elsevier Ltd. All rights reserved. 2017-06-01T10:10:32Z 2017-06-01T10:10:32Z 2005 OCT 2005 article Torres, E., Bertolini, E., Cambra, M., Monton, C., Martin, M.P. (2005). Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX), group. Molecular and cellular probes, 19(5), 334-340. 0890-8508 http://hdl.handle.net/20.500.11939/4609 10.1016/j.mcp.2005.06.002 en openAccess Impreso |
| spellingShingle | Torres, E. Bertolini, Edson Cambra, Mariano Monton, C. Martin, M. P. Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group |
| title | Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group |
| title_full | Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group |
| title_fullStr | Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group |
| title_full_unstemmed | Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group |
| title_short | Real-time PCR for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation (16SrX) group |
| title_sort | real time pcr for simultaneous and quantitative detection of quarantine phytoplasmas from apple proliferation 16srx group |
| url | http://hdl.handle.net/20.500.11939/4609 |
| work_keys_str_mv | AT torrese realtimepcrforsimultaneousandquantitativedetectionofquarantinephytoplasmasfromappleproliferation16srxgroup AT bertoliniedson realtimepcrforsimultaneousandquantitativedetectionofquarantinephytoplasmasfromappleproliferation16srxgroup AT cambramariano realtimepcrforsimultaneousandquantitativedetectionofquarantinephytoplasmasfromappleproliferation16srxgroup AT montonc realtimepcrforsimultaneousandquantitativedetectionofquarantinephytoplasmasfromappleproliferation16srxgroup AT martinmp realtimepcrforsimultaneousandquantitativedetectionofquarantinephytoplasmasfromappleproliferation16srxgroup |