ISHS Acta Horticulturae 530: International Symposium on Methods and Markers for Quality Assurance in Micropropagation
ELIMINATION OF VIRUSES THROUGH MERISTEM CULTURE AND THERMOTHERAPY FOR THE ESTABLISHMENT OF AN IN VITRO COLLECTION OF GARLIC (ALLIUM SATIVUM) |
| Authors: | A. Senula, E.R.J. Keller, D.E. Leseman |
| Keywords: | onion yellow dwarf virus, leek yellow stripe virus, garlic common latent virus, shallot latent virus, mite-borne-filamentous virus, bulbils, apices, ribavirin |
| DOI: | 10.17660/ActaHortic.2000.530.12 |
| Abstract:
An attempt was made to establish a large number of virus- free garlic accessions in vitro. Virus- free in vitro plants of 87 accessions have been obtained via meristem culture. The virus- free status has been determined for the most frequent garlic viruses: OYDV, LYSV, GCLV, SLV and MbFV. In another 7 accessions one or more viruses were still present. The most persistent virus was GCLV. The occurrence of the other viruses was definitely lower. OYDV and LYSV have been eliminated to 85– 95 %. The detection of viruses in the plant was mainly performed by means of the ELISA technique using polyclonal and monoclonal antibodies. ISEM and decoration methods were used for the verification of the ELISA results. The results of ISEM confirmed the virus- free status of plants in all cases, as determined by ELISA. In some cases viruses detected by ELISA could not be found by ISEM or the decoration method. On the other hand, in tests on MbFV with doubtful ELISA results, decoration tests gave a clear indication of virus infection. Small meristems from bulbils with one leaf primordium (0.3– 0.8 mm) were used for the establishment of meristem- tip cultures. The rate of plant regeneration varied between 1 % and 80 % of cultivated meristems and was dependent on the genotype. Meristem culture failed to result in plantlet regeneration in only 6 accessions. The influence of the culture medium was low compared with that of the genotype. The addition of ribavirin to the induction medium reduced the regeneration of plantlets, but virus elimination was significantly increased (from 19.6 to 59.3 %). Hot air treatment of bulbils at 36°C for 6 weeks did not reduce the survival of meristems. It was, however, favourable for virus elimination. Virus elimination was especially high, when thermotherapy and addition of ribavirin to the induction medium were combined. | |
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