For this reason, the most common method for obtaining virus-free plant is meristem culture technique. Different researchers [ 6 , 7 ] have also reported that shoot tip remained insufficient for obtaining the virus-free plants. Biological mechanical inoculation, indexing , serological enzyme-linked immunosorbent assay—ELISA , and molecular polymerase chain reaction—PCR methods are used for detection of viruses in plants and identification of these viruses. One of the main advantages of the real-time PCR is the very low chances of infection and detection of viruses with high efficiency.
Detection of viruses through real-time PCR method has been performed successfully by many researchers such as Roberts et al. The objectives of this study were i comparison of meristem culture technique with shoot tip culture technique to obtain virus-free plants, ii comparison of micropropagation success of two different nutrient media and two different garlic species A. Allium sativum Kastamonu garlic clone and Allium tuncelianum garlic species determined to be infected with viruses were used as plant material.
Meristem and shoot tips from sterile cloves were extracted by sterile forceps and scalpels under a stereobinocular microscope in laminar flow and placed into glass tubes containing MS nutrient medium [ 15 ].
Germinated meristem and shoot tips were transferred to two different nutrient media Medium 1: MS medium containing 0. After 6 weeks, number of shoots per plant were recorded, and the shoots were subcultured. The experiment was designed in a completely randomized experimental design with three replications and thirty explants included per replication. Variance analysis was conducted to evaluate the results, and Tukey test was used for controlling the significance of the differences.
Sixty in vitro plants 15 of them from meristem culture of A. Collection tube was removed and filter tube was placed into a new collection tube. Collection tube was changed with a new one. The collecting tube was changed, and the washing step was repeated. Filter tube was placed into new sterile 1. Filter tubes placed in the Eppendorf tubes were removed, and purified viral nucleic acid was extracted.
This process was completed in two steps. Evaluation was performed by qualitative detection program of this thermocycler. Primers used in the study [ 11 ]. Ninety shoot tips and ninety meristems were used for A. Meristems transformed to plants in eight weeks. While 85 of meristems could grow without any problems, three of them could not germinate, and as infection problem was observed in two meristems.
Cultured shoot tips were turned into plants in weeks. Eighty-four shoot tips germinated healthy; only 1 shoot tip could not grow and 5 of ninety shoot tips were found to be infected. Plants obtained meristems of A. Number of shoots were recorded in both first micropropagation and subculture were higher in Medium 2 compared to Medium 1, and the differences between the two media were found to be significant statistically.
In the first propagation, Similar results were provided from subculture. Shoot tip culture results showed that Medium 2 was better compared to Medium 1. Considered in terms of explants, meristem explants were found to be more successful compared to shoot tip explants in both garlic species and nutrient media.
The experiment for A. While 89 meristems could transform into plant in meristem culture the remaining one meristem was probably damaged , 87 shoot tips could germinate in shoot tip culture the remaining 3 plants were infected.
Meristem and shoot tip culture results of A. As seen in Table 2 , In subculture, These results compared with results of A. In Medium 1, A. In shoot tip culture of A. Viruses were detected in shoot tip culture.
OYDV virus was determined in 12 plants of A. Ten plants of A. Results of this research showed clearly that meristem culture technique was more effective compared to shoot tip culture technique in obtaining virus-free plant. Whereas virus problem could not be solved by shoot tip culture technique, viruses were detected in the majority of plantlets obtained by this technique. In meristem culture, only apical dome and a few leaf primordia are isolated and placed to nutrient media [ 16 ].
According to some researchers, there is a competition between cell proliferation, and the formation of the virus particles in meristem region of plant. Nucleic acid production capacity in meristematic tissue during cell division is used for cell division and this situation prevents the reproduction of virus. According to other researchers, transportation of viruses to the meristem region of the plant is prevented due to lack of transport system in meristem [ 17 ].
Real-time PCR method is more sensitive and specific than other techniques. Therefore, real-time PCR method was used in this study to detect viruses in garlic plants. Also two different nutrient media were tested in terms of micropropagation success. Therefore, effects of these hormones on micropropagation of garlic were tested through using two different nutrient media in this study. One of the most important encountered problems in the production of garlic in Turkey is viruses.
However, there is not any effective chemical application available against virus control. Meristem culture technique is widely used for obtaining virus-free plants.
However, extracting the meristem regions of the plant is difficult, time-consuming and requires expertise. Therefore after achieving certain number of meristem, plants obtained from these meristems are multiplied using tissue culture techniques for commercial production.
In this study, micropropagation capacity of A. The results obtained from the present study are very important for the scientist trying to develop virus-free plants and could also have significance of practical application which will in turn affect positively to increase yield with better quality of crop for the local farmers.
National Center for Biotechnology Information , U. This is the first report of in vitro tuberization becoming feasible for agriculture in seed-potato production. An estimated 36, dormant, miniature tubers were harvested from the aseptic containers incubated on a 10m 2 bench area in a fourmonth period. After three successive plantings in soil, 1, kgs of virusfree seed-potatoes were obtained. This is a preview of subscription content, access via your institution. Rent this article via DeepDyve.
Chen, P. Cheng, T. Chang, W. Google Scholar. Harmey, M. Eur Potato J 9: — Holmes, F. Phytopathology Article Google Scholar. Kassanis, B. Ann Appl Biol — Loo, T. Chinese Biosci — Mingo-Castel, A. PI Physiol — Murashige, T. Physiol Plant — Peterson, R. Bot Gaz — Wang, P. Barz, E. Reinhard, M. Zenk, Eds. Springer-Verleg, Berlin, New York, p. Adv Biochem Engineering —
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