The positive effect of antibiotic paromomycin compared with kanamycin for selection of transgenic plants with NPTII gene on the example of Nicotiana tabacum / Nitovska I.O. et al. 2015
| Рядок 4: | Рядок 4: | ||
{| align=right | {| align=right | ||
| − | |[[Image:The positive effect of antibiotic... Nitovska I.O. 2015 Fig. 1a.jpg| | + | |[[Image:The positive effect of antibiotic... Nitovska I.O. 2015 Fig. 1a.jpg|250px]] |
|} | |} | ||
'''Aims.''' To investigate the efficiency of antibiotic paromomycin as a selective agent for the production of transgenic plants with nptII gene compared with kanamycin sulfate on the example of Nicotiana tabacum. '''Methods.''' a vector containing the nptII gene under the control of 35S promoter of cauliflower mosaic virus was used in Agrobacterium-mediated genetic transformation of leaf explants from Nicotiana tabacum. Following parallel explant selection on media with paromomycin or kanamycin plant regeneration was observed. Total DNa of the regenerated plants was tested for the presence of nptII gene by PCr. '''Results.''' It has been shown that the use of antibiotic paromomycin is effective for obtaining transgenic tobacco plants. Paromomycin revealed a softer effect on plant explants compared with kanamycin sulfate without causing severe depression on regenerative ability of plant tissues, leading to a higher frequency of regeneration on selective medium and transformation. '''Conclusions.''' Transgenic tobacco plants were received on selective media containing paromomycin with high frequency. Due to the mild effects on plant explants, paromomycin is an attractive agent for selection of transgenic plants with nptII gene, especially for those species in which the kanamycin sulfate has a strong negative impact on the regenerative potential. | '''Aims.''' To investigate the efficiency of antibiotic paromomycin as a selective agent for the production of transgenic plants with nptII gene compared with kanamycin sulfate on the example of Nicotiana tabacum. '''Methods.''' a vector containing the nptII gene under the control of 35S promoter of cauliflower mosaic virus was used in Agrobacterium-mediated genetic transformation of leaf explants from Nicotiana tabacum. Following parallel explant selection on media with paromomycin or kanamycin plant regeneration was observed. Total DNa of the regenerated plants was tested for the presence of nptII gene by PCr. '''Results.''' It has been shown that the use of antibiotic paromomycin is effective for obtaining transgenic tobacco plants. Paromomycin revealed a softer effect on plant explants compared with kanamycin sulfate without causing severe depression on regenerative ability of plant tissues, leading to a higher frequency of regeneration on selective medium and transformation. '''Conclusions.''' Transgenic tobacco plants were received on selective media containing paromomycin with high frequency. Due to the mild effects on plant explants, paromomycin is an attractive agent for selection of transgenic plants with nptII gene, especially for those species in which the kanamycin sulfate has a strong negative impact on the regenerative potential. | ||
Версія за 00:01, 21 грудня 2015
The positive effect of antibiotic paromomycin compared with kanamycin for selection of transgenic plants with NPTII gene on the example of Nicotiana tabacum
Nitovska I.O., Avilov I.D., Morgun B.V.
|
Aims. To investigate the efficiency of antibiotic paromomycin as a selective agent for the production of transgenic plants with nptII gene compared with kanamycin sulfate on the example of Nicotiana tabacum. Methods. a vector containing the nptII gene under the control of 35S promoter of cauliflower mosaic virus was used in Agrobacterium-mediated genetic transformation of leaf explants from Nicotiana tabacum. Following parallel explant selection on media with paromomycin or kanamycin plant regeneration was observed. Total DNa of the regenerated plants was tested for the presence of nptII gene by PCr. Results. It has been shown that the use of antibiotic paromomycin is effective for obtaining transgenic tobacco plants. Paromomycin revealed a softer effect on plant explants compared with kanamycin sulfate without causing severe depression on regenerative ability of plant tissues, leading to a higher frequency of regeneration on selective medium and transformation. Conclusions. Transgenic tobacco plants were received on selective media containing paromomycin with high frequency. Due to the mild effects on plant explants, paromomycin is an attractive agent for selection of transgenic plants with nptII gene, especially for those species in which the kanamycin sulfate has a strong negative impact on the regenerative potential.
Keywords: Paromomycin, kanamycin, nptII, Nicotiana tabacum, selection of transgenic plants.
Fig. 1. The effect of kanamycin (a) and paromomycin (B) on leaf explants of the nontransformed tobacco plants. The arrows indicate on regenerated explants
Fig. 2. Electrophoretic analysis of the PCr products from tobacco regenerant lines for the presence of nptII (a) and vir D1 (B) genes. Lanes 25–29, DNa of tobacco regenerants obtained on kanamycin; lanes 30–34, DNa of tobacco pants regenerated at paromomycin; C-, negative control; C+, positive control; M, molecular weight marker λ DNa/ HindIII
Джерела
- Nitovska I.O., Avilov I.D., Morgun B.V. The positive effect of antibiotic paromomycin compared with kanamycin for selection of transgenic plants with NPTII gene on the example of Nicotiana tabacum // Фактори експериментальної еволюції органiзмiв: Зб. наук. пр. / Національна академія наук України, Інститут молекулярної біології і генетикп, Укр. Т-во генетиків і селекціонерів iм. М.I. Вавилова; редкол. / В. А. Кунах (голов. ред.) [та iн.] – К.: Укр. т-во генетиків і селекціонерів ім. М.І. Вавилова, 2015. – Т. 17. – С. 270-273
- Доклад на X Международной научной конференции «Факторы экспериментальной эволюции организмов»
