Incubation was carried out in a growing chamber (24��C, 16h light/8h dark photoperiod) and results www.selleckchem.com/products/Belinostat.html were evaluated on the tenth day of culture.The resistance test was carried out with the transgenic spring wheat line ��T-124�� in the seventh self-pollinated generation (T7). The gene bar had one integration site in this wheat line. Culture conditions during germination of the mature embryos were the same as in the pilot experiment. Media representing fourteen treatments with different concentrations of glufosinate ammonium added to them were as follows: 2, 4, 8, 16, 32, 64, 128, 200, 400, 600, 800, 1000 and 5000mg?L?1. Medium of the control treatment contained no herbicide. One embryo was put into every tube and every treatment was repeated eight times.

After three weeks of culture, plantlets were transferred to pots filled with soil, acclimatized and grown to maturity in the greenhouse. Plants were sprayed with insecticides and fungicides twice during the growing period. Exclusively mechanical weed control was also applied. Spikes were harvested individually and sorted into two groups termed well filled and low filled according to visual qualification. Yield components as number of spikes per plant, number of grains per spike, and yield per plant were measured while thousand kernel weight was calculated after harvesting.2.3. Molecular AssaysPlantlets were analyzed by molecular methods in every transgenic generation. At the seedling stage, 30mg of leaf samples were collected and immediately frozen in liquid nitrogen.

For the purification of total RNA, the ��SV Total RNA Isolation System�� kit (Promega) was applied; the protocol also contained the DNase treatment. To prove not only the presence but also the expression of the bar gene, a fragment 375bp in length derived from its RNA transcript was amplified by RT-PCR (one step reverse transcriptase polymerase chain reaction) with the aid of the specific primers bar5F and bar6R (5��-CAGGAACCGCAGGAGTGGA-3�� and 5��-CCAGAAACCCACGTCATG-3��, resp.). RT-PCR products were detected by electrophoresis on 1% TAE-agarose gel. Only the bar + plants were grown to maturity and harvested in every generation. Concerning the resistance test population, one out of the eight individuals was randomly chosen in each herbicide treatment and analyzed as described above.2.4.

Experimental Conditions of Transgenic ResearchTransgenic experiments were carried out in closed experimental conditions (in vitro growing chamber and closed greenhouse cabin). After the observations destruction of experimental plant material was documented in an official report Dacomitinib for the Hungarian authorities.2.5. Statistical EvaluationResults of well-filled and low-filled groups were evaluated separately. In every treatment, main rates were calculated by averaging of the results of the eight repeats.