Studies On Polyploid Induction Of Blueberry In Vitro

Polyploid induction technique of blueberry, by treating blueberry shoot segments and calluses in vitro with coichicine, inducting anthers callus in vitro and identifying plantlet ploidy of blueberry in vitro, were studied. Mosaic material of ploidy was obtained, which would lay a suitable foundation for blueberry ploidy selection in the future. The main results were as following:1. The methods of polyploid induction of blueberry in vitro with artificial mutation way were studied for the first time. Calli induced from leaves in vitro of three cultivars Blomidon, Darrow and Elliott were obtained efficiently in WPM media with different concentration of ZT, 2,4-D and CPPU. The frequency of callus induction of Elliott was high up to 90% in WPM medium with CPPU 2.0 mg/L, sugar 30 g/L and agar 7 g/L, regeneration frequency at 73.9%. The calli of the three cultivars were getting brown and died seriously after immersing in liquid of 0%-0.2% coichicine for 6h,12h and 24h. The mosaic calli of Elliott were induced in medium with coichicine 0.002%, at 34.3% level of polyploidy cells.The results of polyploidy induced by immersing axillary buds in coichicine liquid or by culturing in culture medium with coichicine showed that induction effect through culturing in medium with coichicine was inferior to that through immersing axillary buds. The rate of survival and variation were closely related to genotype of blueberry, coichicine concentration and treating time in our experiment. Treatment by immersing axillary buds with 0.05% coichicine liquid for 24h got average survival rate at 42.9% and variation rate at 5.7% for the three cultivars. In the treatment of immersing with 0.1% coichicine liquid for 24h, the average survival rate declined to 20.9% and variation rate mounted up at 17.2%. The average rates of survival and variation declined to 9.9% respectively by treating with 0.2% coichicine for 24h. Along with the concentration of coichicine increased, the survival rate declined. But the v ariation rate did not always have positive correlation to the concentration of coichicine. In addition, genotypes of blueberries had different response to the same treatments in our trial. Darrow got better effect than Elliott and Blomidon in the experiment. By immersing axillary buds with 0.1% colchicines liquid for 24h, Darrow's survival rate was 28.9%, and variation rate was 22.6%. The identification results of cell ploidy in the material induced showed that most strains were mosaic with diploid and tetraploid cells at average levels of 79.4% and 20.6% respectively.2. Induction and culture of blueberry anther callus were studied for the first time. The anthers calli...