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以羽衣甘蓝和芥菜型油菜种间杂种(ZH201)(2=3x=ABC=27)为试验材料,以花序轴,果柄和子房为外植体,采用不同处理对其进行再生研究;对获得的再生试管苗用秋水仙素和二甲基亚砜进行处理,比较不同因素对染色体加倍的影响;对获得的三倍体和六倍体植株进行形态学和细胞学比较.结果表明:杂种植株再生频率从高到低依次为果柄、子房、花序轴,将外植体直接接种到分化培养基(MS+0.2 NAA+3.0 mg/L 6-BA 3.0 mg/L+5.0 mg/L AgNO3+0.6 mg/L GA3)上再生频率最高.在染色体加倍处理方面,以100 mg/L秋水仙素处理根尖24 h效果最好,获得的六倍体植株与三倍体相比,花朵颜色较深,花瓣较大,花药饱满,荚果能够正常生长,获得部分种子.在细胞学上,三倍体植株在花粉母细胞减数分裂中期Ⅰ存在着大量单价体,后期Ⅰ、后期Ⅱ发现有大量染色体落后现象.六倍体植株中在花粉母细胞减数分裂后期Ⅰ、后期Ⅱ也发现有染色体落后现象,但是落后染色体比三倍体少.
In order to study the regeneration of rapeseed (Brassica napus L.) and Brassica juncea (ZH201) (2 = 3x = ABC = 27) The regenerated in vitro plantlets were treated with colchicine and dimethyl sulfoxide to compare the effects of different factors on the chromosome doubling.The morphology and cytology of triploid and hexaploid plants were compared.The results showed that hybrid plants The regeneration frequency was in the order of stalk, ovary and inflorescence axis. The explants were inoculated directly into differentiation medium (MS + 0.2 NAA + 3.0 mg / L 6-BA 3.0 mg / L + 5.0 mg / L AgNO3 +0.6 mg / L GA3) .There was the best effect on chromosome doubling treatment when the root tips were treated with 100 mg / L colchicine for 24 h, and compared with the triploid, the flower color Deeper, petal larger, anther full, pod can grow normally, access to some of the seeds in the cytology, triploid plant pollen mother cell meiosis metaphase I there are a large number of monokaryons, late I, late Ⅱ found A large number of chromosome backward phenomenon in the hexaploid plants in pollen mother cell meiosis late Ⅰ, Phase Ⅱ also found chromosomal behind the phenomenon, but behind less than triploid chromosome.