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采用初始能量为100MeV/u的重离子束12C6+对紫花苜蓿下胚轴及子叶外植体进行辐照处理,研究重离子辐照对愈伤组织诱导状态及诱导率、愈伤组织相对生长率、体细胞胚诱导率及植株再生的影响。结果表明,重离子辐照对下胚轴及子叶愈伤组织的诱导具有抑制作用,且出愈率随着辐照剂量的增大而降低;在继代培养过程中,其愈伤组织相对生长率均高于对照组,外植体本身对重离子辐照所造成的损伤具有恢复能力;辐照处理对体细胞胚胎诱导也有影响,30Gy时,下胚轴诱导的体细胞胚胎发生较对照组早,数量多,较早地得到再生植株;而50Gy时,所得到的体细胞胚未能发育成再生植株。同时应用随机扩增多态性DNA(Random amplified polymorphic DNA,RAPD)技术对重离子辐照处理下胚轴所得再生植株进行检测分析,结果表明:所采用的20条随机引物中有11条在对照及处理组所得再生植株之间扩增出差异性多态条带,表明了重离子辐照引起苜蓿再生植株基因组DNA发生变异。
Irradiation of alfalfa hypocotyls and cotyledon explants was performed using heavy ion beam 12C6 + with an initial energy of 100 MeV / u. The effects of heavy ion irradiation on callus induction and callus induction, callus relative growth rate, Somatic Embryo Induction and Plant Regeneration. The results showed that heavy ion irradiation could inhibit the induction of hypocotyl and cotyledon callus, and the rate of callus decreased with the increase of irradiation dose. During the subculture, callus grew relatively The rate of hypocotyl induced somatic embryogenesis was higher than that of the control group, and the explant itself had the ability to restrain the damage caused by heavy ion irradiation. The irradiation also affected somatic embryogenesis, Early, the number of more early regenerated plants; and 50Gy, the resulting somatic embryos failed to develop into regenerated plants. At the same time, RAPD was used to detect the regenerated plants from hypocotyls irradiated by heavy ions. The results showed that 11 out of 20 random primers were used in control And the treatment group obtained between the regenerated plants amplified polymorphic bands showed that heavy ion irradiation led alfalfa plant genomic DNA mutation.