【作者】 齐丽杰；

【导师】 李冠；

【作者基本信息】 新疆大学 ， 植物学， 2005， 硕士

【摘要】 本文主要研究不同剂量的低能氩离子束注入对甜瓜产生的诱变效应,并初步探讨离子束介导外源基因转化的方法。选取伽师瓜和皇后两甜瓜品种进行离子注入,能量为25keV,注入剂量为:0（Ck）, 2×10¹⁶ Ar⁺/cm²,4×10¹⁶ Ar⁺/cm²,5×10¹⁶ Ar⁺/cm²,6×10¹⁶ Ar⁺/cm²,7×10¹⁶ Ar⁺/cm²。处理后的甜瓜主要从生理生化水平和同工酶水平进行分析,其结果如下: （1）发芽率的研究中,离子注入对种子的发芽具有一定的抑制作用, 总的趋势是剂量越大发芽率越低。（2）通过对注入后的两甜瓜品种幼苗的过氧化物酶、过氧化氢酶、超氧化物歧化酶、丙二醛、活性氧自由基、抗坏血酸过氧化物酶、抗坏血酸和类黄酮等生理生化指标的测定,表明低中剂量的离子注入会对细胞造成损伤,产生大量自由基,导致细胞的保护酶活性和非酶类抗氧剂含量的上升,使膜脂过氧化程度受到抑制。但当离子注入剂量过大时,植株的保护酶系统和活性氧非酶促系统都受到抑制,脂质过氧化产物积累。（3）用聚丙烯酰胺凝胶电泳对甜瓜幼叶中的过氧化物同工酶进行分析,结果表明,与对照相比,离子注入使过氧化物同工酶的活性和数量发生了变化。（4）对离子束介导转基因当代幼苗进行NP-1 基因PCR 特异性扩增检测,检测到外源基因转入愈伤组织中。实验证明,皇后品种对离子注入较伽师瓜品种敏感,故在以后的离子束诱变甜瓜育种的工作中,对皇后品种应降低注入剂量减少对其的损伤,而对伽师瓜可再适当的提高注入剂量以期达到更好的刺激效应。更多还原

【Abstract】 In this paper, the biological effects of different doses Ar+ ion implantation on Cucumis melo L were studied in order to search the relationship between ion implantation and Cucumis melo L. Using the ion beam mediated transgenic method to transfer NP-1 gene into seeds of cantaloupe. Seeds of Jiashi cantaloupe and Huanghou cantaloupe were implanted by Ar⁺ ion beam, energy was 25keV, doses were 0, 2×10¹⁶ Ar⁺/cm²,4×10¹⁶ Ar⁺/cm²,5×10¹⁶ Ar+/cm²,6×10¹⁶ Ar⁺/cm²,7×10¹⁶ Ar⁺/cm². The results are as following: （1） After having observed the germination of seeds implanted by ion beams, the results showed that Ar+ implantation restrained the germination of cantaloupe seeds with the doses increasing. （2） Some physiological and biochemical indexes were analyzed, such as the activities of POD、CAT、SOD、ASP 、generating rate of O₂ and the content of H2O2、ASA、MDA, etc. the results indicated that ion implantation induced damnification, produced lots of free radical and lead to the activities of cell defense enzymes and no-enzymatic antioxidants increasing. The increasing activities of cell defense enzymes make the degree of membrane lipid peroxidation decrease. But with the doses increasing, the enzymatic and no-enzymatic protection mechanisms were destroyed and lead to the accumulation of MDA. （3） Analysed the peroxidase isozyme bands of the cantaloupes, we can see that Ar+ implantation caused the mutation of quantity and activity in peroxides isoenzyme. （4） Exogenous NP-1 gene was introduced into cantaloupe seeds by ion implantation. The results of molecular detection proved that exogenous gene had integrated into cantaloupe tissue culture. The experiments proved that Huanghou cantaloupe is more sensitive than Jiashi cantaloupe. So in future we should choose the best implantation parameters according to different cantaloupes in mutation breeding.更多还原