【作者】 吕杰；

【导师】 李冠；

【作者基本信息】 新疆大学 ， 植物学， 2004， 硕士

【摘要】 本文主要研究利用不同剂量氮离子束注入两种紫花苜蓿生物学效应，利用统计学方法及扫描电镜观察研究离子注入两种紫花苜蓿辐射损伤问题。并初步探讨离子束介导外源基因转化的方法。其结果如下：(1)从生理生化角度研究低能N+离子注入对两种紫花苜蓿的影响，用发芽率、发芽势、苗高、百株干重生理生化指标研究紫花苜蓿对低能N+离子注入的敏感性问题，得到种子发芽率和发芽势随注入剂量呈现先降后升再降的“马鞍型”曲线。(2)通过两种紫花苜蓿幼苗的过氧化物酶、过氧化氢酶、超氧化物歧化酶和丙二醛等生理生化指标的研究，发现过氧化物酶活性随着离子注入剂量的递增逐渐增大，过氧化氢酶和超氧化物歧化酶活性随注入剂量增大也呈“马鞍型”曲线。而丙二醛含量和电解质外渗率与过氧化氢酶和超氧化物歧化酶活性变化相反。证明合适剂量的氮离子注入可有效的减弱苜蓿幼苗脂质过氧化作用。(3)通过对两种紫花苜蓿品质的分析，证明合适剂量的N+离子注入，陇东苜蓿粗蛋白含量有了显著的提高。通过对两种苜蓿粗蛋白含量的方差分析以及各剂量组之间差异性的比较，得出当氮离子注入剂量为4×1016 N+/cm2和6×1016 N+/cm2时，能使陇东苜蓿粗蛋白含量显著提高。(4) 通过同工酶谱分析，发现低能氮离子注入能引起紫花苜蓿幼苗过氧化物酶同工酶变异，利用扫描电镜对两品种苜蓿各剂量组处理后种子观察，发现随着剂量的升高，两种苜蓿种子种皮刻蚀程度逐渐加强。（5）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，检测到有外源基因转入苜蓿幼苗中。 实验证明，若以提高苜蓿品质为目的，所选取的剂量范围对陇东苜蓿具有很好的刺激效应，但对新疆大叶苜蓿效果不佳，注入剂量过大，对新疆大叶苜蓿损伤较为严重。更多还原

【Abstract】 In this paper,the biological effects of Nitrogen ion implantation on two alfalfa varieties were studied ,and the difference sensitiveness by low energy N+ implantation were explored with statistical method and observation by means of scanning electron microscope(SEM).Using the ion beam mediated transgenic method to transfer NP-1 gene into seeds of Longdong alfalfa. The experiment was divided into 5 parts:(1)The difference sensitiveness of N+ implantation on two alfalfa varieties has been studied by measuring the germination energy, germination rate ,seedlings height and 100-seedlings weight . “Saddle type” line of germination rate and germination energy was investigated due to the N+ implantation.(2)The physiological and biochemical indexes were also researched. The results showed both activities catalase and superoxides dismutase were “saddle modle”. The high activities of catalase and superoxides dismutase decreased the accumulation of modondiadehyde and electrolyte leakage of cotyledons. It indcates that optimum N+ implantation may weaken the lipid peroxidation in alfalfa seedlings.(3)The quality of different dose of N+ implantation on two kinds of alfalfa were studied .The results showed that the content of coarse protein in Longdong alfalfa increased at optimum dosage of N+ implantation .The content of coarse protein analysis indicated content of coarse protein in Longdong alfalfa was significantly higher than others while the dosages were 4×1016 N+/cm2 and 6×1016 N+/cm2.(4) N+ implantation caused mutation of peroxides isoenzyme. The seed structures of two alfalfa varieties by different dosage of N+ implantation were studied by means of SEM .It showed <WP=7>that with the dosage increasing the structure of seed was seriously damaged.(5)Exogenous NP-1 gene was introduced into alfalfa mature seeds by ion implantation. The results of molecular detection proved that exogenous gene had integrated into alfalfa genome.更多还原