N+离子注入对干旱胁迫条件下甘草幼苗的SOD酶和CAT酶活性及丙二醛含量的影响

以低能氮离子束为诱变源,对甘草种进行注入处理后,以萌发的幼苗为材料对其在干旱胁迫条件下保护酶活性及MDA含量变化进行研究.结果表明注入后甘草种的存活率呈现马鞍形曲线变化,其中在4.1×1016N+/cm2剂量处达到鞍点,在剂量4.6×1016N+/cm2处存活率高于对照.在干旱胁迫下,随着胁迫时间的延长,离子束注入后萌发的幼苗MDA含量明显低于对照.酶活结果显示,在干旱胁迫后期,剂量4.6×1016N+/cm2处理后的甘草幼苗SOD和CAT酶活性都高于对照,说明该注入剂量处理甘草种有助于提高甘草幼苗的抗旱性能,SOD同工酶结果也证明该剂量的离子注入对干旱胁迫下同工酶谱带有所影响.     

Sequence analysis of lacZ~- mutations induced by ion beam irradiation in double-stranded M13mp18DNA

While M13mpl8 double-stranded DNA was irradiated with ion beam, and transfected into E. coli JM103, a decrease of transfecting activity was discovered. The lacZ-mutation frequency at 20% survival could reach (3.6-16.8) × 104, about 2.3-10 times that of unirradiated M13DNA. Altogether, 27 lacZ~ mutants were select-ed, 10 of which were used for sequencing. 7 of the sequenced mutants show base changes in 250-bp region examined (the remaining 3 mutants probably have base changes outside the regions sequenced). 5 of the base-changed mutants contain more than one mutational base sites (some of them even have 5-6 mutational base sites in 250-bp region ex-amined) ; this dense distribution of base changes in polysites has seldom been seen in X-rays, γ-rays or UV induced DNA mutations. Our experiments also showed that the types of base changes include transitions( 50 % ), transversions (45% ) and deletion (5% ); no addition or duplication was observed. The transitions were mainly C→T and A→G; the transversion     

Comparison on the genotoxic effects of nuclear vs cytoplas-mic irradiation from the alteration of CD59 gene locus

Using microbeam to irradiate human-hamster hybrid AL cells with defined number of a particles in a highly localized spatial region, this paper showed that cytoplasmic irradiation induced very little toxicity. For example, the cell killing by 4α particle traversal through the cytoplasm was about 10%, and about 70% cells survived after their cytoplasm was irradiated with 32 a particles. In contrast, the survival fractions for nuclear irradiation at the same doses were 35% and less than 1% respectively. Mutation induction showed that while nuclear irradiation induced 3-4-fold more CD59- mutants than cytoplasmic irradiation at equivalent particle traversal, at an equitoxic dose level of 90% survival, the latter exposure mode induced 3.3-fold more mutants than nuclear irradiation. Moreover, using multiplex PCR to analyze five marker genes on chromosome 11 (WT, CAT, PTH, APO-A1 and RAS), the results showed that the majority of mutants induced by cytoplasmic irradiation had retained all of the marker genes ana     

低能N~+离子束注入香瓜种子引起的变异及后代基因组的RAPD分析

用低能N+离子束对香瓜种子进行不同能量和剂量组合的处理。从各处理组与对照组当代和第三代苗期形态特征观察、考种中发现 :在一定能量和剂量组合下 ,处理组香瓜叶型和果肉厚度发生了明显的变化 ,这种形态学变化是可遗传的。应用随机引物扩增多态性DNA (RandomamplifiedpolymorphicDNA即RAPD)技术分析对照组和注入低能N+离子束处理组的香瓜总DNA的结果显示 :10 0种随机引物中的 2 4种引物扩增出 44条多态性片段 ,且其中一种引物所扩增的条带与叶型变化相连锁。表明低能N+注入香瓜种子可引起其基因组DNA发生变异 ,一定注入剂量和能量的组合 ,可导致特异性的变异。     

离子注入法将外源DNA直接导入小麦的研究

通过离子束介导法将外源GUS基因直接导入小麦成熟种子.组织化学染色结果表明GUS基因的瞬时表达率可以达到70%以上.当代(Ru)PCR分析结果表明,阳性植株的频率与离子注入剂量有关,适宜的注入剂量为7×106离子/cm2.PCR-Southern和Southern B1ot分析结果表明外源基因已整合到小麦基因组中,说明离子束介导外源DNA直接导入小麦是可行的.另外还探讨了用离子束介导创造小麦远缘分子杂种的可能性.     

荞麦组织培养研究

养麦种子含有丰富的碳水化合物、人体所需的各种氨基酸,多种微量元素和维生素等,其营养价值是任何食用作物无法比拟的。同时它也是蜜源植物和医疗与保健食品,更是良好的出口创汇商品之一。但荞麦品种极度退化,严重影响其产量和品质的进一步提高。针对这个问题,从1989年起我们试图用组织培养手段来培育荞麦新品种和改良复壮以促进荞麦生产的发展。现将我们的前期工作初步结果简报如下。     

低能氮离子注入西瓜胚芽的存活率的初步研究

以西瓜胚芽为材料 ,研究了低能氮离子注入植物活体组织的存活率和二甲亚砜 (DMSO)预处理对存活率的影响。结果表明 ,真空冰冻对存活率有一定影响 ,离子注入损伤是植物活组织在离子注入时存活率降低的主要原因 ;在能量为 2 5 ke V,总剂量为 3.9× 10 1 6ions/ cm2 ,脉冲剂量为 1.3× 10 1 4 ions/ cm2 的注入参数下 ,1%的 DMSO预处理 2 5～ 35分钟可以降低失水率并大大提高胚芽的存活率。由此讨论了进一步扩大低能离子注入的应用范围 ,以及在果树、花卉和无性繁殖的作物上进行诱变育种或遗传转化的可能性。     

低能离子束介导的遗传转化研究进展

低能离子束注入对细胞的刻蚀作用提供了外源遗传物质进入细胞的途径,离子束介导的遗传转化技术已在水稻,小麦,烟草,棉花等多种植物上取得成功,对离子束介导遗传转化的原理和最新进展进行了评述。．     

从CD59基因的变化比较细胞核、细胞质受辐射损伤的生物效应差异

通过利用精确数量的α粒子定点定量照射人与中国仓鼠杂交A_L细胞的核或质,显示出细胞质受照射引起的细胞致死率非常小,如4个α粒子的致死率约10%,即使受32个粒子照射,仍有70%以上的细胞存活形成克隆;而在相同的照射剂量下,核受照射的存活率分别是35%和小于1%.诱变试验发现尽管在相同照射剂量下,照射细胞核比照射细胞质在CD59基因位点可多诱发3～4倍的突变子,但在90%相同存活率的情况下,照射细胞质比照射细胞核多诱发3.3倍的突变子.此外,突变子的WT,CAT,APO-A1,PTH和RAS等基因缺失率PCR分析显示,细胞质受照射很少引起基因的大片段丢失,其突变谱类似于自发突变谱,而细胞核受照射的基因缺失率随照射粒子数的增加而增大.     

Alpha粒子和柴油机尾气颗粒提取物共同处理哺乳动物细胞的协同毒性作用

为研究两种有害环境因子同时作用对人类健康的影响,以alpha粒子和柴油机尾气颗粒提取物(d iese lparticu late extract,DPE)共同处理皮肤成纤维细胞(AG1522)为实验模型,对细胞毒性、遗传毒性进行研究。实验发现和未处理的对照组相比,1 cGy alpha粒子和20μg/mL DPE处理24 h,共同作用于细胞能够导致细胞活力下降40%左右,微核形成率上升2倍多;而1 cGy alpha粒子单独辐照时细胞活力下降14.19%,微核形成率上升了36.84%,20μg/mL DPE单独处理24 h时细胞活力下降17.29%,微核形成率上升了73.68%。实验结果表明:这两种环境因子共同作用于哺乳动物细胞时,对于细胞的毒性不是简单的叠加,而是表现出一种协同效应。