【作者】 郑媛；

【导师】 肖娅萍；

【作者基本信息】 陕西师范大学 ， 植物学， 2005， 硕士

【摘要】 烟草（Nicotiana tabacum L.）属茄科（Solanaceae）烟草属（Nicotiana）,原产南美洲,现广植于全世界温带和热带地区。叶为香烟的原料,全株亦可作农业杀虫药,是我国重要经济作物之一。因此,开展对烟草的组织培养研究工作具有重要意义。 环境应力对植物的影响已成为生物力学研究领域的一个热点。超声作为一种环境应力,对植物的生长发育有重要的影响。在长期以来的探索中,烟草作为组织培养中一种繁殖力强、抗病毒能力强的实验品种而成为植物学实验的材料首选,在本实验中选取烟草组培苗为材料从细胞生长、死亡、凋亡、生理、细胞结构五方面进行超声的生物学效应研究。主要研究结果如下: 1.建立了NC89烟草试管苗叶片、根段再生系统,分别确定了叶片不定芽诱导最佳培养基及激素配比为2/3MS+BA0.5mg/l+NAA0.1mg/l,不定芽诱导生根最佳培养基及激素配比为2/3MS+BA1.0mg/l+NAA1.0mg/l。根段不定芽诱导最佳培养基及激素配比为MS+BA0.3mg/l+IBA0.1mg/l,不定芽诱导生根最佳培养基及激素配比为MS+BA0.1mg/l+NAA1.5mg/l。研究表明细胞分裂素与生长素浓度的比值高时,有利于芽的分化,相反细胞分裂素与生长素浓度的比值低时,有利于根的分化。 2.以叶片为外植体,选择培养基2/3MS+BA0.5mg/l+NAA0.1mg/l为愈伤组织诱导培养基,用添加3.33248mg/lCa（NO₃）₂的2/3MS+BA0.5mg/l+NAA0.1mg/l为愈伤组织继代培养基,能迅速获得脆散愈伤组织。可能与这些愈伤组织迅速生长时Ca⁺⁺的需要量及Ca⁺⁺所具有的多种重要生物功能有关。另外利用根段为外植体在一定的激素调节下也可以获得脆散性的愈伤组织,确立了根段最佳愈伤组织诱导培养基MS+ZT0.5mg/l+IAA0.05mg/l,最佳愈伤组织继代培养基MS+KT0.5mg/l+IAA0.05mg/l。 3.超声能够显著诱导烟草细胞超氧化物歧化酶（SOD）活性,采用固定频率为27KHz,分别以不同的时间处理烟草悬浮细胞,以7min超声处理诱导效果最明显;不同时间段都会使SOD、过氧化氢酶（CAT）活性峰推后3天出现,但对CAT和过氧化物酶（POD）的活性诱导作用不明显或表现为抑制作用。说明超声处理刺激细胞产生了防御应答反应,诱导了相关的自由基清除酶类,如SOD酶的活性。可能是超声空化导致了酶分子复合结构的变化,从而有效地改善了酶的性能;或者超声波热学机制导致媒质质点温度升高而提高了酶的活性。 4.低剂量、短时间的温和超声处理能明显加速和诱导植物细胞的分裂,促更多还原

【Abstract】 Abstract: Tabacco（Nicotiana tabacum L.）origined from South America, planted in temperate zone and tropical belt in the world. The leaf is the material of the cigarette, the plant also can be used as agriculture helminthic, and it’s one of the important economic crops in our country. In the long research, tobacco became the preferred material in botany experiment, because of strong propagation ability, strong antivirus ability in tissue culture; some important meaning reseachment results have achieved. The effect of envirmental stresses on the growth and development of plant cells and tissues is one of important fields in plant physiology. In the experiment, we ultilized it to develop ultrasound biology function reseach, on the base of different explant reproduction system and cell suspension culture system establishment, we went on the reseachment in growth, physiology, cell ultrastructure, provided theory support for ultrasound biology effect.1. The experiment went on the reproduction system researchment of NC89 tobacco plantlet leaf and roots. Respectively ensured the best medium of adventitious bud induced by leaf was 2/3MS+BA0.5mg/l+NAA0.1mg/l, the best medium of adventitious root induced by adventitious bud was 2/3MS+BA1.0mg/l+NAA1.0mg/l, the best medium of adventitious bud induced by root was MS+BA0.3mg/l+ IBA0.1mg/1, the best medium of adventitious root induced by adventitious bud was MS+BA0.1mg/l+NAA1.5mg/l. It convinced that the higher concentrations rate of auxins and cytokinins was benefit for the adventitious buds differentiation, on the contrary, the slower concentration of that was benefit for the adventitious roots differentiation.2. The acquisition of friable calli and the establishment of the cell suspension cultures of tobacco NC89 leaf were studied. Firstly, the results indicated that the 2/3MS medium, which contained BA0.5mg/l and NAA0.1mg/l was suitable for calli differentiating from the leaf;the 2/3MS medium,which contained BA0.5mg/l, NAA0.1mg/l, 3.33248mg/l Ca （NO₃）₂ was suitable for calli subculture, so the friable calli can be acquired rapidly and the cell suspension cultures had been successfully established,maybe it’s related to the requirement quantity of the Ca⁺⁺ during these calli mushroom, besides some important biology function of the Ca⁺⁺. Secondly, the results indicated that the MS medium, which contained ZT 0.5mg/l and IAA 0.05mg/l was suitable for calli differentiating from the root; the MS medium, which contained KT0.5mg/l and IAA0.05mg/l was suitable for calli subculture.3. We study the effect of ultrasound on SOD, POD and CAT in cell suspension culture of Nicotiana tabacum L. The results showed that ultrasound could induct the activity of SOD evidently, and the effect of seven minute ultrasound was the most distinction. At the same time the different ultrasound period could make the activity peak of SOD and CAT retardate three days, but the ultrasound induction effect on POD and CAT were controlled and not clearly.We can draw a conclusion that addition of ultrasound to the cell suspension culture of Nicotiana tabacum L.could cause the cells to elicit the defence response and induct the activities of relative enzymes.Maybe更多还原