转BADH基因玉米与非转化受体生长、生理特性及农艺性状比较

发布时间：2018-05-22 11:32

本文选题：BADH基因 + 玉米(Zea　；参考：《山西师范大学》2016年硕士论文

【摘要】：玉米是我国重要的粮食和饲料作物,在我国占据极其重要的位置。由于现如今土地干旱和盐碱化程度越来越严重,因此,研究和培育抗旱耐盐植物迫在眉睫。在干旱胁迫下,甜菜碱作为一种有效的渗透调节物质,可以在调节渗透方面发挥巨大作用。BADH(Betaine aldehydedehydrogenase)是合成甜菜碱的关键酶之一,通过基因工程手段将BADH基因导入玉米植株,从而获得抗旱性玉米,这对提高农作物产量意义重大。本研究所采用的材料为T4代转BADH基因玉米植株,是通过花粉介导法获得的。通过对转基因玉米与非转基因在生长特性、生理和代谢特征、农艺性状以及与环境相互作用等方面的比较研究,为使用该基因转化方法获得的转基因作物商业化生产提供了必须的基础研究,这对全面理解转基因作物与非转化受体二者实质等同意义重大。主要研究结果如下:1、在干旱胁迫下,BADH玉米长势较非转化受体植株好,株高、干重和鲜重比同期非转化受体分别高112.1%,105.4%和54.3%,表现出较好的抗旱性能;在正常水分条件下,两组玉米长势基本一致,株高、干重和鲜重分别比同期非转化受体植株高7.18%、4.26%和0.60%,但是差异均不显著。2、在干旱胁迫下,转BADH基因玉米的可溶性糖、可溶性蛋白、脯氨酸和甜菜碱含量比非转化受体分别高33.45%,92.65%,690%和211.3%,表明其具有更好的渗透调节能力;在正常水分条件下,转BADH基因玉米的可溶性糖、脯氨酸和甜菜碱含量分别比非转化受体分别高4.42%,21.72%和9.5%,可溶性蛋白含量比非转化受体低67.69%,这些物质的变化可能是由于植物通过自我保护机制调节体内代谢所致。3、在干旱胁迫下,BADH玉米的相对电导率、超氧阴离子自由基和丙二醛含量比非转化受体分别低66%、138%和127%,表明在干旱胁迫下叶片受到较轻程度损伤;在正常水分条件下,BADH玉米叶片相对电导率、超氧阴离子自由基比非转化受体分别高185%、91.3%,丙二醛含量比非转化受体低127%,表明BADH玉米叶片受到了一定的损伤,积累了较多的有害产物。4、在干旱胁迫下,BADH玉米的SOD活性和POD活性比非转化受体分别高33%和36%;在正常水分条件下,BADH玉米SOD活性比非转化受体分别高80%和9.8%,表明转基因植株具有较好的活性氧清除能力。5、通过对转BADH基因玉米与非转化对照玉米穗的形态观察和考种发现,两组玉米在主要农艺性状方面无显著差异。6、BADH玉米与非转化受体植株的根部附近土壤细菌、放线菌和真菌数量均没有显著差异,初步表明,两组玉米对微生物多样性的影响相同;7、半定量RT-PCR和Real-time PCR分析的结果表明,BADH基因在转录水平得以表达,而且在干旱逆境中,BADH基因的相对表达量比正常水分供应下高。
[Abstract]:Corn is an important food and feed crop in China, and occupies an extremely important position in our country. Because the soil drought and salinization are becoming more and more serious nowadays, it is urgent to study and cultivate drought-resistant and salt-tolerant plants. Under drought stress, betaine, as an effective osmotic regulator, can play an important role in the regulation of osmotic activity. Betaine Aldehyde dehydrogenase is one of the key enzymes for the synthesis of betaine. The BADH gene was introduced into maize plants by genetic engineering. So as to obtain drought-resistant maize, which is of great significance to improve crop yield. The material used in this study was T 4 transgenic maize with BADH gene, which was obtained by pollen mediated method. By comparing the growth characteristics, physiological and metabolic characteristics, agronomic characters and interaction with environment of transgenic maize and non-transgenic maize, It provides the necessary basic research for the commercial production of transgenic crops obtained by using the method of gene transformation, which is of great significance for the comprehensive understanding of the substantive equivalence between transgenic crops and non-transformed receptors. The main results were as follows: under drought stress, the plant height, dry weight and fresh weight of BADH maize were better than that of non-transformed receptor plants, and the plant height, dry weight and fresh weight were higher than those of non-transformed receptor plants by 105.4% and 54.3%, respectively, under drought stress. The plant height, dry weight and fresh weight of the two groups were 7.18% and 0.60% higher than those of the non-transformed receptor plants, respectively, but the difference was not significant. Under drought stress, the soluble sugar and protein of the transgenic maize with BADH gene were increased. The contents of proline and betaine were 33.45 ~ 92.65% and 211.3% higher than those of non-transforming receptor, respectively, which indicated that proline and betaine had better osmotic regulation ability, and the soluble sugar of BADH transgenic maize was higher than that of normal water. The contents of proline and betaine were 4.42% and 9.5% higher than those of the non-transforming receptor, respectively, and the soluble protein content was 67.69% lower than that of the non-transforming receptor. The changes of these substances may be due to the regulation of metabolism in the body by the self-protective mechanism. The relative conductivity of BADH maize under drought stress, The contents of superoxide anion free radicals and malondialdehyde were 66 138% and 127% lower than those of non-transformed receptors, respectively, indicating that the leaves were slightly damaged under drought stress, and the relative electrical conductivity of leaves of BADH maize was decreased under normal water condition. Superoxide anion free radicals were higher than non-transformant receptors, and malondialdehyde content was 127% lower than that of non-transforming receptors, indicating that BADH maize leaves were damaged to some extent. Under drought stress, the SOD activity and POD activity of Badh maize were 33% and 36% higher than that of non-transformed receptor, respectively, and the SOD activity of BADH maize was 80% and 9.8% higher than that of non-transformed receptor under normal water condition. Because the plants have better scavenging ability of active oxygen species, the ear morphology of transgenic maize with BADH gene and that of non-transformed maize were observed and tested. There was no significant difference in main agronomic characters between the two groups. There was no significant difference in the number of soil bacteria, actinomycetes and fungi between the two groups. The results of semi-quantitative RT-PCR and Real-time PCR analysis showed that the BADH gene was expressed at the transcriptional level, and the relative expression of BADH gene in drought stress was higher than that in normal water supply.
【学位授予单位】：山西师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S513

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