论文标题:人工诱发烟草类根瘤的生理生化研究
The Studies of Physiology and Biochemistry on Artificial Induced Tobacco Para-nodule
论文作者 母波
论文导师 韩善华,论文学位 硕士,论文专业 细胞生物学
论文单位 四川师范大学,点击次数 79,论文页数 94页File Size2513k
2005-05-01论文网 http://www.lw23.com/lunwen_756059587/ 烟草;类根瘤;2、4-D;根瘤菌;超氧化物歧化酶(SOD);影响
tobacco; para-nudule; 2、4-D; Rhizobium; Superoxide dismutase; Effect
我们以只在共生体系中才有固氮活性的突变型豇豆根瘤菌512(R.leguminosarm)和全国推广种植的烟草K326组建共生固氮体系,使用化学合成的植物激素2,4-D人工诱导烟草根系,使其结瘤。我们对烟草K326无菌苗叶片经组织培养产生的再生苗进行了以下4 种处理(CK:使用不含2,4-D和根瘤菌的常规Jensen无氮培养液处理;D:使用只含有0.3mg/l 2,4-D的Jensen无氮培养液处理;R:使用只含有5×108个/L根瘤菌的Jensen无氮培养液处理;D+R:使用同时含有0.3 mg/l 2,4-D和5×108个/L根瘤菌的Jensen无氮培养液处理)。在实验中,我们测定了以上四种处理对不同发育周期烟草不同组织来源的超氧化物歧化酶(SOD)的活性,丙二醛(MDA),可溶性蛋白和叶绿素含量以及根尖细胞染色体的影响。研究结果表明,使用2,4-D 处理后,在处理前期(第一至第三周)抑制了烟草茎和叶细胞中MDA 的积累,诱导其可溶性蛋白含量和SOD 活性的升高;在处理后期(第四至第五周),2,4-D 则加速了烟草茎和叶细胞中MDA 的积累、可溶性蛋白和SOD 活性的下降,并且加快了烟草叶片中叶绿素的降解,但是却抑制烟草根细胞MDA 的积累;仅用根瘤菌处理,在第一至第三周促进烟草根茎叶细胞中MDA 积累,诱导烟草SOD 活性和可溶性蛋白含量的升高,随着时间的延长,根细胞中的MDA 含量下降并且低于对照,而且此时根瘤菌促进烟草根茎叶细胞中可溶性蛋白含量和SOD 活性的下降;并且在处理第一周促进烟草叶片叶绿素的合成,随着处理时间的延长,却加速烟草叶片叶绿素的降解;在我们用2,4-D 和根瘤菌共同处理的烟草幼苗中,其不同发育时期根茎叶的MDA 含量的变化与仅使用2,4-D 或根瘤菌处理的不尽相同。在处理第一和第二周,共同处理加速了烟草各组织中MDA 的积累,并且促进各组织中SOD 活性的升高,然而此时却抑制烟草叶片叶绿素的
We set up the symbiotic nitrogen-fixation system with a mutant of fast-growing strain in R.sp.vigna. Which have nitrogen-fixation activity only in the symbiotic system and the tobacco K326 which is being extended to the whole country. The plant hormone 2,4-D was used to induce artificial tobacco para-nodule.We carried on the following 4 kinds of treatments to the regenerated tobacco K326 plantlets, which were produced from the leaves of the sterile seedling of tobacco.(CK:native Jensen non-nitrogen medium; D:Jensen with 0.3mg/L2,4-D alone; R: Jensen with 5×108/L rhizobium alone; D+R: Jensen with 0.3mg/L 2,4-D and 5 ×108/L rhizobium)In our experiment, effects on the activity of superoxide dismutase(SOD) and the contents of MDA、soluble protein chlorophyll and choromosome in tobacco’s leaf、stem and root at different treatments and for different time were studied. The results show that 2,4-D inhibited accumulation of MDA in the tobacco stem and leaf cell, induced increase content of soluble protein and activity of SOD(from the first week to the third week); later ,2,4-D accelerated the decrease of the content of MDA and soluble protein and activity of SOD of each organization of tobacco.The effect of treatment content of MDA with rhizobium alone is always higher than that of CK at different growth stages in tobacco root ,stem and leaf cells, and rhizobium could induce activity of SOD and content of soluble protein to increase in the first and the second week.At later, the content of MDA descended and was lower than that of CK in the root cell, and rhizobium could promote descrease of soluble protein content and SOD activity of tobacco root, stem and leaf cells, and could still inhibit the tobacco chlorophyll synthesis at the prophase of treatment(from the first week to the second week), but the rhizobium also could decelerate the chlorophyll destroyed with the extension of time; as the tobacco dissimilarity develops, the variety of the content of MDA was treated with 2,4-D and rhizobium didn’t achieve to the utmost as with the use of only either 2,4-D or rhizobium treated. At the prophase treatment, the common processing accelerated each organization of tobacco in the MDA accumulation, and stimulated soluble protein content and SOD activity of each organization to go up.However, at this time it inhibited the tobacco chlorophyll synthesis with the extension of the time. In each organization of tobacco of the common processing, although the content of MDA still presented the up-trend but lower than that of CK. At this time, the activity of SOD was all higher than that of CK in each organization. After different treatment, we found that it was the tobacco root cell that was victimized at the earliest, the stem cell was second to it and the leaf cell was victimized at the latest. The results still show that mitotic index of tobacco root cell was promoted to increase with the use of different treatments in the second week. The mitotic index of tobacco root cell was promoted most obviously with use of 2,4-D and rhizobium treatment. Along with the extension of time, the mitotic index of tobacco root cell was inhibited obviously in different treatments, and caused chromosome mutations in different degrees. Therefore, according to our experiment, when using plant hormone 2,4-D, the artificial induced tobacco para-nodule had certain stimulative function at the early period , then it is harmful to growth of tobacco with the extension of time.
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