转TaCHP基因耐盐碱小麦的理化分析

发布时间：2018-05-05 17:20

本文选题：小麦 + TaCHP　；参考：《山东大学》2016年硕士论文

【摘要】：小麦是甜土作物,土壤盐碱化是限制小麦生长、发育和产量提高的主要因素。随着分子生物学的发展,利用转基因技术进行小麦耐盐碱遗传改良是分子育种的新方向。实验室前期利用不同转化法,经抗生素筛选和分子鉴定获得两组转TaCHP基因小麦株系：以普通小麦济南17(JN 17)为亲本,经茎尖生长点转化法获得的T6代转基因系No.25-2-5和No.33-2-3；以普通小麦周麦22(Zhou 22)为亲本,经愈伤组织转化法获得的T4代转基因系No.21-1-3、No.26-12-5 和 No.27-6-8。本课题以这两组转基因系和各自的亲本对照为研究对象,进行如下实验：1)、实验室中,进行了转基因小麦在盐(NaCl)、碱盐(NaHCO3: Na2CO3=9:1)及混合盐碱[NaCl+(NaHCO3:Na2CO3=9:1)]的水溶液胁迫下种子萌发和早期幼苗生长发育实验；营养土和盐碱土中,种子萌发和早期幼苗的生长发育实验。2)、普通土和盐碱土试验田中,对两组转基因系进行了小麦部分生长发育期(苗期、返青期和灌浆期)形态特征观察记录、理化指标测定和农艺性状统计,并进行产量相关性分析。3)、在梯度盐池和碱池中,对T4代转基因系进行农艺性状和产量相关性分析。主要研究成果：1、实验室中,在萌发期,NaCl水溶液处理的两组转基因系表现出比各自亲本更强的耐盐性；碱性盐水溶液处理下差异不显著；混合盐碱[25 mM NaCl+50 mM (NaHCO3:Na2CO3=9:1)]水溶液处理下,两组转基因系的生长状态与各自的亲本对照差异不显著；混合盐碱[50 mM NaCl+25 mM (NaHCO3: Na2CO3=9:1)]水溶液处理组,两组转基因系的相对胚芽长度大于各自的亲本对照。在苗期,NaCl水溶液处理的转基因系表现出与萌发期一致的强耐盐性；两组转基因系在苗期均表现出高于各自亲本对照的耐混合盐碱胁迫特性。2、实验室中,点种在营养土中的种子,24 h后即开始萌动和快速生长,萌发至第4天时,种子几乎全部发芽；点种在盐碱土中的种子,48 h后才陆续开始萌动,随后几天陆续出芽和生长,且发芽率低于前者；两组转基因系种子的萌动及发芽率都优于各自的亲本对照。生长在营养土中20天的转基因植株生长量与亲本差异不显著,生长在盐碱土中的转基因株系生长量均高于各自的亲本对照,且总体显著低于营养土中。3、通过对生长在盐碱土试验田中的两组转基因系的叶片和根系的理化指标分析发现,各发育期(除返青期根系)的K+/Na+值显著高于各自的亲本对照,且叶片高于根中。两组转基因系各时期叶片和根系的MDA含量都显著低于亲本对照(除返青期叶片)。与K+/Na+值不同的是,随着发育进程的推进,叶片及根中MDA含量都明显升高。4、普通土试验田中,茎尖转化法获得的两个转基因系产量较JN 17分别提高了3.37%和4.53%,但在经愈伤组织法获得的转基因系与亲本Zhou 22的对比发现无显著差异；农艺性状中,小穗数和分蘖数与亩产相关性最大。盐碱土试验田中,两组转基因系的产量较其各自的亲本都有所提高；农艺性状中,株高、旗叶长度和小穗数与亩产相关性最大,其中,株高呈负相关,后两者呈正相关。5、对梯度盐池及碱池的农艺性状分析显示,在一定浓度的盐(0-0.8%NaCl)和碱盐(0-80 mM (NaHCO3:Na2CO3=9:1))胁迫的范围内,小麦产量随处理浓度的增加总体有降低的趋势。但当盐、碱盐处理浓度达到较大值[0.8%NaCl,80 mM碱池(NaHCO3:Na2CO3=9:1)]时,转基因系产量高于亲本对照Zhou 22,此时,在0.8% NaCl盐池中,三个转基因系的产量分别较亲本对照提高了0.03、0.37和0.16倍,80 mM (NaHCO3:Na2CO3=91)碱池中,三个转基因系的千粒重分别较亲本对照提高了的0.18、0.37和0.57倍。综上,实验室中,盐碱胁迫下的两组转基因系,在种子萌发和苗期生长所测的各指标均优于各自的亲本对照；盐碱土试验田中,两组转基因系均通过离子调节和氧化酶活性调节保护质膜免受盐碱伤害,以维持植物体正常生长。研究结果可为小麦转基因系耐盐碱机制的探索提供依据和可参考的技术路线。
[Abstract]:Wheat is a sweet soil crop, and soil salinization is the main factor restricting the growth, development and yield of wheat. With the development of molecular biology, genetic improvement of salt tolerance in wheat is a new direction of molecular breeding with the development of molecular biology. In the early laboratory, two groups of TaCHP were obtained by different transformation methods, through the screening of antibiotics and molecular identification. Gene wheat lines: the T6 generation transgenic lines No.25-2-5 and No.33-2-3 obtained from Ji'nan 17 (JN 17) were obtained by the stem tip growth point transformation method. The T4 generation transgenic lines, No.21-1-3, No.26-12-5 and No.27-6-8., obtained by callus transformation method, were obtained by the transformation method of normal wheat 22 (Zhou 22). The two sets of transgenic lines were given by the two sets of transgenic lines. Compared with their parents, the experiment was carried out as follows: 1), in the laboratory, the seeds germinated and the early seedling growth experiments were carried out under the water solution stress of salt (NaCl), alkali salt (NaHCO3: Na2CO3=9:1) and mixed salt base [NaCl+ (NaHCO3:Na2CO3=9:1)) in the laboratory of Transgenic Wheat; the seed germination and the early young were in the nutrient soil and the saline alkali soil. The growth and development experiment of seedling.2), in the ordinary soil and saline alkali soil test fields, the morphological characteristics of the two groups of transgenic lines were observed, the physical and chemical indexes and the agronomic traits were recorded, and the yield correlation was analyzed by.3). In the gradient salt pool and the alkali pond, the transgenic lines of the T4 generation were introduced into the transgenic lines. The correlation analysis of agronomic traits and yield. 1, 1, in the laboratory, in the germination period, the two groups of transgenic lines treated by NaCl aqueous solution showed better salt tolerance than their parents; the difference was not significant under the alkaline saline solution; the two groups were treated by the mixed saline alkali [25 mM NaCl+50 mM (NaHCO3:Na2CO3=9:1)] solution treatment. The growth state of the gene lines was not significantly different from those of their parents; the relative germ length of the two groups of transgenic lines in the mixed saline alkali [50 mM NaCl+25 mM (NaHCO3: Na2CO3=9:1) treatment group was larger than the parent control. In the seedling stage, the transgenic lines treated by the NaCl aqueous solution showed the strong salt tolerance of the two groups. In the seedling stage, the transgenic lines showed higher resistance to mixed salt stress.2 than their parent controls. In the laboratory, seeds in the nutrient soil began to germinate and grow quickly after 24 h, and the seeds germinated almost all after the germination to fourth days. Seeds in saline alkali soil began to germinate after 48 h, and then several days later. The germination and germination rates were lower than that of the former, and the germination and germination rate of the two transgenic lines were better than those of their parents. The growth of transgenic plants in the 20 days of the vegetative soil was not significantly different from those of their parents, and the transgenic lines growing in saline alkali soil were compared with their parents, and the total of the transgenic lines in the saline alkali soil was significantly lower than that in the two groups. In the nutrient soil,.3, through the analysis of the physical and chemical indexes of the leaves and roots of two groups of transgenic lines growing in saline alkali soil, found that the K+/Na+ value of each development period (except for the root of the green period) was significantly higher than that of the parent control, and the leaves were higher than the root. The MDA content of the leaves and roots of the two groups of transgenic lines were significantly lower than those of the parents. Different from the K+/Na+ value, the MDA content in the leaves and roots increased obviously with the development process, and the yield of two transgenic lines obtained by the stem tip transformation method increased by 3.37% and 4.53% respectively in the ordinary soil test field, but compared with the parent Zhou 22 of the transgenic lines obtained by the callus method. There was no significant difference in the agronomic traits, the number of spikelet and the number of tillers were the most related to the yield per mu. In the saline alkali soil test field, the yield of the two transgenic lines was higher than that of their respective parents; the agronomic traits, the plant height, the flag leaf length and the number of spikes were the most related to the yield, and the plant height was negatively correlated, and the latter two were positively correlated.5, to the ladder. The agronomic characteristics of the salt pond and alkali pond showed that the wheat yield decreased with the increase of the concentration of salt (0-0.8%NaCl) and alkali salt (0-80 mM (NaHCO3:Na2CO3=9:1)), but when salt, alkali salt treatment concentration reached a greater value of [0.8%NaCl, 80 mM alkali pool (NaHCO3:Na2CO3=9:1), the transgenic line The yield of Zhou 22 was higher than that of parental control. At this time, the yield of three transgenic lines increased by 0.03,0.37 and 0.16 times in the 0.8% NaCl salt pond respectively. In the 80 mM (NaHCO3:Na2CO3=91) alkali pool, the 1000 grain weight of three transgenic lines increased by 0.18,0.37 and 0.57 times respectively compared with the parental control. In the laboratory, two groups under saline alkali stress were in the laboratory. The transgenic lines were better than their parents in the seed germination and seedling growth. In the saline alkali soil test fields, the two groups of transgenic lines protected the plasmalemma through ion regulation and oxidase activity to protect the normal growth of the plants. The results could be used to explore the salt tolerance mechanism of the wheat transgenic lines. The cable provides the basis and the technical route that can be referenced.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S512.1

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