青梗菜黄化突变体生理特性及转录组分析

发布时间：2018-08-11 16:12

【摘要】：近年来,随着科研的不断深入,研究者发现植物叶色突变体不仅是研究叶绿素合成代谢等生理机制的理想材料,还能被当做标记性状应用于杂交种生产。本文以青梗菜黄化突变体'564Y'和野生型'564'为试验材料,对突变体和野生型的生长势变化,植物学特征和细胞学结构进行了研究,测定了两者在细胞色素、光合作用参数、叶绿素荧光动力学参数上的差异;利用第二代测序技术对黄化突变体和野生型叶片转录组进行了深度测序,探讨可能参与黄化叶片形成的代谢通路和途径。主要研究结果如下:1.在苗期,黄化突变体'564Y'叶片就表现为黄色,与野生型的绿色形成鲜明对比。在同时期,'564'的植株展开度、第三片真叶的叶长、叶宽均大于'564Y'。'564'的干重、鲜重指标均显著高于'564Y','564Y'下胚轴长度显著高于野生型'564'。2.叶片色素含量分析表明,'564Y'的叶绿素a含量和类胡萝卜素含量都显著低于'564',叶绿素b含量虽也低于'564',但两者相差不显著。在对野生型和突变体植株进行光合参数测定和荧光动力学参数测量发现,'564Y'进行光合作用的能力显著低于'564',在细胞内部进行电子传递能力变弱,把光能转换成能其他能量的效率也显著降低。3.叶绿体超微结构分析表明,相对于野生型,突变体叶肉细胞中的叶绿体排列松散,类囊体发育出现异常,基质和基粒数目减少,堆叠层数减少。4.利用第二代测序技术对'564Y'和'564'叶片转录组进行深度测序,共得到了 2958个差异表达基因,显著富集到78条GO terms,显著富集的Pathway途径有35条。在差异表达基因中,其中有9个为叶绿素合成途径的基因,Bra031690,Bra012595,Bra005677,Bra022628,Bra026410,Bra032155,Bra039206,Bra029875,Bra012511,1个与光合作用相关的基因Bra038011,推测以上基因可能参与了叶片黄化性状的形成。最后选取12个差异表达基因进行qRT-PCR分析,证实了转录组数据的可信度高,能够进行后续的研究分析。
[Abstract]:In recent years, with the deepening of scientific research, the researchers found that leaf color mutants are not only ideal materials for studying physiological mechanisms such as chlorophyll biosynthesis, but also can be used as markers for hybrid production. In this paper, the growth potential, botanical characteristics and cytological structure of the mutant and wild type were studied, and the cytochrome and photosynthesis parameters were determined. The difference of chlorophyll fluorescence kinetic parameters and the deep sequencing of yellowing mutants and wild-type leaf transcriptome were carried out by using the second generation sequencing technique to explore the metabolic pathways and pathways that might be involved in the formation of yellowing leaves. The main results are as follows: 1. At seedling stage, yellow leaves of yellow mutants were yellow, contrasted with wild type green. At the same time, the length and width of the third true leaf were larger than the dry weight of the wild type, and the fresh weight index was significantly higher than that of the wild type. The analysis of pigment content in leaves showed that the content of chlorophyll a and carotenoid were significantly lower than that of H564Y', and the content of chlorophyll b was lower than that of C564Y', but there was no significant difference between them. The photosynthetic parameters and fluorescence kinetic parameters of wild type and mutant plants were measured. It was found that the ability of photosynthesis was significantly lower than that of H564Y', and the ability of electron transport in the cells became weaker. The efficiency of converting light energy into other energies is also significantly reduced by .3. The ultrastructural analysis of chloroplast showed that the chloroplast arrangement in mesophyll cells was loosely arranged, thylakoid development was abnormal, the number of matrix and grana was decreased, and the number of stacking layers was decreased. A total of 2958 differentially expressed genes were obtained by deep sequencing of the transcriptome of 564 'and 564' leaves by the second generation sequencing technique. The differentially expressed genes were significantly enriched to 78 go termsand 35 Pathway pathways were significantly enriched. Among the differentially expressed genes, there are 9 genes Bra012595- Bra012595- Bra012595- Bra00567- Bra026410- Bra026410- Bra032155- Bra039206Bra039206Bra029875- Bra01212511, one of which is related to photosynthesis, Bra03808011. it is suggested that the above genes may be involved in the formation of leaf yellowing traits. Finally, 12 differentially expressed genes were selected for qRT-PCR analysis, which confirmed that the transcriptome data were reliable and could be used for further analysis.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S634.3

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