马铃薯转亚精胺合成酶基因（SPDS）植株低温抗性的初步研究

发布时间：2018-04-15 06:09

本文选题：亚精胺合成酶(SPDS) + 马铃薯　；参考：《湖南农业大学》2016年硕士论文

【摘要】：马铃薯品种逆境抗性的改善,是马铃薯育种的主要目标之一,多胺类物质与植物逆境抗性关系密切,植物亚精胺合成酶是多胺生物合成代谢中的关键酶之一。本研究以转植物亚精胺合成酶基因SPDS的马铃薯植株为材料,对转SPDS马铃薯植株导入基因稳定性和表达情况,低温胁迫期间转SPDS马铃薯植株的低温响应表现、冷冻处理叶片相对电导率、多胺含量、生理抗性指标,低温胁迫遭遇前后的转录组测序的差异表达基因进行了检测、观察和初步的生物信息学分析。主要研究结果如下：1、对14个转SPDS马铃薯株系的PCR、RT-PCR检测和多胺HPLC定量分析,择优筛选、初步获得6个导入基因遗传及表达稳定、多胺代谢强化显著的转SPDS马铃薯株系。2、初步筛选的6个转SPDS马铃薯株系的低温胁迫应答响应、低温胁迫处理后回复常温生育状况、人工冷冻低温处理叶片相对电导率三方面的变化均优于对照植株,说明SPDS的导入提高了马铃薯植株的低温抗性。3、遭遇低温胁迫转SPDS马铃薯植株的多胺含量、光合作用和生理抗性指标的变化均优于对照种,从生理水平对转SPDS马铃薯植株低温抗性改善的机理机制进行了初步探讨。4、以转SPDS马铃薯株系02-42及其对照种东农303为材料,运用DGE技术对低温胁迫遭遇前后马铃薯植株的表达差异基因进行了初步的研究。基于对照和转SPDS植株所特有的差异表达基因的比较分析,初步发现有14个表达差异达显著水平的基因,其中明显上调的7个基因有利于转SPDS马铃薯植株低温抗性改善。
[Abstract]:The improvement of stress resistance of potato varieties is one of the main objectives of potato breeding. Polyamines are closely related to plant stress resistance. Spermidine synthase is one of the key enzymes in polyamine biosynthesis.In this study, the transgenic potato plants with spermatine synthase gene SPDS were used as materials to study the gene stability and expression of SPDS transgenic potato plants, and the low temperature response of the transgenic potato plants to SPDS during low temperature stress.The relative electrical conductivity, polyamine content, physiological resistance index and differentially expressed genes of transcriptome sequencing before and after cold stress were detected, observed and preliminarily analyzed by bioinformatics.The main results were as follows: 1. PCR and polyamine HPLC quantitative analysis were performed on 14 potato lines transferred to SPDS, and 6 introduced genes were selected for genetic analysis and stable expression.The polyamine metabolism was enhanced significantly in SPDS potato strain. 2. The response to low temperature stress of 6 SPDS potato lines was preliminarily screened, and the growth status at room temperature was restored after low temperature stress treatment.The changes of relative electrical conductivity in leaves of artificial cryopreservation treatment were better than that of control plants, indicating that the introduction of SPDS increased the low temperature resistance of potato plants, and the polyamines content of potato plants transferred to SPDS under low temperature stress.The changes of photosynthesis and physiological resistance indexes were better than that of the control species. The mechanism of improving the low temperature resistance of transgenic SPDS potato plants was preliminarily discussed from physiological level. The transgenic potato strain 02-42 and its control Dongnong 303 were used as materials.DGE technique was used to study the differentially expressed genes in potato plants before and after exposure to low temperature stress.Based on the comparative analysis of the specific differentially expressed genes between the control and transgenic SPDS plants, 14 genes with significant difference were found, among which 7 genes upregulated were beneficial to the improvement of low temperature resistance of transgenic potato plants with SPDS.
【学位授予单位】：湖南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S532

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