黑心菊耐铅基因的发掘与克隆
发布时间：2018-11-06 19:38
【摘要】：土壤中的铅污染存在分布范围广、持续时间长、不易在物质循环和能量交换中分解、易蓄积等特点,给人类的生存环境和人类健康造成严重威胁。近年来,植物修复技术以成本低,不引起二次污染的优点,受到各国科学家的关注。在已发现的铅超富集植物中往往受到生物量小、生长周期长或地理环境等因素的限制,导致在土壤植物修复中得不到实际的应用。随着转基因技术的的迅速发展,通过外源耐性基因在植物体内的过量表达的方法,可以培育出生物量大、生长周期短的超富集转基因植物。根据以往的报道,关于耐铅基因的甚少。本研究利用新一代高通量Illumina测序技术、以筛选出的铅超富集植物黑心菊为研究对象,通过转录组测序,得到了大量的原始序列。对得到的高质量序列进行拼接、组装和功能注释,发掘其与耐铅相关的基因,为土壤Pb污染植物修复提供更多的参考。同时,通过RT-PCR法克隆了黑心菊的Rh NRAMP1基因并构建了双元克隆载体,为进一步验证其基因功能和深入了解黑心菊耐铅机制提供了基础材料。
[Abstract]:Lead pollution in soil has the characteristics of wide distribution, long duration, not easy to decompose in material cycle and energy exchange, easy to accumulate, and so on, which poses a serious threat to human living environment and human health. In recent years, phytoremediation technology has attracted the attention of scientists all over the world because of its advantages of low cost and no secondary pollution. It is often limited by such factors as small biomass, long growth period or geographical environment in the discovered lead enrichment plants, which lead to no practical application in soil phytoremediation. With the rapid development of transgenic technology, super-enriched transgenic plants with large biomass and short growth cycle can be obtained by overexpression of exogenous tolerance genes in plants. According to previous reports, there are very few lead-tolerant genes. In this study, a new generation of high-throughput Illumina sequencing technology was used to select the lead super-enriched plant, chrysanthemum nigra, and a large number of primordial sequences were obtained by transcriptome sequencing. The high quality sequences were spliced, assembled and annotated, and the genes related to lead tolerance were discovered, which provided more references for phytoremediation of soil Pb contamination. At the same time, the Rh NRAMP1 gene was cloned by RT-PCR method and the binary cloning vector was constructed, which provided a basic material for further verification of its gene function and further understanding of the lead tolerance mechanism of the chrysanthemum nigra.
【学位授予单位】：贵州师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2;X173
本文编号：2315264
[Abstract]:Lead pollution in soil has the characteristics of wide distribution, long duration, not easy to decompose in material cycle and energy exchange, easy to accumulate, and so on, which poses a serious threat to human living environment and human health. In recent years, phytoremediation technology has attracted the attention of scientists all over the world because of its advantages of low cost and no secondary pollution. It is often limited by such factors as small biomass, long growth period or geographical environment in the discovered lead enrichment plants, which lead to no practical application in soil phytoremediation. With the rapid development of transgenic technology, super-enriched transgenic plants with large biomass and short growth cycle can be obtained by overexpression of exogenous tolerance genes in plants. According to previous reports, there are very few lead-tolerant genes. In this study, a new generation of high-throughput Illumina sequencing technology was used to select the lead super-enriched plant, chrysanthemum nigra, and a large number of primordial sequences were obtained by transcriptome sequencing. The high quality sequences were spliced, assembled and annotated, and the genes related to lead tolerance were discovered, which provided more references for phytoremediation of soil Pb contamination. At the same time, the Rh NRAMP1 gene was cloned by RT-PCR method and the binary cloning vector was constructed, which provided a basic material for further verification of its gene function and further understanding of the lead tolerance mechanism of the chrysanthemum nigra.
【学位授予单位】：贵州师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2;X173
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