【摘要】：土壤中的鉛污染存在分布范圍廣、持續(xù)時間長、不易在物質(zhì)循環(huán)和能量交換中分解、易蓄積等特點,給人類的生存環(huán)境和人類健康造成嚴重威脅。近年來,植物修復技術(shù)以成本低,不引起二次污染的優(yōu)點,受到各國科學家的關(guān)注。在已發(fā)現(xiàn)的鉛超富集植物中往往受到生物量小、生長周期長或地理環(huán)境等因素的限制,導致在土壤植物修復中得不到實際的應用。隨著轉(zhuǎn)基因技術(shù)的的迅速發(fā)展,通過外源耐性基因在植物體內(nèi)的過量表達的方法,可以培育出生物量大、生長周期短的超富集轉(zhuǎn)基因植物。根據(jù)以往的報道,關(guān)于耐鉛基因的甚少。本研究利用新一代高通量Illumina測序技術(shù)、以篩選出的鉛超富集植物黑心菊為研究對象,通過轉(zhuǎn)錄組測序,得到了大量的原始序列。對得到的高質(zhì)量序列進行拼接、組裝和功能注釋,發(fā)掘其與耐鉛相關(guān)的基因,為土壤Pb污染植物修復提供更多的參考。同時,通過RT-PCR法克隆了黑心菊的Rh NRAMP1基因并構(gòu)建了雙元克隆載體,為進一步驗證其基因功能和深入了解黑心菊耐鉛機制提供了基礎(chǔ)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.
【學位授予單位】：貴州師范大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：Q943.2;X173

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