| Summary: | 碩士 === 國立成功大學 === 生物科技研究所碩博士班 === 93 === Heat stress is detrimental to both the vegetative and reproductive stages of plants. The ability for plants to withstand heat stress may be of great importance in agricultural productivity. Genetically engineering approach to introduce the thermotolerant traits into crops has several advantages over traditional breeding program.
Heat shock proteins are induced when plants subject to high temperature stress. Particularly, HSP101 plays an important function in acquired thermotolerance of plants. One of the major functions of HSP101 is to promote the re-solubilization of aggregated proteins, and reactivate protein activity in cooperation with other molecular chaperones as a refolding machine under heat stress. In addition, HSP101 can bind to the 5’ leader sequence (Ω) of tobacco mosaic virus, and enhance the translational activity of RNA transcripts. To take advantage of dual activities of HSP101, We overexpressed OsHsp101 in the cytoplasm or chloroplasts of tobacco by nuclear transformation technology.
Two tobacco homozygous lines with Ω-regulating the expression of OsHsp101 in cytoplasm were obtained. One homozygous line with Ω-regulating the expression of OsHsp101 in chloroplasts was obtained. In addition, two homozygous lines with expression of His-tag OsHsp101 fusion protein in chloroplasts were obtained. The HSP101 expression levels are much higher in transgenic lines than wild type plants. Thermotolerant assay indicated that transgenic plants have better recovery ability than wild type after subjecting to heat treatment.
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