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過表現 AtFd1 可提升阿拉伯芥之高溫逆境耐受能力 = Overexp...

國立高雄大學生物科技研究所碩士班

過表現 AtFd1 可提升阿拉伯芥之高溫逆境耐受能力 = Overexpression of AtFd1 enhances heat tolerance in transgenic Arabidopsis thaliana

紀錄類型:	書目-語言資料,印刷品 : 單行本
並列題名:	Overexpression of AtFd1 enhances heat tolerance in transgenic Arabidopsis thaliana
作者:	黃麗子,
其他團體作者:	國立高雄大學
出版地:	[高雄市]
出版者:	撰者;
出版年:	民102[2013]
面頁冊數:	41葉部分彩圖，表格 : 30公分;
標題:	過氧化物傷害
標題:	ROS damage
電子資源:	http://hdl.handle.net/11296/ndltd/81858016613882590207
附註:	106年10月31日公開
附註:	參考書目：葉34-40
附註:	附錄：Primers used in this study
摘要註:	高溫逆境會對植物造成傷害，影響植物的生長以及產量。植物葉綠體中的光合作用系統II (photosystem II, PSII)對高溫極為敏感，易受到高溫的影響而導致電子傳遞中斷，促使大量過氧化物(reactive oxygen species, ROS)的生合成，造成植物的損傷。為了抵抗此逆境的傷害，植物會在高溫逆境下誘導抗逆境相關基因的表現，如heat shock transcription factors (Hsfs)、heat shock proteins (HSPs)、ascorbate peroxidases (APXs)及glutathione-S- transferase (GST)。硫鐵蛋白(ferredoxin, Fd)是一群負責傳遞電子的蛋白，參與了植物體內許多的代謝反應。前人的研究指出AtFd1為阿拉伯芥中負責進行光合作用循環式電子傳遞之蛋白，此電子傳遞鏈被認為可以幫助植物抵抗逆境。此外文獻中也指出過表現藍綠藻中的Fd可提升菸草抵抗高溫、強光、殺草劑等傷害之能力。然而，AtFd1此阿拉伯芥中之Fd在植物對高溫逆境反應時所扮演之角色仍尚未明瞭。為解決此問題，本篇論文之目的為探討過表現AtFd1是否能增強阿拉伯芥的高溫耐受能力及其調控機制。研究結果發現atfd1會受到高溫而誘導表現，顯示其可能在植物面對高溫逆境時扮演重要角色。而經由高溫處理後，atfd1轉殖株具有較高之耐熱性，其體內之H2O2及malondialdehyde (MDA)含量也少於未轉殖株(WT)，可得知轉殖株所受到之ROS傷害較少。高溫處理後，WT體內的葡萄糖含量較轉殖株多，推測可能原因為WT所受到之高溫傷害程度較大，必須合成較多的滲透性保護物質以抵抗逆境。另外與WT相比，轉殖株之apx2、hsfA2、hsp101在高溫處理時有較高之表現量。綜合上述，本研究結果指出過表現AtFd1可能調控hsfA2及其目標基因的表現，以及降低ROS的傷害，藉此增強其耐熱性。 Heat stress belongs to abiotic stress that affects the growth and yield of plants. For chloroplast, photosystem II (PSII) is the most heat-sensitive site and the electron transport may be blocked and results in accumulation of reactive oxygen species (ROS) after heat stress. Several genes are also induced by heat stress, such as heat shock transcription factors (Hsfs), heat shock proteins (HSPs), ascorbate peroxidases (APXs) and glutathione-S- transferase (GST). Ferredoxin (Fd) is a group of electron transport proteins which involves in many metabolic processes. Study indicates AtFd1 in Arabidopsis is responsible for the cyclic electron flow, the electron transport which is suggested to act in stress. It's indicated that replacement of Fd with a cyanobacterial flavodoxin in tobacco enhanced tolerance to stresses. However, function of AtFd1 in tolerance of heat stress is unclear. In order to reveal this issue, atfd1-transgenic Arabidopsis thaliana (ecotypes Col-0) were used to investigate heat tolerance and possible mechanism. For 40°C-treatment, the atfd1-expression pattern showed that atfd1 is a heat-inducible gene which may play an important role in heat response. Overexpression of atfd1 enhanced the heat tolerance in transgenic Arabidopsis under 10 h of 45°C- treatment. The H2O2 and malondialdehyde (MDA) content after heat treatment indicated that WT exhibited more severe damage than transgenic lines. Additionally, after 8 d-treatment in 38°C, the Fv/Fm and electron transport rate (ETR) shown no significant difference between WT and transgenic lines, while the Fo of WT was higher than those of transgenic lines, indicating the heat damage in WT was more severe than transgenic lines. The expression of stress-related genes (apx2, hsfA2, hsp101) in WT were higher than those in transgenic lines after 45°C heat treatment, whereas the expression of apx1 and gst had the opposite result. Furthermore, soluble sugar accumulation was observed after 5 d-heat treatment, and the glucose data showed that heat treatment induced more severe threat to WT. In this study, we demonstrated overexpression of AtFd1 confers heat tolerance through regulating ROS content and expression of hsfA2 and its target genes in heat response.

過表現 AtFd1 可提升阿拉伯芥之高溫逆境耐受能力 = Overexpression of AtFd1 enhances heat tolerance in transgenic Arabidopsis thaliana
黃, 麗子

過表現 AtFd1 可提升阿拉伯芥之高溫逆境耐受能力 = Overexpression of AtFd1 enhances heat tolerance in transgenic Arabidopsis thaliana / 黃麗子撰 - [高雄市] : 撰者, 民102[2013]. - 41葉 ; 部分彩圖，表格 ; 30公分.
106年10月31日公開參考書目：葉34-40附錄：Primers used in this study.
過氧化物傷害ROS damage

過表現 AtFd1 可提升阿拉伯芥之高溫逆境耐受能力 = Overexpression of AtFd1 enhances heat tolerance in transgenic Arabidopsis thaliana
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330 $a 高溫逆境會對植物造成傷害，影響植物的生長以及產量。植物葉綠體中的光合作用系統II (photosystem II, PSII)對高溫極為敏感，易受到高溫的影響而導致電子傳遞中斷，促使大量過氧化物(reactive oxygen species, ROS)的生合成，造成植物的損傷。為了抵抗此逆境的傷害，植物會在高溫逆境下誘導抗逆境相關基因的表現，如heat shock transcription factors (Hsfs)、heat shock proteins (HSPs)、ascorbate peroxidases (APXs)及glutathione-S- transferase (GST)。硫鐵蛋白(ferredoxin, Fd)是一群負責傳遞電子的蛋白，參與了植物體內許多的代謝反應。前人的研究指出AtFd1為阿拉伯芥中負責進行光合作用循環式電子傳遞之蛋白，此電子傳遞鏈被認為可以幫助植物抵抗逆境。此外文獻中也指出過表現藍綠藻中的Fd可提升菸草抵抗高溫、強光、殺草劑等傷害之能力。然而，AtFd1此阿拉伯芥中之Fd在植物對高溫逆境反應時所扮演之角色仍尚未明瞭。為解決此問題，本篇論文之目的為探討過表現AtFd1是否能增強阿拉伯芥的高溫耐受能力及其調控機制。研究結果發現atfd1會受到高溫而誘導表現，顯示其可能在植物面對高溫逆境時扮演重要角色。而經由高溫處理後，atfd1轉殖株具有較高之耐熱性，其體內之H2O2及malondialdehyde (MDA)含量也少於未轉殖株(WT)，可得知轉殖株所受到之ROS傷害較少。高溫處理後，WT體內的葡萄糖含量較轉殖株多，推測可能原因為WT所受到之高溫傷害程度較大，必須合成較多的滲透性保護物質以抵抗逆境。另外與WT相比，轉殖株之apx2、hsfA2、hsp101在高溫處理時有較高之表現量。綜合上述，本研究結果指出過表現AtFd1可能調控hsfA2及其目標基因的表現，以及降低ROS的傷害，藉此增強其耐熱性。 Heat stress belongs to abiotic stress that affects the growth and yield of plants. For chloroplast, photosystem II (PSII) is the most heat-sensitive site and the electron transport may be blocked and results in accumulation of reactive oxygen species (ROS) after heat stress. Several genes are also induced by heat stress, such as heat shock transcription factors (Hsfs), heat shock proteins (HSPs), ascorbate peroxidases (APXs) and glutathione-S- transferase (GST). Ferredoxin (Fd) is a group of electron transport proteins which involves in many metabolic processes. Study indicates AtFd1 in Arabidopsis is responsible for the cyclic electron flow, the electron transport which is suggested to act in stress. It's indicated that replacement of Fd with a cyanobacterial flavodoxin in tobacco enhanced tolerance to stresses. However, function of AtFd1 in tolerance of heat stress is unclear. In order to reveal this issue, atfd1-transgenic Arabidopsis thaliana (ecotypes Col-0) were used to investigate heat tolerance and possible mechanism. For 40°C-treatment, the atfd1-expression pattern showed that atfd1 is a heat-inducible gene which may play an important role in heat response. Overexpression of atfd1 enhanced the heat tolerance in transgenic Arabidopsis under 10 h of 45°C- treatment. The H2O2 and malondialdehyde (MDA) content after heat treatment indicated that WT exhibited more severe damage than transgenic lines. Additionally, after 8 d-treatment in 38°C, the Fv/Fm and electron transport rate (ETR) shown no significant difference between WT and transgenic lines, while the Fo of WT was higher than those of transgenic lines, indicating the heat damage in WT was more severe than transgenic lines. The expression of stress-related genes (apx2, hsfA2, hsp101) in WT were higher than those in transgenic lines after 45°C heat treatment, whereas the expression of apx1 and gst had the opposite result. Furthermore, soluble sugar accumulation was observed after 5 d-heat treatment, and the glucose data showed that heat treatment induced more severe threat to WT. In this study, we demonstrated overexpression of AtFd1 confers heat tolerance through regulating ROS content and expression of hsfA2 and its target genes in heat response.
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