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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 金洛仁(Laurent Zimmerli) | |
dc.contributor.author | Pin-Yao Huang | en |
dc.contributor.author | 黃品堯 | zh_TW |
dc.date.accessioned | 2021-06-16T17:14:49Z | - |
dc.date.available | 2017-08-20 | |
dc.date.copyright | 2012-08-20 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-08-19 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/63613 | - |
dc.description.abstract | 病原菌相關分子模式(pathogen-associated molecular patterns, PAMPs)
會被植物細胞表面的模式辨識受體(pattern-recognition receptors, PRR)所辨 識而引發植物的免疫反應。模式誘發免疫反應(pattern-triggered immunity, PTI)是植物免疫中最初步限制病原菌入侵增生的機制。先前研究顯示,利用基因 轉殖技術,將阿拉伯芥的EFR(一種PRR)轉殖到菸草(Nicotiana benthamiana) 及番茄(Solanum lycopersicum)中,會使得該基因轉殖植物能廣泛地抵禦不同種 的病原菌,這顯示著PTI 的訊息傳遞機制,即使在不同科的植物中仍是保守的。 除此之外,這項研究也為利用PTI 來進行基因改造,培育抗病作物設立了典範。 我們先前發現阿拉伯芥的凝集素受體激酶VI.2(AtLecRK-VI.2)是PTI 的正向調 控者,為了瞭解AtLecRK-VI.2 在其他植物物種中是否也是PTI 的正向調控者, 我培育了過度表現AtLecRK-VI.2 的菸草及蕃茄。初步的實驗結果顯示,菸草轉 殖株較野生型能抵抗病菌Pseudomonas syringae pv. syringae B728a (Pss)。 進一步分析顯示,轉殖菸草的抗性可能來自於較強的PTI反應,如較胝質的累積、 活性氧自由基的產生及PTI 標的基因的表現等。這些初步的結果,顯示了菸草可 能帶有AtLecRK-VI.2 訊息傳遞的相關元件,來正向調控PTI。 在阿拉伯芥的類受體蛋白激酶中,富含半胱胺酸類受體激酶(Cysteine-rich receptor-like kinases, CRKs)被認為參與植物抵抗病原菌的調控。藉由結合阿拉伯芥凝集素受體激酶VI.2的過度表現轉殖株的轉錄組學分析及公開的微陣列資料,我們發現有12個CRK可能參與植物的防禦反應。我們藉由基因功能喪失的篩選,這些CRK並沒有顯現與野生種相異的表現型,因此,我們將藉由讓基因過度表現的方式來對這些CRK進行分析。 | zh_TW |
dc.description.abstract | Recognition of pathogen-associated molecular patterns (PAMPs) by host cell
surface-localized pattern-recognition receptors (PRRs) activates plant innate immunity. PAMP-triggered immunity (PTI) is the first layer of plant immunity. Crucial experiments demonstrated that the transfer of the Arabidopsis PRRs EFR to N. benthamiana and tomato (S. lycopersicum) gave the transgenic plants broad-spectrum resistance against pathogens, suggesting PTI signaling machinery is conserved across different plant families and setting an example utilizing PTI to confer disease resistance in crops. We have recently identified Arabidopsis LecRK-VI.2 as a positive regulator of PTI. To test whether AtLecRK-VI.2 can also be a positive regulator of PTI in other plant species, I generated transgenic N. benthamiana and tomato plants over-expressing AtLecRK-VI.2. Preliminary test on transgenic N. benthamiana T2 plants showed that they are resistant against bacteria P. syringae pv. syringae (Pss) B728a. Additional analyses suggest their resistance is due to enhanced PTI responses, such as callose deposition, ROS production, and expression of PTI marker genes. These preliminary findi ngs indicate N. benthamiana contains the components required for AtLecRK-VI.2 signaling to positively modulate the PTI response. Of the receptor-like kinases (RLKs) in Arabidopsis, cysteine-rich receptor-like kinases (CRKs) have been suggested to play important roles in the regulation of pathogen defense. Combining transcriptome analysis from a AtLecRK-VI.2 overexpression line and publicly available microarray data revealed 12 CRKs that could potentially be involved in plant defense. Our results show that loss-of-function analysis on these CRKs did not reveal an altered phenotype; thus further analysis is being conducted using gain-of-function analysis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T17:14:49Z (GMT). No. of bitstreams: 1 ntu-101-R99b42024-1.pdf: 988821 bytes, checksum: 9aebbbc275bb75a7c9d6e80453c841a5 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | Acknowledgements i
摘要一 ii Abstract I iii Introduction I 1 Pathogen-associated molecular patterns and pattern recognition receptors 1 Pattern-triggered immunity responses (PTI) 1 Lectin receptor kinases 3 The characteristic of AtLecRK-VI.2 in plant defense 4 PTI in N. benthamiana and tomato 5 From models to crops 6 Objectives I 8 Results I 9 Generation of N. benthamiana and tomato transgenics 9 Identification of N. benthamiana transgenic lines with single T-DNA insertion 9 Transgenic N. benthamiana lines overexpressing AtLecRK-VI.2 exhibit enhanced resistance to P. syringae pv. syringae B728a. 10 PTI-mediated callose deposition is increased in transgenic N. benthamiana lines overexpressing AtLecRK-VI.2 after Pst DC3000 hrcC inoculation or flg22 treatment 11 flg22-induced ROS burst is increased in transgenic N. benthamiana lines overexpressing AtLecRK-VI.2 11 MAMP-responsive genes expression is increased in transgenic N. benthamiana lines overexpressing AtLecRK-VI.2 after flg22 treatment 12 Discussion I 13 AtLecRK-VI.2 plays a role in N. benthamiana PTI 13 Future perspectives I 15 Transgenic N. benthamiana and tomato plants overexpressing AtLecRK-VI.2 15 Figures I 16 Figure 1. Identification of transgenic N. benthamiana lines with single T-DNA insertion 16 Figure 2. Transgenic expression of AtLecRK-VI.2 in N. benthamiana confers resistance against virulent bacteria Pss B728a 17 Figure 3. Transgenic N. benthamiana lines overexpressing AtLecRK-VI.2 demonstrate increased callose deposition after Pst DC3000 hrcC or flg22 treatment 18 Figure 4. Transgenic N. benthamiana lines overexpressing AtLecRK-VI.2 show more ROS production after flg22 treatment. 19 Figure 5. Transgenic N. benthamiana lines overexpressing AtLecRK-VI.2 show increased expression levels of MAMP responsive genes after flg22 treatment 21 Material and Methods I 22 Plant growth conditions 22 Transgene construction 22 Transformation of Tomato Plants 23 Transformation of N. benthamiana 25 Disease assays 26 Callose deposition assays 26 ROS evaluation 27 RNA extraction and gene expression analysis 27 Appendixes I 29 Appendix 1. Summary of transgenics generation 29 Appendix 2. Media used for tomato transformation 29 Appendix 3. Media used for N. benthamiana transformation 31 Appendix 4. Primers used in RT-PCR or real-time PCR 32 Screening of Cysteine-rich Receptor-like Kinases Involved in Arabidopsis Defense Responses 33 Abstract II 33 摘要二 34 Introduction II 35 Cysteine-rich receptor-like kinases 35 Objectives II 36 Results II 37 Disease screening of T-DNA insertion mutant lines of CRKs 37 Discussion II 38 Members of the CRK family may have functional redundancy 38 Future perspectives II 39 Functional analysis of CRKs 39 Figures II 40 Figure 1. Phenotype of Pst DC3000 infected Col-0 and crks. 40 Material and methods II 41 Plant growth 41 Disease assays 41 Appendixes II 42 Appendix 1. List of primers used to screen for homozygous crks mutants 42 References 43 | |
dc.language.iso | en | |
dc.title | 阿拉伯芥凝集素受體激酶VI.2在菸草轉殖植物的功能性分析及篩選參與阿拉伯芥防禦反應之富含半胱胺酸類受體激酶 | zh_TW |
dc.title | Functional Characterization of AtLecRK-VI.2 in Nicotiana benthamiana and Tomato Plants and Screening of Cysteine-rich Receptor-like Kinases Involved in Arabidopsis Defense Responses | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳昭瑩(Chao-Ying, Chen),林乃君(Nai-Chun, Lin) | |
dc.subject.keyword | 阿拉伯芥,菸草,模式誘發免疫反應,基因轉殖植物,凝集素受體激酶,防禦反應,細菌,富含半胱胺酸類受體激酶,凝集素受體激酶,VI.2, | zh_TW |
dc.subject.keyword | Arabidopsis,Nicotiana benthamiana,PTI,transgenic plants,lectin receptor-like kinase,defense response,bacteria,cysteine-rich receptor-like kinase,lectin receptor-like kinase-VI.2, | en |
dc.relation.page | 49 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-08-20 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 植物科學研究所 | zh_TW |
顯示於系所單位: | 植物科學研究所 |
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ntu-101-1.pdf 目前未授權公開取用 | 965.65 kB | Adobe PDF |
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