本文選題：細(xì)胞極性 + AN�。� 參考：《福建農(nóng)林大學(xué)》2017年碩士論文

【摘要】：細(xì)胞極性,廣義上是細(xì)胞內(nèi)的不對稱分布,是細(xì)胞實現(xiàn)發(fā)育和環(huán)境調(diào)節(jié)功能的基本特征之一。擬南芥作為模式植物,其單細(xì)胞的花粉管和多細(xì)胞表皮系統(tǒng)已經(jīng)成為研究植物細(xì)胞極性的重要模式系統(tǒng)。早先有研究表明在擬南芥的子葉表皮細(xì)胞中,生長素可以激活植物細(xì)胞極性通路下游特有的ROP6(Rho-related GTPase from plants 6)-RIC1(ROP-interactive CRIB motif-containing protein 1)信號轉(zhuǎn)導(dǎo)途徑,進(jìn)而影響細(xì)胞骨架的排列來調(diào)控細(xì)胞極性生長和形態(tài)建成。ROP6-RIC1信號途徑可以促進(jìn)細(xì)胞凹陷區(qū)(Neck Region)周質(zhì)微管的有序排列,進(jìn)而維持細(xì)胞的生長極性。本論文通過正向遺傳學(xué)方法篩選ROP6-RIC1途徑中的新成員。在rop6-2缺失背景下,本實驗通過EMS(ethylmethane-sulfonate,甲基磺酸乙酯)誘變篩選擬南芥表皮鋪板細(xì)胞中凸起數(shù)量(Lobe Number)明顯降低的突變體—507-2。經(jīng)遺傳學(xué)方法驗證該突變是一個隱性純和的突變體。通過BSA-重測序(Bulked Sergeant Analysis)方法找到了可能導(dǎo)致該突變的四個候選基因。在對四個候選基因進(jìn)行鑒定分析后,確認(rèn)突變基因為Angustifolia(AN)。突變發(fā)生于AN基因上的第877脫氧核糖核酸上,其中的G突變成了 A并導(dǎo)致其編碼的第293個氨基酸谷氨酸(E)突變?yōu)橘嚢彼?K)。根據(jù)突變基因的特點,將去掉rop6-2背景的單基因突變體507-2-SM命名為ANE293K。之前的研究證實AN在細(xì)胞質(zhì),細(xì)胞核以及高爾基體等上表達(dá)。本實驗用煙草做AN和ANE293K的瞬時表達(dá),發(fā)現(xiàn)其在細(xì)胞質(zhì)以及類似片層結(jié)構(gòu)的高爾基體上皆有表達(dá)。AN和AN E293K蛋白在細(xì)胞內(nèi)都能運動,但是AN的運動速率明顯高于ANE293K。觀察ANE293K的微管排列分布,發(fā)現(xiàn)較野生型相比,AN E293K的微管束較粗且其分布密度較大,說明突變的AN影響到微管的組織動態(tài)。而且在ANE293K里也沒有觀察到在野生型里面垂直于細(xì)胞生長方向抑制細(xì)胞生長的微管束。507-2的細(xì)胞表型與ANE293K更為相似,因此AN是位于ROP6信號通路途徑的下游來調(diào)控細(xì)胞極性生長,但具體調(diào)控機制尚需進(jìn)一步實驗進(jìn)行解析。由于AN和ANE293K在野生型和突變體里面的轉(zhuǎn)錄水平?jīng)]有發(fā)生變化,本論文推測是ANE293K蛋白生化功能上的改變,導(dǎo)致其影響了細(xì)胞微管的組織動態(tài)并最終影響其細(xì)胞的極性建成。
[Abstract]:Cell polarity, in a broad sense, is the asymmetric distribution of cells, which is one of the basic characteristics of cell development and environmental regulation. Arabidopsis thaliana (Arabidopsis thaliana) as a model plant, its single cell pollen tube and multicellular epidermal system have become an important model system for studying the polarity of plant cells. Previous studies have shown that in Arabidopsis thaliana cotyledon epidermis cells, auxin activates the ROP6(Rho-related GTPase from plants 6)-RIC1(ROP-interactive CRIB motif-containing protein 1) signal transduction pathway that is unique downstream of the plant cell polarity pathway. Furthermore, the regulation of cell polarity growth and morphogenesis by affecting the arrangement of cytoskeleton. ROP6-RIC1 signaling pathway can promote the ordered arrangement of periplasmic microtubules in the cell depression area check region, and then maintain the growth polarity of the cells. In this paper, the new members of ROP6-RIC1 pathway were screened by forward genetics. Under the background of rop6-2 deletion, the mutant -507-2was screened by mutagenesis of ethylmethane-sulfonate (ethyl methane-sulfonate) in Arabidopsis thaliana epidermal paving cells. The mutation was proved to be a recessive pure sum mutant by genetic methods. Four candidate genes for the mutation were identified by BSA-re sequencing and barked Sergeant analysis. After identification and analysis of four candidate genes, it was confirmed that the mutant gene was Angustifolia angustifolia. The mutation occurred in the 877 deoxyribonucleic acid (DNA) of an gene, in which the G process became A and led to the mutation of the 293rd amino acid glutamate E) to Lysine Ko. According to the characteristics of the mutant gene, the single gene mutant 507-2-SM, which removed the background of rop6-2, was named ANE293K. Previous studies have confirmed an expression in cytoplasm, nucleus and Golgi apparatus. In this study, tobacco was used for transient expression of an and ANE293K. It was found that both an and an E293K proteins could be expressed in the cytoplasm and Golgi bodies similar to lamellar structure, but the movement rate of an was significantly higher than that of ANE293K. The distribution of microtubules in ANE293K was observed. The results showed that the microtubule bundles of ANE293K were thicker and denser than those of wild type, indicating that the mutant an affected the tissue dynamics of microtubules. Moreover, the phenotypes of microtubules .507-2, which are perpendicular to the growth direction of cells, were not observed in ANE293K, and the phenotype of microtubules .507-2 was more similar to that of ANE293K, so an is located downstream of the ROP6 signaling pathway to regulate cell polar growth. However, the specific regulatory mechanisms need further experimental analysis. Since the transcription levels of an and ANE293K in wild type and mutant have not changed, this paper speculated that the changes of biochemical function of ANE293K protein resulted in its influence on the cellular microtubule tissue dynamics and ultimately on the cell polarity formation.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：Q943.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 胡軼紅,徐永華;細(xì)胞極性的形成[J];細(xì)胞生物學(xué)雜志;2004年04期

2 ;陳正軍組關(guān)于上皮細(xì)胞極性建立機制的研究取得突破[J];生命科學(xué);2006年06期

3 陳勵;辛天池;李明發(fā);;非典型蛋白激酶C復(fù)合物在細(xì)胞極性建立中的作用[J];細(xì)胞生物學(xué)雜志;2007年04期

4 曹景利;朱學(xué)良;;上皮細(xì)胞極性的建立和維持[J];中國細(xì)胞生物學(xué)學(xué)報;2010年02期

5 關(guān)默;戚中田;;細(xì)胞極性與丙型肝炎病毒受體介導(dǎo)的細(xì)胞入侵[J];第二軍醫(yī)大學(xué)學(xué)報;2010年08期

6 衛(wèi)旭彪;劉厚奇;;PAR3在上皮細(xì)胞緊密連接形成中作用和分子機制研究[J];生命科學(xué);2008年05期

7 蔣輝;;蛋白激酶GSK-3β控制了神經(jīng)細(xì)胞極性的形成[J];中國基礎(chǔ)科學(xué);2006年02期

8 王繼英;饒青;;RHO蛋白家族與細(xì)胞極性[J];細(xì)胞生物學(xué)雜志;2008年01期

9 ;細(xì)胞極性蛋白Occludin在上皮細(xì)胞遷移過程中的調(diào)節(jié)機制[J];臨床合理用藥雜志;2010年07期

10 邊文杰;陳曉萍;;細(xì)胞極性蛋白復(fù)合物PAR/aPKC的研究進(jìn)展[J];生物技術(shù)通訊;2007年06期

相關(guān)會議論文 前7條

1 張蕾;李雁潔;馮德勤;蔣鶴春;歐陽浩淼;周慧;羅元明;金城;;蛋白質(zhì)糖基化修飾對煙曲霉(Aspergillus fumigatus)細(xì)胞極性生長的影響[A];2008年中國微生物學(xué)會學(xué)術(shù)年會論文摘要集[C];2008年

2 高向東;;Rho GTP酶與酵母細(xì)胞極性[A];基因開啟未來：新時代的遺傳學(xué)與科技進(jìn)步——湖北省遺傳學(xué)會第八次代表大會暨學(xué)術(shù)討論會論文摘要匯編[C];2009年

3 蘇榮健;陳譽華;宋今丹;;非折疊蛋白質(zhì)應(yīng)答對293A細(xì)胞遷移特性的影響[A];中國細(xì)胞生物學(xué)學(xué)會第八屆會員代表大會暨學(xué)術(shù)大會論文摘要集[C];2003年

4 趙艷梅;苗龍;;胞吐作用在Ascaris線蟲精子細(xì)胞激活過程中調(diào)控細(xì)胞極性建立及細(xì)胞運動的功能研究[A];第十一次中國生物物理學(xué)術(shù)大會暨第九屆全國會員代表大會摘要集[C];2009年

5 高向東;Erfei Bi;;釀酒酵母Rho-家族GTP-結(jié)合蛋白Cdc42p在細(xì)胞極性建立過程中調(diào)控的遺傳分析[A];第二屆中國青年學(xué)者微生物遺傳學(xué)學(xué)術(shù)研討會論文集[C];2006年

6 高向東;Erfei Bi;;釀酒酵母Rho-家族GTP-結(jié)合蛋白Cdc42p在細(xì)胞極性建立過程中調(diào)控的遺傳分析[A];2006中國微生物學(xué)會第九次全國會員代表大會暨學(xué)術(shù)年會論文摘要集[C];2006年

7 韓冰;楊寧;王臺;;應(yīng)用定量蛋白質(zhì)組技術(shù)理解花粉管極性建立的蛋白質(zhì)基礎(chǔ)[A];2011全國植物生物學(xué)研討會論文集[C];2011年

相關(guān)重要報紙文章 前1條

1 徐亞靜;我國原創(chuàng)性論文首次被《Cell》點評[N];中國醫(yī)藥報;2007年

相關(guān)博士學(xué)位論文 前2條

1 李林璋;ROS對乳腺癌上皮細(xì)胞極性變化及單核細(xì)胞浸潤調(diào)控機制的研究[D];吉林大學(xué);2017年

2 王芊藝;JAK1對平面細(xì)胞極性通路的調(diào)控及機制的研究[D];中國人民解放軍軍事醫(yī)學(xué)科學(xué)院;2014年

相關(guān)碩士學(xué)位論文 前5條

1 蔡常青;Triciribine在dHL-60細(xì)胞極性化中的作用研究[D];南方醫(yī)科大學(xué);2015年

2 穆小云;Reelin缺失對小鼠皮質(zhì)片層化及細(xì)胞極性化的影響[D];河南大學(xué);2015年

3 張晶晶;擬南芥ROP6信號通路調(diào)控細(xì)胞極性發(fā)育的突變體篩選和突變基因克隆[D];福建農(nóng)林大學(xué);2017年

4 李傳武;低濃度毒死蜱對發(fā)育期神經(jīng)細(xì)胞極性復(fù)合體的影響[D];浙江工業(yè)大學(xué);2010年

5 符星;海馬微環(huán)境下Reelin對細(xì)胞分化和極性的影響[D];河南大學(xué);2013年

，

本文編號：1906109