本文關鍵詞：外源ABA處理下擬南芥微管結合蛋白AtMAP18功能研究　出處：《沈陽農業(yè)大學》2017年碩士論文　論文類型：學位論文

  更多相關文章： 微管結合蛋白AtMAP18 ABA 微管骨架 ABA響應基因 擬南芥

【摘要】：ABA是植物激素中重要的一種,它參與調控植物生長發(fā)育的各個階段。它作為"逆境激素",在植物應對各種脅迫環(huán)境的過程中均發(fā)揮著非常關鍵作用。因此研究植物ABA脅迫的機制具有十分重要的科學意義。在生物體的許多細胞活動中均有微管骨架動態(tài)變化的參與,并在其中起到重要的作用。已經有研究表明,微管的動態(tài)變化參與ABA信號轉導途徑中。微管的動態(tài)變化主要是由微管結合蛋白來調節(jié),但是,目前關于調控植物適應ABA的微管結合蛋白作用的研究報道仍比較罕見。AtMAP18是一種去穩(wěn)定微管的微管結合蛋白,本研究以擬南芥野生型、AtMAP18過表達(AtMAP18-OE)、突變體(map18)和AtMAP18-RNAi植物為實驗材料,通過明確ABA處理下微管結合蛋白AtMAP18的表達模式,觀察ABA處理下擬南芥野生型、AtMAP18-OE、map18和AtMAP18-RNAi的形態(tài)以及微管的動態(tài)變化,測定外源ABA處理下ABA脅迫響應基因的表達量變化,從而揭示ABA下微管結合蛋白AtMAP18的作用。主要結果如下:1.外源ABA處理下,可以明顯誘導AtMAP18的表達。2.在正常條件下,AtMAP18-OE、AtMAP18-RNAi,map18與WT的種子萌發(fā)率基本達到一致。隨著外源ABA處理濃度的增加,AtMAP18-RNAi、map18種子萌發(fā)受到抑制,與WT種子萌發(fā)率差異逐漸增大;AtMAP18-OE與WT種子萌發(fā)率差異不顯著。而AtMAP18-OE種子萌發(fā)受到的抑制明顯小于AtMAP18-RNAi和map18。3.在正常條件下,除AtMAP18-RNAi植株的側根數(shù)略低于野生型外,AtMAP18-OE、map18和AtMAP18-RNAi植株在葉片數(shù)、葉面積、主根長和側根數(shù)上與野生型相比無顯著差異。在外源ABA處理下,AtMAP18-OE植株的長勢明顯好于野生型,其側根數(shù)更多、主根長更長、葉面積更大;而AtMAP18-RNAi和map18植株的長勢則與AtMAP18-OE植株相反,相比較野生型而言側根數(shù)更少、主根長更短、葉面積更小。4.在外源ABA處理下,與野生型相比,AtMAP18-OE植株中ABF2、ABI3、KIN2和RD29A這4個基因在AtMAP18-OE植株中表達量升高,而在map18和AtMAP18-RNAi植株中表達水平降低。5.在外源ABA處理下,map18植株與WT植株子葉中的微管相比較發(fā)現(xiàn),map18植株的微管更加穩(wěn)定,而WT植株微管解聚更加明顯。綜上,ABA可以誘導微管結合蛋白AtMAP18的表達,進而調節(jié)微管的動態(tài)變化。在外源ABA處理下,AtMAP18的過表達可以誘導ABA響應基因的表達,也對植株的生長起到促進作用,并提高了植株適應ABA的能力。反之,AtMAP18的缺失無法誘導ABA響應基因的表達,抑制了植株的生長,降低了擬南芥對ABA的適應能力,研究結果表明微管結合蛋白AtMAP18參與擬南芥ABA信號途徑,并起重要的作用。
[Abstract]:ABA is an important plant hormone, which participates in the regulation of plant growth and development. As a "adversity hormone", it plays a very important role in the process of plant coping with various coercive environments. Therefore, it is of great scientific significance to study the mechanism of plant ABA stress. The involvement of the microtubule framework dynamic changes in many cell activities of the organism and plays an important role in it. Studies have shown that the dynamic changes in microtubules are involved in the ABA signal transduction pathway. The dynamic changes of microtubules are mainly regulated by microtubule binding proteins. However, the current studies on the role of microtubule binding proteins regulating plant adaptation to ABA are still rare. AtMAP18 is a microtubule stabilizing microtubule binding to proteins, in this study, the overexpression of AtMAP18 in Arabidopsis wild type (AtMAP18-OE), and AtMAP18-RNAi (map18) mutant plants as the experimental material, through the clear ABA treatment microtubule binding protein expression patterns of AtMAP18, to observe the dynamic changes of ABA treatment of Arabidopsis wild type and AtMAP18-OE and map18 the morphology of AtMAP18-RNAi and microtubule, determination of exogenous ABA treatment of ABA stress response gene expression, thus revealing ABA AtMAP18 protein function. The main results are as follows: 1. the expression of AtMAP18 can be obviously induced by exogenous ABA treatment. 2. under normal conditions, the seed germination rate of AtMAP18-OE, AtMAP18-RNAi, map18 and WT basically reached the same. With the increase of ABA concentration, the germination of AtMAP18-RNAi and map18 seeds was inhibited, and the seed germination rate of WT increased gradually. There was no significant difference in seed germination rate between AtMAP18-OE and WT. The inhibition of AtMAP18-OE seed germination was less than that of AtMAP18-RNAi and map18. 3. under normal conditions, except for the lateral root number of AtMAP18-RNAi plants was slightly lower than that of wild type, AtMAP18-OE, map18 and AtMAP18-RNAi plants had no significant difference in leaf number, leaf area, main root length and lateral root number compared with wild type. Under the treatment of exogenous ABA, AtMAP18-OE plants were obviously better than that of the wild type, the lateral root number and root length more longer, the greater leaf area; while AtMAP18-RNAi and map18 plants and AtMAP18-OE plants growing on the contrary, compared to wild type, length of taproot and lateral roots and fewer shorter, smaller leaf area. 4., under exogenous ABA treatment, compared with wild type, AtMAP18-OE, ABF2, ABI3, KIN2 and RD29A 4 genes increased in AtMAP18-OE plants, but decreased in map18 and AtMAP18-RNAi plants. 5. under exogenous ABA treatment, map18 plants and WT microcotyledon microtubules showed that map18 microtubule was more stable, while WT plant microtubule depolymerization was more obvious. In conclusion, ABA can induce the expression of microtubule binding protein AtMAP18, and then regulate the dynamic changes of microtubules. Under exogenous ABA treatment, AtMAP18 overexpression can induce ABA response gene expression, and also promote the growth of plants, and improve the ability of plants to adapt to ABA. Conversely, the deletion of AtMAP18 could not induce the expression of ABA responsive genes, inhibit the growth of plants, and reduce the adaptability of Arabidopsis to ABA. The results showed that AtMAP18 was involved in ABA signaling pathway and played an important role in Arabidopsis thaliana.
【學位授予單位】：沈陽農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：Q946

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