本文選題：馬鈴薯　切入點(diǎn)：擬南芥　出處：《西南大學(xué)》2017年碩士論文

【摘要】：鉀離子是植物生長(zhǎng)所必需的營(yíng)養(yǎng)元素之一,參與到多種生理過(guò)程中,包括滲透調(diào)節(jié),糖類(lèi)的協(xié)同運(yùn)輸以及光合作用等。TOR是在真核生物中進(jìn)化上保守的絲氨酸/蘇氨酸蛋白激酶,屬于PI3K蛋白激酶家族,可以響應(yīng)能量、營(yíng)養(yǎng)等信號(hào)調(diào)控細(xì)胞的生長(zhǎng)發(fā)育,TOR激酶功能缺失時(shí),導(dǎo)致植物葉片發(fā)黃,同時(shí)伴隨有光合效率下降以及葉綠素降解。TOR信號(hào)通路是否通過(guò)調(diào)控鉀素營(yíng)養(yǎng)的吸收代謝從影響植物的光合作用、葉綠素生物合成等還未見(jiàn)報(bào)道,因此在本研究中通過(guò)藥理學(xué)和遺傳學(xué)試驗(yàn)研究了TOR信號(hào)通路與鉀離子吸收的關(guān)系,得出以下四點(diǎn)結(jié)論:1)在低鉀培養(yǎng)基上,馬鈴薯外植體迅速地變黃且不能正常完成根系再生過(guò)程;這種表型與在培養(yǎng)基中加入TOR抑制劑(KU、AZD)后植物的表型相似,均出現(xiàn)黃化的表型。同時(shí)通過(guò)鉀離子吸收實(shí)驗(yàn),測(cè)定了在TOR抑制時(shí)馬鈴薯對(duì)于鉀離子的吸收能力,結(jié)果顯示TOR的二代抑制劑能夠明顯抑制馬鈴薯中鉀離子的吸收。2)本研究發(fā)現(xiàn)擬南芥低鉀敏感型突變體cbl1cbl9,lks1-2,lks1-3以及akt1均對(duì)TOR抑制劑表現(xiàn)出敏感性。TOR抑制劑可以有效抑制野生型擬南芥和突變體的生長(zhǎng),這種抑制作用在突變體植株中表現(xiàn)尤為明顯。測(cè)定鉀離子濃度后發(fā)現(xiàn),TOR抑制劑可以顯著下調(diào)擬南芥對(duì)于鉀離子的吸收;然而對(duì)比發(fā)現(xiàn)在鉀離子吸收方面,TOR抑制劑對(duì)于野生型擬南芥植株的影響比突變體植株要更顯著。3)本研究擬南芥中擴(kuò)增了At CBL9,At CIPK23,At AKT1基因,構(gòu)建了攜帶GFP標(biāo)簽的過(guò)表達(dá)載體,將此載體在擬南芥轉(zhuǎn)基因植株BP12-2(雷帕霉素敏感的擬南芥轉(zhuǎn)基因材料)中過(guò)表達(dá),獲得了At CBL9-GFP/BP12,At CIPK23-GFP/BP12以及At AKT1-GFP/BP12過(guò)表達(dá)株系。通過(guò)雷帕霉素敏感性試驗(yàn)確定了TOR與At CBL9,At CIPK23,At AKT1之間的關(guān)系。發(fā)現(xiàn)在雷帕霉素處理下At CIPK23-GFP/BP12過(guò)表達(dá)株系的生長(zhǎng)情況要優(yōu)于擬南芥BP12-2株系。在雷帕霉素和KU雙抑制處理下,At CIPK23-GFP/BP12過(guò)表達(dá)株系可以繼續(xù)生長(zhǎng),而擬南芥BP12-2株系則表現(xiàn)為葉片發(fā)黃,停止生長(zhǎng)。通過(guò)鉀離子濃度測(cè)定及葉綠素含量測(cè)定試驗(yàn)發(fā)現(xiàn),在TOR抑制劑處理下,與BP12-2相比,過(guò)表達(dá)At CIPK23基因可以提高植株對(duì)于鉀離子的吸收和葉綠素含量,這些結(jié)果表明CIPK23可能在TOR信號(hào)通路影響鉀素吸收過(guò)程中具有重要的作用。4)在馬鈴薯基因組中,通過(guò)序列比對(duì),發(fā)現(xiàn)存在At CBL1,At CIPK23以及At AKT1的同源基因;同時(shí)通過(guò)q PCR試驗(yàn)確定了基因在馬鈴薯中的表達(dá)模式為組成性表達(dá)。擴(kuò)增了馬鈴薯中St CIPK23基因,并構(gòu)建了馬鈴薯St CIPK23-GFP過(guò)表達(dá)株系;通過(guò)試驗(yàn)發(fā)現(xiàn)在幼苗生長(zhǎng)中St CIPK23過(guò)表達(dá)株系對(duì)于TOR抑制劑具有抗性。這些數(shù)據(jù)表明馬鈴薯中CBL1-CIPK23-AKT1信號(hào)通路同樣參與植物對(duì)低鉀環(huán)境的響應(yīng)。綜合這些結(jié)果表明TOR信號(hào)通路可能通過(guò)調(diào)控CIPK23的功能來(lái)調(diào)節(jié)植物鉀離子的吸收。之前以及最近的研究都表明TOR信號(hào)通路作為核心調(diào)控元件來(lái)調(diào)控植物的光合生長(zhǎng),細(xì)胞增殖以及細(xì)胞伸長(zhǎng)等生理過(guò)程,然而這些研究大多均集中與對(duì)擬南芥的研究。在本研究中發(fā)現(xiàn)在馬鈴薯中TOR的功能同在擬南芥中同樣保守,通過(guò)對(duì)馬鈴薯中TOR信號(hào)通路功能的研究將有助于提高馬鈴薯的產(chǎn)量。
[Abstract]:Potassium is one of the essential nutrients for plant growth, to participate in a variety of physiological processes, including osmotic adjustment, sugar transport and Photosynthesis of collaborative.TOR is an evolutionary conserved in eukaryotes on serine / threonine protein kinase, belongs to the PI3K protein kinase family, can respond to energy and nutrition regulation of cellular signal growth of TOR kinase function loss, resulting in plant leaf yellow, accompanied by the photosynthetic efficiency decreased and chlorophyll degradation of.TOR signaling pathway by regulating potassium absorption and metabolism from affecting plant photosynthesis, chlorophyll biosynthesis has not been reported, so in this study by pharmacological and genetic tests on the relationship between TOR signal pathway and potassium uptake, draw the following four conclusions: 1) in low potassium medium, potato explants quickly turn yellow and not just To complete the regeneration process of root; this phenotype and the TOR inhibitor was added into the culture medium (KU, AZD) after the phenotypic similarity of plants showed chlorosis phenotype. The potassium ion absorption experiments in TOR inhibition of potato for K + absorption was measured, the results showed the uptake of.2 TOR two generation inhibitor can obviously inhibit the growth of potassium ions in Arabidopsis) this study found that low potassium sensitive mutants cbl1cbl9, lks1-2, lks1-3 and AKT1 were on TOR inhibitors showed sensitivity to.TOR inhibitors can effectively inhibit the Arabidopsis wild-type and mutant growth, the inhibition performance in mutant plants is particularly obvious. Determination of potassium ion concentration found TOR inhibitors can significantly down regulated in Arabidopsis for potassium ion absorption; however, compared the absorption of potassium ion, TOR inhibitors for the wild-type Arabidopsis plants. Ring than the mutant plants more significant.3) this study in Arabidopsis thaliana was amplified At CBL9, At CIPK23, At AKT1 gene, constructed over expression vector carrying the GFP tag, this vector in Arabidopsis transgenic plants BP12-2 (rapamycin sensitive Arabidopsis transgenic materials) in over expression, won the At CBL9-GFP/BP12, At CIPK23-GFP/BP12 and At over expression of AKT1-GFP/BP12 and At strains. TOR CBL9 was determined by At CIPK23 At rapamycin sensitivity test, AKT1. The relationship of overexpression lines in rapamycin treated CIPK23-GFP/BP12 growth of At is superior to BP12-2 in Arabidopsis lines. Rapamycin and KU double inhibition, At CIPK23-GFP/BP12 overexpression strains can continue to grow, and Arabidopsis BP12-2 strains showed the yellow leaves, stop growing. Through the determination of the chlorophyll content and the potassium ion concentration test found in TOR The inhibitor treatment, compared with BP12-2, expression of At CIPK23 gene can improve plant for absorption and chlorophyll content of potassium ions, these results suggest that CIPK23 may affect the potassium in the signal pathway of TOR uptake has an important role for.4 in the process) in potato genome, found by sequence alignment, At CBL1, At CIPK23 and At AKT1 homologous genes; at the same time through the Q test to determine the expression pattern of PCR gene in potato is constitutively expressed in St. The amplification of potato CIPK23 gene, and constructed the potato St CIPK23-GFP overexpression strains; the experiment found that overexpression strains with resistance to TOR inhibitors in seedling growth in St CIPK23 these data. Show that the CBL1-CIPK23-AKT1 signaling pathway in potato is also involved in plant response to low potassium environment. These results indicated that TOR signaling pathway may be regulated by CIPK2 3 functions to regulate plant potassium absorption. Before, recent studies have indicated that TOR signaling pathway as a key regulatory element to regulate plant photosynthesis and growth, cell proliferation, cell elongation and other physiological processes, however, most of these studies are focused on the research and the Arabidopsis. In this study, found in potato TOR the function in Arabidopsis as conservative through the research on the function of TOR pathway in potato will help to increase the yield of potato.

【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S532;Q943.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

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本文編號(hào)：1654680