發(fā)布時間：2018-05-19 12:17

  本文選題：PLATZ5 + SOS3��； 參考：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：土壤鹽漬化是影響當今農(nóng)作物產(chǎn)量和質(zhì)量的一個重要因素。土壤鹽分濃度過高會導(dǎo)致植物代謝紊亂,影響植物對必需元素的吸收,造成滲透脅迫及氧化脅迫等次生脅迫,進而影響植物正常的生長發(fā)育。如何提高植物的耐鹽性已成為農(nóng)業(yè)發(fā)展急需解決的問題。PLATZ(Plant A/T rich sequence-and zinc-binding protein)是一類植物特有的轉(zhuǎn)錄因子,2002年豌豆PLATZ1首次被報道能非特異地結(jié)合富含A/T堿基的序列并發(fā)揮轉(zhuǎn)錄抑制的作用。擬南芥中共有12個PLATZ成員,本研究對擬南芥AtPLATZ5進行了表達分析和功能鑒定,結(jié)果如下:(1)利用半定量RT-PCR和GUS染色的方法檢測了AtPLATZ5的組織表達模式,結(jié)果表明AtPLATZ5在各個組織中均有表達,且表達差異不大。RT-PCR和qRT-PCR的結(jié)果同時證明了AtPLATZ5的表達量受鹽脅迫誘導(dǎo)。(2)利用瞬時轉(zhuǎn)化煙草的方法對PLATZ5的亞細胞定位進行了分析,結(jié)果表明PLATZ5除在細胞核中表達外,在細胞質(zhì)中也有表達,這說明PLATZ5可能發(fā)揮多種功能。(3)利用雙熒光素酶報告系統(tǒng)對PLATZ5的轉(zhuǎn)錄活性進行了分析,結(jié)果表明PLATZ5具有轉(zhuǎn)錄抑制活性。(4)對AtPLATZ5超表達株系進行100mM和150mM NaCl處理,發(fā)現(xiàn)AtPLATZ5超表達株系地上部鮮重比對照分別降低了30%與50%,而根部的鮮重比對照分別降低了20%與18%,這表明鹽脅迫對AtPLATZ5超表達株系地上部抑制更明顯。提高NaCl濃度到200mM時,超表達株系葉片白化,存活率顯著低于野生型。這說明AtPLATZ5負調(diào)控幼苗對鹽脅迫的抗性。AtPLATZ5超表達株系地上部也會對100mM NaNO3、100mM KCl和100mM KNO3脅迫處理表現(xiàn)出微弱敏感的表型,同時超表達株系對10mM LiCl超敏感而對甘露醇不敏感。以上結(jié)果表明AtPLATZ5主要響應(yīng)離子脅迫。(5)插入到外顯子區(qū)的platz5突變體對鹽脅迫處理沒有表型,表明PLATZ家族成員可能存在著功能冗余。(6)鈉離子探針染色結(jié)果顯示超表達株系在受到鹽脅迫時根與葉片部積累的鈉離子增多,表明超表達株系體內(nèi)鈉離子的吸收轉(zhuǎn)運受到了影響。(7)qRT-PCR分析鹽脅迫后超表達株系體內(nèi)信號通路marker基因的變化,發(fā)現(xiàn)SOS途徑的SOS3/CBL4及其同源基因CBL10表達量下調(diào),而染色質(zhì)免疫共沉淀實驗證明PLATZ5能靶向SOS3啟動子,這表明PLATZ5通過抑制SOS3的表達來響應(yīng)鹽脅迫。
[Abstract]:Soil salinization is an important factor affecting crop yield and quality. The high concentration of soil salt will lead to the disorder of plant metabolism, which will affect the absorption of essential elements, osmotic stress, oxidative stress and other secondary stresses, and then affect the normal growth and development of plants. How to improve plant salt tolerance has become an urgent problem in agricultural development. The plant A / T rich sequence-and zinc-binding protein is a kind of plant-specific transcription factor. In 2002, pea PLATZ1 was first reported to be able to combine the sequence with rich in A / T base and play an important role. Transcriptional inhibition. There are 12 PLATZ members in Arabidopsis thaliana. In this study, the expression and function of Arabidopsis AtPLATZ5 were analyzed. The results are as follows: 1) the expression pattern of AtPLATZ5 was detected by semi-quantitative RT-PCR and GUS staining. The results showed that AtPLATZ5 was expressed in all tissues, and the difference was not significant. The results of RT-PCR and qRT-PCR also proved that the expression of AtPLATZ5 was induced by salt stress. (2) the subcellular localization of PLATZ5 was analyzed by transient transformation of tobacco. The results showed that PLATZ5 was expressed not only in the nucleus, but also in the cytoplasm, which suggested that PLATZ5 might play a variety of functions.) the transcriptional activity of PLATZ5 was analyzed by using double luciferase report system. The results showed that PLATZ5 had transcriptional inhibitory activity. 4) 100mM and 150mM NaCl were used to treat AtPLATZ5 overexpression lines. The results showed that the fresh weight of AtPLATZ5 superexpression lines was 30% and 50% lower than that of the control, while the fresh weight of roots was 20% and 18% lower than that of the control, respectively, which indicated that salt stress had more obvious inhibition on the shoot of AtPLATZ5 overexpressed lines. When the concentration of NaCl was increased to 200mM, the overexpressed lines were albino, and the survival rate was significantly lower than that of wild type. This indicated that the shoots of the seedlings with negative AtPLATZ5 regulation showed weak phenotypic sensitivity to 100mM NaNO3100mM KCl and 100mM KNO3 stress, but the overexpressed lines were not sensitive to mannitol but hypersensitive to 10mM LiCl. The results indicated that AtPLATZ5 was mainly responsive to ion stress. The platz5 mutant inserted into the exon region had no phenotype under salt stress. The results indicated that the PLATZ family members might have functional redundancy. 6) the results of sodium probe staining showed that the accumulation of sodium ions in roots and leaves of the overexpression lines increased under salt stress. The results showed that the absorption and transport of sodium ions in the overexpression lines were affected by QRT-PCR. The changes of marker gene in the signaling pathway of the overexpression lines after salt stress were analyzed. It was found that the expression of SOS3/CBL4 and its homologous gene CBL10 in the SOS pathway was down-regulated. The chromatin immunoprecipitation assay showed that PLATZ5 could target the SOS3 promoter, which suggested that PLATZ5 responded to salt stress by inhibiting the expression of SOS3.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：Q943.2

【參考文獻】

相關(guān)期刊論文 前10條

1 林抗雪;劉修杰;孫石;陳莉;韓天富;侯文勝;;轉(zhuǎn)TaNHX2大豆的耐鹽性分析[J];中國農(nóng)業(yè)科學(xué);2015年20期

2 董連紅;史素娟;S M Nuruzzaman Manik;蘇玉龍;劉朝科;馮祥國;胡曉明;王倩;劉好寶;;植物CBL基因家族的研究進展[J];核農(nóng)學(xué)報;2015年05期

3 于瑞;胡月;黃旭新;申澤丹;韓彥莎;鄧澍榮;趙瑞;沈昕;陳少良;;過表達秋茄C2H2型鋅指蛋白基因KcZFP提高煙草耐鹽性[J];基因組學(xué)與應(yīng)用生物學(xué);2013年02期

4 尤佳;王文瑞;盧金;賈鵬翔;繆建順;楊穎麗;;鹽脅迫對鹽生植物黃花補血草種子萌發(fā)和幼苗生長的影響[J];生態(tài)學(xué)報;2012年12期

5 宋旭麗;侯喜林;胡春梅;孟靜靜;何啟偉;李新國;;NaCl脅迫對超大甜椒種子萌發(fā)及幼苗生長的影響[J];西北植物學(xué)報;2011年03期

6 劉e，

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