本文關(guān)鍵詞： 桑樹 MAPK 非生物脅迫 脅迫響應(yīng) 啟動(dòng)子　出處：《林業(yè)科學(xué)》2017年09期 　論文類型：期刊論文

【摘要】：【目的】植物促分裂原活化蛋白激酶(MAPK)在響應(yīng)生物與非生物脅迫中發(fā)揮重要作用。本研究通過分析桑樹(川桑)B家族MAPK基因MnMAPK5的亞細(xì)胞定位和啟動(dòng)子活性,并將其轉(zhuǎn)入擬南芥中進(jìn)行過表達(dá)研究,探討MnMAPK5在逆境脅迫下的作用機(jī)制�！痉椒ā繉nMAPK5的上游啟動(dòng)子連接到p BI121載體上,通過花粉介導(dǎo)法轉(zhuǎn)入擬南芥中,經(jīng)高溫、低溫、NaCl和PEG處理后進(jìn)行GUS染色分析。通過構(gòu)建過表達(dá)載體,將MnMAPK5轉(zhuǎn)化到擬南芥中,用T3轉(zhuǎn)基因植株進(jìn)行抗逆境脅迫分析。【結(jié)果】MnMAPK5基因的編碼蛋白定位于細(xì)胞質(zhì)和細(xì)胞核中。在擬南芥中檢測(cè)MnMAPK5啟動(dòng)子驅(qū)動(dòng)的GUS活性的結(jié)果顯示,MnMAPK5啟動(dòng)子活性受到了高溫、低溫、高鹽和干旱脅迫的誘導(dǎo)。在高鹽、干旱、低溫和H_2O_2處理?xiàng)l件下,過表達(dá)MnMAPK5抑制了轉(zhuǎn)基因擬南芥的萌發(fā)和生長(zhǎng)。在鹽脅迫下,過表達(dá)MnMAPK5提高了轉(zhuǎn)基因植株中的MDA含量和POD活性,降低了CAT活性、可溶性糖含量和H_2O_2含量;在干旱脅迫下,過表達(dá)MnMAPK5提高了轉(zhuǎn)基因擬南芥中的MDA和H_2O_2含量,降低了CAT和POD活性;在低溫下,轉(zhuǎn)基因擬南芥相比野生型植株表現(xiàn)出更低的CAT活性、POD活性、脯氨酸含量和可溶性糖含量;在氧化環(huán)境脅迫下,轉(zhuǎn)基因擬南芥的MDA含量顯著高于野生型擬南芥,而脯氨酸含量和可溶性糖含量要低。此外,在各個(gè)脅迫環(huán)境條件下,AtPOD、AtCAT和AtRD22等脅迫相關(guān)基因的表達(dá)量在轉(zhuǎn)基因植株低于野生型擬南芥。【結(jié)論】MnMAPK5啟動(dòng)子在轉(zhuǎn)基因擬南芥中的活性受到了不同非生物脅迫的誘導(dǎo)。過量表達(dá)MnMAPK5降低了擬南芥對(duì)非生物脅迫的抵抗能力,表明MnMAPK5在逆境脅迫下起著負(fù)調(diào)控作用。
[Abstract]:[objective] Plant mitogen-activated protein kinase (MAPK) plays an important role in response to biotic and abiotic stress. Subcellular localization and promoter activity of B family MAPK gene MnMAPK5. It was transferred into Arabidopsis thaliana to investigate the mechanism of MnMAPK5 under stress. [methods] the upstream promoter of MnMAPK5 was ligated to p BI121 vector. Arabidopsis thaliana was transformed into Arabidopsis thaliana by pollen-mediated method. GUS staining was carried out after high temperature, low temperature and PEG treatment. MnMAPK5 was transformed into Arabidopsis thaliana by construction of overexpression vector. T3 transgenic plants were used to analyze the stress resistance. [results] the coding protein of MnMAPK5 gene was located in cytoplasm and nucleus. Detection of gu driven by MnMAPK5 promoter in Arabidopsis thaliana. The results of S-activity showed that. The activity of MnMAPK5 promoter was induced by high temperature, low temperature, high salt and drought stress. Overexpression of MnMAPK5 inhibited the germination and growth of transgenic Arabidopsis thaliana. Under salt stress, overexpression of MnMAPK5 increased MDA content and POD activity in transgenic plants. The activity of CAT, the content of soluble sugar and the content of H _ 2O _ 2 were decreased. Under drought stress, overexpression of MnMAPK5 increased the contents of MDA and H _ 2O _ 2 and decreased the activities of CAT and POD in transgenic Arabidopsis thaliana. At low temperature, transgenic Arabidopsis thaliana showed lower CAT activity, proline content and soluble sugar content than wild-type plants. Under oxidative stress, the MDA content of transgenic Arabidopsis thaliana was significantly higher than that of wild type Arabidopsis thaliana, while the content of proline and soluble sugar was lower. The expression of stress related genes such as AtCAT and AtRD22 in transgenic plants was lower than that in wild type Arabidopsis thaliana. [conclusion] the activity of MnMAPK5 promoter in transgenic Arabidopsis thaliana was different from that in transgenic Arabidopsis. Overexpression of MnMAPK5 reduced the resistance of Arabidopsis thaliana to abiotic stress. The results suggest that MnMAPK5 plays a negative regulatory role under stress.
【作者單位】： 西南大學(xué)家蠶基因組生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室農(nóng)業(yè)部蠶桑生物學(xué)與遺傳育種重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系建設(shè)專項(xiàng)(CARS-18-ZJ0201) 國(guó)家農(nóng)業(yè)部公益性行業(yè)(農(nóng)業(yè))科研專項(xiàng)(201403064)
【分類號(hào)】：Q943.2
【正文快照】： 促分裂原活化蛋白激酶(mitogen-activatedprotein kinases,MAPK)在植物響應(yīng)逆境脅迫中發(fā)揮著重要的作用,可將細(xì)胞外的信號(hào)轉(zhuǎn)入胞內(nèi),影響其下游基因的表達(dá),從而調(diào)節(jié)生物體內(nèi)各種生理反應(yīng),響應(yīng)外界環(huán)境。植物中的MAPK基因最早在紫花苜蓿(Medicago sativa)中被鑒定,此后陸續(xù)在擬，

本文編號(hào)：1453703