本文關(guān)鍵詞： 紫花苜蓿 胚胎晚期富集蛋白(LEA) 逆境脅迫 遺傳轉(zhuǎn)化　出處：《草業(yè)學(xué)報》2017年12期 　論文類型：期刊論文

【摘要】：胚胎晚期富集蛋白(LEA)廣泛參與植物對多種逆境脅迫的反應(yīng)。本研究利用同源克隆的方法,從紫花苜蓿中克隆了一個LEA4類基因的開放閱讀框(ORF),命名為Ms LEA4-4。該基因編碼512個氨基酸,結(jié)構(gòu)分析顯示MsLEA4-4包含5個重復(fù)的由11個氨基酸TAQAAKEKTQQ組成的序列特征。利用實(shí)時熒光定量PCR檢測了Ms LEA4-4在不同逆境下的表達(dá)量,結(jié)果顯示,該基因受干旱、NaCl、Cu~(2+)、Zn~(2+)和外源ABA誘導(dǎo)表達(dá)上調(diào),其中Na Cl脅迫2h、Cu2+和Zn2+脅迫8 h,Ms LEA4-4基因表達(dá)量最高;冷脅迫和干旱脅迫下,該基因的表達(dá)量隨處理時間的延長呈逐漸上升趨勢,表明該基因可能參與了紫花苜蓿的抗逆性調(diào)控。構(gòu)建植物超表達(dá)載體p CAMBIA3301-Ms LEA4-4,采用農(nóng)桿菌介導(dǎo)法侵染擬南芥花序,通過草銨膦(PPT)篩選和分子檢測,7株抗性苗呈陽性,表明目的基因已成功導(dǎo)入擬南芥基因組中。本研究為進(jìn)一步探索Ms LEA4-4基因在紫花苜�？鼓嫘哉{(diào)控中的作用奠定了基礎(chǔ)。
[Abstract]:The late embryo enriched protein Lea was widely involved in the response of plants to various stresses. The method of homologous cloning was used in this study. An open reading frame of LEA4 gene was cloned from alfalfa, named MS LEA4-4. the gene encodes 512 amino acids. Structural analysis showed that MsLEA4-4 contained five repetitive sequence features consisting of 11 amino acid TAQAAKEKTQQ. Ms was detected by real-time fluorescence quantitative PCR. The expression of LEA4-4 under different stress. The results showed that the expression of the gene was up-regulated by drought induced ABA and NaCl stress for 2 hours. The expression of Ms LEA4-4 gene was the highest under the stress of Cu2 and Zn2 for 8 h. Under cold stress and drought stress, the expression of the gene increased with the prolongation of treatment time. It was suggested that the gene might be involved in the regulation of resistance to stress in alfalfa. The plant overexpression vector p CAMBIA3301-Ms LEA4-4 was constructed and infected with Arabidopsis thaliana inflorescence by Agrobacterium tumefaciens. Seven resistant seedlings were tested positive by PPTs screening and molecular detection. The results indicated that the target gene had been successfully introduced into Arabidopsis thaliana genome. This study laid a foundation for further exploring the role of Ms LEA4-4 gene in regulation of resistance to stress in alfalfa.
【作者單位】： 山西省農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所;中國農(nóng)業(yè)科學(xué)院北京畜牧獸醫(yī)研究所;北京綠京華園林工程有限公司;山西農(nóng)業(yè)大學(xué)動物科技學(xué)院;
【基金】：現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系專項資金(CARS-35-25) 物種資源保護(hù)費(fèi)(2130135) 山西省農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所項目(xms201506)資助
【分類號】：S541.9
【正文快照】： 由于植物的不可移動性,在生長發(fā)育過程中往往會遭受干旱、高溫、低溫、高鹽等各種非生物脅迫,這些脅迫都會引起植物內(nèi)部的水分缺失。缺水會引起植物體內(nèi)的呼吸作用和光合作用等一系列生理活動失調(diào)[1]。在長期的進(jìn)化過程中,植物演化出一系列從細(xì)胞到生理水平的應(yīng)答反應(yīng),如基因

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