本文關(guān)鍵詞：抗逆相關(guān)基因GmAREB轉(zhuǎn)基因小麥的獲得與鑒定，，由筆耕文化傳播整理發(fā)布。

抗逆相關(guān)基因GmAREB轉(zhuǎn)基因小麥的獲得與鑒定

Molecular and Functional Analysis of Transgenic Wheat Transformed with Stress-related Gene GmAREB

[1] [2] [3] [4] [5] [6] [7] [8]

CHEN Hong-min,CHEN Ming,WEI An-zhi,XU Zhao-shi,LI Lian-cheng,XU Hui-jun,DU Li-pu,MA You-zhi（1 College of Agronomy,Northwest A＆F University,Yangling Shaanxi 712100;2 National Key Faci

[1]西北農(nóng)林科技大學(xué)林學(xué)院,陜西楊凌712100; [2]中國農(nóng)業(yè)科學(xué)院作物科學(xué)研究所/國家農(nóng)作物基因資源與基因改良重大科學(xué)工程/農(nóng)業(yè)部作物遺傳改良與育種重點(diǎn)開放實驗室,北京100081; [3]河南省濮陽市種子管理站,濮陽457000

文章摘要：從大豆中克隆一個抗逆相關(guān)的bZIP類轉(zhuǎn)錄因子基因GmAREB,功能分析表明：GmAREB基因的過表達(dá)可以顯著提高轉(zhuǎn)基因擬南芥和煙草的抗旱、耐鹽和耐寒性。為了獲得抗逆轉(zhuǎn)基因小麥,本研究利用玉米的Ubiqutin啟動子控制GmAREB基因表達(dá),構(gòu)建了用于小麥轉(zhuǎn)化的載體pSK-GmAREB。采用基因槍共轉(zhuǎn)化法轉(zhuǎn)化小麥品種鄭147和濟(jì)麥22。通過PCR檢測共獲得T0代的陽性植株70株,轉(zhuǎn)化率為1.37%。其中,鄭147陽性植株共31株,轉(zhuǎn)化率為2.14%;濟(jì)麥22陽性植株39株,轉(zhuǎn)化率為1.08%。目前,已經(jīng)獲得T1代轉(zhuǎn)基因株系18個,其中以鄭147為受體的株系4個,以濟(jì)麥22為受體的株系14個。對部分株系進(jìn)行Southern blotting分析,進(jìn)一步證實GmAREB基因已經(jīng)整合到小麥基因組中。在低溫脅迫條件下,3個以濟(jì)麥22為受體的轉(zhuǎn)基因株系體內(nèi)脯氨酸的積累與受體小麥相比有顯著增加,初步證明在小麥中過表達(dá)GmAREB基因,可以促進(jìn)滲透調(diào)節(jié)物質(zhì)脯氨酸的積累,可能有助于轉(zhuǎn)基因小麥抗逆性的提高。本研究為進(jìn)一步篩選抗逆轉(zhuǎn)基因小麥新材料奠定了基礎(chǔ)。

Abstr：We isolated stress-related bZIP transcription factor gene,GmAREB from soybean previously.The functional analysis indicated that overexpression of GmAREB can significantly improve the stress resistance of transgenic plants.In this study,GmAREB was inserted in downstream of maize Ubiqutin promoter to construct vector,pSK-GmAREB for wheat transformation.The vectors were transformed into wheat varieties Zheng 147 and Jimai 22 by using biolistic method.After transformation,70 T0 transgenic plants were identified using PCR and the transformational efficiency was 1.37%.Among transgenic plants,31 plants came from host Zheng 147 and the transformational efficiency was 2.14%.39 plants came from host Jimai 22 and the transformational efficiency was 1.08%.At present,18 T1 generation of transgenic line had been identificated,and 4 and 14 transgenic plants came from Zheng 147 and Jimai 22,respectively.The southern blot proved that GmAREB was inserted into wheat genome.The stress-tolerance analysis showed that under low temperature stress condition,proline accumulation in three transgenic plants host in Jimai 22 were higher than that of wild type wheat plants,which suggested that overexpression of GmAREB increased accumulation of proline in transgenic wheat.This higher level of proline content might be contributed for stress-tolerance of transgenic wheat plants.We hope that our study can pave the way for selecting stress-tolerance transgenic wheat lines in the future.

文章關(guān)鍵詞：

Keyword：:Wheat Biolistic Transcription factor GmAREB gene Proline

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課題項目：國家高技術(shù)研究發(fā)展計劃（863計劃）項目（2008AA10Z124）; 國家自然科學(xué)基金項目（30700508）

  本文關(guān)鍵詞：抗逆相關(guān)基因GmAREB轉(zhuǎn)基因小麥的獲得與鑒定，由筆耕文化傳播整理發(fā)布。