本文選題：農(nóng)桿菌　切入點(diǎn)：轉(zhuǎn)基因棉花　出處：《中國農(nóng)業(yè)科技導(dǎo)報(bào)》2016年04期

【摘要】：利用農(nóng)桿菌介導(dǎo)法將耐旱耐鹽轉(zhuǎn)錄因子基因Pe DREB2a和Kc ERF導(dǎo)入受體材料陸地棉R15中,獲得12個(gè)株系的轉(zhuǎn)基因植株,通過硫酸卡那霉素初篩以及PCR分子檢測(cè)最終獲得7個(gè)轉(zhuǎn)Kc ERF-Pe DREB2a基因的棉花株系。通過測(cè)定轉(zhuǎn)基因棉花的抗逆相關(guān)生理指標(biāo)以及對(duì)基因相對(duì)表達(dá)量測(cè)定分析,結(jié)果表明,200mmol/L的Na Cl和15%的PEG-6000脅迫處理后,轉(zhuǎn)基因棉花幼苗的CAT、SOD活性,游離脯氨酸含量均高于對(duì)照組,MDA含量較對(duì)照組棉花明顯下降。實(shí)時(shí)定量RT-PCR結(jié)果表明,干旱脅迫下,轉(zhuǎn)基因棉花幼苗葉片中Pe DREB2a基因的表達(dá)量高于Kc ERF基因的表達(dá)量;高鹽脅迫下,轉(zhuǎn)基因棉花葉片中的Pe DREB2a,Kc ERF基因的相對(duì)表達(dá)量持平。本研究結(jié)果表明,在干旱、高鹽脅迫下,Kc ERF-Pe DREB2a基因有助于提高棉花的耐旱耐鹽能力。
[Abstract]:Agrobacterium tumefaciens were used to transfer the drought and salt tolerant transcription factor genes Pe DREB2a and Kc ERF into Upland Cotton R15, and 12 transgenic plants were obtained.Through the primary screening of kanamycin sulfate and PCR molecular analysis, 7 cotton lines with Kc ERF-Pe DREB2a gene were obtained.By determining the physiological indexes related to stress resistance and the relative expression of genes in transgenic cotton, the results showed that the activity of CATD-SOD in transgenic cotton seedlings was increased after NaCl and 15% PEG-6000 stress.The content of free proline was significantly lower than that of control cotton.The results of real-time quantitative RT-PCR showed that, under drought stress, the expression of Pe DREB2a gene was higher than that of Kc ERF gene in transgenic cotton seedling leaves, and the relative expression of Pe DREB2a Kc ERF gene in transgenic cotton leaves was the same under high salt stress.The results showed that Kc ERF-Pe DREB2a gene was helpful to improve drought tolerance and salt tolerance of cotton under drought and high salt stress.
【作者單位】： 四川農(nóng)業(yè)大學(xué)生命科學(xué)學(xué)院;中國農(nóng)業(yè)科學(xué)院生物技術(shù)研究所;深圳市鐵漢生態(tài)環(huán)境股份有限公司;
【基金】：國家轉(zhuǎn)基因生物新品種培育重大專項(xiàng)(2014ZX08005-004) 廣東省省級(jí)科技計(jì)劃項(xiàng)目(2015B090904008)資助
【分類號(hào)】：S562
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本文編號(hào)：1702202