本文選題：棉鈴蟲 + cryAh-M�。� 參考：《中國農(nóng)業(yè)科技導(dǎo)報(bào)》2017年04期

【摘要】：轉(zhuǎn)Bt基因抗蟲棉經(jīng)過20多年的研究和生產(chǎn)應(yīng)用,雖然取得了巨大的生態(tài)和經(jīng)濟(jì)效益,但棉鈴蟲的抗性風(fēng)險(xiǎn)也逐漸增加,因此,通過發(fā)掘不同殺蟲機(jī)理的Bt基因,不僅可以提高殺蟲效率,而且有效的避免或延遲昆蟲的抗性。依據(jù)棉花遺傳密碼子偏好性,對(duì)來源于蘇云金芽胞桿菌(Bacillus thuringiensis,Bt)新型Bt基因cry2Ah-like進(jìn)行了密碼子優(yōu)化改造,利用農(nóng)桿菌介導(dǎo)法導(dǎo)入煙草。通過熒光定量PCR方法,Southern blot和Western blot方法,證明了轉(zhuǎn)基因煙草中外源基因在RNA和蛋白水平上均穩(wěn)定表達(dá),并且已整合到煙草基因組中。生物活性檢測(cè)結(jié)果表明,飼喂轉(zhuǎn)基因煙草葉片4 d后,棉鈴蟲幼蟲校正死亡率達(dá)86.59%,達(dá)到高抗水平,進(jìn)一步證明密碼子優(yōu)化后的cry2Ah-M基因能夠高效表達(dá)且具有較高的殺蟲活性,是有效防治棉鈴蟲的新型基因。
[Abstract]:Transgenic BT cotton has been studied and applied for more than 20 years. Although it has achieved great ecological and economic benefits, the resistance risk of Helicoverpa armigera has also gradually increased. Therefore, the BT gene of different insecticidal mechanism was explored. It can not only improve insecticidal efficiency, but also effectively avoid or delay insect resistance. Based on the preference of cotton genetic codon, a new BT gene cry2Ah-like from Bacillus thuringiensis Btwas optimized and introduced into tobacco by Agrobacterium tumefaciens. Southern blot and Western blot methods were used to confirm the stable expression of transgenic tobacco genes at the RNA and protein levels, and they were integrated into the tobacco genome. The results of bioactivity test showed that the corrected mortality of cotton bollworm larvae reached 86.59 and reached a high level of resistance after feeding transgenic tobacco leaves for 4 days. It was further proved that the codon optimized cry2Ah-M gene could be expressed efficiently and had higher insecticidal activity. It is a new gene to control cotton bollworm effectively.
【作者單位】： 新疆農(nóng)業(yè)大學(xué)農(nóng)學(xué)院;中國農(nóng)業(yè)科學(xué)院生物技術(shù)研究所;中國農(nóng)業(yè)科學(xué)院植物保護(hù)研究所;
【基金】：國家轉(zhuǎn)基因生物新品種培育重大專項(xiàng)(2014ZX08005-004)資助
【分類號(hào)】：S435.62
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本文編號(hào)：1914789