本文選題：抗蟲性 + cryAb基因�。� 參考：《華南農(nóng)業(yè)大學(xué)學(xué)報(bào)》2016年05期

【摘要】：【目的】構(gòu)建包含抗鱗翅目害蟲基因cry1Ab13的重組植物表達(dá)載體,并利用其創(chuàng)制對(duì)玉米螟Ostrinia furnacalis具有優(yōu)良抗性的轉(zhuǎn)基因玉米Zea may L.新種質(zhì)�！痉椒ā坷猛粗亟M法將cry1Ab13基因連接到表達(dá)載體pCAMBIA3300-Bar上,獲得以抗除草劑Bar基因?yàn)楹Y選標(biāo)記的植物表達(dá)載體pCAMBIA3300-cry1Ab13-Bar。通過農(nóng)桿菌介導(dǎo)法轉(zhuǎn)化玉米自交系H99幼胚,對(duì)再生植株進(jìn)行逐代除草劑篩選、PCR檢測(cè)及T2代植株的Southern blotting檢測(cè)、實(shí)時(shí)熒光定量PCR檢測(cè),并對(duì)轉(zhuǎn)基因植株進(jìn)行田間及室內(nèi)抗蟲性鑒定�！窘Y(jié)果】構(gòu)建了cry1Ab13基因的植物表達(dá)載體,轉(zhuǎn)化玉米獲得3株高抗玉米螟和1株抗玉米螟的T2代轉(zhuǎn)基因植株。Southern blotting證明cry1Ab13基因已經(jīng)整合到玉米基因組中,實(shí)時(shí)熒光定量PCR結(jié)果表明cry1Ab13基因已經(jīng)在玉米植株內(nèi)表達(dá)�？瓜x性鑒定結(jié)果表明,與對(duì)照相比轉(zhuǎn)基因植株對(duì)玉米螟的抗性顯著提高�！窘Y(jié)論】將cry1Ab13基因?qū)胗衩撞⒊晒Ρ磉_(dá),顯著提高了轉(zhuǎn)基因玉米對(duì)玉米螟的抗性,為抗蟲玉米新種質(zhì)的創(chuàng)制奠定了基礎(chǔ)。
[Abstract]:[objective] to construct a recombinant plant expression vector containing the Lepidoptera pest resistance gene cry1Ab13, and to create a transgenic maize may L- Zea may with excellent resistance to Ostrinia furnacalis. [methods] the cry1Ab13 gene was ligated to the expression vector pCAMBIA3300-Bar by homologous recombination method, and the plant expression vector pCAMBIA3300-cry1Ab13-Barwas obtained by screening marker pCAMBIA3300-Bar. The immature embryos of maize inbred line H99 were transformed by Agrobacterium tumefaciens. The regenerated plants were screened by herbicides and detected by Southern blotting and real-time fluorescence quantitative PCR. The plant expression vector of cry1Ab13 gene was constructed. Three transgenic plants with high resistance to corn borer and one plant resistant to corn borer were obtained. Southern blotting showed that the cry1Ab13 gene had been integrated into the maize genome. The results of real-time fluorescence quantitative PCR showed that the cry1Ab13 gene had been expressed in maize plants. The results of insect-resistance identification showed that the resistance of transgenic plants to corn borer was significantly higher than that of control. [conclusion] cry1Ab13 gene was introduced into maize and expressed successfully, which significantly improved the resistance of transgenic maize to corn borer. It lays a foundation for the creation of new insect-resistant maize germplasm.
【作者單位】： 吉林農(nóng)業(yè)大學(xué)生命科學(xué)學(xué)院;
【基金】：吉林省省級(jí)糧食生產(chǎn)發(fā)展專項(xiàng)(2015001) 吉林農(nóng)業(yè)大學(xué)科研啟動(dòng)基金(201242)
【分類號(hào)】：S513
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本文編號(hào)：2007005