【摘要】：為確定擬南芥抗灰霉病相關(guān)基因AtSEC14的功能,本試驗(yàn)構(gòu)建了AtSEC14基因的反義RNA載體;通過(guò)農(nóng)桿菌介導(dǎo)的遺傳轉(zhuǎn)化方法,將其轉(zhuǎn)化擬南芥野生型Col-0中;利用潮霉素抗性篩選和PCR檢測(cè),獲得了陽(yáng)性轉(zhuǎn)基因植株。利用半定量RT-PCR技術(shù),在轉(zhuǎn)基因株系中未檢測(cè)到AtSEC14基因的表達(dá),說(shuō)明該基因反義RNA載體的轉(zhuǎn)入能特異影響AtSEC14基因的表達(dá),表明試驗(yàn)所獲得的轉(zhuǎn)基因植株是AtSEC14基因的反義RNA轉(zhuǎn)基因植株。對(duì)所獲得的反義RNA轉(zhuǎn)基因植株進(jìn)行抗病性鑒定,發(fā)現(xiàn)反義RNA轉(zhuǎn)基因植株對(duì)灰葡萄孢的敏感性增強(qiáng),表明AtSEC14基因在擬南芥抗灰葡萄孢過(guò)程中起正調(diào)控的作用。
[Abstract]:In order to determine the function of Arabidopsis cinerea resistance gene AtSEC14, the antisense RNA vector of AtSEC14 gene was constructed, transformed into Arabidopsis wild-type Col-0 by Agrobacterium tumefaciens mediated genetic transformation, hygromycin resistance screening and PCR detection were used. Positive transgenic plants were obtained. The expression of AtSEC14 gene was not detected in transgenic lines by semi-quantitative RT-PCR technique, which indicated that the transfer of antisense RNA vector could specifically affect the expression of AtSEC14 gene. The results showed that the transgenic plants were antisense RNA transgenic plants of AtSEC14 gene. The antisense RNA transgenic plants were identified for disease resistance, and it was found that the antisense RNA transgenic plants were more sensitive to grapevine spores, indicating that the AtSEC14 gene plays a positive role in the process of resistance of Arabidopsis thaliana to grapevine spores.
【作者單位】： 河北農(nóng)業(yè)大學(xué)真菌毒素與植物分子病理學(xué)實(shí)驗(yàn)室;河北北方學(xué)院農(nóng)林科技學(xué)院;
【基金】：河北省自然科學(xué)基金項(xiàng)目(C2014405010)
【分類號(hào)】：S432.4
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本文編號(hào)：2215835