馬鈴薯StNCED1基因過表達(dá)載體的構(gòu)建及其遺傳轉(zhuǎn)化
發(fā)布時間:2018-03-12 12:08
本文選題:NCED基因 切入點:植物表達(dá)載體 出處:《分子植物育種》2017年01期 論文類型:期刊論文
【摘要】:馬鈴薯塊莖的休眠和發(fā)芽對于塊莖生產(chǎn)和加工極為重要,延長塊莖休眠期有助于降低塊莖水分和養(yǎng)分的消耗,從而提高其應(yīng)用價值。本研究用PCR方法從馬鈴薯中克隆了9-順式-環(huán)氧類胡蘿卜素雙氧合酶(NCED)編碼基因St NCED1,并構(gòu)建了馬鈴薯塊莖特異表達(dá)啟動子CIPP驅(qū)動的St NCED1基因的植物過表達(dá)載體p BIC-St NCED1,然后通過根癌農(nóng)桿菌介導(dǎo)法轉(zhuǎn)化馬鈴薯栽培品種"隴薯3號"獲得轉(zhuǎn)化植株,經(jīng)卡那霉素抗性篩選得到5株抗性苗,PCR檢測證明St NCED1基因已整合到馬鈴薯的基因組中。qRT-PCR檢測表明St NCED1基因在轉(zhuǎn)基因植株試管薯中的表達(dá)量分別比未轉(zhuǎn)基因的對照增加2.10~8.09倍。該研究有助于培育塊莖休眠期延長的馬鈴薯新種質(zhì)。
[Abstract]:The dormancy and germination of potato tubers are very important for tuber production and processing. Prolonging the dormancy period of tubers is helpful to reduce the consumption of water and nutrients in tubers. In order to improve its application value, we cloned 9-cis-epoxy-carotenoid dioxygenase (NCED1) encoding gene St NCED1 from potato by PCR method, and constructed St NCED1 gene driven by potato tuber specific expression promoter CIPP. The plant overexpression vector p BIC-St NCED1 was transformed into potato cultivar "Longshu 3" by Agrobacterium tumefaciens mediated by Agrobacterium tumefaciens. Five resistant seedlings were screened by kanamycin resistance. The results showed that St NCED1 gene had been integrated into potato genome. QRT-PCR analysis showed that St NCED1 gene expression in transgenic tuber was higher than that in untransgenic control. This study was helpful to cultivate new potato germplasm with prolonged dormancy of tuber.
【作者單位】: 甘肅省作物遺傳改良與種質(zhì)創(chuàng)新重點實驗室甘肅省干旱生境作物學(xué)省部共建國家重點實驗室培育基地;甘肅農(nóng)業(yè)大學(xué)生命科學(xué)技術(shù)學(xué)院;
【基金】:國家自然科學(xué)基金(31160298;31560413) 甘肅省杰出青年科學(xué)基金項目(1308RJDA011) 甘肅農(nóng)業(yè)大學(xué)“伏羲人才”計劃項目(FXRC20130102)共同資助
【分類號】:S532;Q943.2
【相似文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 馬瑞;馬鈴薯StNCED1基因的克隆及其功能研究[D];甘肅農(nóng)業(yè)大學(xué);2016年
,本文編號:1601493
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