Prd29A与CaMV 35S启动子调控转胡萝卜AFP基因提高烟草抗寒性

发布时间：2018-07-04 08:05

  本文选题：抗冻蛋白(AFPs) + PrdA启动子　； 参考：《农业生物技术学报》2017年08期

【摘要】：抗冻蛋白(antifreeze proteins,AFPs)是耐寒生物为适应冷环境而产生的特殊小分子蛋白质,具有降低冰点、抑制重结晶的作用。本实验研究了在环境诱导型启动子(Prd29A(responsive to desiccation29A))和组成型启动子(花椰菜花叶病毒35S(Cauliflower mosaic virus 35S,Ca MV 35S))驱动下胡萝卜(Daucus carota)AFP在烟草(Nicotiana tabacum)中的表达。结果表明,Dc AFP基因已整合到烟草基因组中且可以稳定遗传,T1转基因烟草花序发育不良,蒴果容易从果柄基部脱落,极少数果实能够正常发育;4℃低温驯化10 d,-1℃处理24 h,室温恢复生长1周,T1代转Prd29A:Dc AFP基因烟草存活率比转Ca MV35S:Dc AFP基因烟草高34.6%,比野生型烟草高61.1%,且长势良好,而野生型烟草大多数枯萎死亡;低温胁迫处理导致所有烟草叶绿素荧光参数Fv/Fm均有所降低,但25℃恢复生长后,转Prd29A:Dc AFP基因烟草Fv/Fm接近正常水平,而转CaMV35S:DcAFP基因烟草Fv/Fm仅比野生型略高;与野生型对比,两种不同启动子驱动的转Dc AFP基因烟草在4℃低温驯化(0~15 d)期间积累了较多的脯氨酸、可溶性蛋白及可溶性糖等渗透调节物质,抗氧化酶(超氧化物歧化酶(superoxide dismutase,SOD)和过氧化物酶(peroxidase,POD))酶活有所提高,同工酶谱带增多;特别是转Prd29A:DcAFP基因烟草比转CaMV35S:DcAFP基因烟草中脯氨酸含量、可溶性糖含量、可溶性蛋白含量、SOD酶活酶活及POD酶活分别增加45.1%、12.1%、20%、19.7%、8.6%,而且转入Prd29A启动子驱动的DcAFP基因可以更有效的提高转基因烟草的抗寒性,并且其对转基因烟草生长发育影响相对较小。研究结果对于抗冻转基因植物的培育具有重要参考价值。
[Abstract]:Antifreeze proteins (antifreeze proteinsAFPs) are special small molecular proteins produced by cold-tolerant organisms to adapt to cold environment, which can reduce freezing point and inhibit recrystallization. In this study, the expression of Daucus carota AFP in tobacco (Nicotiana tabacum) induced by environmental inducible promoter (Prd29A (responsive to desiccation29A) and constitutive promoter (Cauliflower mosaic virus 35s (Cauliflower mosaic virus 35S) was studied. The results showed that the DcAFP gene had been integrated into the tobacco genome and could be stably inherited as the inflorescence of T 1 transgenic tobacco inflorescence was poor, and the capsule could easily be removed from the base of the stalk. A few fruits could develop normally at 4 鈩，

本文编号：2095393