中文摘要：

抗冻蛋（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 virus35S，CaMV35S））驱动下胡萝卜（Daucuscarota）A即在烟草（Nicotiana tabacum）中的表达。结果表明，DcAFP基因己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10d，-1℃处理24h，室温恢复生长1周，T1代转Prd29A：DcAFP基因烟草存活率比转醌MV35S：DcAFP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25qC恢复生长后，转Prd29A：DcAFP基因烟草FWFm接近正常水平，而转CaMV35S：DeAFP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DcAFP基因烟草在4℃低温驯化（0～15d）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（superoxidedismutase，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基因烟草比转能M妇5S？D叫FP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。

英文摘要：

Antifreeze proteins （AFPs） is a kind of special small molecule protein produced by cold-resistant organisms for adapting low temperature environment, and it can decrease ice point and inhibit ice recrystalization. In this research, the expression level of DcAFP from carrot （Daucus carota） driven by environment inducible promoter Prd29A （responsive to desiccation 29A） and constitutive promoter Cauliflower mosaic virus 35S （CaMV35S） in transgenic tobacco （Nicotiana tabacum） were detected. The results showed that the DcAFP gene has been integrated into tobacco genome and could express stably in progeny. The phenotype of inflorescence in T℃ transgenic tobacco was dysplasia, and its capsule was easily to detach from the carpopodium, and few fruits could develop normally. After 4 ℃ acclimation for 10 d, - 1 ℃ low temperature treatment for 24 h and 25 ℃ recover for 7 d, the survival rate of T℃ tobacco plants transformed with Prd29A：AFP was 34.6% higher than that transformed with CaMV35S：AFP, and was 61.1% higher than wild type tobacco plants. Tl tobacco plants transformed with Prd29A：AFP grew well, while wild type tobacco was most dead. The chlorophyll fluorescence parameters （Fv/Fm） of all kinds of tobacco plants were decreased under cold stress, but after 25 ℃ recover, the Fv/Fm in Prd29A：AFP transgenic tobacco restored to the normal level, but the Fv/Fm in CaMV35S：AFP transgenic tobacco was just a little higher than that in wild type tobacco. Compared with wide type tobacco, proline content, soluble sugar content and soluble protein content in the two DcAFP transgenic tobacco under cold acclimation at 4 ℃ were increased, and the activity of superoxide dismutase （SOD） and peroxidase （POD） enhanced, and isozyme bands were increased. At the same condition, proline content, soluble sugar content, soluble protein content as well as SOD and POD activity in Prd29A：AFP transgenic tobacco were 45.1%, 12.1%, 20%, 19.7% and 8.6% higher than that in CaMV35S：AFP transgen