木薯海藻糖-6-磷酸酯酶MeTPP6基因克隆及其表达分析
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摘要
海藻糖-6-磷酸酯酶(TPP)负责海藻糖生物合成催化反应的最后一步,是植物海藻糖生物合成途径的关键酶。采用RT-PCR的方法从木薯叶片中克隆了1个TPP基因,命名为MeTPP6,该基因含有1个1 122 bp的开放阅读框,编码373个氨基酸,具有TPP家族保守结构域。系统进化树分析表明,MeTPP6与杞柳和杨树中同源基因的亲缘关系较近,序列相似性高达89.7%和89.0%。启动子分析表明,MeTPP6含有干旱、低温、热胁迫、激素(如ABA)和光响应等相关元件。荧光定量PCR分析表明,MeTPP6在叶片和叶柄中表达量最低;在须根和储藏根中表达量最高,分别为叶片表达量的4.2倍和4.5倍。而且,MeTPP6基因的表达能被干旱、低温和ABA处理显著诱导。这些结果表明,MeTPP6通过依赖于ABA的信号通路在转录水平参与木薯干旱和低温胁迫,可作为重要候选基因进一步研究其在木薯非生物逆境中的功能。
引文
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[14]丁泽红,付莉莉,铁韦韦,等. 木薯MeNCED3基因克隆、结构变异及其表达分析[J]. 生物技术通报,2016,32(10):148-153.
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[16]Garg A K,Kim J K,Owens T G,et al. Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses[J]. Proceedings of the National Academy of Sciences of the United States of America,2002,99(25):15898-15903.
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[18]Shinozaki K,Yamaguchi-Shinozaki K. Gene networks involved in drought stress response and tolerance[J]. Journal of Experimental Botany,2007,58(2):221-227.
[2]Okogbenin E,Setter T L,Ferguson M,et al. Phenotypic approaches to drought in cassava:review[J]. Frontiers in Physiology,2013,4(1):93.
[3]卢赛清,盘欢,马崇熙,等. 2008年广西木薯低温冻害情况及应对措施[J]. 广西热带农业,2009(1):21-22.
[4]张雯,王宇斐,郭延平.高等植物6-磷酸海藻糖信号调控研究进展[J]. 植物生理学报,2016,52(4):394-400.
[5]张建波,王莎莎,郝大海,等. 干旱和低温胁迫影响烟草幼苗海藻糖代谢的差异比较[J]. 生物技术通报,2015,31(10):111-118.
[6]史健志,徐燕,纪德华,等. 坛紫菜6-磷酸海藻糖合成酶(TPS)家族基因的克隆及表达特征分析[J]. 水产学报,2015,39(4):485-495.
[7]Ge L F,Chao D Y,Shi M,et al. Overexpression of the trehalose-6-phosphate phosphatase gene OsTPP1 confers stress tolerance in rice and results in the activation of stress responsive genes[J]. Planta,2008,228(1):191-201.
[8]Vandesteene L,Lopez-Galvis L,Vanneste K,et al. Expansive evolution of the trehalose-6-phosphate phosphatase gene family in Arabidopsis[J]. Plant Physiology,2012,160(2):884-896.
[9]Li P H,Ma S S,Bohnert H J. Coexpression characteristics of trehalose-6-phosphate phosphatase subfamily genes reveal different functions in a network context[J]. Physiologia Plantarum,2008,133(3):544-556.
[10]Pramanik M R,Imai R. Functional identification of a trehalose 6-phosphate phosphatase gene that is involved in transient induction of trehalose biosynthesis during chilling stress in rice[J]. Plant Molecular Biology,2005,58(6):751-762.
[11]Nuccio M L,Wu J,Mowers R,et al. Expression of trehalose-6-phosphate phosphatase in maize ears improves yield in well-watered and drought conditions[J]. Nature Biotechnology,2015,33(8):862.
[12]Kretzschmar T,Pelayo M A,Trijatmiko K R,et al. A trehalose-6-phosphate phosphatase enhances anaerobic germination tolerance in rice[J]. Nature Plants,2015,1(9):15124.
[13]Vogel G,Aeschbacher R A,Muller J,et al. Trehalose-6-phosphate phosphatases from Arabidopsis thaliana:identification by functional complementation of the yeast tps2 mutant[J]. Plant Journal,1998,13(5):673-683.
[14]丁泽红,付莉莉,铁韦韦,等. 木薯MeNCED3基因克隆、结构变异及其表达分析[J]. 生物技术通报,2016,32(10):148-153.
[15]Fu L L,Ding Z H,Han B Y,et al. Physiological investigation and transcriptome analysis of polyethylene glycol (PEG)-induced dehydration stress in cassava[J]. International Journal of Molecular Sciences,2016,17(3):283.
[16]Garg A K,Kim J K,Owens T G,et al. Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses[J]. Proceedings of the National Academy of Sciences of the United States of America,2002,99(25):15898-15903.
[17]Krasensky J,Broyart C,Rabanal F A. The redox-sensitive chloroplast trehalose-6-phosphate phosphatase AtTPPD regulates salt stress tolerance[J]. Antioxidants & Redox Signaling,2014,21(9):1289-1304.
[18]Shinozaki K,Yamaguchi-Shinozaki K. Gene networks involved in drought stress response and tolerance[J]. Journal of Experimental Botany,2007,58(2):221-227.