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| Study on Transgenic Potato Contained AtCDPK1 Gene Drived by Rd29A Promoter |
| NIE Li-zhen1, YU Xiao-xia1, LI Guo-jing1, SUN Jie2, JIANG Chao1, YU Zhuo1 |
1. Agronomy College, Inner Mongolia Agricultural University, Huhhot 010019, China;
2. Inner Mongolia Academy of Agricultural AnimalHusbandry Sciences, Hohhot 010031, China |
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Abstract In order to obtain normal growth and resistant drought transgenic potato, using Arabidopsis thaliana (Columbia ecotype) as material, DNA sequence of AtRd29A gene ATG upstream from +83bp to -1 441bp is cloned with PCR and recombinant DNA technologies. The sequence of bases was entirely consistent with that of promoter of the known gene. The plant expression vector pCHFRd-CDPK1 of AtCDPK1 gene drived by Rd29A promoter was constructed, and transformed pCHFRd-CDPK1 into virus-free miniature of potato cultivar‘Favorita’using Agrobacterium-mediated method. After plant selection and regeneration, regenerated plants with resistance were obtained successfully. The PCR and Southern blotting analysis proved that the AtCDPK1 gene has been integrated in the genome of the potato. After drought stress using 30% PEG, RT-PCR analysis proved that the transcript levels of AtCDPK1 gene drived by Rd29A promoter in transgenic potato plants were significantly higher. AtCDPK1 gene drived by Rd29A promoter in transgenic potato plants is barely detectable in the water treatment control. However, expression of AtCDPK1 gene drived by 35S promoter did not make significant difference in transgenic potato plants between PEG stress treatment and water treatment. Morphological observation showed that transgenic plants can grow normally after 30% PEG stress, the plants growing was better than non-transgenic plants, and control plants occur slightly wilt. This research will lay the foundation for further improved stress tolerance of crops by expression of resistant gene drived by stress-inducible promoter in the crop.
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Received: 11 June 2015
Published: 24 November 2015
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[1] 刘玲. 内蒙古马铃薯产业化问题研究. 呼和浩特:内蒙古农业大学,农学院,2003. Liu L. Study on Industrialization of Potato in Inner Mongolia Autonomous Region. Huhhot: Inner Mongolia Agricultural University, Agricultural College,2003.
[2] 孙国凤,马鑫. 农业生物技术发展现状与展望. 农业展望,2010,(11):56-59. Sun G F, Ma X. Development status and prospect of agricultural biotechnology. Agricultural Outlook, 2010, (11): 56-59.
[3] Yeo E T, Kwon H B, Han S E, et al. Genetic engineering of drought-resistant potato plants by introduction of the trehalose-6-phosphate synthase ( TPS1 ) gene from Saccharomyces cerevisiae. Molecular Cells, 2000, 10(3): 263-268.
[4] 张宁. 应用甜菜碱醛脱氢酶基因工程提高马铃薯抗逆性的研究. 兰州:甘肃农业大学,农学院,2004. Zhang N. Studies on Increasing Stress Resistance of Potato by Genetic Engineering of Betaine Aldehyde Dehydrogenase. Lanzhou: Gansu Agricultural University, Agricultural College,2004.
[5] Roberts D M, Harmon A C. Calcium-modulated proteins: targets of intracellular calcium signals in higher plants. Annual Review of Physiol Plant Molecular Biology, 1992, 43: 375-414.
[6] Hrabak E M. Plant Protein Kinases: Calcium-dependent Protein Kinases and Their Relatives. In: Kreis M, Walker J C. Advances in Botanical Sciences. New York: Academic Press, 2000: 185-223.
[7] Harmon A C, Gribskov M, Gubrum E, et al. The CDPK superfamily of protein kinases. New Phytologist, 2001, 151: 175-183.
[8] Harper J F, Breton G, Harmon A C. Decoding Ca2+ signals through plant protein kinases. Annual Review of Plant Biology, 2004, 55: 263-288.
[9] Hetherington A, Trewavas A. Calcium-dependent protein kinase in pea shoot membranes. FEBS Letters, 1982, 145(1): 67-71.
[10] Sheen J. Ca2+-dependent protein kinases and stress signal transduction in plants. Science, 1996, 274(5294): 1900-1902.
[11] 卢慧星. 拟南芥 AtCDPK1 参与非生物胁迫响应和ABA信号途径的证据. 呼和浩特: 内蒙古农业大学,生命科学院,2009. Lu H X. The Evidence of AtCDPK1 Involved in Abiotic Stresses and ABA Signaling Pathway. Huhhot: Inner Mongolia Agricultural University, College of Life Science,2009.
[12] Zou J J, Wei F J, Wang C, et al. Arabidopsis calcium-dependent protein kinase CPK10 functions in abscisic acid and Ca2+-mediated stomatal regulation in response to drought stress. Plant Physiol, 2010, 154(3): 1232-1243.
[13] 路静,赵华燕,何奕昆,等. 高等植物启动子及其应用研究进展. 自然科学进展,2004,14( 8):856-862. Lu J, Zhao H R, He Y K, et al. Research and application of promoter in higher plants . Progress of Natural Science, 2004, 14(8): 856-862.
[14] Gittins J R1, Pellny T K, Hiles E R, et al. Transgene expression driven by heterologous ribulose-1, 5-bisphosphate carboxylase/oxygenase small-subunit gene promoters in the vegetative tissues of apple (Malus pumila Mill.) . Planta, 2000, 210(2): 232-240.
[15] 纪巍,李杰,朱延明,等.不同启动子调控的 DREB1A 基因对黄瓜的遗传转化.东北农业大学学报,2005,36(4):442-447. Ji W, Li J, Zhu Y M,et al. DREB1A gene drived by different promoters transformation into Cucumissativus. Journal of Northeast Agricultural University, 2005, 36(4): 442-447.
[16] 聂利珍,于肖夏,李国婧,等. 拟南芥 AtCDPK1 基因克隆与转化马铃薯的研究.西北植物学报,2015,35(3):447-453. Nie L Z, Yu X X, Li G J, et al. Cloning and Transformation into potato of AtCDPK1 gene from Arabidopsis thaliana. Acta Botanica Boreall-occidentalia Sinica, 2015, 35(3): 447-453.
[17] Xiong L, Schumaker K S, Zhu J K. Cell signaling during cold, drought, and salt stress. Plant Cell, 2002, 14: S165-S183.
[18] ZHu J K, Hasengawa P M, Bressan R A, et al. Molecular aspects of osmotic stress in plants. Critical Reviews in Plant Sciences, 1997, 16(3): 253-277.
[19] Hasegawa P M, Bressan R A, Zhu J K, et al. Plant cellular and molecular responses to high salinity. Annual Review of Plant Physiology and Plant Molecular Biology, 2000, 51: 463-499.
[20] 聂利珍. 拟南芥 AtCDPK1 在响应丁香假单胞菌和非生物胁迫中的功能分析. 呼和浩特:内蒙古农业大学,生命科学院,2008. Nie L Z. Functional Analysis of AtCDPK1 in Arabidopsis Response to Pseudomonas syringae and Abiotic Stresses. Huhhot: Inner Mongolia Agricultural University, College of Life Science,2008.
[21] Yamaguchi, Shinozaki K, Shinozaki K. Arabidopsis DNA encoding two desiccation responsive rd29A genes. Plant Physiol, 1993, 101: 1119-1120.
[22] Shinozaki K, Yamaguchi, Shinozaki K. Gene expression and signal transduction in water stress response. Plant Physiol, 1997, 115: 327-334.
[23] Lin Q, Li J,Zhang G Y, et al. Isolation of cDNAs encoding two distinct transcription factors binding to DRE cis-acting element involved in cold-and drought-induced expression of Arabidopsis rd29A Gene . Tsinghua Science and Technology. 1999, 4(3):256-260.
[24] 刘强,赵南明,Yamaguch S K,等.DREB类转录因子在提高植物抗逆性中的作用.科学通报,2000,45(11):11-17. Liu Q, Zhao N M, Yamaguch S K, et al. The role of DREB transcription factors in improving plant resistance. Chinese Science Bulletin, 2000, 45(11): 11-17. |
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