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Journal of Integrative Agriculture  2020, Vol. 19 Issue (4): 941-952    DOI: 10.1016/S2095-3119(19)62714-0
Special Issue: 麦类遗传育种合辑Triticeae Crops Genetics · Breeding · Germplasm Resources
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Heterologous expression of the ThIPK2 gene enhances drought resistance of common wheat
ZHANG Shu-juan*, LI Yu-lian*, SONG Guo-qi, GAO Jie, ZHANG Rong-zhi, LI Wei, CHEN Ming-li, LI Gen-ying
Crop Research Institute, Shandong Academy of Agricultural Sciences/Key Laboratory of Wheat Biology and Genetic Improvement on North Yellow and Huai River Valley, Ministry of Agriculture/National Engineering Laboratory for Wheat and Maize, Jinan 250100, P.R.China
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ThIPK2 is an inositol polyphosphate kinase gene cloned from Thellungiella halophila that participates in diverse cellular processes.  Drought is a major limiting factor in wheat (Triticum aestivum L.) production.  The present study investigated whether the application of the ThIPK2 gene could increase the drought resistance of transgenic wheat.  The codon-optimized ThIPK2 gene was transferred into common wheat through Agrobacterium-mediated transformation driven by either a constitutive maize ubiquitin promoter or a stress-inducible rd29A promoter from Arabidopsis.  Molecular characterization confirmed the presence of the foreign gene in the transformed plants.  The transgenic expression of ThIPK2 in wheat led to significantly improve drought tolerance compared to that observed in control plants.  Compared to the wild type (WT) plants, the transgenic plants showed higher seed germination rates, better developed root systems, a higher relative water content (RWC) and total soluble sugar content, and less cell membrane damage under drought stress conditions.  The expression profiles showed different expression patterns with the use of different promoters.  The codon-optimized ThIPK2 gene is a candidate gene to enhance wheat drought stress tolerance by genetic engineering.
Keywords:  codon-optimized        drought        rd29A        ThIPK2        Triticum aestivum L  
Received: 29 December 2018   Accepted:
Fund: This research was supported by the Youth Foundation of Shandong Academy of Agricultural Science, China (2016YQN01), the National Science and Technology Major Project of the Ministry of Agriculture of China (2018ZX08009-10B), the National Natural Science Foundation of China (31601301, 31501312, 31401378), the Natural Science Foundation of Shandong Province, China (ZR2014CM006), the Youth Foundation of Crop Research Institute, Shandong Academy of Agricultural Sciences, China (3201-04), and the Key Research and Development Plan of Shandong Province, China (2017GNC10113).
Corresponding Authors:  Correspondence LI Gen-ying, Tel: +86-531-66658122, E-mail:   
About author:  ZHANG Shu-juan, E-mail:; LI Yu-lian, E-mail:; * These authors contributed equally to this study.

Cite this article: 

ZHANG Shu-juan, LI Yu-lian, SONG Guo-qi, GAO Jie, ZHANG Rong-zhi, LI Wei, CHEN Ming-li, LI Gen-ying. 2020. Heterologous expression of the ThIPK2 gene enhances drought resistance of common wheat. Journal of Integrative Agriculture, 19(4): 941-952.

Budak H, Kantar M, Kurtoglu K Y. 2013. Drought tolerance in modern and wild wheat. Scientific World Journal, 2013, 548246.
Communi D, Vanweyenberg V, Erneux C. 1995. Molecular study and regulation of D-myo-inositol 1,4,5-trisphosphate 3-kinase. Cell Signal, 7, 643–650.
Fan L, Zheng S, Wang X. 1997. Antisense suppression of phospholipase D alpha retards abscisic acid- and ethylene-promoted senescence of postharvest Arabidopsis leaves. The Plant Cell, 9, 2183–2396.
Fleury D, Jefferies S, Kuchel H, Langridge P. 2010. Genetic and genomic tools to improve drought tolerance in wheat. Journal of Experimental Botany, 61, 3211–3222.
Hill T D, Dean N M, Boynton A L. 1988. Inositol 1,3,4,5-tetrakisphosphate induces Ca2+ sequestration in rat liver cells. Science, 242, 1176–1178.
Inan G, Zhang Q, Li P, Wang Z, Cao Z, Zhang H, Zhang C, Quist T M, Goodwin S M, Zhu J, Shi H, Damsz B, Charbaji T, Gong Q, Ma S, Fredricksen M, Galbraith D W, Jenks M A, Rhodes D, Hasegawa P M, et al. 2004. Salt cress. A halophyte and cryophyte Arabidopsis relative model system and its applicability to molecular genetic analyses of growth and development of extremophiles. Plant Physiology, 135, 1718–1737.
Kasuga M, Miura S, Shinozaki K, Yamaguchi-Shinozaki K. 2004. A combination of the Arabidopsis DREB1A gene and stress-inducible rd29A promoter improved drought- and low-temperature stress tolerance in tobacco by gene transfer. Plant and Cell Physiology, 45, 346–350.
Kong X, Zhou S, Yin S, Zhao Z, Han Y, Wang W. 2016. Stress-inducible expression of an F-box gene TaFBA1 from wheat enhanced the drought tolerance in transgenic tobacco plants without impacting growth and development. Frontiers in Plant Science, 7, 1295.
Landi S, Hausman J F, Guerriero G, Esposito S. 2017. Poaceae vs. abiotic stress: Focus on drought and salt stress, recent insights and perspectives. Frontiers in Plant Science, 8, 1214.
Lata C, Muthamilarasan M, Prasad M. 2015. Drought stress responses and signal transduction in plants. In: Pandey G, ed., Elucidation of Abiotic Stress Signaling in Plants. Speinger, New York, NY.
Lawlor D W. 2013. Genetic engineering to improve plant performance under drought: Physiological evaluation of achievements, limitations, and possibilities. Journal of Experimental Botany, 64, 83–108.
Li F, Han Y, Feng Y, Xing S, Zhao M, Chen Y, Wang W. 2013. Expression of wheat expansin driven by the RD29 promoter in tobacco confers water-stress tolerance without impacting growth and development. Journal of Biotechnology, 163, 281–291.
Liu M, Li D M, Wang Z K, Meng F L, Li Y G, Wu X X, Teng W L, Han Y P, Li W B. 2012. Transgenic expression of ThIPK2 gene in soybean improves stress tolerance, oleic acid content and seed size. Plant Cell Tissue and Organ Culture, 111, 277–289.
Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(–Delta Delta C(T)) Method. Method, 25, 402–408.
Lopes M S, Araus J L, van Heerden P D, Foyer C H. 2011. Enhancing drought tolerance in C(4) crops. Journal of Experimental Botany, 62, 3135–3153.
Mignery G A, Johnston P A, Sudhof T C. 1992. Mechanism of Ca2+ inhibition of inositol 1,4,5-trisphosphate (InsP3) binding to the cerebellar InsP3 receptor. Journal of Biological Chemistry, 267, 7450–7455.
Odom A R, Stahlberg A, Wente S R, York J D. 2000. A role for nuclear inositol 1,4,5-trisphosphate kinase in transcriptional control. Science, 287, 2026–2029.
Oztur Z N, Talame V, Deyholos M, Michalowski C B, Galbraith D W, Gozukimizi N, Tuberosa R, Bohnert H J. 2002. Monitoring large-scale changes in transcript abundance in drought- and salt-stressed barley. Plant Molecular Biology, 48, 551–573.
Passioura J. 2007. The drought environment: Physical, biological and agricultural perspectives. Journal of Experimental Botany, 58, 113–117.
Pattni K, Banting G. 2004. Ins(1,4,5)P3 metabolism and the family of IP3-3Kinases. Cell Signal, 16, 643–654.
Pellegrineschi A, Reynolds M, Pacheco M, Brito R M, Almeraya R, Yamaguchi-Schinozaki K, Hoisington D. 2004. Stress-induced expression in wheat of the Arabidopsis thaliana DREB1A gene delays water stress symptoms under greenhouse conditions. Genome, 47, 493–500.
Pennisi E. 2008. Plant genetics. The blue revolution, drop by drop, gene by gene. Science, 320, 171–173.
Perera I Y, Heilmann I, Boss W F. 1999. Transient and sustained increases in inositol 1,4,5-trisphosphate precede the differential growth response in gravistimulated maize pulvini. Proceedings of the National Academy of Scieces of the United States of America, 96, 5838–5843.
Risacher T, Craze M, Bowden S, Paul W, Barsby T. 2009. Highly efficient Agrobacterium-mediated transformation of wheat via in planta inoculation. Methods in Molecular Biology, 478, 115–124.
Schonfeld M A, Johnson R C, Carver B F, Mornhinweg D W. 1988. Water relations in winter wheat as drought resistance indicator. Crop Science, 28, 526–531.
Shi H, Lee B H, Wu S J, Zhu J K. 2003. Overexpression of a plasma membrane Na+/H+ antiporter gene improves salt tolerance in Arabidopsis thaliana. Nature Biotechnology, 21, 81–85.
Stevenson-Paulik J, Bastidas R J, Chiou S T, Frye R A, York J D. 2005. Generation of phytate-free seeds in Arabidopsis through disruption of inositol polyphosphate kinases. Proceedings of the National Academy of Scieces of the United States of America, 102, 12612–12617.
Sun S, Zhou J. 2017. Molecular mechanisms underlying stress response and adaptation. Thoracic Cancer, 9, 218–227.
Volkov V, Amtmann A. 2006. Thellungiella halophila, a salt-tolerant relative of Arabidopsis thaliana, has specific root ion-channel features supporting K+/Na+ homeostasis under salinity stress. The Plant Journal, 48, 342–353.
Wang P, Yang Q, Sang S, Chen Y, Zhong Y, Wei Z. 2017. Arabidopsis inositol polyphosphate kinase AtIpk2beta is phosphorylated by CPK4 and positively modulates ABA signaling. Biochemical and Biophysical Research Communications, 490, 441–446.
Xia H J, Brearley C, Elge S, Kaplan B, Fromm H, Mueller-Roeber B. 2003. Arabidopsis inositol polyphosphate 6-/3-kinase is a nuclear protein that complements a yeast mutant lacking a functional ArgR-Mcm1 transcription complex. The Plant Cell, 15, 449–463.
Xiong L, Schumaker K S, Zhu J K. 2002. Cell signaling during cold, drought, and salt stress. The Plant Cell, 14(Suppl.), S165–S183.
Xu J, Brearley C A, Lin W H, Wang Y, Ye R, Mueller-Roeber B, Xu Z H, Xue H W. 2005. A role of Arabidopsis inositol polyphosphate kinase, AtIPK2 alpha, in pollen germination and root growth. Plant Physiology, 137, 94–103.
Yamaguchi-Shinozaki K, Shinozaki K. 1993. Characterization of the expression of a desiccation-responsive rd29 gene of Arabidopsis thaliana and analysis of its promoter in transgenic plants. Molecular and General Genetics, 236, 331–340.
Yamaguchi-Shinozaki K, Shinozaki K. 2006. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Annual Reveiw of Plant Biology, 57, 781–803.
Yang L, Tang R, Zhu J, Liu H, Mueller-Roeber B, Xia H, Zhang H. 2008. Enhancement of stress tolerance in transgenic tobacco plants constitutively expressing AtIpk2 beta, an inositol polyphosphate 6-/3-kinase from Arabidopsis thaliana. Plant Molecualr Biology, 66, 329–343.
Yang Q, Sang S, Chen Y, Wei Z, Wang P. 2018. The role of Arabidopsis inositol polyphosphate kinase AtIPK2 beta in glucose suppression of seed germination and seedling development. Plant and Cell Physiology, 59, 343–354.
Yemm E W, Willis A J. 1954. The estimation of carbohydrates in plant extracts by the anthrone. Biochemical Journal, 57, 508–514.
Zhang S J, Song G Q, Li Y L, Gao J, Liu J J, Fan Q Q, Huang C Y, Sui X X, Chu X S, Guo D, Li G Y. 2014. Cloning of a 9-cis-epoxycarotenoid dioxygenase gene (TaNCED1) in common wheat and its expression in response to abiotic stresses. Biologia Plantarum, 58, 89–98.
Zhu J Q, Zhang J T, Tang R J, Lv Q D, Wang Q Q, Yang L, Zhang H X. 2009. Molecular characterization of ThIPK2, an inositol polyphosphate kinase gene homolog from Thellungiella halophila, and its heterologous expression to improve abiotic stress tolerance in Brassica napus. Physiologia Plantarum, 136, 407–425.
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