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Journal of Integrative Agriculture  2018, Vol. 17 Issue (10): 2204-2214    DOI: 10.1016/S2095-3119(17)61897-5
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Overexpression of AmDUF1517 enhanced tolerance to salinity, drought, and cold stress in transgenic cotton
HAO Yu-qiong*, LU Guo-qing*, WANG Li-hua, WANG Chun-ling, GUO Hui-ming, LI Yi-fei, CHENG Hong-mei
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
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Abstract  As abiotic stresses become more severe as a result of global climate changes, the growth and development of plants are restricted. In the development of agricultural crops with greater stress tolerance, AmDUF1517 had been isolated from the highly stress-tolerant shrub Ammopiptanthus mongolicus, and can significantly enhance stress tolerance when inserted in Arabidopsis thaliana. In this study, we inserted this gene into cotton to analyze its potential for conferring stress tolerance. Two independent transgenic cotton lines were used. Southern blot analyses indicated that AmDUF1517 was integrated into the cotton genome. Physiological analysis demonstrated that AmDUF1517-transgenic cotton had stronger resistance than the control when treated with salt, drought, and cold stresses. Further analysis showed that trans-AmDUF1517 cotton displayed significantly higher antioxidant enzyme (superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and glutathione S-transferase (GST)) activity and less reactive oxygen species (ROS) accumulation, which suggests that overexpression of AmDUF1517 can improve cotton resistance to stress by maintaining ROS homeostasis, as well as by alleviating cell membrane injury. These results imply that AmDUF1517 is a candidate gene in improving cotton resistance to abiotic stress. 
Keywords:  transgenic cotton        stress tolerance        AmDUF1517        Ammopiptanthus mongolicus        reactive oxygen species  
Received: 27 October 2017   Accepted:
Fund: This work was financially supported by the Key Project for Breeding Genetic Modified Organisms, China (2016ZX08005004) and the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences.
Corresponding Authors:  Correspondence CHENG Hong-mei, Tel/Fax: +86-10-82106125,   
About author:  HAO Yu-qiong, E-mail:; * These authors contributed equally to this study.

Cite this article: 

HAO Yu-qiong, LU Guo-qing, WANG Li-hua, WANG Chun-ling, GUO Hui-ming, LI Yi-fei, CHENG Hong-mei. 2018. Overexpression of AmDUF1517 enhanced tolerance to salinity, drought, and cold stress in transgenic cotton. Journal of Integrative Agriculture, 17(10): 2204-2214.

Abrahám E, Hourton-Cabassa C, Erdei L, Szabados L. 2010. Methods for determination of proline in plants. Methods in Molecular Biology, 639, 317–331.
Ahuja I, de Vos R C, Bones A M, Hall R D. 2010. Plant molecular stress responses face climate change. Trends in Plant Science, 15, 664–674.
Arnon D I. 1949. Copper enzymes in isolated chloroplasts.polyphenoloxidase in Beta vulgaris. Plant Physiology, 24, 1.
Ashraf M. 2002. Salt tolerance of cotton: Some new advances. Critical Reviews in Plant Sciences, 21, 1–30.
Ashraf M, Foolad M R. 2007. Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environmental and Experimental Botany, 59, 206–216.
Azarabadi S, Abdollahi H, Torabi M, Salehi Z, Nasiri J. 2017. ROS generation, oxidative burst and dynamic expression profiles of ROS-scavenging enzymes of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) in response to Erwinia amylovora in pear (Pyrus communis L.). European Journal of Plant Pathology, 147, 279–294.
Bouaziz D, Pirrello J, Ben A H, Hammami A, Charfeddine M, Dhieb A, Bouzayen M, Gargouri-Bouzid R. 2012. Ectopic expression of dehydration responsive element binding proteins (StDREB2) confers higher tolerance to salt stress in potato. Plant Physiology and Biochemistry, 60, 98–108
Cao X, Yang K Z, Xia C, Zhang X Q, Chen L Q, Ye D. 2010. Characterization of DUF724 gene family in Arabidopsis thaliana. Plant Molecular Biology, 72, 61–73.
Chen J, Wan S, Liu H, Fan S, Zhang Y, Wang W, Xia M, Yuan R, Deng F, Shen F. 2016. Overexpression of an Apocynum venetum DEAD-Box helicase gene (AvDH1) in cotton confers salinity tolerance and increases yield in a saline field. Frontiers in Plant Science, 6, 1227.
Clarke J D. 2009. Cetyltrimethyl ammonium bromide (CTAB) DNA miniprep for plant DNA isolation. Cold Spring Harbor Protocols, 3, doi: 10.1101/pdb.prot5177
Ding L Y, Zhang W, Wang J C, Tian L L, LI N N, Guo Q, Yang S M, He M L, Guo W Z. 2014. Overexpression of a Gossypium hirsutum stress-associated protein gene (GhSAP1) improves salt stress tolerance in transgenic tobacco. Scientia Agricultura Sinica, 47, 1458–1470. (in Chinese)
Divya K, Kirti P B. 2008. Expression of a mustard annexin confers abiotic stress tolerance and improved fiber quality in transgenic cotton. Journal of Biotechnology, 136, S236–S237.
Esim N, Atici O, Mutlu S. 2014. Effects of exogenous nitric oxide in wheat seedlings under chilling stress. Toxicology and Industrial Health, 30, 268–274.
Fujimoto S Y, Ohta M, Usui A, Shinshi H, Ohme-Takagi M. 2000. Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box-mediated gene expression. The Plant Cell, 12, 393–404.
Gebbie L. 2014. Genomic Southern blot analysis. Methods in Molecular Biology, 1099, 159–177.
Giannopolitis C N, Ries S K. 1977. Superoxide dismutases. 1. Occurrence in higher plants [Corn, oats, peas]. Plant Physiology, 59, 309–314.
Gil R, Lull C, Boscaiu M, Bautista I, Vicente O. 2011. Soluble carbohydrates as osmolytes in several halophytes from a Mediterranean salt marsh. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 39, 9–17.
Gilmour S J, Thomashow M F. 2000. Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation. Plant Physiology, 124, 1854–1865.
González L, González-Vilar M. 2001. Determination of relative water content. Handbook of Plant Ecophysiology Techniques. Springer, The Netherlands. University of Vigo, Spain. pp. 207–212.
Greenway A, Munns R. 1980. Mechanisms of salt tolerance in nonhalophytes. Annual Review of Plant Biology, 31, 61–69.
Gu L, Cheng H. 2014. Isolation, molecular cloning and characterization of a cold-responsive gene, AmDUF1517, from Ammopiptanthus mongolicus. Plant Cell Tissue and Organ Culture, 117, 201–211.
Guo L, Yu Y H, Xia X L, Yin W L. 2010. Identification and functional characterisation of the promoter of the calcium sensor gene CBL1 from the xerophyte Ammopiptanthus mongolicus. BMC Plant Biology, 10, 18.
Hansen S F, Harholt J, Oikawa A, Scheller H V. 2012. Plant glycosyltransferases beyond CAZy. Frontiers in Plant Science, 3, 379–411.
Hu X W, Zhang R, Min D D, Fan S G, Wang Y R, Baskin J M. 2016. Factors restricting seed germination and seedling recruitment of Ammopiptanthus mongolicus: An evergreen shrub endemic to cold deserts in China. International Seed Testing Association, 44, 1–14.
Jambunathan N. 2010. Determination and detection of reactive oxygen species (ROS), lipid peroxidation, and electrolyte leakage in plants. Methods in Molecular Biology, 639, 291–297.
Jiang Z, Zhu S, Ye R, Xue Y, Chen A, An L, Pei Z M. 2013. Relationship between NaCl- and H2O2-induced cytosolic Ca2+ increases in response to stress in Arabidopsis. PLoS ONE, 8, e76130.
Jin L G, Li H, Liu J Y. 2010. Molecular characterization of three ethylene responsive element binding factor genes from cotton. Journal of Integrative Plant Biology, 52, 485–495.
Jonesrhoades M W, Borevitz J O, Preuss D. 2007. Genome-wide expression profiling of the Arabidopsis female gametophyte identifies families of small, secreted proteins. PLoS Genetics, 3, e171.
Kizis D, Lumbreras V, Pagès M. 2001. Role of AP2/EREBP transcription factors in gene regulation during abiotic stress. FEBS Letters, 498, 187–189.
Kurepa J, Smalle J, Van M M, Inzé D. 1998. Oxidative stress tolerance and longevity in Arabidopsis: The late-flowering mutant gigantea is tolerant to paraquat. Plant Journal for Cell & Molecular Biology, 14, 759–764.
Lee J A. 1984. Cotton as a world crop. Agronomy Monograph-American Society of Agronomy, 24, 1–25.
Li X, Wang X D, Zhao X, Dutt Y. 2004. Improvement of cotton fiber quality by transforming the acsA and acsB genes into Gossypium hirsutum L. by means of vacuum infiltration. Plant Cell Reports, 22, 691–697.
Li X X, Li J Z. 2013. Determination of the content of soluble sugar in sweet corn with optimized anthrone colorimetric method. Storage & Process, doi: 10.3969/j.issn.1009-6221. 2013.04.006
Liu M, Shi J, Lu C. 2013. Identification of stress-responsive genes in Ammopiptanthus mongolicus using ESTs generated from cold- and drought-stressed seedlings. BMC Plant Biology, 13, 88.
Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔCt method. Methods, 25, 402–408.
Meng C, Zhang T, Guo W. 2009. Characterization of six novel NAC genes and their responses to abiotic stresses in Gossypium hirsutum L. Plant Science, 176, 352–359.
Miller G, Suzuki N, Ciftci-Yilmaz S, Mittler R. 2010. Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant, Cell & Environment, 33, 453–467.
Neill S J, Desikan R, Clarke A, Hurst R D, Hancock J T. 2002. Hydrogen peroxide and nitric oxide as signalling molecules in plants. Journal of Experimental Botany, 53, 1237–1247.
Ogawa A, Yamauchi A. 2006. Root osmotic adjustment under osmotic stress in maize seedlings: 1. Transient change of growth and water relations in roots in response to osmotic stress. Plant Production Science, 9, 27–38.
Ohmetakagi M, Shinshi H. 1995. Ethylene-inducible DNA binding proteins that interact with an ethylene-responsive element. The Plant Cell, 7, 173–182.
Ohta M, Ohmetakagi M, Shinshi H. 2000. Three ethylene-responsive transcription factors in tobacco with distinct transactivation functions. The Plant Journal, 22, 29–38.
Schmedes A, Hølmer G. 1989. A new thiobarbituric acid (TBA) method for determining free malondialdehyde (MDA) and hydroperoxides selectively as a measure of lipid peroxidation. Journal of the American Oil Chemists Society, 66, 813–817.
Shah S T, Pang C, Fan S, Song M, Arain S, Yu S. 2013. Isolation and expression profiling of GhNAC transcription factor genes in cotton (Gossypium hirsutum L.) during leaf senescence and in response to stresses. Gene, 531, 220–234.
Shan D P. 2004. Cloning and functional analysis of three stressinduced genes (GhDREB1A/1B, GhBADH ) in cotton. MSc thesis, Shandong Agricultural University, Tai’an. (in Chinese)
Shan D P, Huang J G, Yang Y T, Guo Y H, Wu C A, Yang G D, Gao Z, Zheng C C. 2007. Cotton GhDREB1 increases plant tolerance to low temperature and is negatively regulated by gibberellic acid. The New Phytologist, 176, 70–81.
Shangguan X X, Wu X, Liang Y S, Zhang L S, Li Y E. 2008. Improvement of cotton (Gossypium hirsutum L.) fiber quality by rabbit keratin gene transformation. Chinese Journal of Eco-Agriculture, 16, 451–454. (in Chinese)
Trujillo L E, Borras Hidalgo O, Sotolongo M, Coll Y, Hernandez L, Thomma B P H J, Ochogavia M E, Menendez C, Vera P, Hernandez I. 2008. SodERF3, a novel sugarcane [Saccharum officinarum] ethylene responsive factor (ERF), enhances salt and drought tolerance when overexpressed in tobacco [Nicotiana tabacum] plants. Plant & Cell Physiology, 49, 512–525.
Wang C, Lu G, Hao Y, Guo H, Guo Y, Zhao J, Cheng H. 2017. ABP9, a maize bZIP transcription factor, enhances tolerance to salt and drought in transgenic cotton. Planta, 6, 1–17.
Wang Q, Guan Y, Wu Y, Chen H, Chen F, Chu C. 2008. Over expression of a rice OsDREB1F gene increases salt, drought, and low temperature tolerance in both Arabidopsis and rice. Plant Molecular Biology, 67, 589–602.
Wang Z, Li J, Jia C, Xu B, Jin Z. 2016. Molecular cloning and expression analysis of eight calcium-dependent protein kinase (CDPK) genes from banana (Musa acuminata L. AAA group, cv. Cavendish). South African Journal of Botany, 104, 134–141.
Yue Y, Zhang M, Zhang J, Tian X, Duan L, Li Z. 2012. Overexpression of the AtLOS5 gene increased abscisic acid level and drought tolerance in transgenic cotton. Journal of Experimental Botany, 63, 3741–3748.
Zeng D E, Hou P, Xiao F, Liu Y. 2015. Overexpression of Arabidopsis XERICO gene confers enhanced drought and salt stress tolerance in rice (Oryza sativa L.). Journal of Plant Biochemistry and Biotechnology, 24, 56–64.
Zhang B. 2013. Agrobacterium-mediated transformation of cotton. Methods in Molecular Biology, 958, 31–45.
Zhou L, Wang N N, Gong S Y, Lu R, Li Y, Li X B. 2015. Overexpression of a cotton (Gossypium hirsutum) WRKY gene, GhWRKY34, in Arabidopsis enhances salt-tolerance of the transgenic plants. Plant Physiology and Biochemistry, 96, 311–320.
Zong J M, Li X W, Zhou Y H, Wang F W, Wang N, Dong Y Y, Yuan Y X, Chen H, Liu X M, Yao N. 2016. The AaDREB1 transcription factor from the cold-tolerant plant adonis amurensis enhances abiotic stress tolerance in transgenic plant. International Journal of Molecular Sciences, 17, 611.
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