ORIGINAL PAPER A Gene Encoding Cold-Circadian Rhythm-RNA Binding-Like Protein (CCR-Like) from Upland Cotton (Gossypium hirsutum L.) Confers Tolerance to Abiotic Stresses in Transgenic Tobacco Gurusamy Dhandapani & Mogilicherla Kanakachari & Kethireddy Venkata Padmalatha & Mullapudi Lakshmi Venkata Phanindra & Vivek Kumar Singh & Sanagala Raghavendrarao & Narayanasamy Jayabalan & Azhagiyamanavalan Lakshmi Prabha & Polumetla Ananda Kumar # Springer Science+Business Media New York 2014 Abstract Water-deficit is the major abiotic stress factor that limits the yield and quality of cotton produced around the world. We observed earlier that a CCL (CCR-like; cold- circadian rhythm-RNA binding like) gene has been differen- tially expressed during boll development in cotton under drought stress in the field. Isolation and functional character- ization of GhCCL from upland cotton (Gossypium hirsutum L. cv. Bikaneri Narma) was carried out in the present study. We studied the GhCCL gene structure and organization and demonstrated for the first time that GhCCL may be involved in abiotic stress tolerance response in plants. RT-PCR analysis indicated that GhCCL is differentially regulated in cotton seedlings by abiotic stresses such as salt, mannitol, cold, heat, dehydration, wounding and jasmonic acid, salicylic acid, and hydrogen peroxide. In silico and subcellular localization anal- ysis suggested that GhCCL is localized in the chloroplasts. Constitutive expression of GhCCL in tobacco (Nicotiana tabacum var. Petit Havana) conferred tolerance to water- deficit stress and salt stresses during seed germination on amended MS media. The transgenic plants showed better growth performance and increased fresh weight under long- term stress. The transgenic plants grown in the glass house tolerated water-deficit stress (by withholding water) and re- covered upon rewatering (45 days) whereas WT plants did not survive. Characterization of the GhCCL promoter sequence by in silico analysis showed various light, stress, tissue spe- cific, and hormone responsive cis-elements. The present study suggested that GhCCL positively regulates the response to abiotic stresses, especially water-deficit stress in transgenic plants and that the overexpression of GhCCL may enhance the stability of mRNA thereby conferring tolerance to abiotic stresses. Keywords Cold-circadianrhythm-RNAbinding . Gossypium hirsutum . Water-deficit stress . Transgenic tobacco Introduction Cotton is the major source of natural fibers used in the textile industry and is cultivated across the globe (Smith and Cothren 1999). Among the four cultivated species, Gossypium arboreum and G. herbaceum are diploids with A genome and G. hirsutum and G. barbadense are allotetraploid species with AD genome. G. hirsutum represents more than 95 % of the cultivated cotton worldwide (National Cotton Council, 2012, http:/www.cotton.org/econ/cropinfo/index.cfm). Cotton fiber development includes four distinct, but overlapping stages: fiber cell initiation, elongation/primary Electronic supplementary material The online version of this article (doi:10.1007/s11105-014-0729-x) contains supplementary material, which is available to authorized users. G. Dhandapani : M. Kanakachari : K. V. Padmalatha : M. L. V. Phanindra : V. K. Singh : S. Raghavendrarao : P. A. Kumar (*) National Research Centre on Plant Biotechnology, LBS Building, Pusa Campus, New Delhi 110012, India e-mail: polumetla@hotmail.com G. Dhandapani : M. Kanakachari : N. Jayabalan : A. Lakshmi Prabha Department of Plant Science, Bharathidasan University, Tiruchirappalli, Tamil Nadu, India Present Address: P. A. Kumar Institute of Biotechnology, Acharya NG Ranga Agricultural University, Rajendranagar, Hyderabad 500030, Andhra Pradesh, India Plant Mol Biol Rep DOI 10.1007/s11105-014-0729-x