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Bulletin of Botanical Research ›› 2016, Vol. 36 ›› Issue (5): 721-729.doi: 10.7525/j.issn.1673-5102.2016.05.013

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Overexpression of DmDREBa Gene Significantly Enhances Low Temperature Tolerance in Transgenic Tobacco

YANG Yan-Fang1, WU Jian2, ZHU Kai2, LIU Li-Qing4, CHEN Fa-Di3, YU De-Yue2   

  1. 1. Research Institute of Forestry, Chinese Academy of Forestry, State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of State Forestry Administration, Beijing 100091;
    2. National Center for Soybean Improvement, National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095;
    3. College of Horticulture, Nanjing Agricultural University, Nanjing 210095;
    4. Fujian Provincial Key Lab of Subtropic Plant Physiology and Biochemistry, Fujian Institute of Subtropical Botany, Xiamen 361009
  • Received:2016-04-19 Online:2016-09-15 Published:2016-09-27
  • Supported by:
    Partly supported by National Natural Science Foundation of China(31300567);National non-profit Research Institutions of Chinese Academy of Forestry(CAFYBB2014QB001)

Abstract: Many environmental factors such as drought, salt stress, hormones and low/high temperature influence the growth and development of plants. It has been found that DREB transcription factors play important roles in plants against different abiotic stresses. In this study, DmDREBa(GenBank accession No.EF490996), a DREB-like gene isolated from chrysanthemum was overexpressed in tobacco plants by mediation of Agrobacterium tumefaciens under the control of the constitutive promoter CaMV 35S, and 43 transgenic tobacco plants were obtained by PCR method. 9 transgenic tobacco plants were randomly selected to perform RT-PCR investigation and 7 plants indicated RNA transcripts accumulation. The southern blot result showed that 1-3 copies of DmDREBa were randomly inserted. The tolerance of transgenic tobacco plants was increased comparing with the wild type tobacco plants under low temperature condition. The water loss of transgenic tobacco was less than wild type plants after natural transpiration. Moreover, the density of the stomata in the epidermis was observed. The result showed that the numbers of stomata in upper or lower epidermis of transgenic plants were reduced comparing to wild type tobacco. The result of t test demonstrated that the differences between the upper and lower epidermis of transgenic plants and wild type were significant. Furthermore, the content of malondialdehyde(MDA) in transgenic plants was lower than that in wild type tobacco plants at lower temperature condition, which indicated the tolerance ability of transgenic plants was increased. All these results indicated that DmDREBa improved tolerance ability of trangenic tobacco plants to the lower temperature stress. This research will provide a very useful reference for functional analysis of DREB genes in chrysanthemum in the future.

Key words: chrysanthemum, DREB, Agrobacterium-mediated transformation, transgenic tobacco(Nicotiana tabacum ‘Xantbine’), tolerance

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