Enhanced Oxidative Stress Tolerance of Transgenic Tobacco Plants by Overexpression of Brachypodium distachyon BdDREB1-like Gene

doi:10.7668/hbnxb.20190485

Abstract

Abstract: In order to explore the function of BdDREB1-like gene in transgenic tobacco, it was cloned and analyzed through bioinformatics technology. The open reading frame of this gene was 684 bp, encoded 228 amino acids, and the molecular weight of its encoding protein was 24.178 ku. It contained only one typical AP2 domain and belonged to the DREB family of AP2/EREBP superfamily. Multiple sequence alignment and evolutionary analysis showed that it was closely related to Rye ScCBFI. Subcellular localization analysis showed that BdDREB1-like was located in the nucleus. It was then constructed into plant expression vector and transformed into tobacco by Agrobacterium-mediated method to further confirm its function. PCR and RT-PCR screening showed that BdDREB1-like gene was not only integrated into tobacco genome, but also expressed stably. Transgenic tobacco seedlings with BdDREB1-like gene had higher germination rate under hydrogen peroxide stress. In order to further determine the relationship between BdDREB1-like gene and oxidative stress, we treated the one-month-old transgenic tobacco plants under oxidative stress with MV (Methyl viologen). The results showed that there were no significant differences in the contents of hydrogen peroxide, catalase and other physiological indicators except for SOD between wild type and T3 transgenic line, while there were only two physiological indexes of POD and chloroplast content of T1 transgenic line, which were significantly different from those of wild type under normal growth condition. Under oxidative stress, transgenic tobacco plants with BdDREB1-like gene had lower malondialdehyde and hydrogen peroxide content, higher peroxidase activity, catalase, superoxide dismutase and chlorophyll content compared with wild type. These results suggested that overexpression of BdDREB1-like gene enhanced the tolerance of plant to oxidative stress.

Key words: Brachypodium distachyon, BdDREB1-like, Expression vector construction, Genetic transformation

CLC Number:

S572.01

LI Lun, KE Jie, HUANG Gang, LIU Hongshi, SONG Yanni, ZHANG Yingjie, CHEN Lihong. Enhanced Oxidative Stress Tolerance of Transgenic Tobacco Plants by Overexpression of Brachypodium distachyon BdDREB1-like Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(2): 1-8. doi: 10.7668/hbnxb.20190485.

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