Drought is a major environmental factor that limits crop growth and productivity. Flue-cured tobacco (Nicotiana tabacum) is one of the most important commercial crops worldwide and its productivity is vulnerable to drought. However, comparative analyses of physiological, biochemical and gene expression changes in flue-cured tobacco varieties differing in drought tolerance under long-term drought stress are scarce. In this study, drought stress responses of two flue-cured tobacco varieties, LJ851 and JX6007, were comparatively studied at the physiological and transcriptional levels. After exposing to progressive drought stress, the drought-tolerant LJ851 showed less growth inhibition and chlorophyll reduction than the drought-sensitive JX6007. Moreover, higher antioxidant enzyme activities and lower levels of H2O2, Malondialdehyde (MDA), and electrolyte leakage after drought stress were found in LJ851 when compared with JX6007. Further analysis showed that LJ851 plants had much less reductions than the JX6007 in the net photosynthesis rate and stomatal conductance during drought stress; indicating that LJ851 had better photosynthetic performance than JX6007 during drought. In addition, transcriptional expression analysis revealed that LJ851 exhibited significantly increased transcripts of several categories of drought-responsive genes in leaves and roots under drought conditions. Together, these results indicated that LJ851 was more drought-tolerant than JX6007 as evidenced by better photosynthetic performance, more powerful antioxidant system, and higher expression of stress defense genes during drought stress. This study will be valuable for the development of novel flue-cured tobacco varieties with improved drought tolerance by exploitation of natural genetic variations in the future.
Drought is one of the most important limiting factors for plant growth and development. To identify genes required for drought stress response in tobacco, one highly induced mRNA encoding a RING-H2 Finger gene (RHF1) was isolated by mRNA differential display. The full-length NtRHF1 encodes a protein of 273 amino acids and contains a single C3H2C3-type RING motif in its C-terminal region. NtRHF1 is an ortholog of Arabidopsis SDIR1 (salt- and drought-induced RING finger 1) (73 % identity to AtSDIR1). The recombinant NtRHF1 protein purified from E. coli exhibited an in vitro E3 ubiquitin ligase activity. Real-time quantitative PCR analysis indicated that the transcript levels of NtRHF1 were higher in aerial tissues and were markedly up-regulated by drought stress. Overexpression of NtRHF1 enhanced drought tolerance in transgenic tobacco plants while RNA silencing of NtRHF1 reduced drought tolerance. Further expression analysis by real-time PCR indicated that NtRHF1 participates in drought stress response possibly through transcriptional regulation of downstream stress-responsive genes NtLEA5, NtERD10C, NtAREB, and NtCDPK2 in tobacco. Together, these results demonstrated that NtRHF1 plays a positive role in drought stress tolerance possibly through transcriptional regulation of several stress-responsive marker genes in tobacco. This study will facilitate to improve our understanding of molecular and functional properties of plant RING-H2 finger proteins and to provide genetic evidence on the involvement of the RING-H2 E3 ligase in drought stress response in Nicotiana tabacum plants.
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