Opposing Control by Transcription Factors MYB61 and MYB3 Increases Freezing Tolerance by Relieving C-Repeat Binding Factor Suppression.

Cold acclimation is an important process by which plants respond to low temperature and enhance their winter hardiness. C-REPEAT BINDING FACTOR1 (CBF1), CBF2, and CBF3 genes were shown previously to participate in cold acclimation in Medicago truncatula In addition, MtCBF4 is transcriptionally induced by salt, drought, and cold stresses. We show here that MtCBF4, shown previously to enhance drought and salt tolerance, also positively regulates cold acclimation and freezing tolerance. To identify molecular factors acting upstream and downstream of the MtCBF4 transcription factor (TF) in cold responses, we first identified genes that are differentially regulated upon MtCBF4 overexpression using RNAseq Digital Gene Expression Profiling. Among these, we showed that MtCBF4 directly activates the transcription of the COLD ACCLIMATION SPECIFIC15 (MtCAS15) gene. To gain insights into how MtCBF4 is transcriptionally regulated in response to cold, an R2R3-MYB TF, MtMYB3, was identified based on a yeast one-hybrid screen as binding directly to MYB cis-elements in the MtCBF4 promoter, leading to the inhibition of MtCBF4 expression. In addition, another MYB TF, MtMYB61, identified as an interactor of MtMYB3, can relieve the inhibitory effect of MtMYB3 on MtCBF4 transcription. This study, therefore, supports a model describing how MtCBF4 is regulated by antagonistic MtMYB3/MtMYB61 TFs, leading to the up-regulation of downstream targets such as MtCAS15 acting in cold acclimation in M. truncatula.

Zhang Z ，Hu X ，Zhang Y ，Miao Z ，Xie C ，Meng X ，Deng J ，Wen J ，Mysore KS ，Frugier F ，Wang T ，Dong J ... -  《-》

Transcriptional profiling of Medicago truncatula under salt stress identified a novel CBF transcription factor MtCBF4 that plays an important role in abiotic stress responses.

Li D ，Zhang Y ，Hu X ，Shen X ，Ma L ，Su Z ，Wang T ，Dong J ... -  《BMC PLANT BIOLOGY》

Cold acclimation-induced freezing tolerance of Medicago truncatula seedlings is negatively regulated by ethylene.

To evaluate the role of ethylene in cold acclimation and cold stress, freezing tolerance and characteristics associated with cold acclimation were investigated using legume model plant Medicago truncatula Gaertn Jemalong A17. There was a rapid suppression of ethylene production during cold acclimation in A17 plants. Ethylene level was negatively correlated with freezing tolerance as inhibition of ethylene biosynthesis by inhibitors of ethylene biosynthesis enhanced freezing tolerance, while exogenous application of ethylene reduced cold acclimation-induced freezing tolerance. The involvement of ethylene signaling in modulation of freezing tolerance and cold acclimation was further studied using ethylene-insensitive mutant sickle skl. Although skl mutant was more tolerant to freezing than its wild-type counterpart A17 plants, cold acclimation enhanced freezing tolerance in 17 plants, but not in skl mutant. Expression of several ethylene response genes including EIN3, EIN3/EIL and ERFs was suppressed in skl mutant compared to A17 plants under non-cold-acclimated conditions. Cold acclimation downregulated expression of EIN3, EIN3/EIL and ERFs in A17 plants, while expression patterns of these genes were relatively constant in skl mutant during cold acclimation. Cold acclimation-induced increases in transcription of MtCBFs and MtCAS15 were suppressed in skl mutant compared with A17 plants. These results suggest that MtSKL1 is required for perception of the change of ethylene level in M. truncatula plants for the full development of the cold acclimation response by suppressing expression of MtEIN3 and MtEIN3/EIL1, which in turn downregulates expression of MtERFs, leading to the enhanced tolerance of M. truncatula to freezing by upregulating MtCBFs and MtCAS15.

Zhao M ，Liu W ，Xia X ，Wang T ，Zhang WH ... -  《-》

Comparative studies on tolerance of Medicago truncatula and Medicago falcata to freezing.

Zhang LL ，Zhao MG ，Tian QY ，Zhang WH ... -  《-》

Comparative genomic sequence and expression analyses of Medicago truncatula and alfalfa subspecies falcata COLD-ACCLIMATION-SPECIFIC genes.

Pennycooke JC ，Cheng H ，Stockinger EJ 《-》