Elucidating the role of a basic helix loop helix (bHLH) transcription factor (TF) during bud break in hybrid aspen

In extreme weather conditions, perennial trees have developed mechanisms that help them to survive these environmental cues. Temperature and day length regulates the bud burst in Aspen in the spring. The molecular basis of basic helix loop helix (bHLH) transcription factors mediated control of bud break is poorly understood. Here we identify and elucidate a transcription factor involved in the control of bud break in hybrid aspen. The transcription factor T89CIB is closely related to Arabidopsis Floral inducer AtCIB1 acting, activating, and regulated in a similar manner as we can infer until now. To understand where is localized, how it is regulated, and the effect on bud break. Two constructs 35S: T89CIB and 35S: AtCIB1 were made, and subcellular localization of the transcription factors was performed by Confocal Laser Scanning Microscopy. Furthermore, to explain the regulation of the proteins, Nicotiana benthamiana plants were infiltrated and exposed under dark and blue light conditions, finally, COL-O and cry2 lines were transformed with T89 CIB using Floral dip as a transformation method in Arabidopsis Thaliana. We elucidate the nuclear localization and demonstrate the role of blue light and dark as positive and negative regulators respectively. And the possible interaction and phenotype of T89CIB and Cryptochrome 2 (CRY2). Thus, our results reveal the basis of the T89CIB transcription factor and support the idea of the importance of this protein as part of the pathways activated during bud break in Hybrid Aspen. Elucidating these pathways will give a better understanding of the survival of Perennial trees under harsh conditions.