Professor Gao Hongbo's team from the College of Biological Science and Biotechnology has recently made a significant breakthrough in the field of gene regulation analysis. Their research paper, entitled "Quantitative RUBY Reporter Assay for Gene Regulation Analysis," was published in the prestigious journal Plant, Cell & Environment. Doctoral students Liu Jia and Li Hao, also from the College of Biological Science and Biotechnology, are the co-first authors of this important study. Professor Gao Hongbo serves as the corresponding author, with Beijing Forestry University proudly associated as the affiliation of the first author. This groundbreaking research opens new avenues for understanding gene regulation mechanisms, potentially leading to advancements in related fields.

The regulation of gene expression is a process of activating or inhibiting gene's expression to synthesize mRNA and proteins, and it plays an important role in an organism's normal development, routine cellular functions, and response to environment. This process is involved in the response to various environmental factors, such as light, temperature, and diseases, which is essential for plant's adaption to the environment. During the process, transcription factors (TFs) respond to the upstream signals and bind to the specific elements in the promoter regions, to regulate the expression of downstream genes.

Various reporter genes have been developed to study gene expression pattern and gene regulation. The RUBY reporter gene was recently developed and widely used, because of its visible and noninvasive advantages. However, quantitative analysis of RUBY gene expression levels was lacking. In this study, a novel betalain quantification method in combination with the tobacco transient expression system is introduced. The betalain produced in tobacco leaves was extracted and purified, and its concentration was quantitatively measured. This approach in studying the transcriptional regulation of ARC5 gene is successfully applied by transcription factors CPD25 and CPD45. Furthermore, with this method, is shows that the gene expression of RCA and Rbcs1A gene were regulated by light, transcription factors HY5 and PIFs through G-box and I-box elements. The development of this betalain quantification approach with the tobacco transient expression system offers a cost-effective and intuitive strategy for studying the regulatory mechanism of gene expression.

This work was supported by the National Natural Science Foundation of China (Grant No. 32070696).

Paper link: https://doi.org/10.1111/pce.14947

Written by Liu Jia, Gao Hongbo
Translated and edited by Song He
Reviewed by Yu Yangyang