Loss of GLK1 transcription factor function reveals new insights in chlorophyll biosynthesis and chloroplast development

Birch (Betula platyphylla x B. pendula) is an important tree for landscaping due to its attractive white bark and straight trunk. In this study, we characterized a T-DNA yellow-green leaf mutant, yl. We identified six insertion sites (ISs) in the mutant by genome resequencing and found a 40-kb deletion containing BpGLK1 around IS2 on chromosome 2. Complementation experiments with the yl mutant and repression of BpGLK1 in wild-type plants confirmed that BpGLK1 was responsible for the mutated phenotype. Physiological and ultrastructural analyses showed that the leaves of the yl mutant and BpGLK1-repression lines had decreased chlorophyll content and defective chloroplast development compared to the wild-type. Furthermore, the loss function of BpGLK1 also affected photosynthesis in leaves. Transcriptomics, proteomics, and ChIP-PCR analysis revealed that BpGLK1 directly interacted with the promoter of genes related to antenna proteins, chlorophyll biosynthesis, and photosystem subunit synthesis, and regulated their expression. Overall, our research not only provides new insights into the mechanism of chloroplast development and chlorophyll biosynthesis regulated by BpGLK1, but also provides new transgenic birch varieties with various levels of yellowing leaves by repressing BpGLK1 expression.