J Integr Plant Biol

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  • 收稿日期:2025-03-20 接受日期:2025-05-29

The horizontally transferred gene, CsMTAN, rewired purine traffic to build caffeine factories in tea leaves

Xinxin Jia1†, Xiaoliang Zhang1†, Xueli Chen1, Alisdair R. Fernie2 and Weiwei Wen1*   

  1. 1. National Key Laboratory for Germplasm Innovation and Utilization of Horticultural Crops, College of Horticulture and Forestry Sciences, Hubei Hongshan Lab, Huazhong Agricultural University, Wuhan 430070, China
    2. Max‐Planck‐Institute of Molecular Plant Physiology, Am Muehlenberg 1Potsdam‐Golm 14476, Germany

    †These authors contributed equally to this work.
    *Correspondence: Weiwei Wen (wwwen@mail.hzau.edu.cn)
  • Received:2025-03-20 Accepted:2025-05-29
  • Supported by:
    The research was financed by the National Natural Science Foundation of China (32494781, 32161133017) to Weiwei Wen.

Abstract: Purine-related metabolites are central to primary metabolic pathways in plants and serve as precursors for purine alkaloid biosynthesis in caffeinated species such as tea plants (Camellia sinensis). In this study, metabolite profiling of two tissues (young and mature leaves) was performed across 183 genetically diverse tea accessions, identifying and quantifying 10 purine alkaloid-related metabolites. Metabolite genome-wide association studies revealed 17 significant loci associated with these metabolites, including both known loci such as caffeine synthase and 16 novel loci (P < 1.05 × 10−5). Through functional annotation and in vitro enzymatic assay, we characterized 5′-methylthioadenosine/S-adenosylhomocysteine nucleosidase (CsMTAN) as the causal gene underlying natural variation in adenosine and adenine content. CsMTAN can catalyze the degradation of both 5ʹ-methylthioadenosine and S-adenosylhomocysteine to release adenine. The T → A nucleotide substitution at SNP55151898, which leads to a phenylalanine → tyrosine substitution at residue 179 (F179Y), resulted in a significant alteration of enzyme activity in vitro, as evidenced by an approximately 50% reduction in adenine abundance (P < 0.05). Transient overexpression of CsMTAN-A and CsMTAN-T in Nicotiana benthamiana both significantly increased adenine content and dramatically decreased adenosine content, providing direct evidence for the functional involvement of CsMTAN in plant purine metabolism. CsMTAN-T overexpression resulted in significantly lower adenosine level than CsMTAN-A (P < 0.05). Phylogenetic analysis across 115 species and protein structural modeling revealed a distinct evolutionary divergence between plant MTAN evolution and species phylogeny, strongly suggesting the occurrence of horizontal gene transfer events in the evolutionary history of plant MTANs. This study thus furthered our understanding of the genetics and molecular mechanisms regulating purine metabolism and purine alkaloid biosynthesis in tea plants and provided novel targets for molecular breeding and synthetic biology applications.

Key words: Camellia sinensis, mGWAS, purine alkaloid‐related metabolite

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