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Plant synthetic biology: new opportunities for large-scale culture of plant cells
YAN Zhaotao, ZHOU Pengfei, WANG Yangzhong, ZHANG Xin, XIE Wenyan, TIAN Chenfei, WANG Yong
Synthetic Biology Journal    2025, 6 (5): 1107-1125.   DOI: 10.12211/2096-8280.2024-095
Abstract   (1366 HTML82 PDF(pc) (1622KB)(2139)  

Plant Cell Culture (PCC) has emerged as a highly promising chassis for synthetic biology, offering a range of advantages such as short growth cycles, cost-effectiveness, absence of pathogenic risks, and abundant secondary metabolites. These features make PCC an attractive alternative for applications in medicine, food, and health. However, insufficient production efficiency due to difficulties in genetic transformation, complex regulatory networks, cell aggregation, and poor genetic stability remains a major obstacle that limits the commercialization of PCC. Synthetic biology, with its bottom-up engineering design approach, provides a powerful toolkit to address these challenges. By enabling the precise design and modification of native plant cells, synthetic biology offers innovative strategies to develop efficient and economically viable plant cell factories. In this paper, we first review the current status of PCC in synthesizing high-value compounds, particularly recombinant proteins and secondary metabolites. Recent advancements have demonstrated the potential of PCC to produce therapeutic proteins, vaccines, industrial enzymes and bioactive compounds such as alkaloids, flavonoids, and terpenoids. These successes underscore the versatility of PCC as a bioproduction platform. We then explore the role of synthetic biology in advancing PCC industrialization. Key developments include the creation of high-quality plant cell lines through genome editing tools like CRISPR/Cas9, enhancing genetic stability and metabolic efficiency. Additionally, synthetic biology has improved genetic transformation systems, overcoming a critical bottleneck in PCC. Enhanced expression systems, incorporating synthetic promoters and regulatory elements, have significantly boosted target compound yields. Furthermore, synthetic biology has expanded PCC applications by enabling the biosynthesis of heterologous compounds beyond their native metabolic pathways. Finally, we discuss future prospects, emphasizing the potential of synthetic biology to overcome current technical challenges. Emerging technologies including multi-omics integration, machine learning, and synthetic organelle development are anticipated to further enhance PCC’s scalability and efficiency. By addressing these challenges, synthetic biology will pave the way for large-scale plant cell cultivation, thereby facilitating its widespread adoption in industrial bioproduction. The convergence of PCC and synthetic biology holds immense potential for the sustainable, cost-effective, and scalable production of high-value compounds.


植物物种拉丁名蛋白名称细胞材料产量参考文献
水稻Oryza sativa L.人骨形态发生蛋白2(BMP2)水稻愈伤组织细胞21.5 μg/mL培养液[33]
水稻Oryza sativa L.人类生长激素(hGH)水稻愈伤组织细胞57 mg/L培养液[34]
水稻Oryza sativa L.合成牛胰蛋白酶原(synthetic bovine trypsinogen)水稻愈伤组织细胞15 mg/L培养液[35]
水稻Oryza sativa L.酸性葡萄糖苷酶(GAA)水稻愈伤组织细胞37 mg/L培养液[36]
水稻Oryza sativa L.血管内皮生长因子(VEGF)水稻愈伤组织细胞19 mg/L培养液[37]
水稻Oryza sativa L.包膜糖蛋白(envelope glycoprotein)水稻愈伤组织细胞18.5 μg/g[38]
水稻Oryza sativa L.贝伐单抗(Bevacizumab monoclonal antibody)水稻愈伤组织细胞160.7~242.8 mg/kg[39]
烟草Nicotiana tabacum L.人抗胰蛋白酶(human α1-antitrypsin)BY-2悬浮细胞34.7 mg/L培养液[40]
烟草Nicotiana tabacum L.人生长激素(human growth hormone)BY-2悬浮细胞5.2%总可溶蛋白[41]
Table 2 Heterologous synthesis of recombinant proteins in plant cells
Extracts from the Article
植物细胞独特的结构赋予其优越的生物制造特性。坚韧的细胞壁提供支持和保护,巨大的液泡不仅能储存潜在的毒性物质,还能调节渗透压以维持稳态,使植物细胞具备卓越的机械强度和抗逆性。此外,用于疾病治疗的功能蛋白在哺乳动物细胞中往往具有生理活性,从而影响细胞生长甚至产生细胞毒性,而植物细胞通常不受这些限制,这使其在复杂蛋白的安全、稳定、高效生产上具有显著优势。PCC生产重组蛋白技术已经被广泛用于药用蛋白(如抗原、疫苗、抗菌蛋白)、胶原蛋白和工业酶的商业化生产[13](表2)。
调控元件决定了基因的表达模式与表达强度,主要包括启动子、5′和3′非编码区、终止子、增强子、转录因子、顺式作用元件等DNA序列和蛋白序列。在植物细胞体系中,通过将调控元件序列和报告基因构建成基本表达盒或简单遗传回路,转化到植物细胞中进行定量表征,可用于优化植物细胞表达系统。例如,通过双荧光素酶报告系统(萤火虫荧光素酶/海肾荧光素酶),对不同植物启动子在BY2愈伤细胞中进行定量表征,可获得不同梯度强度的定量数据,用于精细控制蛋白表达水平和目标产物产量[85]。稳定的转基因植物细胞培养体系通常需要经过长时间的筛选过程,因此,将愈伤组织或悬浮细胞经酶解制备成原生质体是元件快速表征的一种高效替代方法。Ko Kato团队[86]通过制备水稻(O. sativa)悬浮培养细胞、黑麦(Secale cereale L.)叶片和拟南芥(A. thaliana)悬浮细胞的原生质体,在三个体系中统一验证了15个5′UTR序列的增强子活性。这种方法显著提升了元件表征效率,为调控元件在植物细胞中的应用提供了重要依据。
利用植物合成生物学能够突破植物合成能力的限制,让植物细胞得以合成原本无法产生的高附加值产物。例如,在胡萝卜愈伤组织细胞中异源表达六个拟南芥来源的苄基硫代葡萄糖苷生物合成途径基因,成功在细胞中实现了苄基硫代葡萄糖苷的积累,产量达到2.5 nmol/g FW[102]。
绿原酸1799.69 ng/mL培养液; ...
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