Ajayo Babatope Samuel, Li Yangping, Wang Yayun, Dai Chengdong, Gao Lei, Liu Hanmei, Yu Guowu, Zhang Junjie, Huang Yubi, Hu Yufeng
State Key Laboratory of Crop Gene Resource Exploration and Utilization in Southwest China, Chengdu, China.
College of Agronomy, Sichuan Agricultural University, Chengdu, China.
Front Plant Sci. 2022 Jul 15;13:943050. doi: 10.3389/fpls.2022.943050. eCollection 2022.
The process of starch biosynthesis is a major developmental event that affects the final grain yield and quality in maize ( L.), and transcriptional regulation plays a key role in modulating the expression of the main players in the pathway. , which encodes the small subunits of AGPase, is a rate-controlling gene of the pathway; however, much remains unknown about its transcriptional regulation. Our earlier study identifies a short functional fragment of promoter (394-bp), and further shows it contains multiple putative -acting regulatory elements, demonstrating that several transcription factors may govern expression. Here, we identified a novel TCP transcription factor (TF), ZmTCP7, that interacted with the functional fragment of the promoter in a yeast one hybrid screening system. We further showed that ZmTCP7 is a non-autonomous TF targeted to the nucleus and predominantly expressed in maize endosperm. Using promoter deletion analyzes by transient expression in maize endosperm protoplasts combined with electrophoretic mobility shift assays, we found that ZmTCP7 bound to GAACCCCAC elements on the promoter to suppress its expression. Transgenic overexpression of in maize caused a significant repression of transcription by ~77.58%, resulting in a 21.51% decrease in AGPase activity and a 9.58% reduction in the endosperm starch content of transgenic maize. Moreover, the expressions of , , and were increased, while those of and reduced significantly in the endosperm of the transgenic maize. Overall, this study shows that ZmTCP7 functions as a transcriptional repressor of and a negative regulator of endosperm starch accumulation, providing new insights into the regulatory networks that govern expression and starch biosynthesis pathway in maize.
淀粉生物合成过程是影响玉米最终籽粒产量和品质的主要发育事件,转录调控在调节该途径中主要参与者的表达方面起着关键作用。编码AGPase小亚基的基因是该途径的限速基因;然而,其转录调控仍有许多未知之处。我们早期的研究鉴定了基因启动子的一个短功能片段(394bp),并进一步表明它包含多个推定的顺式作用调控元件,这表明几种转录因子可能调控基因的表达。在这里,我们在酵母单杂交筛选系统中鉴定了一个新的TCP转录因子ZmTCP7,它与基因启动子的功能片段相互作用。我们进一步表明,ZmTCP7是一种靶向细胞核的非自主转录因子,主要在玉米胚乳中表达。通过在玉米胚乳原生质体中瞬时表达结合电泳迁移率变动分析进行启动子缺失分析,我们发现ZmTCP7与基因启动子上的GAACCCCAC元件结合以抑制其表达。在玉米中过表达转基因导致基因转录显著抑制约77.58%,导致转基因玉米中AGPase活性降低21.51%,胚乳淀粉含量降低9.58%。此外,在转基因玉米的胚乳中,基因、、和的表达增加,而和的表达显著降低。总体而言,本研究表明ZmTCP7作为基因的转录抑制因子和胚乳淀粉积累的负调控因子,为调控玉米中基因表达和淀粉生物合成途径的调控网络提供了新的见解。