Xing Mengyang, Li Hongli, Liu Gangshuai, Zhu Benzhong, Zhu Hongliang, Grierson Donald, Luo Yunbo, Fu Daqi
Laboratory of Fruit Biology, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China; Key Laboratory of Plant Resources, Institute of Botany, The Innovative Academy of Seed Design, Chinese Academy of Sciences, Beijing 100093, China.
Laboratory of Fruit Biology, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100083, China.
Plant Sci. 2022 Sep;322:111366. doi: 10.1016/j.plantsci.2022.111366. Epub 2022 Jun 30.
In flowering plants, sepals play important roles in the development of flowers and fruit, and both processes are regulated by MADS-box (MADS) transcription factors (TFs). SlMADS1 was previously reported to act as a negative regulator of fruit ripening. In this study, expression analysis shown that its transcripts were very highly expressed during the development of sepals. To test the role of SlMADS1, we generated KO-SlMADS1 (knock-out) tomato mutants by CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9) technology and over-expression of SlMADS1 (OE-SlMADS1). The sepals and individual cells of KO-SlMADS1 mutants were significantly elongated, compared with the wild type (WT), whereas the sepals of OE-SlMADS1 tomatoes were significantly shorter and their cells were wider. RNA-seq (RNA-sequencing) of sepal samples showed that ethylene-, gibberellin-, auxin-, cytokinin- and cell wall metabolism-related genes were significantly affected in both KO-SlMADS1 and OE-SlMADS1 plants with altered sepal size. Since SlMACROCALYX (MC) is known to regulate the development of tomato sepals, we also studied the relationship between SlMC and SlMADS1 and the result showed that SlMADS1 interacts directly with SlMC. In addition, we also found that manipulating SlMADS1 expression alters the development of tomato plant leaves, roots and plant height. These results enrich our understanding of sepal development and the function of SlMADS1 throughout the plant.
在开花植物中,萼片在花和果实的发育过程中发挥着重要作用,这两个过程均受MADS盒(MADS)转录因子(TFs)调控。此前有报道称SlMADS1是果实成熟的负调控因子。在本研究中,表达分析表明其转录本在萼片发育过程中高度表达。为了测试SlMADS1的作用,我们通过CRISPR/Cas9(成簇规律间隔短回文重复序列及其相关蛋白9)技术构建了KO-SlMADS1(敲除)番茄突变体,并对SlMADS1进行了过表达(OE-SlMADS1)。与野生型(WT)相比,KO-SlMADS1突变体的萼片和单个细胞显著伸长,而OE-SlMADS1番茄的萼片显著缩短,其细胞更宽。萼片样本的RNA测序(RNA-seq)表明,萼片大小改变的KO-SlMADS1和OE-SlMADS1植株中,与乙烯、赤霉素、生长素、细胞分裂素和细胞壁代谢相关的基因均受到显著影响。由于已知SlMACROCALYX(MC)可调控番茄萼片的发育,我们还研究了SlMC与SlMADS1之间的关系,结果表明SlMADS1可直接与SlMC相互作用。此外,我们还发现,调控SlMADS1的表达会改变番茄植株叶片、根的发育以及株高。这些结果丰富了我们对萼片发育以及SlMADS1在整个植株中功能的理解。