Wang Yudan, Song Shiwei, Hao Yanwei, Chen Changming, Ou Xi, He Bin, Zhang Jiewen, Jiang Zhehao, Li Chengming, Zhang Shuaiwei, Su Wei, Chen Riyuan
Key Laboratory of Horticultural Crop Biology and Germplasm Innovation in South China, Ministry of Agriculture, College of Horticulture, South China Agricultural University, Guangzhou 510642, China.
Hortic Res. 2023 Jun 2;10(8):uhad119. doi: 10.1093/hr/uhad119. eCollection 2023 Aug.
Gibberellin (GA) plays a major role in controlling stalk development. As an essential negative regulator of GA signal transduction, DELLA proteins may exert significant effects on stalk development. However, the regulatory mechanisms underlying this regulation remain unclear. In this study, we report highly efficient and inheritable mutagenesis using the CRISPR/Cas9 gene editing system in (phytoene desaturase) and (key DELLA protein) genes. We observed a loss-of-function mutation in due to two amino acids in GRAS domain. The flower bud differentiation and bolting time of mutants were significantly advanced. The expression of GA-regulatory protein (), flowering related genes (, ), expansion protein () and xyloglucan endotransferase () genes was also significantly upregulated in these mutants. -overexpressing plants displayed the contrasting phenotypes. mutants were more sensitive to GA signaling. BraRGL1 interacted with BraSOC1, and the interaction intensity decreased after GA treatment. In addition, BraRGL1 inhibited the transcription-activation ability of for and genes, but the presence of GA enhanced the activation ability of , suggesting that the BraRGL1-BraSOC1 module regulates bolting and flowering of through GA signal transduction. Thus, we hypothesized that BraRGL1 is degraded, and BraSOC1 is released in the presence of GA, which promotes the expression of and , thereby inducing stalk development in . Further, the mutant promoted the flower bud differentiation without affecting the stalk quality. Thus, can serve as a valuable target for the molecular breeding of early maturing varieties.
赤霉素(GA)在控制茎发育中起主要作用。作为GA信号转导的重要负调控因子,DELLA蛋白可能对茎发育产生显著影响。然而,这种调控背后的机制仍不清楚。在本研究中,我们报道了利用CRISPR/Cas9基因编辑系统在(八氢番茄红素去饱和酶)和(关键DELLA蛋白)基因中进行高效且可遗传的诱变。我们观察到由于GRAS结构域中的两个氨基酸,导致功能丧失突变。突变体的花芽分化和抽薹时间显著提前。在这些突变体中,GA调节蛋白()、开花相关基因(,)、扩张蛋白()和木葡聚糖内转糖基酶()基因的表达也显著上调。过表达植物表现出相反的表型。突变体对GA信号更敏感。BraRGL1与BraSOC1相互作用,GA处理后相互作用强度降低。此外,BraRGL1抑制了对于和基因的转录激活能力,但GA的存在增强了的激活能力,表明BraRGL1-BraSOC1模块通过GA信号转导调节的抽薹和开花。因此,我们推测在GA存在下,BraRGL1被降解,BraSOC1被释放,这促进了和的表达,从而诱导的茎发育。此外,突变体促进了花芽分化而不影响茎的质量。因此,可作为早熟品种分子育种的有价值靶点。