College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China.
Sichuan Provincial Academy of Natural Resources Sciences, Chengdu 610015, China.
Int J Biol Macromol. 2023 Jul 1;242(Pt 3):124928. doi: 10.1016/j.ijbiomac.2023.124928. Epub 2023 May 22.
Fruits provide abundant carotenoid nutrients for humans, whereas the understanding of the transcriptional regulatory mechanisms of carotenoids in fruits is still limited. Here, we identified a transcription factor AcMADS32 in kiwifruit, which was highly expressed in the fruit, correlated with carotenoid content and localized in the nucleus. The silencing expression of AcMADS32 significantly reduced the content of β-carotene and zeaxanthin and expression of β-carotene hydroxylase gene AcBCH1/2 in kiwifruit, while transient overexpression increased the accumulation of zeaxanthin, suggesting that AcMADS32 was an activator involved in the transcriptional regulation of carotenoid in fruit. When AcMADS32 was further stably transformed into kiwifruit, the content of total carotenoid and components in the leaves of transgenic lines significantly increased, and the expression level of carotenogenic genes was up-regulated. Moreover, Y1H and dual luciferase reporter experiments confirmed that AcMADS32 directly bound the AcBCH1/2 promoter and activated its expression. Through Y2H assays, AcMADS32 can interact with other MADS transcription factor AcMADS30, AcMADS64 and AcMADS70. These findings will contribute to our understanding of the transcriptional regulation mechanisms underlying carotenoid biosynthesis in plants.
水果为人类提供了丰富的类胡萝卜素营养物质,但人们对水果中类胡萝卜素的转录调控机制仍知之甚少。本研究在猕猴桃中鉴定到一个转录因子 AcMADS32,该基因在果实中高表达,与类胡萝卜素含量相关,并定位于细胞核中。AcMADS32 的沉默表达显著降低了猕猴桃中β-胡萝卜素和玉米黄质的含量以及β-胡萝卜素羟化酶基因 AcBCH1/2 的表达,而过表达则增加了玉米黄质的积累,表明 AcMADS32 是参与果实类胡萝卜素转录调控的激活子。进一步将 AcMADS32 稳定转化到猕猴桃中,发现转基因株系叶片中的总类胡萝卜素及其组分含量显著增加,类胡萝卜素生物合成基因的表达水平上调。此外,酵母单杂交(Y1H)和双荧光素酶报告基因实验证实 AcMADS32 可直接结合 AcBCH1/2 启动子并激活其表达。通过酵母双杂交(Y2H)实验发现,AcMADS32 可以与其他 MADS 转录因子 AcMADS30、AcMADS64 和 AcMADS70 相互作用。这些发现将有助于我们理解植物中类胡萝卜素生物合成的转录调控机制。