摘要
为了分析丝氨酸乙酰基转移酶(serine acetyltransferase, SAT)在含硫化合物代谢中的功能,利用大豆毛状根体系进行了 LsSAT2的异源表达,检测转基因大豆毛状根株系中O-乙酰丝氨酸及半胱氨酸、甲硫氨酸等代谢物的含量变化,分析LsSAT2的蛋白互作网络。结果表明:利用发根农杆菌K599实现了 LsSAT2在大豆毛状根株系中的异源表达;转 LsSAT2基因大豆毛状根株系中的O-乙酰丝氨酸及含硫氨基酸半胱氨酸、甲硫氨酸的含量均获得显著上调;与LsSAT2互作的蛋白有催化蛋白以及多种分子结合蛋白,这些蛋白主要定位于胞质内蛋白复合体上,参与次生代谢以及碳代谢途径,氨基酸代谢及糖酵解途径等新陈代谢途径及相关分子生物合成等途径。
The neuroactiveβ-N-oxalyl-L-α,β-diaminopropionicacid(β-ODAP)is the causative agent to cause lathyrism,and the deficiency of sulfur-containing amino acid in Lathyrus sativus is closely correlated withβ-ODAP biosynthesis and lathyrism.To analyze the functions of serine acetyltransferases in sulfur metabolism,LsSAT2 in L.sativus was ectopically expressed in soybean.Subsequently,the content of O-acetyl serine,cysteine and methionine in soybean hairy root lines overexpressing LsSAT2 was examined via LC-MS.Proteins interacting with LsSAT2 were also identified and characterized via IP-MS.The results showed that a hairy root transformation system in soybean was established successfully using Agrobacterium rhizogenes K599,and soybean hairy root lines ectopically expressing LsSAT2 were obtained.The contents of O-acetyl serine,cysteine and methionine in transgenic hairy root lines were significantly increased.The proteins interacting with LsSAT2,including catalytic proteins and various molecular binding proteins,were mainly located in the cytoplasmic protein complex and were involved in secondary metabolites and carbon metabolism,as well as in amino acid metabolism and glycolysis.These results provide valuable insight into the regulatory mechanisms ofβ-ODAP biosynthesis mediated by LsSAT2 and further improvement of L.sativus germplasm.
作者
唐艳妮
尚天雪
苗智博
张雪萍
曹宁
连荣芳
王磊
徐全乐
TANG Yanni;SHANG Tianxue;MIAO Zhibo;ZHANG Xueping;CAO Ning;LIAN Rongfang;WANG Lei;XU Quanle(College of Life Sciences,Northwest A&F University,Yangling Shaanxi 712100,China;Dingxi Academy of Agricultural Sciences,Dingxi Gansu 743000,China;School of Ecology and Environment,Northwestern Polytechnical University,Xi’an 710072,China)
出处
《西北农业学报》
北大核心
2025年第8期1484-1490,共7页
Acta Agriculturae Boreali-occidentalis Sinica
基金
陕西省自然科学基金(2023-JC-YB-152)
国家食用豆产业技术体系(CARS-08-Z21)
河南省教育厅高等学校重点科研项目(25B180013)
西北农林科技大学大学生创新创业训练计划(202401360B1)。
关键词
山黧豆
大豆毛状根
丝氨酸乙酰基转移酶
异源表达
含硫氨基酸
三七素
Lathyrus sativus
Hairy root of soybean
Serine acetyltransferase
Ectopic expression
Sulfur-containing amino acids
Dencichine
