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作 者:徐纪明 吕秋萍 胡晗[2] XU Jiming;LV Qiuping;HU Han(Institute of Plant Biology,College of Life Science,Zhejiang University,Hangzhou 310058,China;Agricultural Experiment Station,Zhejiang University,Hangzhou 310058)
机构地区:[1]浙江大学生命科学学院植物生物学研究所,杭州310058 [2]浙江大学农业试验站,杭州310058
出 处:《食品与发酵科技》2024年第4期53-58,共6页Food and Fermentation Science & Technology
基 金:国家重点研发计划(2021YFF1000403)。
摘 要:为了确定甜菜红素作为报告基因检测基因表达时的定量分析方法,采用单因素实验和正交实验对转甜菜红素合成基因水稻叶片中甜菜红素提取工艺和稳定性进行研究。结果表明,水稻叶片甜菜红素最佳提取溶剂为蒸馏水(pH值为5.0),最佳提取工艺为:料液比1∶30 g/mL、提取温度20℃、提取时间30 min。水稻叶片甜菜红素经氯仿进行二次抽提后在538 nm处出现明显的最大吸收峰,与甜菜红素标准品的吸收峰值一致。稳定性实验结果表明,温度、光照和存放时间对甜菜红素稳定性都有影响,室温自然光条件下存放1 d,甜菜红素OD值下降37.3%,而存放3 d后,所有储存条件下甜菜红素OD538值都显著下降,说明按照该研究提取工艺提取的甜菜红素需低温避光保存,并且保存时间较短。In order to determine the quantitative analysis method of betacyanins as a reporter gene for detecting gene expression,this study investigated the extraction conditions and stability of betacyanins in rice leaves with the synthetic betacyanins synthesis gene using single-factor experiments and orthogonal experiments.The results indicated that the optimal extracting solvent for betacyanins in rice leaves is distilled water(pH 5.0),with the best extraction conditions being a material-liquid ratio of 1∶30 g/mL,extraction temperature of 20℃,and extraction time of 30 minutes.After adding chloroform for secondary extraction,a clear maximum absorption peak appeared at 538 nm in rice leaves,which was consistent with the absorption peak of the standard beet red pigment.Stability tests showed that temperature,light exposure,and storage time all affect the stability of betacyanins.When stored at room temperature under natural light conditions for 1 day,the OD538 of betacyanins concentration decreased 37.3%.After 3 days of storage,the betacyanins concentration significantly decreased under all conditions,indicating that the betacyanins extracted under the conditions of this study should be stored at low temperature and protected from light,and the storage time should not be too long.
分 类 号:TS264.4[轻工技术与工程—发酵工程]
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