大花金挖耳培养细胞总黄酮提取工艺及抑菌活性研究  

Study on Extraction Technology and Antifungal Activity of Total Flavonoids from Cultured Cells of Carpesium macrocephalum

作　　者：李玉平[1,2,3] 龚宁 LI Yu-ping;GONG Ning(College of Life Sciences/Northwest A&F University,Shaanxi 712100,China;Institute for New Rural Development/Qinghai University,Xining 810016,China;Technology and Engineering Center of Biopesticide of Shaanxi Province/Northwest A&F University,Shaanxi 712100,China;College of Chemistry&Pharmacy/Northwest A&F University,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100 [2]青海大学新农村发展研究院,青海西宁810016 [3]西北农林科技大学陕西省生物农药工程技术研究中心,陕西杨凌712100 [4]西北农林科技大学化学与药学院,陕西杨凌712100

出　　处：《山东农业大学学报（自然科学版）》2020年第6期1038-1043,共6页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家公益性行业(农业)科研专项(200903052);国家重点研发计划项目(2016YFC0402904)。

摘　　要：为优化大花金挖耳细胞培养物总黄酮提取条件并初步测试细胞培养物抑菌活性。本文在考察不同干燥方式、溶剂、粉碎度和固液比等单因素对培养物总黄酮提取效果的基础上,采用L9(34)正交试验优化了振荡提取时间、超声波功率、超声波处理时间对培养物总黄酮提取工艺,并采用菌丝生长速率法测定了细胞培养物抑菌活性。结果表明,大花金挖耳培养细胞的黄酮含量显著高于培养液;室温下以丙酮为最佳提取溶剂;烘干和冷冻干燥对大花金挖耳胞内总黄酮的得率没有显著差异;样品粒度40~60目,固液比1:10~1:15(g/mL),超声提取(功率600 W、60min)1次后,再摇床振荡提取(18 h)1次,胞内黄酮的得率达1.22%,该方法简单,耗时短。抑菌试验表明,大花金挖耳悬浮培养细胞的乙醇粗提物对供试8种病原菌有不同程度的抑菌作用,在药液浓度50 mg/mL时,抑制率达到50%以上的供试病原菌有辣椒疫霉、苹果炭疽和小麦纹枯等3种病原菌。本研究为大花金挖耳细胞培养抑菌活性物质的提取工艺提供了参考。The extraction conditions of total flavonoids were optimized and the antifungal activity of the cell cultures was preliminarily tested.The effects of different drying methods,solvents,grinding degree and ratio of solid to liquid on the extraction of total flavonoids from cultural materials were investigated.Based on single factor experiment,the L9(34)orthogonal test was used to optimize the extraction process of total flavonoids from cultural materials by vibration extraction time,ultrasonic power and ultrasonic treatment time.The antifungal activity of the culture materials on pathogenic fungi tested were also determined using the mycelial growth rate method.The results showed that the content of flavonoids in the cultured cells was significantly higher than that in the culture medium.The optimal solvent was acetone at 25℃.There was no significant difference between the two methods included the freeze-dry and drying means.The optimal extraction conditions were as follows:grinding degree 40~60 mesh,solid-to-liquid ratio 1:10~1:15(g/mL),after vibration extracting the cultural materials with acetone for 18 h at 25℃,then extracting with ultrasonic wave for 60 min at power of 600 W.Under these conditions,the extraction yield of total flavonoids was 1.22%.The method is simple and takes less time.Also,the ethanol extracts of cultural materials from Carpesium macrocephalum had different antifungal activity against 8 pathogenic fungi.The fungicidal activities against Phytophthora capsici,Glomerella cingulata,Rhizoctonia cerealis were above 50%under the medicament concentration of 50 mg/mL.The results would be used to provide reference for the development and utilization of the antibiological substances in large-scale cell culture.

关 键 词：大花金挖耳 总黄酮提取 抑菌活性 

分 类 号：Q813.1[生物学—生物工程]