摘要
为探究不同温度培养对黄芩愈伤组织生长过程中黄芩苷含量及基因相对表达量的影响,该研究设置4个培养温度对黄芩愈伤组织暗培养40 d,每5 d取样1次,测定生长量和黄芩苷含量,并以18S RNA为内参基因,实时荧光定量PCR分析黄芩苷合成途径中5个关键酶苯丙氨酸解氨酶（PAL）、肉桂酸-4-羟化酶（C4H）、查尔酮合成酶（CHS）、β-葡糖醛酸酶（GUS）、黄芩素-7-O-葡萄糖醛酸转移酶（UBGAT）基因表达量。结果表明,生物量、黄芩苷含量及积累量均随温度的升高呈增加趋势,25,30℃较适宜黄芩愈伤组织生长,25℃培养30 d黄芩苷含量与积累量最高,达到2.75%和12.44 mg;15℃培养条件下C4H,CHS,GUS,UGBAT基因相对表达量较高,25℃培养30 d至35 d PAL基因相对表达量显著高于其他处理组。各关键酶基因相对表达量与黄芩苷含量的相关性随培养温度的升高,由负相关转变为正相关,25,30℃培养条件下PAL,C4H,CHS基因相对表达量与黄芩苷含量达到极显著或显著水平。相对低温条件不利于黄芩愈伤组织生长和黄芩苷的积累,合成途径的上游酶基因PAL,C4H对黄芩苷的积累作用显著。
The study is aimed to explore the effect of different temperature on the content of baicalin and gene expression in the growth of Scutellaria baicalensis. Four culture temperatures were used to establish the callus culture of S. baicalensis under dark conditions for 40 days and once every 5 days. The growth and baicalin contents were determined. 18 S RNA was used as a reference gene to analyze the five key factors in baicalin biosynthesis pathway（PAL）,cinnamic acid 4-hydroxylase（C4 H）,chalcone synthase（CHS）,β-glucuronidase（GUS）,baicalein-7-O-glucuronosyltransferase（UBGAT） gene expression levels. The results showed that biomass,baicalin content and accumulation increased with the increase of temperature. 25 ℃ and 30 ℃ were more suitable for the growth of S. baicalensis. The content and accumulation of baicalin at 25 ℃ reached the highest level at 30 days,reaching 2. 75% and 12. 44 mg,respectively. The relative expression levels of C4 H,CHS,GUS and UGBAT genes at 15 ℃ were higher than those at other treatments. The correlation between the relative expression levels of each key enzyme and the content of baicalin was negatively correlated with the increase of incubation temperature. The relative expression levels of PAL,C4 H and CHS genes at 25 ℃ and 30 ℃ were significantly correlated with the contents of baicalin and reached a highly significant or significant level. Relative low temperature conditions were not conducive to the growth of S. baicalensis and the accumulation of baicalin. The accumulation of baicalin by PAL and C4 H in the upstream of the synthetic pathway was significant.
作者
侯云亮
韩梅
杨利民
程林
罗燕
HOU Yun-liang;HAN Mei;YANG Li-min;CHENG Lin;LUO Yan(College of Chinese materials,Jilin Agricultural University,Changchun 130118,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2018年第13期2670-2675,共6页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(31570327)
国家中药材产业技术体系专项资金项目(CARS-21)
吉林省科技厅自然科学基金项目(20170101029JC)
关键词
愈伤组织
温度
黄芩苷
荧光定量PCR
基因表达
callus
temperature
baicalin
fluorescence quantitative PCR
gene expression
