稻曲病菌厚垣孢子中cAMP提取条件优选  

Extraction Conditions Optimization of cAMP in Chlamydospores of Ustiloginoidea virens

作　　者：李媛[1] 任佐华[1] 刘敏捷[2] 杨浪[1] 陈娟芳[3] 刘二明[1] 

机构地区：[1]湖南农业大学生物安全科学技术学院,湖南长沙410128 [2]湖南农业大学信息科学技术学院,湖南长沙410128 [3]湖南农业大学东方科技学院,湖南长沙410128

出　　处：《微生物学杂志》2013年第2期12-16,共5页Journal of Microbiology

基　　金：国家自然科学基金(30871615);湖南省大学生研究性学习和创新性实验计划项目(SCX1219)

摘　　要：探究稻曲病菌(Ustiloginoidea virens(Cooke.)Takahashi)黑色(休眠)与黄色(非休眠)厚垣孢子中的环磷酸腺苷(cAMP)最佳提取条件,为进一步的研究cAMP功能奠定基础。采用超声-水浴法对cAMP进行浸提,按3因素3水平正交设计,用高效液相色谱法检测cAMP含量;在设定V(甲醇)V(0.05 mol/L KH2PO4)=2080、流速为0.8 mL/min、检测波长为254 nm、进样量为20μL的条件下,以黄绿色厚垣孢子为提取样品,其提取cAMP效果最佳组合条件:超声破碎时间10 min(功率400 W、间歇时间2 s),水浴温度80℃,物料比为1100,提取的cAMP为6.827 6μg/mL。在此最佳条件下,测定出黄色厚垣孢子的cAMP为12.805 0±0.533 2μg/mL,黑色厚垣孢子的cAMP为4.171 7±0.097 1μg/mL。此结果表明,由黄色转换为黑色,其厚垣孢子的cAMP含量显著降低。The extraction condition of 3＇-5＇-cyclic adenosine monophosphate （cAMP） from black （dormancy） and yellow （non-dormancy） chlamydospore in Ustiloginoidea virens was probed deeply into to provide foundation for further research on its function. In this study ultrasonic-bath method was adopted to extract cAMP. The orthogonal design was used according to 3 factors at 3 levels, using high-performance liquid chromatography （HPLC） to test the content of cAMP. When setting V （methanol） ： V （0.05 mol/L KH2PO4） = 20 ： 80, testing wave at 254 nm, under the con-dition of sample injection at 20 μL, taking yellow-green chlamydospore as extraction sample, the optimal combination condition of cAMP extraction result was ultrasonic breaking time for 10 min （400 W, 2 see interval） , water bath at 80 ℃ , and the material ratio at 1 ： 100 （V/V） , and extracted cAMP at 6. 827 6 μg/mL. Under the above optimal ex-traction condition, the contents of cAMP in the yellow ehlamydospore was 12. 805 0 ± 0. 533 2 μg/mL, and the black 4. 171 7 ±0. 097 1 μg/mL respectively. The results suggested that the content of chlamydospore cAMP significantly decreased when changed from yellow into black.

关 键 词：稻曲病菌 厚垣孢子 环磷酸腺苷 提取条件 

分 类 号：Q939.95[生物学—微生物学]