摘要
利用血球计数法测定藻密度,研究不同浓度硫酸铈铵诱导子对三角褐指藻细胞生长的影响。采用高效液相色谱（HPLC）测定经不同浓度诱导子处理后三角褐指藻岩藻黄素的含量。通过荧光定量PCR研究岩藻黄素生物合成途径中相关基因转录差异,并利用SAS9.0统计软件对荧光定量PCR结果做主成分分析。结果显示：在一定浓度范围内硫酸铈铵（0.2-1.6 mg·L-1）抑制三角褐指藻细胞的生长。HPLC结果表明：随着硫酸铈铵浓度的增加,岩藻黄素含量呈先上升后下降的趋势,当硫酸铈铵浓度为0.2 mg·L-1时,岩藻黄素含量最高,达1.28 mg·g-1DW,比对照组提高了60%。定量PCR分析结果表明,硫酸铈铵浓度在0.2 mg·L-1时,岩藻黄素生物合成相关基因玉米黄素环氧酶基因（zep）、八氢番茄红素合成酶基因（pys）、ζ-胡萝卜素脱氢酶基因（zds）、番茄红素β-环化酶基因（lcyb）、胡萝卜素异构酶基因（rtiso）和八氢番茄红素脱氢酶基因（pds）转录水平与对照组相比极显著提高（P〈0.01）,与岩藻黄素含量变化相一致。主成分分析（PCA）结果表明：这些基因与岩藻黄素生物合成显著相关,其中,lcyb基因和pds基因对岩藻黄素生物合成贡献率最大。
The cell growth of algae induced by different concentration of ammonium cerous sulfate was researched by blood count to measure alga density. The content of fucoxanthin was measured by high performance liquid chro- matography. Using quantitative PCR, variant expressions of genes, which were related to the pathway of biosynthe- sis of fucoxanthin were detected. Furthermore, principal component analysis （PCA） was used to analyze quantita-rive PCR by the statistic software SAS 9.0. The results showed that certain concentrative （0.2-1.6 mg·L-1 ） of ammonium cerous sulfate inhibited the biomass of the algae. HPLC showed that when the concentration of ammoni- um cerous sulfate reached 0.2 mg·L-1 , the content of fucoxanthin got the maximum optimum. The content of fu- coxanthin reached 1.28 mg·g-1 DW and was higher than the control by 60%. The quantitive PCR results showed that when the concentration of ammonium cerous sulfate reached at 0.2 mg·L - 1, the relevant genes, phytoene syn- thase （psy）, carotenoid isomerase （rtiso）, phytoene desaturase （pals）, lycopene β-cyclase （lcyb）, zeaxanthin ep- oxidase （zep） and X-carotene desaturase （zds） involved in the pathway of biosynthesis of fucoxanthin, were ex- pressed higher strikingly than the control （ P 〈 0.01 ）. The result was consisted with the variation of the content of fucoxanthin. The result of PCA suggested that these genes were all related to the biological synthesis of fucoxanthin and the cumulative proportions of lcyb gene and pds gene were the best.
出处
《中国稀土学报》
CAS
CSCD
北大核心
2014年第6期750-757,共8页
Journal of the Chinese Society of Rare Earths
基金
浙江省科技厅重点科技创新团队项目(2012R10029-07
2010R50029)资助
关键词
硫酸铈铵
三角褐指藻
岩藻黄素
转录
主成分分析
稀土
ammonium cerous sulfate
Phaeodactylum tricornutum
fucoxanthin
transcription
principle component analysis
rare earths
