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本研究以生长在3个不同温度(均温18.5℃,均温23.5℃,均温28.5℃)人工气候室中的“云烟87”为材料,运用液相色谱质谱联用法、分光光度法和荧光定量PCR法,研究了不同生长温度对烟叶生长发育过程中黄酮类化合物芸香苷和山奈酚-3-O-芸香苷含量、代谢相关酶活性及相关基因表达的影响。结果表明:随着烟草叶片的生长发育,黄酮类化合物代谢相关基因表达呈先降低后升高的趋势,从而调控了黄酮类化合物代谢相关酶活性的变化趋势为先降低后升高,使得黄酮类化合物中的芸香苷含量变化趋势为先降低后升高,山奈酚-3-O-芸香苷的含量则呈逐渐降低的趋势。此外,与生长在较高生长温度(均温23.5℃和均温28.5℃)下的叶片相比,较低生长温度(均温18.5℃)上调了烟草叶片类黄酮代谢相关基因PAL、C4H、4CL、CHS、F3H、F3’H和FLS的基因表达,提高了类黄酮代谢关键酶PAL、C4H、4CL和CHI的活性,促进了黄酮类化合物芸香苷和山奈酚-3-O-芸香苷的积累(p<0.05),表明较低的生长温度(均温18.5℃)有利于黄酮类化合物在烟草叶片中的积累。
In this study, “Yunyan 87” grown in three different temperature (average temperature 18.5 ℃, average temperature 23.5 ℃, average temperature 28.5 ℃) artificial climate chamber, using liquid chromatography-mass spectrometry, spectrophotometry And fluorescence quantitative PCR method were used to study the effect of different growth temperature on flavonoid rutin and kaempferol-3-O- rutin content, metabolism related enzyme activity and related gene expression during the growth and development of tobacco leaves. The results showed that with the growth and development of tobacco leaves, the expression of flavonoid metabolism-related genes first decreased and then increased, and thus the regulation of flavonoid metabolism-related enzyme activity trends first and then decreased, making flavonoids The content of rutin in the compounds changed first and then decreased, and the content of kaempferol-3-O-rutin gradually decreased. In addition, the lower growth temperature (average temperature 18.5 ℃) up-regulated the flavonoid metabolism-related genes PAL, C4H, 4CL in tobacco leaves compared with those grown at higher growth temperature (average temperature 23.5 ℃ and average temperature 28.5 ℃) , CHS, F3H, F3’H and FLS increased the activities of flavonoid metabolism key enzymes PAL, C4H, 4CL and CHI, promoted the accumulation of flavonoid rutin and kaempferol-3-O-rutin (p <0.05), indicating that the lower growth temperature (average temperature 18.5 ℃) is conducive to the accumulation of flavonoids in tobacco leaves.