CAJ | 학술논문

对离体培养青蒿试管苗中3个青蒿素合成相关基因的非生物胁迫诱导表达模式进行了初步研究.半定量RT-PCR测定结果表明,当暴露于冷、热和紫外光后,青篙植株的紫穗槐-4,11-二烯合酶基因(ADS)和细胞色素P450单加氧酶基因(CYP71AV1)转录上调;相反,在未经胁迫处理的情况下,ADS和CYP71AV1基因的表达水平较低,而在胁迫处理前后细胞色素P450还原酶基因(CPR)转录所产生的mRNA均保持恒定.同时,冷胁迫的诱导效果也得到实时荧光定量RT-PCR测定数据的支持.经低温锻炼的青蒿试管苗,其ADS和CYP71AV1基因的转录产物拷贝数比对照分别提高11倍和7倍,而CPR基因的mRNA拷贝数与对照基本持平.短暂预冷处理还可显著提高青蒿试管苗的青蒿素含量,达到7.5～8.8 mg/g干重,比对照提高66.7%～95.6%,为进一步探索利用环境胁迫促进青蒿素高产的新途径提供了可能性.图2表3参21
대리체배양청호시관묘중3개청호소합성상관기인적비생물협박유도표체모식진행료초보연구.반정량RT-PCR측정결과표명,당폭로우랭、열화자외광후,청고식주적자수괴-4,11-이희합매기인(ADS)화세포색소P450단가양매기인(CYP71AV1)전록상조;상반,재미경협박처리적정황하,ADS화CYP71AV1기인적표체수평교저,이재협박처리전후세포색소P450환원매기인(CPR)전록소산생적mRNA균보지항정.동시,랭협박적유도효과야득도실시형광정량RT-PCR측정수거적지지.경저온단련적청호시관묘,기ADS화CYP71AV1기인적전록산물고패수비대조분별제고11배화7배,이CPR기인적mRNA고패수여대조기본지평.단잠예랭처리환가현저제고청호시관묘적청호소함량,체도7.5～8.8 mg/g간중,비대조제고66.7%～95.6%,위진일보탐색이용배경협박촉진청호소고산적신도경제공료가능성.도2표3삼21
The abiotie stress-induced expression pattern of three genes responsible for artemisinin accumulation was preliminarily elucidated in cultured Artemisia annua plants．The semi-quantitative RT-PCR determination showed that upon exposure to chilling,heat shock or UV light,the transcription levels of amorpha-4,11-diene synthase gene(ADS)and cytochrome P450 monooxygenase gene(CYP71AV1)were up-regulated;In contrast,under the circumstance without stress treatments,the expression levels of ADS and CYP71AV1 genes were relatively low,while the mRNA generated by the transcription of cytochrome P450 reductase gene(CPR)remained stable under pre-and post-treatments．The induced outcome by chilling stress Was also supported by the measurement data from real-time fluorescent quantitative RT-PCR．Consequently,for A．annua shoots acclaimed by low temperature,the transcripts of ADS and CYP71AV1 genes were 11 folds and 7 folds higher than those of the control,but the CPR mRNA copy was almost equivalent to the contro1．Furthermore,a transient pre-chilling treatment led to an elevated artemisinin content up to 7．5～8．8 mg/g dry weight(DW),i．e．,increased by 66．7％～95．6％as compared with the control,thereby providing an open space for enhanced artemisinin production by the environmental stresses．Fig 2,Tab 3,Ref 21