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在含有拟南芥HSP18.2-欧芹PAL2嵌合基因的转基因矮牵牛中,热激后苯丙氨酸解氨酶活性受到抑制。

Suppressed phenylalanine ammonia-lyase activity after heat shock in transgenic Nicotiana plumbaginifolia containing an Arabidopsis HSP18.2-parsley PAL2 chimera gene.

作者信息

Moriwaki M, Yamakawa T, Washino T, Kodama T, Igarashi Y

机构信息

Department of Biotechnology, University of Tokyo, 1-chome, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.

出版信息

J Biosci Bioeng. 1999;87(5):588-93. doi: 10.1016/s1389-1723(99)80119-4.

Abstract

The activity of phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) after heat shock (HS) in leaves and buds of transgenic Nicotiana plumbaginifolia containing an Arabidopsis HSP18.2 promoter-parsley phenylalanine ammonia-lyase 2 (HSP18.2-PAL2) chimera gene was examined. Immediately after HS treatment at 44 degrees C for 5 h, the PAL activity in both transgenic and normal (untransformed) plants was 35-38% lower than that before HS. At normal temperature (25-26 degrees C), the PAL activity recovered within 5 h of ending the HS treatment in normal plants, but not until 12-24 h in transgenic plants containing the HSP18.2-PAL2 gene. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed the presence of parsley PAL2 mRNA in transgenic plants, which remained for 8-12 h following 5-h HS at 44 degrees C; the mRNA was not observed before HS. The content of chlorogenic acid (CGA; 3-caffeoylquinic acid) decreased drastically 8-12 h after HS in transgenic plants, but only slightly in normal plants. Thus, the decrease in PAL activity accompanied by expression of the parsley PAL2 gene after HS treatment corresponded to the decrease in CGA synthesis. These results might be attributed to post-transcriptional degradation of endogenous PAL mRNA triggered by transcription of the transgene.

摘要

对含有拟南芥热激蛋白18.2启动子-欧芹苯丙氨酸解氨酶2(HSP18.2-PAL2)嵌合基因的转基因矮牵牛叶片和芽在热激(HS)处理后苯丙氨酸解氨酶(PAL;EC 4.3.1.5)的活性进行了检测。在44℃热激处理5小时后,转基因植株和正常(未转化)植株中的PAL活性立即比热激前降低了35%-38%。在常温(25-26℃)下,正常植株在热激处理结束后的5小时内PAL活性恢复,但含有HSP18.2-PAL2基因的转基因植株直到12-24小时后才恢复。逆转录-聚合酶链反应(RT-PCR)分析显示转基因植株中存在欧芹PAL2 mRNA,在44℃热激5小时后该mRNA持续存在8-12小时;热激前未观察到该mRNA。在转基因植株中,热激8-12小时后绿原酸(CGA;3-咖啡酰奎尼酸)含量急剧下降,但在正常植株中仅略有下降。因此,热激处理后PAL活性的降低伴随着欧芹PAL2基因的表达,这与CGA合成的减少相对应。这些结果可能归因于转基因转录引发的内源性PAL mRNA的转录后降解。

相似文献

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