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JXB Advance Access originally published online on June 25, 2008
Journal of Experimental Botany 2008 59(11):3027-3037; doi:10.1093/jxb/ern152
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© The Author [2008]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Phenylalanine ammonia lyase functions as a switch directly controlling the accumulation of calycosin and calycosin-7-O-β-D-glucoside in Astragalus membranaceus var. mongholicus plants

Haiyun Pan1, Yuguo Wang1, Yongfeng Zhang2, Tongshui Zhou1, Changming Fang1, Peng Nan1, Xiaoqiang Wang3, Xiaobing Li4, Yinlong Wei1 and Jiakuan Chen1,*

1Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, Institute of Biodiversity Science, Fudan University, Shanghai 200433, China
2Department of Genetics, School of Life Sciences, Anhui Agricultural University, 130 Changjiang Road, Hefei, Anhui 230061, China
3Plant Biology Division, Samuel Roberts Noble Foundation 2510 Sam Noble Parkway Ardmore, OK 73401, USA
4School of Pharmacy, Anhui College of Traditional Chinese Medicine, 45 Shihe Road, Hefei, Anhui 230031, China

* To whom correspondence should be addressed. E-mail: jkchen{at}fudan.edu.cn

Previously it had been shown that calycosin and calycosin-7-O-β-D-glucoside (CGs) accumulate in whole plants, mainly in leaves, of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao (A. mongholicus) plants in response to low temperature. In this work, it was demonstrated that the influences of different conditions on CGs biosynthesis, by examining the changes in CGs content, as well as the expression of related genes, including phenylalanine ammonia lyase (PAL1), cinnamic acid 4-hydroxylase (C4H), chalcone synthase (CHS), chalcone reductase (CHR), chalcone isomerase (CHI), isoflavone synthase (IFS), and isoflavone 3'-hydroxylase (I3'H). The seven gene mRNAs accumulated in leaves of A. mongholicus upon exposure to low temperature in a light-dependent manner, though they exhibited different expression patterns. Transcriptions of CHS, CHR, CHI, IFS, and I3'H of the calycosin-7-O-β-D-glucoside pathway were all up-regulated when plants were transferred from 16 °C to 2 °C or 25 °C or from 2 °C (kept for 24 h) to 25 °C. However, fluctuations in temperature influenced differently the transcriptions of PAL1 and C4H of the general phenylpropanoid pathway in leaves. Moreover, the amount of PAL1 expression changed sharply up and down, consistent with the variation of the content of CGs. PAL enzyme activity appears to be the limiting factor in determining the CGs levels. The inhibitor of PAL enzyme, L-{alpha}-aminooxy-β-phenylpropionic acid, almost entirely shut down CGs accumulation at low temperature. All these results confirmed that PAL1, as a smart gene switch, directly controls the accumulation of CGs in A. mongholicus plants, in a light-dependent manner, during low temperature treatment.

Key words: Accumulation, enzyme inhibitor, gene switch, isoflavonoid, phenylalanine ammonia lyase

Received 18 February 2008; Revised 18 April 2008 Accepted 8 May 2008


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