Phenylalanine ammonia lyase functions as a switch directly controlling the accumulation of calycosin and calycosin-7-O-beta-D-glucoside in Astragalus membranaceus var. mongholicus plants.
Previously it had been shown that calycosin and calycosin-7-O-beta-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-beta-D-glucoside pathway were all up-regulated when plants were transferred from 16 degrees C to 2 degrees C or 25 degrees C or from 2 degrees C (kept for 24 h) to 25 degrees 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-beta-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.
Pan
Haiyun
H
Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, Institute of Biodiversity Science, Fudan University, Shanghai 200433, China.
Wang
Yuguo
Y
Zhang
Yongfeng
Y
Zhou
Tongshui
T
Fang
Changming
C
Nan
Peng
P
Wang
Xiaoqiang
X
Li
Xiaobing
X
Wei
Yinlong
Y
Chen
Jiakuan
J
D000429:Alcohol Oxidoreductases / Q000235:genetics; D027883:Astragalus Plant / Q000201:enzymology* / Q000235:genetics; D003080:Cold Temperature; D018076:DNA, Complementary / Q000302:isolation & purification; D018506:Gene Expression Regulation, Plant; D005960:Glucosides / Q000096:biosynthesis*; D019753:Intramolecular Lyases / Q000235:genetics; D007529:Isoflavones / Q000096:biosynthesis*; D008027:Light; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000494:pharmacology; D010650:Phenylalanine Ammonia-Lyase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D050564:Trans-Cinnamate 4-Monooxygenase / Q000235:genetics
D018076:DNA, Complementary; D005960:Glucosides; D007529:Isoflavones; D012333:RNA, Messenger; C526426:calycosin-7-O-beta-D-glucoside; C121707:7,3'-dihydroxy-4'-methoxyisoflavone; C019145:alpha-aminooxy-beta-phenylpropionic acid; D010649:Phenylalanine; D000429:Alcohol Oxidoreductases; C093070:chalcone reductase; D050564:Trans-Cinnamate 4-Monooxygenase; D010650:Phenylalanine Ammonia-Lyase; D019753:Intramolecular Lyases; C019865:chalcone isomerase