Psychoactive Plant Database - Neuroactive Phytochemical Collection

1. Plants (Basel). 2024 Sep 27;13(19):2704. doi: 10.3390/plants13192704.

FtMYB163 Gene Encodes SG7 R2R3-MYB Transcription Factor from Tartary Buckwheat 
(Fagopyrum tataricum Gaertn.) to Promote Flavonol Accumulation in Transgenic 
Arabidopsis thaliana.

Du H(1), Ke J(2), Sun X(2), Tan L(1), Yu Q(1), Wei C(1), Ryan PR(3), Wang A(1), 
Li H(2).

Author information:
(1)Panxi Featured Crops Research and Utilization Key Laboratory of Sichuan 
Province, Xichang University, Xichang 615000, China.
(2)Research Center of Buckwheat Industry Technology, College of Life Sciences, 
Guizhou Normal University, Guiyang 550025, China.
(3)Division of Plant Sciences, Research School of Biology, The Australian 
National University, Canberra, ACT 2601, Australia.

Tartary buckwheat (Fagopyrum tataricum Gaertn.) is a coarse grain crop rich in 
flavonoids that are beneficial to human health because they function as 
anti-inflammatories and provide protection against cardiovascular disease and 
diabetes. Flavonoid biosynthesis is a complex process, and relatively little is 
known about the regulatory pathways involved in Tartary buckwheat. Here, we 
cloned and characterized the FtMYB163 gene from Tartary buckwheat, which encodes 
a member of the R2R3-MYB transcription factor family. Amino acid sequence and 
phylogenetic analysis indicate that FtMYB163 is a member of subgroup 7 (SG7) and 
closely related to FeMYBF1, which regulates flavonol synthesis in common 
buckwheat (F. esculentum). We demonstrated that FtMYB163 localizes to the 
nucleus and has transcriptional activity. Expression levels of FtMYB163 in the 
roots, stems, leaves, flowers, and seeds of F. tataricum were positively 
correlated with the total flavonoid contents of these tissues. Overexpression of 
FtMYB163 in transgenic Arabidopsis enhanced the expression of several genes 
involved in early flavonoid biosynthesis (AtCHS, AtCHI, AtF3H, and AtFLS) and 
significantly increased the accumulation of several flavonoids, including 
naringenin chalcone, naringenin-7-O-glucoside, eriodictyol, and eight flavonol 
compounds. Our findings demonstrate that FtMYB163 positively regulates flavonol 
biosynthesis by changing the expression of several key genes in flavonoid 
biosynthetic pathways.

DOI: 10.3390/plants13192704
PMCID: PMC11478641
PMID: 39409574

Conflict of interest statement: The authors declare no conflict of interest.


2. J Hazard Mater. 2024 Oct 6;480:136074. doi: 10.1016/j.jhazmat.2024.136074. 
Online ahead of print.

GmSTOP1-3 regulates flavonoid synthesis to reduce ROS accumulation and enhance 
aluminum tolerance in soybean.

Liu G(1), Li D(2), Mai H(3), Lin X(4), Lu X(5), Chen K(6), Wang R(7), Riaz M(8), 
Tian J(9), Liang C(10).

Author information:
(1)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: liuguoxuanscau@163.com.
(2)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: 738137794@qq.com.
(3)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: 1570767334@qq.com.
(4)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: 3249839641@qq.com.
(5)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: xinglu@scau.edu.cn.
(6)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: chenkang@scau.edu.cn.
(7)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: 1575828734@qq.com.
(8)College of Resources and Environment, Zhongkai University of Agriculture and 
Engineering, Guangzhou 510225, PR China. Electronic address: 
riaz1480@hotmail.com.
(9)Root Biology Center, State Key Laboratory for Conservation and Utilization of 
Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: jtian@scau.edu.cn.
(10)Root Biology Center, State Key Laboratory for Conservation and Utilization 
of Subtropical Agro-bioresources, College of Natural Resources and Environment, 
South China Agricultural University, Guangzhou 510642, PR China. Electronic 
address: liangcy@scau.edu.cn.

Aluminum (Al) toxicity is a significant limiting factor for crop production in 
acid soils. The functions and regulatory mechanisms of transcription factor 
STOP1 (Sensitive to Proton Rhizotoxicity 1) family genes in Al-tolerance have 
been widely studied in many plant species, except for soybean. Here, expression 
of GmSTOP1-3 was significantly enhanced by Al stress in soybean roots. 
Overexpression of GmSTOP1-3 resulted in enhanced root elongation and decreased 
Al content, which was accompanied by increased antioxidant capacity under Al 
treatment. Furthermore, RNA-seq identified 498 downstream genes of GmSTOP1-3, 
including genes involved in flavonoid biosynthesis. Among them, the expression 
of chalcone synthase (GmCHS) and isoflavone synthase (GmIFS) were highly 
enhanced by GmSTOP1-3 overexpression. Further quantitative flavonoid metabolome 
analysis showed that overexpression of GmSTOP1-3 significantly increased the 
content of naringenin chalcone, naringenin, and genistein in soybean roots under 
Al treatment, which positively correlated with the expression level of the genes 
relative to flavonoid biosynthesis. Notably, genistein had a significant 
positive correlation with the expression levels of GmIFS. Combination of Dual 
Luciferase Complementation (LUC) and Electrophoretic Mobility Shift Assays 
(EMSA) revealed that GmSTOP1-3 directly bound to the promoters of GmCHS/GmIFS 
and activated both genes' transcription. Taken together, these results suggest 
that GmSTOP1-3 enhances soybean Al tolerance partially through regulating the 
flavonoid synthesis.

Copyright © 2024 Elsevier B.V. All rights reserved.

DOI: 10.1016/j.jhazmat.2024.136074
PMID: 39383698

Conflict of interest statement: Declaration of Competing Interest The authors 
declare that they have no known competing financial interests or personal 
relationships that could have appeared to influence the work reported in this 
paper.


3. Plant Physiol. 2024 Sep 29:kiae515. doi: 10.1093/plphys/kiae515. Online ahead
of  print.

Naringenin chalcone carbon double-bond reductases mediate dihydrochalcone 
biosynthesis in apple leaves.

Yauk YK(1), Dare AP(1), Cooney JM(2), Wang Y(3), Hamiaux C(1), McGhie TK(4), 
Wang MY(1), Li P(3), Atkinson RG(1).

Author information:
(1)The New Zealand Institute for Plant and Food Research Limited (Plant & Food 
Research), Auckland 1142, New Zealand.
(2)Plant & Food Research, Hamilton 3240, New Zealand.
(3)State Key Laboratory for Crop Stress Resistance and High-Efficiency 
Production/Shaanxi Key Laboratory of Apple, College of Horticulture, Northwest 
A&F University, Yangling, Shaanxi 712100, China.
(4)Plant & Food Research, Palmerston North 4442, New Zealand.

Dihydrochalcones (DHCs) are flavonoids produced as a side branch of the 
phenylpropanoid pathway. DHCs are found at high concentrations in apples (Malus 
spp.) but not in pears (Pyrus spp.) or other members of the Rosaceae. 
Biosynthesis of DHCs in apple has been hypothesized to occur via reduction of 
p-coumaroyl CoA by a Malus × domestica hydroxycinnamoyl CoA double-bond 
reductase (MdHCDBR) followed by the action chalcone synthase to produce 
phloretin or via direct reduction of naringenin chalcone to phloretin via an 
unknown enzyme. In this study, we report that genetic downregulation of MdHCDBR 
does not reduce DHC concentrations in apple leaves. We used comparative 
transcriptome analysis to identify candidate naringenin chalcone reductases 
(NCRs), designated MdNCR1a-c, expressed in apple leaves but not fruit. These 
MdNCR1 genes form an expanded gene cluster found exclusively in apple. Transient 
expression of MdNCR1 genes in Nicotiana benthamiana leaves indicated they 
produced DHCs at high concentrations in planta. Recombinant MdNCR1 utilized 
naringenin chalcone to produce phloretin at high efficiency. Downregulation of 
NCR genes in transgenic apple reduced foliar DHC levels by 85-95%. Reducing DHC 
production redirected flux to the production of flavonol glycosides. In situ 
localization indicated that NCR proteins were likely found in the vacuolar 
membrane. Active site analysis of AlphaFold models indicated that MdNCR1a-c 
share identical substrate binding pockets, but the pockets differ substantially 
in related weakly active/inactive NCR proteins. Identifying the missing enzyme 
required for DHC production provides opportunities to manipulate DHC content in 
apple and other fruits and has other applications, e.g., in biofermentation and 
biopharming.

© The Author(s) 2024. Published by Oxford University Press on behalf of American 
Society of Plant Biologists.

DOI: 10.1093/plphys/kiae515
PMID: 39343732


4. Genes (Basel). 2024 Sep 20;15(9):1228. doi: 10.3390/genes15091228.

Flavonoid Synthesis Pathway Response to Low-Temperature Stress in a Desert 
Medicinal Plant, Agriophyllum Squarrosum (Sandrice).

Zhao P(1)(2)(3), Yan X(1)(4), Qian C(1)(2), Ma G(5), Fan X(1)(2), Yin X(1)(2), 
Liao Y(1)(2)(3), Fang T(1)(2), Zhou S(1)(2), Awuku I(1)(2), Ma XF(1)(2).

Author information:
(1)Key Laboratory of Ecological Safety and Sustainable Development in Arid 
Lands, Northwest Institute of Eco-Environment and Resources, Chinese Academy of 
Sciences, Lanzhou 730000, China.
(2)Key Laboratory of Stress Physiology and Ecology in Cold and Arid Regions of 
Gansu Province, Northwest Institute of Eco-Environment and Resources, Chinese 
Academy of Sciences, Lanzhou 730000, China.
(3)University of Chinese Academy of Sciences, Beijing 100049, China.
(4)Key Laboratory of Inland River Ecohydrology, Cold and Arid Regions 
Environmental and Engineering Research, Northwest Institute of Eco-Environment 
and Resources, Chinese Academy of Sciences, Lanzhou 730000, China.
(5)Gulang County Sand Prevention and Control Technology Promotion Center, Wuwei 
733100, China.

Background/Objectives:Agriophyllum squarrosum (L.) Moq. (A. squarrosum), also 
known as sandrice, is an important medicinal plant widely distributed in dunes 
across all the deserts of China. Common garden trials have shown content 
variations in flavonoids among the ecotypes of sandrice, which correlated with 
temperature heterogeneity in situ. However, there have not been any 
environmental control experiments to further elucidate whether the accumulation 
of flavonoids was triggered by cold stress; Methods: This study conducted a 
four-day ambient 4 °C low-temperature treatment on three ecotypes along with an 
in situ annual mean temperature gradient (Dulan (DL), Aerxiang (AEX), and 
Dengkou (DK)); Results: Target metabolomics showed that 12 out of 14 flavonoids 
in sandrice were driven by cold stress. Among them, several flavonoids were 
significantly up-regulated, such as naringenin and naringenin chalcone in all 
three ecotypes; isorhamnetin, quercetin, dihydroquercetin, and kaempferol in DL 
and AEX; and astragalin in DK. They were accompanied by 19 structural genes of 
flavonoid synthesis and 33 transcription factors were markedly triggered by cold 
stress in sandrice. The upstream genes, AsqAEX006535-CHS, AsqAEX016074-C4H, and 
AsqAEX004011-4CL, were highly correlated with the enrichment of naringenin, 
which could be fine-tuned by AsqAEX015868-bHLH62, AsqAEX001711-MYB12, and 
AsqAEX002220-MYB1R1; Conclusions: This study sheds light on how desert plants 
like sandrice adapt to cold stress by relying on a unique flavonoid biosynthesis 
mechanism that regulating the accumulation of naringenin. It also supports the 
precise development of sandrice for the medicinal industry. Specifically, 
quercetin and isorhamnetin should be targeted for development in DL and AEX, 
while astragalin should be precisely developed in DK.

DOI: 10.3390/genes15091228
PMCID: PMC11431328
PMID: 39336819 [Indexed for MEDLINE]

Conflict of interest statement: The authors declare no conflicts of interest.


5. Appl Microbiol Biotechnol. 2024 Aug 10;108(1):435. doi: 
10.1007/s00253-024-13271-7.

Step-by-step optimization of a heterologous pathway for de novo naringenin 
production in Escherichia coli.

Gomes D(1), Rodrigues JL(2)(3), Rodrigues LR(1)(4).

Author information:
(1)CEB-Centre of Biological Engineering, Universidade Do Minho, Campus de 
Gualtar, 4710-057, Braga, Portugal.
(2)CEB-Centre of Biological Engineering, Universidade Do Minho, Campus de 
Gualtar, 4710-057, Braga, Portugal. joanarodrigues@deb.uminho.pt.
(3)LABBELS - Associate Laboratory, Braga, Guimarães, Portugal. 
joanarodrigues@deb.uminho.pt.
(4)LABBELS - Associate Laboratory, Braga, Guimarães, Portugal.

Naringenin is a plant polyphenol, widely explored due to its interesting 
biological activities, namely anticancer, antioxidant, and anti-inflammatory. 
Due to its potential applications and attempt to overcome the industrial demand, 
there has been an increased interest in its heterologous production. The 
microbial biosynthetic pathway to produce naringenin is composed of tyrosine 
ammonia-lyase (TAL), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), and 
chalcone isomerase (CHI). Herein, we targeted the efficient de novo production 
of naringenin in Escherichia coli by performing a step-by-step validation and 
optimization of the pathway. For that purpose, we first started by expressing 
two TAL genes from different sources in three different E. coli strains. The 
highest p-coumaric acid production (2.54 g/L) was obtained in the 
tyrosine-overproducing M-PAR-121 strain carrying TAL from Flavobacterium 
johnsoniae (FjTAL). Afterwards, this platform strain was used to express 
different combinations of 4CL and CHS genes from different sources. The highest 
naringenin chalcone production (560.2 mg/L) was achieved by expressing FjTAL 
combined with 4CL from Arabidopsis thaliana (At4CL) and CHS from Cucurbita 
maxima (CmCHS). Finally, different CHIs were tested and validated, and 
765.9 mg/L of naringenin was produced by expressing CHI from Medicago sativa 
(MsCHI) combined with the other previously chosen genes. To our knowledge, this 
titer corresponds to the highest de novo production of naringenin reported so 
far in E. coli. KEY POINTS: • Best enzyme and strain combination were selected 
for de novo naringenin production. • After genetic and operational 
optimizations, 765.9 mg/L of naringenin was produced. • This de novo production 
is the highest reported so far in E. coli.

© 2024. The Author(s).

DOI: 10.1007/s00253-024-13271-7
PMCID: PMC11316701
PMID: 39126431 [Indexed for MEDLINE]

Conflict of interest statement: The authors declare no competing interests.