Mechanisms and effective control of physiological browning phenomena in plant cell cultures.
Browning phenomena are ubiquitous in plant cell cultures that severely hamper scientific research and widespread application of plant cell cultures. Up to now, this problem still has not been well controlled due to the unclear browning mechanisms in plant cell cultures. In this paper, the mechanisms were investigated using two typical materials with severe browning phenomena, Taxus chinensis and Glycyrrhiza inflata cells. Our results illustrated that the browning is attributed to a physiological enzymatic reaction, and phenolic biosynthesis regulated by sugar plays a decisive role in the browning. Furthermore, to confirm the specific compounds which participate in the enzymatic browning reaction, transcriptional profile and metabolites of T. chinensis cells, and UV scanning and high-performance liquid chromatography-mass spectrometry (HPLC-MS) profile of the browning compounds extracted from the brown-turned medium were analyzed, flavonoids derived from phenylpropanoid pathway were found to be the main compounds, and myricetin and quercetin were deduced to be the main substrates of the browning reaction. Inhibition of flavonoid biosynthesis can prevent the browning occurrence, and the browning is effectively controlled via blocking flavonoid biosynthesis by gibberellic acid (GA3 ) as an inhibitor, which further confirms that flavonoids mainly contribute to the browning. On the basis above, a model elucidating enzymatic browning mechanisms in plant cell cultures was put forward, and effective control approaches were presented.
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Yan-Shan
YS
Dong
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Chun-Hua
CH
Fu
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Peng
P
Su
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Xiang-Ping
XP
Xu
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Jie
J
Yuan
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Sheng
S
Wang
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Meng
M
Zhang
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.
Chun-Fang
CF
Zhao
Institute of Resource Biology and Biotechnology, Department of Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.|Key Laboratory of Molecular Biophysics Ministry of Education, Huazhong University of Science and Technology, Wuhan, China.|Wuhan Institute of Biotechnology, Wuhan Biolake, Wuhan, China.
Long-Jiang
LJ
Yu
D019149:Bioreactors; D004156:Catechol Oxidase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018929:Cell Culture Techniques; D002463:Cell Membrane Permeability; D005419:Flavonoids / Q000302:isolation & purification / Q000378:metabolism; D006035:Glycyrrhiza / Q000737:chemistry / Q000201:enzymology / Q000502:physiology*; D015416:Maillard Reaction; D010100:Oxygen / Q000378:metabolism; D010636:Phenols / Q000302:isolation & purification / Q000378:metabolism*; D059828:Plant Cells / Q000737:chemistry / Q000201:enzymology / Q000502:physiology*; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011794:Quercetin / Q000302:isolation & purification / Q000378:metabolism; D020946:Taxus / Q000737:chemistry / Q000201:enzymology / Q000502:physiology*; D046509:Tissue Culture Techniques
D005419:Flavonoids; D010636:Phenols; D010940:Plant Proteins; C040015:myricetin; D011794:Quercetin; D004156:Catechol Oxidase; D010100:Oxygen