[A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].
OBJECTIVE
To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions.


METHODS
To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA.


RESULTS
Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration.


CONCLUSIONS
We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.
Southern Medical University, Guangzhou, China. qianzhen.love@163.com
Qianzhen
Q
Mo
Rongjia
R
Mai
Zhixiao
Z
Yang
Minfang
M
Chen
Tiezhao
T
Yang
Huafang
H
Lai
Peiliang
P
Yang
Qiang
Q
Chen
Xiaohong
X
Zhou
D018506:Gene Expression Regulation, Plant; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D018527:Hydroponics / Q000379:methods*; D030821:Plants, Genetically Modified / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000254:growth & development
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins