[The analysis of rbcS gene function by post-transcription gene silencing in Nicotiana benthamiana].
A system of virus-induced post-transcriptional gene silencing for studying rbcS gene function was established and optimized using tobacco rattle virus vector and Nicotiana benthamiana as experimental materiaes. The following analyses were conducted: phenotypic characterization of rbcS gene silenced plants, transcription levels of rbcS gene by RT-PCR; protein levels of rbcS by the antibodies of rbcS and rbcL and photosynthetic pigments wntents in rbcS silenced plants by HPLC method. The results showed that the seedlings at 21-24-day-old and Agrobacterium concentration at OD600 = 1-1.5 gave the best results for gene silencing. The expression level of rbcL was very likely regulated by rbcS, and rbcS gene did not relate to the collection of photosynthetic energy. Probability analysis showed that the tobacco rattle virus vector system is a useful and effective technique to study rbcS gene function via post-transcriptional gene silencing.
Zhou
Xiao-Fu
XF
The Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China.
Ma
Peng-Da
PD
Wang
Ren-Hou
RH
Zhu
Xiao-Juan
XJ
Liu
Bao
B
Wang
Xing-Zhi
XZ
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D015153:Blotting, Western; D002734:Chlorophyll / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D014975:Lutein / Q000032:analysis; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D010942:Plant Viruses / Q000235:genetics; D010957:Plasmids; D034622:RNA Interference; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics* / Q000378:metabolism; D036226:Seedlings / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D010940:Plant Proteins; D011994:Recombinant Proteins; D002734:Chlorophyll; C101625:RbcL protein, plastid; D012273:Ribulose-Bisphosphate Carboxylase; D014975:Lutein