[Increasing the content of active constituents in Polygonum cuspidatum hairy root by gene transformation technology].
To increase the content of active constituent--RE and PD of Polygonum cuspidatum hairy root, through Ri-mediated gene transformation technology, modified high salt low pH method was used to distill genome DNA of grapevine (Vitis raparia). Primer was designed according to sequence of Genebank (AF128861). Through PCR amplification obtain RS gene sequence was obtained. Binary vector pCAMBIA1300-35S-RS was constructed. Frost thawing method was used to transform Agrobacterium rhizogenes ATCC11325. Scratched aseptic seedling leaf of Polygonum cuspidatum was contaminated subsequently. DNA conformity and mRNA expression of RS gene were investigated by PCR and RT-PCR respectively. RE and PD in transgenic hairy root were determined by HPLC. For the first time successfully inducement acquires transformed RS gene hairy root of Polygonum cuspidatum. Content of active constituents--RE and PD were 17 - 187 microg x g(-1) DW and 836 - 1 970 microg x g(-1) DW, respectively, the non-transgenic hairy root was 0 - 130 microg x g(-1) DW and 190 - 320 microg x g(-1) DW. In the different root selected, the content of PD was much higher than that in non-transformed hairy roots of Polygonum cuspidatum, the highest content is 5 times, but the content of RE has not increased apparently.
Miao
Xiao-Yan
XY
College of Life Science, Hebei Normal University, Shijiazhuang 050016, China.
Yu
Shu-Hong
SH
Shen
Yin-Zhu
YZ
Huang
Zhan-Jing
ZJ
D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D004365:Drugs, Chinese Herbal / Q000032:analysis; D045468:Fallopia japonica / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D005960:Glucosides / Q000032:analysis*; D008969:Molecular Sequence Data; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D010946:Plants, Medicinal / Q000235:genetics / Q000378:metabolism; D000077185:Resveratrol; D012231:Rhizobium / Q000235:genetics; D013267:Stilbenes / Q000032:analysis*; D014170:Transformation, Genetic
D017931:DNA Primers; D018744:DNA, Plant; D004365:Drugs, Chinese Herbal; D005960:Glucosides; D013267:Stilbenes; D000217:Acyltransferases; C055200:resveratrol synthase; D000077185:Resveratrol; C058229:polydatin