[Studies on population difference of Dendrobium officinale II establishment and optimization of the method of ISSR fingerprinting marker].
OBJECTIVE
To establish and optimize ISSR-PCR system of Dendrobiwn officinale according to the ISSR-PCR characters of D. officinale.


METHODS
The effects of ISSR-PCRs was examined by selecting primers and designing different concentrations of the factors in the ISSR-PCRs, the reliable ISSR-PCR systems for D. officinale populations researching was established by analyzing the reasons for occurrence of differential bands and optimizing reaction conditions.


CONCLUSIONS
The optimal ISSR-PCR system in D. officinale was reported for the first time, and 25 15327012 microL ISSR-PCR system contained 10 x Taq buffer, 1.5 U Taq DNA polymerase, 1.2-1.8 mmol x L(-1) MgCl2, 80 micromol x L(-1) dNTP, 0.2 micro mol x L(-1) primer and 20 ng template DNA. The appropriate annealing temperature was among 52-60 degrees C. ISSR PCRs were significantly influenced by Taq DNA polymerase, template DNA quantity and annealing temperature, etc. The ISSR-PCR systems, which were established in this paper for studying D. officinale, could provide clear reliable abundant polymorphisms molecular markers and were proved suitable for studying population authentication and population molecular ecology of D. officinale.
Shen
Jie
J
Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210097, China.
Ding
Xiao-Yu
XY
Ding
Ge
G
Liu
Dong-Yang
DY
Tang
Feng
F
He
Jia
J
D016172:DNA Fingerprinting; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics*; D031667:Dendrobium / Q000235:genetics*; D005819:Genetic Markers; D005828:Genetics, Population; D010946:Plants, Medicinal / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D015203:Reproducibility of Results
D017931:DNA Primers; D018744:DNA, Plant; D005819:Genetic Markers