Genotyping and species identification of Fritillaria by DNA chips.
OBJECTIVE
To investigate the genetic polymorphism of several species of Fritillaria and to develop a DNA chip for the genotyping and identification of the origin of various species of Fritillaria at molecular level.


METHODS
Genomic DNA from bulbs of several Fritillaria species was extracted and the polymorphisms of the D2 and D3 regions inside the 26S rDNA gene were identified by direct sequencing. Oligonucleotide probes specific for these polymorphisms were designed and printed on the poly-lysine coated slides to prepare the DNA chip. PCR products from the Fritillaria species were labeled with fluorescence by incorporation of dye-labeled dideoxyribonucleotides and hybridized to the immobilized probes on the chip.


RESULTS
The polymorphisms were used as markers for discrimination among various species. Specific oligonucleotide probes were designed and immobilized on a DNA chip. Differentiation of the various Fritillaria species was accomplished based on hybridization of fluorescent labeled PCR products with the DNA chip.


CONCLUSIONS
The results demonstrated the reliability of using DNA chips to identify different species of Fritillaria, and the DNA chip technology can provide a rapid, high throughput tool for genotyping and quality assurance of the plant species verification.
Tsoi
Pui-yan
PY
Applied Research Centre for Genomic Technologies, Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong, China.
Woo
Hok-sin
HS
Wong
Man-sau
MS
Chen
Shi-lin
SL
Fong
Wan-fung
WF
Xiao
Pei-gen
PG
Yang
Meng-su
MS
D001483:Base Sequence; D018744:DNA, Plant / Q000032:analysis*; D031410:Fritillaria / Q000145:classification / Q000235:genetics; D005838:Genotype; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010946:Plants, Medicinal / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D012335:RNA, Ribosomal / Q000235:genetics; D013045:Species Specificity
D018744:DNA, Plant; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S