Construction of differential cDNA library on the roots of Dendrobium candidum induced by a symbiotic mycorrhizal fungus
- Author:
Biao LI
1
Author Information
1. Institute of Medicinal Plant Development
- Publication Type:Journal Article
- Keywords:
CDNA library;
Dendrobium candidum Wall.;
Mycorrhizal fungus;
Suppressive subtraction hybridization (SSH);
Symbiosis
- From:
Chinese Pharmaceutical Journal
2012;47(22):1790-1796
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE: To screen differentially expressed genes on the roots of Dendrobium candidum induced by a symbiotic mycorrhizal fungus, the differential cDNA library was constructed. METHODS: The ds cDNAs were synthesized and enriched by SMARTer PCR cDNA synthesis kit using total RNA of D. candidum roots symbiotic with the fungus as templates. The differential cDNA library was constructed by using suppressive subtraction hybridization (SSH). RESULTS: The differential cDNA library, containing 1975 clones in the storage capacity, was constructed successfully. It was detected that above 90% clones could be amplified effective products. The lengths of the differential cDNA fragments cloned were 150 bp to 1 kb by electrophoresis detection to 20 randomly selected clones. Then, the 20 ESTs similarity analysis based on BLASTx software was carried on and most of the cloned genes were the defensive genes of plant responding to environmental stresses. CONCLUSION: Using this technology system could construct a fine differential cDNA library and be operated easily, especially suitable to the rare species.
