Gene amplification from cryopreserved Arabidopsis thaliana shoot tips.
Cryopreservation is a way to store elite quality plant germplasms. The exact mechanism of stress tolerance during cryopreservation is unknown. Unavavailability of a detailed protocol for understanding the molecular genetics of plant cryostress is a major obstacle in plant cryobiology research. This paper describes the methods of extraction of total RNA from cryogenically stored plant tissues accompanied by successful amplication of cDNAs by reverse transcriptase PCR. The whole process can be completed in two to three days. Through this protocol, several genes were identified which were differentially expressed during cryostress. This protocol will help researchers to pursue further research in the field of molecular genetics of plant cryostress. Interesting genes identified via these processes can be cloned and plants can be transformed for the purpose of trait enhancement and modification.
Basu
Chhandak
C
School of Biological Sciences, University of Northern Colorado, 501 20th Street, Greeley, CO 80639, USA. chhandak.basu@unco.edu
D017360:Arabidopsis / Q000235:genetics*; D015925:Cryopreservation; D005784:Gene Amplification; D017343:Genes, Plant; D018519:Meristem / Q000235:genetics*; D018520:Plant Shoots / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction