ABSTRACT
Objective: To clone and analyze the coding region of gibberellin 20-oxidase (GA20ox) gene of Panax quinquefolium in seed dormancy and germination process. Methods: According to P. quinquefolium seeds transcriptome annotation, one transcript coding of GA20ox was obtained by comparative analysis of nine related unigenes. The full-length cDNA of PqGA20ox was determined by using RT-PCR method. Then the bioinformatic analysis of this gene and encoded protein was performed. The expression level of PqGA20ox in the roots, stems, leaves, and several seed dormancy periods was detected by real time fluorescent quantitative PCR (RT-qPCR). Results: Bioinformatic analysis showed that PqGA20ox contained 1092 bp and encoded 363 amino acids. PqGA20ox encoded protein had neither transmembrane nor signal peptide. The expression level of PqGA20ox was higher in the inset periods of morphological and physiological dormancy than that in the other periods based on RT-qPCR analysis. Conclusion: The PqGA20ox gene is cloned for the first time, and will provide a foundation for the dormancy release molecular mechanism of P. quinquefolium.
ABSTRACT
Objective: To clone and analyze the gibberellin 2-oxidase (GA2ox) gene of Panax quinquefolium in seed germination. Methods: Gene sequences about gibberellins synthesis and catabolism were found out from annotation information of 78 207 unigenes obtained by high-throughput sequencing in the early study. Then one transcript coding GA2ox was obtained from 11 unigenes related to GA2ox. Primers were designed according to selected sequence to get the full-length cDNA of P. quinquefolium using PCR method. Predictive analysis and expression analysis of PqGA2ox were obtained by bioinformatics and real-time PCR. Results: A GA2ox gene containing 987 bp encoding 328 amino acids was cloned and named as PqGA2ox. Bioinformatics analysis showed that PqGA2ox had no transmembrane domain or signal peptide, but had the 20G-Fell_Oxy conserved domains. The expression level of PqGA2ox was lower in the metaphase of morphological dormancy and physiological dormancy than that in the intitial period and release period based on real-time PCR analysis. Conclusion: The PqGA2ox gene from the seeds of P. quinquefolium is cloned for the first time, which will provide a foundation for the molecular mechanism of dormancy release of P. quinquefolium.