ABSTRACT
Objective To provide a basis for further study and utilization of the functional gene family of Pht1 in Salviae miltiorrhizae, the Pht1 gene family members from S. miltiorrhizae genome should be explored, identified and functionally predicted. Methods By utilizing the bioinformatics methods, based on the genomic and NCBI database of S. miltiorrhiza, the sequences of Pht1 family candidate genes of S. miltiorrhiza were obtained firstly by screening, and then carried on the multiple sequence alignment, conserved structure lookup as well as phylogenetic tree construction. Finally, the characteristic analysis and biological function prediction were carried out. Results The results showed that 12 Pht1 candidate genes were extracted from the genome of Salvia miltiorrhiza, together with the reported Smpht1, there are 13 Pht1 members in total, their homology as high as 74.34%, and nine members of them could provide more reliable functional annotation. Among the gene family members, Smpht1 & Sm1, Sm4 & Sm6, Sm5 & Sm11, in which these three pairs may share similar function as well as coevolution relation between each other; Smpht1, Sm1, Sm3, Sm5, Sm7, and Sm11, all of the six members can be only expressed in the roots of S. miltiorrhiza. furthermore, except Sm7 induced by p-deficiency and Sm3 member unaffected by the phosphorus supply factor, the other four probably belong to the P-deficiency induced up-regulated expressive genotype, which play an important role in phosphorus absorption of S. miltiorrhiza; Moreover, both Sm7 and Sm16 are possible to expressed in the flower organs of S. miltiorrhiza and then participate in the regulation of phosphorus balance in the flowering stage of plants. In addition, Sm14 is likely to be a mycorrhizal-induced specific expressive gene with the function of phosphorus transfer in Pht1 family members. Conclusion The Pht1 gene family was screened and identified systematically from the genome of S. miltiorrhiza for the first time, in addition, the members with characteristic and biological function were found out, which will provide dependable background support and ideas reference for the further research on the role of Pht1 in the growth and development of S. miltiorrhiza.
ABSTRACT
The use of optimal regulatory sequences for simultaneous expression of the transgenes might play a significant role in engineering plants with increased disease and insect resistance.The plant expression vector pOMS-GUS,which contained the GUS gene under the control of a chimeric promoter based upon the mannopine synthase(mas)promoter and the octopine synthase(ocs)enhancer,was constructed.Used as control,another vector pMAS-GUS,carried the GUS gene driven by only the mas promoter.The two vectors were introduced into tobacco plants by Agrobacterium-mediated transformation.Fluorometric assays for GUS activity and reverse transcription-polymerase chain reaction(RT-PCR)analysis revealed that GUS gene expressed weakly with untreated transgenic tobacco while the level of GUS activity increased steadily after 1 h subjected to wounding.The expression of the mas and ocs/mas promoters was induced a further 1.8-fold and 5.7-fold,respectively.SA(1 mmol/L)or MJ(250 ?mol/L)treatment also caused a large induction of the ocs/mas chimeric promoter;And the application of SA in combination with MJ(1 mmol/LSA & 250 ?mol/L MJ)produced an additive effect that exceeded the wounding response.The results showed that the ocs/mas chimeric promoter is a strong inducible promoter that can be activated by various stresses.The chimeric promoter should have utility in development of disease and insect resistant transgenic crops.