Cloning, space-time expression and bioinformatics analysis of ACS3 gene from different species of Lonicera macranthoides / 中草药

ABSTRACT

Objective To clone ACS3 gene from wild type and ‘Xianglei’ cultivar of Lonicera macranthoides, respectively, followed by bioinformatics analysis and detection of spatio-temporal expression pattern. Methods Unigene sequence which is highly homologous with ACS protein from the transcriptome database of L. macranthoides was screened, the primers were designed based on it to amplify the full length of the Unigene by qRT-PCR and RACE techniques; Bioinformatics tools were used to analyze and identify the physicochemical property, conserved domain and gene homology of ACS3 proteins; Finally, qRT-PCR technique was used to detect the gene expression patterns of different species of L. macranthoides. Results Lm-ACS3 (GenBank MH724196) and Lm-XL-ACS3 (GenBank MH724197) were isolated from wild type and ‘Xianglei’ cultivar of L. macranthoides, respectively. The length of open reading frame (ORF) were all 1 452 bp, encoding 483 amino acids, containing the conserved Aminotran_1_2 structural domain, which were highly similar to the ACC synthase of other plants; And qRT-PCR results showsed that the expression quantity of ACS3 gene in wild L. macranthoides changed significantly at different blossoming stages, the overall trend was upward from flowering stage 3, while the expression difference between the flowering stages was relatively small in “Xianglei” cultivar. Conclusion Lm-ACS3 and Lm-XL-ACS3 gene were separately obtained from L. macranthoides and L. macranthoides ‘Xianglei’ cultivar, the expression patterns of ACS3 in this two varieties were different; It’s speculated that ACS3 gene might be a possible functional gene that causing different phenotypes of two strains of L. macranthoides, it provides theoretical basis for further verifying the biological function of ACS3 gene in regulating flower’s bud duration and phenotype.