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| Cloning and Expression Analysis of ACC Oxidase Gene ( HaACO1) from Sunflower (Helianthus annuus L.) |
| SUN Rui-fen, ZHANG Yan-fang, GUO Shu-chun, YU Hai-feng, LI Su-ping, QIAO Hui-lei, NIE Hui, AN Yu-lin |
| Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences, Huhhot 010031, China |
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Abstract Objective: In an effort to understand the physiological functions of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase and exploit ACO gene more profitably, an ACC oxidase gene (HaACO1) was cloned from sunflower and analyzed by bioinformatics method, and expression pattern of HaACO1 gene in sunflower was revealed under salt stress. Method: The full length cDNA sequence of ACO gene in sunflower was obtained by RT-PCR and 5'/3'RACE methods based on the sequence of 4-4-7TDF (accession No. KM823963), a gene fragment of ACC oxidase from Inner Mongolia Sunflower Hybrid No.4. The cDNA sequence and its encoded protein was analyzed by bioinformatics method. The genemic DNA (gDNA) of ACO sequence was cloned by PCR. The expression level of HaACO1 gene in roots, hypocotyl and leaves under NaCl stress was tested by quantitative Real-time PCR (qRT-PCR). Results: The full length sequence of HaACO1 gene in sunflower was 1 135bp, and its open reading frame (ORF) was 942bp, then the protein sequence analysis results showed that the HaACO1 gene encoded 313 amino acids with molecular weight 35.84kDa and pI 5.13. The sequence accession No. in GenBank was KP966508. The nucleotide sequence and the amino acid sequence of HaACO1 was highly homologous to other species (75%~83%, 77%~88% respectively). The full length of the coding region of gDNA was 1 018bp, comprised of two exons separated by one intron. The sequence has been submitted to GenBank (Accession No. KP988289). The results of qRT-PCR showed that the expression level of HaACO1 had specific expression differences in different organs. Different concentration of NaCl treatmet or different induction time with NaCl also lead to the specific expression of HaACO1 in sunflower. Conclusion: HaACO1 belongs to ACO gene family in plants, and this gene has distinctive expression pattern when plants response to salt stress.
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Received: 02 April 2015
Published: 25 September 2015
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