Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (9): 114-124.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1482

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Expression Analysis and Functional Verification of the HaACO1 Gene in Sunflower

SUN Rui-fen1(), ZHANG Yan-fang2, NIU Su-qing1, GUO Shu-chun1, LI Su-ping1, YU Hai-feng1, NIE Hui1, MOU Ying-nan1   

  1. 1. Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Huhhtot 010031
    2. College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Huhhtot 010011
  • Received:2020-12-07 Online:2021-09-26 Published:2021-10-25

Abstract:

To exploit genes encoding ACC oxidase in sunflower more profitably,we studied one ACC oxidase gene HaACO1(GenBank accession number:KP966508)cloned from previous salt-induced sunflower plants. The expression patterns of HaACO1 under different stress types were investigated. Additionally,the overexpression of this gene in tobacco was carried out. The results showed that the expression of HaACO1 in sunflower was induced by pathogens,mechanical damage,low temperature,NaCl and salicylic acid stress,and expression patterns were distinct under these types of stress. HaACO1 expressed in the roots,hypocotyls and leaves of sunflower plants,with the highest expression in the leaves and the lowest expression in the roots. By taking advantage of a transient expressive vector,the subcellular localization of HaACO1-GFP was found to be largely in the cytoplasm of onion epidermal cells. A plant vector of expressing HaACO1 was constructed to analyze the overexpressing of HaACO1. The results indicated the decrease in green color of leaves of transgenic tobacco was smaller than that of the wild plants placed on differentiation medium with NaCl,while the leaf differentiation was higher than that of the wild plants. Under low temperature,drought and NaCl stresses,the relative expression of HaACO1 in the transgenic tobacco plants was higher than that of the wild plants. During NaCl stress,the contents of total soluble protein,proline and chlorophyll and the activities of peroxidase and superoxide dismutase in the transgenic tobacco plants increased;and the expression of a key gene P5CS for proline biosynthesis and three antioxidant related genes POD,MnSOD and CuZnSOD were all up-regulated. These results indicate that overexpression of HaACO1 in tobacco plants enhances the plants’ tolerance to high concentrations of NaCl,providing foundations for further understanding the molecular mechanism of salt tolerance in sunflower,and laying a foundation for using this gene in crop stress resistance improvement.

Key words: sunflower, HaACO1, stress response, subcellular localization, overexpression