Scientific Reports of the Faculty of Agriculture, Okayama University
Published by the Faculty of Agriculture, Okayama University
ONLINE ISSN : 2186-7755

メロンとキュウリのACC合成酵素遺伝子の構造的特徴とGUSトランジェントアッセイによるプロモーター活性

Shiomi, Shinjiro
Emi, Ogura
Yamamoto, Mikihiro Kaken ID publons researchmap
Nakamura, Reinosuke
Inaba, Akitsugu
Published Date
2001-02
Abstract
In orader to clarify the differences in regulatory mechanism(s) of the expression of 1-aminocyclopropane-1-carboxylate(ACC) synthase(ACS)genes during ripening in climacteric melon fruit and non-climacteric cucumber fruit, two sets of their genomic DNA sequences, including ca. 2kb of the promoter regions were determined, using PCR-based methods. ACS genes from melon (CMe-ACS1,2) were structurally similar to their counterpart from cucumber (CS-ACS1,2) in terms of size and position of exons and introns, restriction map, and sequencd identity of exeons, introns, proximal 5'-flanking promoter regions and splice junction. Southern blot analysis indicated that each ACS gene is present as a single copy. Transient promoter activity was investigated with two constructs of promoter-β-glucuronidase (GUS) fusion, CMe-ACS1:GUS and CS-ACS1:GUS, in mature mesocarp tissue of the two fruits. In melon disks, GUS activities conferred by the promoters of both CS-ACS1 (-2098~+42) and CMe-ACS-1(-2187~+67) were detected, which were decreased by treatment with 1-methylcyclopropene(1-MCP), an ethylene action inhibitor. In cucumber disks, however, only CS-ACS1:GUS was expressed; the activity was decreased with 1-MCP, and it was not affected by propylene. These results suggest that the promoter of CS-ACS1 has a potential to be expressed in the mesocarp tissue of ripening melon fruit, and that the difference in ethylene biosynthesis between melon and cucumber during ripening may be due to the difference in capability of forming trans-acting factor(s), not due to their ACS1 promoter activities.
Keywords
Cucumis sativus L.
Cucumis melo L
fruit ripening
GUS transient assay
ISSN
0474-0254
NCID
AN00033029