DOI: 10.3724/SP.J.1006.2018.00218

Acta Agronomica Sinica (作物学报) 2018/44:2 PP.218-226

Cloning and Expression Analysis of Galactosyltransferase Gene GhGalT1 Promoter in Cotton

Glycosytransferases (GTs) transfer an activated sugar donor to a specific acceptor to form glucosidic bond, which are regulated by various abiotic and biotic stresses, and may play a role in plant responses to changes in living conditions. In this study, a 539 bp fragment of GhGalT1 5'-flanking sequence was isolated from upland cotton Coker 312 by PCR, designated pGhGalT1. Analysis of pGhGalT1 sequence by PlantCARE revealed it contained not only putative CAAT box, TATA box sequence, but also MBS, HSE, TC-rich repeats, MYCCONSE and CGTCA-motif cis-acting element which involved in drought, heat, dehydration, defense and stress responsiveness. Thus, we constructed it into pBI101-GUS vector and formed pGhGalT1::GUS fusion expression vector (pBI101-pGhGalT1-GUS), then transferred the vector into Arabidopsis by the Agrobacterium-mediated floral dip method, and successfully obtained positive transgenic plants by screening test of resistance to kanamycin and PCR detection. Histochemical assay of T3 generation of transgenic Arabidopsis revealed that GUS activities were mainly accumulated in root tips of primary and lateral roots in 5-to 15-day-old seedlings, and less strongly in cotyledons and rosette leaves. The histochemical staining results and the assay of quantitative GUS activity and GUS gene expression under abiotic stresses and hormone treatments revealed that the GhGalT1 promoter was salt-/osmotic-/6-BA-/MeJA-/BL-inducible. These findings contribute to the selection of a suitable promoter for crop molecular improvement.

Key words:cotton (Gossypium hirsutum L.),glycosytransferase,promoter,cis-acting element,GUS histochemical staining,quantitative GUS assay

ReleaseDate:2018-03-08 10:13:39

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