Isolation and characterization of the CYP71D16 trichome-specific promoter from Nicotiana tabacum L.

Abstract

Trichomes are specialized epidermal cells that produce secretions that are thought to provide a first line of defence against pests and pathogens. Many trichome-secreted compounds are used commercially as flavourings, medicines, etc. Described here is the cloning and characterization of the promoter of a tobacco trichome-specific P450 gene, CYP71D16. This promoter is shown to direct the specific expression of the reporter gene, beta-glucuronidase (GUS), in glandular trichomes of Nicotiana tabacum cv. T.I. 1068 at all developmental stages. With the full promoter, GUS activity was predominantly in the gland cell, with less in the stalk cell adjacent to the gland, and in lower stalk cells. GUS staining was also observed in the most distal trichome stalk cells of non-glandular trichomes found on variety T.I. 1112. Promoter deletion analysis revealed that the region from -223 to +111 bp is sufficient to direct trichome-specific expression, but not strong gland expression. Examination of the literature suggests that this is the first characterized trichome-specific-promoter shown to function at all stages of plant development. This promoter may provide efficient bioengineering to enhance pest and pathogen resistance, and for molecular farming based on the trichome gland system.

Photosynthetic Trichomes Contain a Specific Rubisco with a Modified pH-Dependent Activity

“…2A). CYP71D16, which has been demonstrated to be trichome specific (Wang et al, 2002), was used as a positive control, and ATP2, coding for the b-subunit of mitochondrial ATP synthase, was used as a loading control (Boutry and Chua, 1985). A strong NtRbcS-T signal was observed in the trichome sample.…”

Trichome Specific Expression: Promoters and Their Applications

“…The first tobacco trichome specific promoter identified was that of the CYP71D16 gene, which encodes the CBT-ol hydroxylase (Wang et al, 2002). The gene was itself identified through subtractive cDNA library construction which was followed by the cloning of the promoter.…”

Functional analysis of a cotton glucuronosyltransferase promoter in transgenic tobaccos

“…Several motifs including AGAAA, AAATGA and GTGA, and six MYB npg response elements are also present in the promoter region ( Table 1). The AGAAA, AAATGA and GTGA motifs were reported to relate to pollen grain development [32][33][34], while MYB response elements were related to trichome development [35,36]. Additionally, the promoter region also contains several potential motifs with homology to inducible regulatory elements such as the W-box and AuxRE ( Table 1).…”