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A 920 bp fragment of the ACC oxidase gene promoter from tomato (LEACO1) was used to drive GUS gene expression. The LEACO1 _0.92kb fragment contained two stress-responsive short motifs; a 10 bp TCA motif (5'-TCATCTTCTT-3') twice (allowing two substitutions) and an 8 bp element (5'-AA/TTTCAAA-3') once. The TCA motif is found in over 30 stress- and pathogen-inducible genes while the 8 bp element is necessary for ethylene-response in the carnation GST1 and the tomato E4 gene promoters. Previously in chrysanthemum, cytokinin regulation with LEACO1 _0.92kb produced dramatic increases in lateral branching and bud initiation. Tobacco plants carrying LEACO1 _0.92kb –GUS were used to examine the response of the LEACO1 _0.92kb promoter to various hormones and hormone inhibitors. GUS activity in LEACO1 _0.92kb –GUS plants was detected in leaves and stems, but not roots. High expression was detected in shoots with the apical bud intact, but GUS activity decreased with the apical bud removed. Applying IAA to the shoot apex after removing the apical bud, restored GUS activity. However, the IAA transport inhibitor TIBA reduced GUS activity in shoots with intact apical buds, and in IAA-treated shoots with excised buds. In shoots with excised apical buds, GUS activity increased when the ethylene precursor ACC was applied, but decreased in intact shoots when the ethylene biosynthesis inhibitor AOA was applied. These data suggest that auxins produced in the apical meristem are capable of regulating LEACO1 _0.92kb activity, probably through auxin-induced ethylene biosynthetic pathway activity.

Euonymus alatus (Thunb.) Sieb., commonly known as “burning bush,” is an extremely popular landscape plant in the United States as a result of its brilliant showy red leaves in fall. However, E. alatus is also seriously invasive because of its prolific seed production and effective seed dispersal by birds. Thus, development of sterile, non-invasive, seedless triploid E. alatus is in high demand. In this article, we report successful production of triploid E. alatus using endosperm tissues as explants. In our study, ≈50% of immature endosperm explants and 14% of mature endosperm explants formed compact, green calli after culture in the dark for 8 weeks and then under light for 4 weeks on Murashige and Skoog (MS) medium supplemented with 2.2 μM BA and 2.7 μM α-naphthaleneacetic acid (NAA). Approximately 5.6% of the immature endosperm-derived calli and 13.4% of mature endosperm-derived calli initiated shoots within 8 weeks after they were cultured on MS medium with 4.4 μM benzyladenine (BA) and 0.5 μM indole-3-butyric acid (IBA). Eighty-five percent of shoots rooted after culture on woody plant medium (WPM) containing 4.9 μM IBA for 2 weeks and then on hormone-free WPM medium containing 2.0 g·L⁻¹ activated charcoal for 4 weeks. Eight independently regenerated triploid plants have been identified. Triploid plant regeneration rates observed were 0.42% from immature endosperm explants and 0.34% from mature endosperm explants, respectively, based on the number of endosperm explants cultured. Because triploid plants cannot produce viable seeds, and thus are sterile and non-invasive, some triploid E. alatus plant lines reported here can be used to replace the currently used invasive counterparts. Chemical names used: benzyladenine (BA), indole-3-butyric acid (IBA), and α-naphthaleneacetic acid (NAA).