Cytokinins play an important role in regulating plant growth and development. The cytokinin gene, isopentenyl transferase (ipt), was placed under the control of the ACC oxidase promoter from the LEACO1 gene from Lycopersicon esculentum and introduced into Nicotiana tabacum (cv. Havana) and chrysanthemum (Dendranthema × grandiflorum `Iridon'). Transformants were confirmed by PCR reaction and Southern blot and analyzed for phenotypical changes under both greenhouse and growth chamber conditions. With both species, LEACO1-ipt transgenic plants displayed a wide range of vegetative and generative phenotypes. With plants growing in the vegetative state, some LEACO1-ipt transgenic lines appeared similar to the non-transgenic wild-type cultivars while other lines showed excessive lateral branch development and short internodes. With plants grown under generative conditions, several LEACO1-ipt transgenic lines showed a 2 to 10-fold increase in the number of flower buds relative to the wild-type cultivars. With chrysanthemum, dramatic increases in bud count were observed on transgenic lines that otherwise displayed a morphology similar to the non-transgenic lines. Analysis of ipt expression indicated a marked change in gene expression between the most extreme phenotypes observed in this study. LEACO1-ipt lines that express normal vegetative development but increased flower bud counts appear to have great potential for ornamental crop improvement.
Richard J. McAvoy*, Mariya V. Khodakovskaya, Hong Liu, and Yi Li
Mariya V. Khodakovskaya, Richard J. McAvoy*, Keming Luo, and Yi Li
The bacterial isopentenyl transferase (ipt) gene involved in cytokinin biosynthesis was fused with a promoter from the serine proteinase inhibiter (win3.12) gene of Populus x generosa and introduced into Nicotiana tabacum (cv. Havana). Transformants were confirmed by PCR reaction and Southern blot analysis, and then analyzed for phenotypic characteristics. RT-PCR analysis detected transcripts of the ipt gene following the wounding of win3.12:ipt transgenic plants. In win3.12: ipt transgenic plants, lateral shoot number and the diameter of lateral branches that developed following apical shoot removal increased relative to wild-type plants. However, the increase in cytokinin production following wounding appeared to be short lived. The potential utility of this construct in enhancing lateral branching in ornamental crops will be discussed.
Mariya V. Khodakovskaya, Richard J. McAvoy*, Hao Wu, and Yi Li
It has been reported that constitutive expression of the fatty acid desaturase enzyme increased the trienoic fatty acid content of thylakoid membranes in transgenic tobacco, allowing the membranes to remain fluid under cold conditions. While increased cold tolerance resulted from this genetic modification, plants with a constitutively expressed desaturase enzyme would not be particularly well suited for growth under warm temperatures. To increase the ability of plants to tolerate prolonged cold-storage and still perform under greenhouse production conditions (25 °C), a unique cold-inducible genetic construct was cloned and tested. The FAD7 gene, which encodes an omega-3-fatty acid desaturase enzyme, was put under the control of a cold-inducible promoter (cor15a) from Arabidopsis thaliana. Transgenic petunia plants (cv, Marco Polo Odyssey) harboring cor15a:FAD7 were established and conformed by PCR and Southern analysis. Therefore in our study, FAD7 gene expression was induced by exposure to cold temperatures and down regulated under normal growing conditions. RT-PCR indicated a marked increase in FAD7 expression between transgenic plants exposed to a short (3 days) cold treatment prior to long-term cold storage and those that did not receive a cold induction treatment. Transgenic and wild-type plants were induced in cold (3 °C) for 3 days, returned for normal greenhouse conditions for 5 days and then subjected 3 weeks of continuous cold storage. It was observed that two out of eight transgenic lines showed superior cold tolerance relative to wild-type petunia plants. Additionally, plants that showed cold tolerance completely recovered; growing and flowering normally when returned to the 25 °C greenhouse conditions.