Functional analysis and localization of auxin transporters in Momordica charantia L.
Date Issued
2009
Date
2009
Author(s)
Shih, Wan-Yun
Abstract
Bitter gourd is one of the most important vegetables in Taiwan whose growth and development are mediated by plant hormone auxin. Recent studies showed that the distributions and transportation of auxin between cells are affected by the auxin carriers. Plasmids for over-expression and antisense auxin carriers from bitter gourd were constructed. Subsequently, the plasmids were introduced into tobaccos by Agrobacterium-mediated transformation. Each five transgenic plants for 35S::McPIN3 and 35S::anti-McPIN3 were confirmed by GUS staining and Southern analysis. McPIN3 was expressed in transgenic tobaccos analyzed by reverse transcription polymerase chain reaction and Northern hybridization. In contrast to the untranformed plant, the height, weight of fresh leaves and root length in transgenic plants were all much heiger. Besides, the transgenic plants bloomed earlier. The cell divisions of transgenic tobacco plants were much more active than the untranform tobacco plants by flow cytometry analysis. Forthermore, the cell numbers of vascular bundles in shoot of transgenic tobacco through transverse section were more than the untranformed tobacco observed under the anatomic microscopy. After gene gun transformation with gene fusion plasmid, green fluorescence was observed on the plasma membrane in onion epidermal cells under the scanning confocal microscopy. The cell membrane in lateral root cap, cortex, epidermis, endodermis, vasculature, columella and quenscent centre ofransgenic tobaccos carring the auxin efflux transporters McPIN1 expressed green fluorescence proteins examined under scanning confocal microscopy. Auxin efflux carriers McPIN3 expressed on the cell membranes of epidermis, endodermis, columella, cortex and vasculature of transgenic tobaccos. Auxin influx carriers McLAX1 localized on the cell membranes of endodermis, cortex, vasculature and quenscent centre. Auxin influx carriers McLAX2 expressed on the cell membranes of endodermis, cortex, vasculature and epidermis. Moreover, the strengths of green fluorescence in transgenic plant root are much stronger in apical and basal cell membrane than the lateral layers which may due to the polar localization of auxin transporters.
Subjects
auxin transporters
polar transport
cell membrane
green fluorescent protein
Southern analysis
gene gun
scanning confocal microscopy
Type
thesis
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