<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of protocorm-like bodies in <Emphasis Type="Italic">Cattleya</Emphasis> |
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Authors: | Lin Zhang Dong Poh Chin Masahiro Mii |
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Institution: | (1) College of Agriculture and Bio-Engineering, Tianjin University, 300072 Tianjin, Pepole’s Republic of China;(2) Jinling Institute of Technology, 210038 Nanjing, Pepole’s Republic of China |
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Abstract: | A protocol for in vitro induction of crape myrtle tetraploids using nodes from in vitro-grown shoots (2n = 48) was established. Nodal buds were excised from in vitro-grown shoots, maintained on proliferation medium containing
Murashige and Skoog medium supplemented with 4.44 μM 6-benzyladenine , 0.54 μM α-naphthaleneacetic acid, and treated with
a range of concentrations of colchicine under three different conditions. Nodal bud explants treated in liquid proliferation
medium supplemented with either 15 or 20 mM colchicine for 24 h turned necrotic and died; whereas, those cultured on solid
proliferation medium supplemented with either 125 or 250 μM colchicine for 30 days survived, but no tetraploid plants were
obtained. However, when explants were cultured in liquid proliferation medium containing 250, 500 or 750 μM colchicine for
10 days, tetraploid plants (2n = 96) were obtained. Incubation of explants in medium containing 750 μM colchicine promoted the highest frequency of survival
(40%) of explants and of recovered tetraploids (60%). Morphological and anatomical characteristics of leaves, including leaf
index, stomata size and number, stomata index (length/width), and number of chloroplasts in guard cells correlated with ploidy
of crape myrtle plants. The number of chloroplasts in guard cells of stomata was a stable and reliable marker in discriminating
plants of different ploidy levels. Chromosome counts and flow cytometry confirmed these findings. |
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