Establishment of an efficient <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated leaf disc transformation of <Emphasis Type="Italic">Thellungiella halophila</Emphasis> |
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Authors: | Hong-Qing Li Jie Xu Lei Chen Mei-Ru Li |
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Institution: | (1) Guangdong Provincial Key Lab of Biotechnology for Plant Development, South China Normal University, Guangzhou, 510631, People’s Republic of China;(2) South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, 510650, People’s Republic of China |
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Abstract: | Thellungiella halophila is a salt-tolerant close relative of Arabidopsis, which is adopted as a halophytic model for stress tolerance research. We established an Agrobacterium tumefaciens-mediated transformation procedure for T. halophila. Leaf explants of T. halophila were incubated with A. tumefaciens strain EHA105 containing a binary vector pCAMBIA1301 with the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene as a reporter gene. Following
co-cultivation, leaf explants were cultured on selective medium containing 10 mg l−1 hygromycin and 500 mg l−1 cefotaxime. Hygromycin-resistant calluses were induced from the leaf explants after 3 weeks. Shoot regeneration was achieved
after transferring the calluses onto fresh medium of the same composition. Finally, the shoots were rooted on half strength
MS basal medium supplemented with 10 mg l−1 hygromycin. Incorporation and expression of the transgenes were confirmed by PCR, Southern blot analysis and GUS histochemical
assay. Using this protocol, transgenic T. halophila plants can be obtained in approximately 2 months with a high transformation frequency of 26%. |
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Keywords: | Thellungiella halophila Callus Plant regeneration Transformation |
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