In vitro plantlet regeneration system from rhizomes and mannose-binding lectin analysis of <Emphasis Type="Italic">Polygonatum cyrtonema</Emphasis> Hua. |
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Authors: | Qi Zhao Chuanfang Wu Wenguo Wang Shu Yuan Jinku Bao Fang Chen |
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Institution: | (1) School of Life Sciences, Sichuan University, 29 Wangjiang Road, 610064 Chengdu, People’s Republic of China;(2) Faculty of Biotechnology Industry, Chengdu University, 1 Shiling Street, Chengluo Road, 610106 Chengdu, People’s Republic of China |
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Abstract: | Polygonatum cyrtonema Hua. lectins (PCLs) were extracted from plantlets regenerated from rhizome explants of P. cyrtonema. Rhizome explants demonstrated a high frequency of callus induction (72.5%) and adventitious shoots differentiation (83.7%)
on Murashige Skoog (MS) medium supplemented with 2.0 mg l−1 2,4-dichlorophenoxyacetic acid and 1.0 mg l−1 6-benzyladenine. The adventitious shoots could root readily on 1/2 MS medium + 0.5 mg l−1 α-naphthaleneacetic acid and regenerate plantlets with a survival rate of 75.0%. Regenerated rhizomes were freeze-dried,
macerated and prepared for total RNAs and proteins extraction. The PCL gene was cloned and its expression level was measured by RT-PCR. Western blot using a lectin-specific antibody revealed a
similar amount in regenerated rhizomes compared to wild rhizomes, Furthermore, lectin derived from regenerated rhizomes retained
its ability to haemagglutinate rabbit blood cells. |
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