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Acquisition and Salt-tolerance Evaluation of Transgenic Tobacco (Nicotiana tabacum) with β-carotene Hydroxylase Gene (chyb) from Dunaliella viridis |
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Abstract β-carotene hydroxylase (chyb) catalyzes the conversion of β-carotene to zeaxanthin through β-cryptoxanthin in the pathway of carotenoid biosynthesis. The salt-tolerance of chyb gene in Dunaliella viridis has important research value in the aspect of plant resistance. chyb gene (GenBank accession No. JN118489) cloned in previous research was used to construct the plant expression vector pCAMBIA3301-chyb and transferred into tobacco (Nicotiana tabacum) by Agrobacterium tumefaciens LBA4404, and 16 regenerated tobaccos resisted to glufosinate ammonium were obtained. The results of PCR and RT-PCR indicated that chyb gene had been transferred successfully into 13 of them. Besides, salt tolerance of transgenic tobacco was detected through measuring plant height, leaf area, chlorophyll content, leaf relative conductivity and proline content. The result showed that plant height, leaf area and chlorophyll content were extremely significantly increased (P<0.01) compared with the control group. The average height of transgenic plant was 37.82 cm, which was 1.37 times higher than that of the control group. Transgenic tobaccos were treated with different concentrations of NaCl. The results demonstrated that the average survival rate of transgenic tobacco was 1.58 times higher than that of control group under 100 mmol/L NaCl stress. Transgenic tobacco could grow under the treatment of 200 mmol/L NaCl, while the growth of control tobacco was inhibited. The leaf relative conductivity increased slightly, and the proline content of transgenic plants were improved extremly significantly than the control group (P<0.01). It was concluded that chyb gene could improve salt-tolerance in transgenic tobacco plants and all the results help to lay the foundation for further exploring salt-tolerance of chyb gene.
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Received: 14 January 2015
Published: 09 July 2015
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