转盐生杜氏藻β-胡萝卜素羟化酶基因(chyb)烟草的获得及耐盐性鉴定

Abstract
Figure/Table
References
Related Citation (15)

Download: PDF (1683 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

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.

Key words： β-carotene hydroxylase gene (chyb) Vector construction Transformation Salt tolerance

Received: 14 January 2015 Published: 09 July 2015

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	XU Feng
	GONG Yi-Fu
	LIU Lin
	ZHOU Jing
	XUN Jiao-Lu
	YU He-Yu

Cite this article:

XU Feng,GONG Yi-Fu,LIU Lin, et al. Acquisition and Salt-tolerance Evaluation of Transgenic Tobacco (Nicotiana tabacum) with β-carotene Hydroxylase Gene (chyb) from Dunaliella viridis [J]. , 2015, 23(9): 1149-1156.

URL:

http://journal05.magtech.org.cn/Jwk_ny/EN/ OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2015/V23/I9/1149

丁林云, 张微, 王晋成, 田亮亮, 李妮娜, 郭琪, 杨淑明, 何曼林, 郭旺珍, 2014. 过量表达棉花GhSAP1提高转基因烟草的耐盐性[J]. 中国农业科学, 47(8): 1458-1470.（Ding L Y, Zhang W, Wang J C, Tian L L, Li N N, Guo Q, Yang S M, He M L and Guo W Z, 2014. Overexpression of a Gossypium hirsutum Stress Associated Protein Gene (GhSAP1) Improves Salt Stress Tolerance in Transgenic Tobacco[J]. Scientia Agricultura Sinica, 47(8): 1458-1470.）
冯唐锴, 李思光, 汪艳璐, 罗玉萍, 2007. 南丰蜜橘β-胡萝卜素羟化酶基因的克隆和序列分析[J]. 西北植物学报, 27(2): 238-243.（Feng T K, Li S G, Wang Y L and Luo Y P, 2007. Cloning and Sequence Analysis of β-carotene Hydroxylase Gene from Nanfeng Orange (Citrus reticulate Blanco var.kinokuni (Tanaka) H. H. Hu)[J]. Acta Botanica Boreali- Occidentalia Sinica, 27(2): 238-243.）
贺庆华, 张庆莲, 2008. 盐藻的耐盐机理的研究. 西南民族大学学报(自然科学版)[J], 34(3): 487-490.（He Q H and Zhang Q L, 2008. Research on the Salt tolerance mechanism of Dunaliella[J]. Journal of SouthWest University for Nationalities (Natural Science Edition), 34(3): 487-490.）
季静, 杨海兰, 王罡, 武卫党, 金超, 吴疆, 2014. AtchyB基因过表达对转基因洋桔梗抗氧化性的影响. 天津大学学报(自然科学与工程技术版)[J], 47(8): 735-740.（Ji J, Yang H L, Wang G, Wu W D, Jin C and Wu J, 2014. Effect of Over-Expression of AtchyB on the Oxidation Resistance of Transgenic Eustoma Grandiflorum. Journal of Tianjin University (Science and Technology)[J], 47(8): 735-740.）
李合生, 主编, 2000. 植物生理生化实验原理和技术[M]. 中国, 北京：高等教育出版社,（Li H S, 2000. The experiment principle and technique on plant physiology and biochemistry[M]. China, Beijing: Higher Education Press.）
吕靖, 蒿若超, 张文英, 武美燕, 2012. 烟草转基因研究进展[J]. 黑龙江农业科学, 2012(7): 148-152. （Lv J, Hao R C, Zhang W Y and Wu M Y, 2012. Progress in Transgenic Tobacco[J]. Heilongjiang Agricultural Sciences, 2012(7): 148-152）
孙辉, 2005. 盐藻的生长特性及逆境下β-胡萝卜素积累规律和机理研究[D]. 硕士学位论文. 四川大学.（Sun H, 2005. Biological Properties, β-carotene accumulation rule and mechanism under the stress of Dunaliella salina[D]. Thesis for M.S., Sichuan University.）
舒展, 张晓素, 陈娟, 陈根云, 许大全, 2010. 叶绿素含量测定的简化[J]. 植物生理学通讯, 46(4): 399-402.（Shu Z, Zhang X S, Chen J, Chen G Y and Xu D Q, 2010. The Simplification of Chlorophyll Content Measurement[J]. Plant Physiology Communications, 46(4): 399-402.）
王冬梅, 王玮蔚, 孙雪, 徐年军, 2014. 一株杜氏藻的分子鉴定与耐盐特性[J]. 植物生理学报, 50(3): 315-323.（Wang D M, Wang W W, Sun X and Xu N J, 2014. Molecular Identification and Halophilic Charactristics of a Strain of Dunaliella D3[J]. Plant Physiology Journal, 50(3): 315-323.）
王敏华, 丁明全, 陈爱群, 张毓婷, 戎均康, 2014. 过量表达棉花GaSus3基因增强拟南芥的耐盐性[J]. 植物生理学报, 50(7): 901-908.（Wang M H, Ding M Q, Chen A Q, Zhang Y T, Rong J K, 2014. Over Expression of a Cotton GaSus3 Gene Enhancing the Salt Stress Tolerance of Arabidopsis thaliana[J]. Plant Physiology Journal, 50 (7): 901-908.）
王先艳, 单晓映, 杨爱芳, 张举仁, 2003. 植物表达载体构建和转基因烟草抗除草剂及耐盐性的分析[J]. 农业生物技术学报, 11(6): 554-560.（Wang X Y, Shan X Y, Yang A F and Zhang J R, 2003. Construction of Plant Expression Vector and Analysis of Herbicide-resistance and Salt-tolerance of Transgenic Tobacco[J]. Journal of Agricultural Biotechnology, 11(6): 554-560.）
杨劲松, 2008. 中国盐渍土研究的发展历程与展望[J]. 土壤学报, 45(5): 837-845.（Yang J S, 2008. Development and prospect of the research on salt-affected soils in China[J]. Acta Pedologica Sinica, 45(5): 837-845.）
章丽, 龚一富, 刘晓丹, 张文青, 2013. 盐生杜氏藻β-胡萝卜素羟化酶基因(chyb)的克隆及表达分析[J]. 农业生物技术学报, 21(8): 920-930.（Zhang L, Gong Y F, Liu X D and Zhang W Q, 2013. Identification and expression analysis of beta-carotene hydroxylase from Dunaliella salina[J]. Journal of Agricultural Biotechnology, 21(8): 920-930.）
张俊莲, 王丽, 秦舒浩, 司怀军, 刘玉汇, 王蒂, 2011. 利用拟南芥rd29A启动子驱动AtNHX1基因提高烟草耐盐性[J]. 农业生物技术学报, 19(4): 669-676.（Zhang J L, Wang L, Qin S H, Si H J, Liu Y H and Wang D, 2011. The AtNHX1 Gene Driven by Stress-inducible rd29A Promoter from Arabidopsis thaliana Can Increase Tobacco Saline Tolerance[J]. Journal of Agricultural Biotechnology, 19(4): 669-676.）
郑凯静, 胡章立, 黎双飞, 雷安平, 王潮岗, 2007. 雨生红球藻β-胡萝卜素羟化酶基因的克隆与序列分析[J]. 水生生物学报, 3l(5): 666-670.（Zheng K J, Hu Z L, Li S F, Lei A P and Wang C G, 2007. Cloning and Sequence of β-carotene hydroxylase Gene from Haematococcus Pluvialis[J]. Acta Hydrobiologica Sinica, 3l(5): 666-670.）
Cui H L, Yu X N, Wang Y, Cui Y L, Li X Q, Liu Z P and Qin S, 2014.Gene cloning and expression profile of a novel caroteniod hydroxylase (CYP97C) from the green alga Haematococcus pluvialis[J]. Journal of Applied Phycology, 26: 91-103.
Davison P A, Hunter C N and Horton P, 2002. Overexpression of betacarotene hydroxylase enhances stress tolerance in Arabidopsis[J]. Nature, 418: 203-206.
Du H, Wang N, Cui F, Li X, Xiao J and Xiong L, 2010. Characterization of the β-carotene hydroxylase gene DSM2 conferring drought and oxidative stress resistance by increasing xanthophyIIs and abscisic acid snthesis in rice[J]. Plant Physiology, 154 (3): 1304-1318.
G?tz T, Sandmann G and R?mer S, 2002. Expression of a bacterial carotene hydroxylase gene (crtZ) enhances UV tolerance in tobacco[J]. Plant Molecular Biology, 50(1): 127-140.
Kato M, Ikoma Y and Matsumoto H, 2004. Accumulation of carotenoids and expression of carotenoid biosynthetic genes during maturation in citrus fruit[J]. Plant Physiology, 134(2): 824-837.
Kim S H, Ahn Y O, Ahn M J, Lee H S and Kwak S S, 2012. Down-regulation of β-carotene hydroxylase increases β-carotene and total carotenoids enhancing salt stress tolerance in transgenic cultured cells of sweetpotato. Phytochemistry[J], 74:69-78.
Liu J and Zhu J K, 1997. Proline accumulation and salt-stress induced gene expression in a salt-hyper sensitive mutant of Arabidopsis[J]. Plant Physiology, 11(4): 591-596.
Nathalie V and Christian H, 2008. Proline accumulation in plants: a review[J]. Amino Acids, 35: 753-759.
Oren A, 2005.A hundred years of Dunaliella research: 1905-2005[J]. Saline Systems, 1: 2.
Rania B S, Nabil Z, Walid B R, Jalel A, Donaldo M, Emmanuel G and Afif H, 2010. Improved drought and salt stress tolerance in transgenic tobacco overexpressing a novel A20/AN1 zinc-finger“AlSAP” gene isolated from the halophyte grass Aeluropus littoralis[J]. Plant Molecular Biology, 72: 171-190.
Sun Y S, Chen S, Huang H J, Jiang J, Bai S and Liu G F, 2014. Improved salt tolerance of Populus davidiana × P. bolleana overexpressed LEA from Tamarix androssowii[J]. Journal of Forestry Research, 25(4): 813-818.
Stephen F C and Cory S, 2012. Genetic modification; the development of transgenic ornamental plant varieties[J]. Plant Biotechnology Journal, 10: 891-903
Yu B Y, Derek J L, Ulrike A S and Abdelali H, 2007. Characterization of a β-carotene hydroxylase of Adonis aestivalis and its expression in Arabidopsis thaliana[J]. Planta, 226: 181-192.
Zhao Q, Wang G, Ji J, Jin C, Wu W D and Zhao J, 2014. Over-expression of Arabidopsis thaliana β-carotene hydroxylase(chyB) gene enhances drought tolerance in transgenic tobacco[J]. Journal of Plant Biochemistry and Biotechnology, 23(2): 190-198.