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植物研究 ›› 2010, Vol. 30 ›› Issue (4): 448-454.doi: 10.7525/j.issn.1673-5102.2010.04.012

• 论文 • 上一篇    下一篇

过量表达水稻细胞质型OsAPXs基因提高转基因烟草的耐盐性

鲁振强1;刘大丽2,3;柳参奎4   

  1. 1.黑龙江大学生命科学学院生物化学与分子生物学重点实验室,哈尔滨 150080;2.黑龙江大学农作物研究院遗传育种重点实验室,哈尔滨 150080;3.东北林业大学森林植物生态学教育部重点实验室,哈尔滨 150040;4.东北林业大学盐碱地生物资源环境研究中心,哈尔滨 150040
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-07-20 发布日期:2010-07-20
  • 基金资助:
     

Elevated Salt Tolerance in Transgenic Tobacco Plants Expressing Cytosolic Ascorbate Peroxidase Genes from Rice(Oryza sativa L.)

LU Zhen-Qiang;LIU Da-Li;LIU Shen-Kui   

  1. 1.Key Laboratory of Biochemistry and Molecular Biology,College of Life Sciences,Heilongjiang University,Harbin 150080;2.Key Laboratory of Genetics and Breeding,Academy of Crop Sciences,Heilongjiang University,Harbin 150080;3.Key Laboratory of Forest Plant Ecology,the Ministry of Education of China,Northeast Forestry University,Harbin 150040;4.Alkali Soil Natural Environmental Science Center(ASNESC),Northeast Forestry University,Harbin 150040
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-07-20 Published:2010-07-20
  • Supported by:
     

摘要: 为了验证水稻(Oryza sativa L.)细胞质型APXs与细胞耐盐性的关系,实验分别将OsAPXaOsAPXb(基因登录号:D45423、AB053297)转化到烟草(Nictiana tabacum,N.plum)植株中。Southern结果表明,二基因分别整合到烟草的基因组;Northern分析表明,外源基因在转基因烟草中得到高效表达;在碳酸盐逆境下,T2代转基因植株与野生型对照相比,其APX活性呈现显著的提高,T2代品系的H2O2含量和叶片受害程度显著低于野生型;T2代品系分别在含有10 mmol·L-1 NaHCO3、5 mmol·L-1 Na2CO3的MS培养基上生长,根的生长受到抑制,叶片产生黄化;野生型烟草则难以存活。水稻细胞质型OsAPXs基因的过量表达提高了转基因烟草的耐盐性,揭示出OsAPXaOsAPXb在碳酸盐逆境应答过程中发挥着重要的作用。

关键词: 细胞质型抗坏血酸过氧化物酶, 碳酸盐, 水稻, 耐盐性, 转基因烟草

Abstract: To identify cell salt tolerant ability generated by cytosolic ascorbate peroxidase (cAPX) from rice(Oryza sativa L.), two encoding genes, OsAPXa and OsAPXb(GenBank accession nos. D45423 and AB053297, respectively) were introduced into tobacco(Nictiana tabacum, N.plum), respectively. Southern blot confirmed that the two genes were integrated into genome of N.tabacum, respectively. Moreover, Northern blot further revealed that foreign genes effectively expressed in vitro. OsAPXa/b transgenic N.tabacum progeny T1, T2 generation were identified and assayed further. Under the carbonate stress, the T2 independent homozygous transgenic lines showed the elevated activity of APX, the lower H2O2, and the lower ion leakage than that in wild type plants. Surprisingly, T2 generation seedlings could survive on MS medium containing either 10 mmol·L-1 NaHCO3 or 5 mmol·L-1 Na2CO3, just along with the retarded growth of root and yellowish leaf, whereas the wild type plants even could not grow under lower carbonate stress. Our results suggested that transgenic tobacco expressing cytosolic OsAPXs improved cell salt tolerance; both cytosolic OsAPXs played the crucial roles, especially in response to carbonate stress.

Key words: cytosolic ascorbate peroxidase(cAPX), carbonate, rice(Oryza sativa L.), salt tolerance, transgenic tobacco(Nictiana tabacum, N.plum)

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