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寻找转化标记:对由稗草基因组DNA转化产生的水稻种质的差异组织蛋白质组进行分析。

Seeking transformation markers: an analysis of differential tissue proteomes on the rice germplasm generated from transformation of Echinochloa crusgalli genomic DNA.

作者信息

Zhao Caifeng, Zhao Bingran, Ren Yan, Tong Wei, Wang Jingqiang, Zhao Kang, Shu Shaokun, Xu Ningzhi, Liu Siqi

机构信息

Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101300, China.

出版信息

J Proteome Res. 2007 Apr;6(4):1354-63. doi: 10.1021/pr0605015. Epub 2007 Feb 28.

Abstract

The transformation of distally related genomic DNAs into plant was proposed as a novel technique to breed new cultivars. For example, a restorer rice line, RB207, was successfully developed and stabilized through the transformation of genomic DNAs of Echinochloa crusgalli (E. crusgalli) into a rice line, R207. Although the phenotypes of this variant line are apparently different from its receptor, the molecular bases are not elucidated yet. Herein, we have systematically studied the differential proteomes from the tissues of E. crusgalli, R207, and RB207 in an attempt to find an explanation regarding the phenotypic changes of RB207. The 2-DE method was employed to separate the leaf and embryo proteins of these plants followed by protein identification with mass spectrometry. In the leaf, 953 +/- 15, 1084 +/- 11, and 1091 +/- 11 silver-stained spots were detected, whereas in the embryo, 986 +/- 3, 884 +/- 10, and 892 +/- 14 spots were found from E. crusgalli, R207, and RB207, respectively. In comparison to the 2-DE images of the two rice lines, which showed many similarities, the ones of the E. crusgalli and rice were found to be so different that they were incomparable. There were some differentially expressed 2-DE spots between the two rice cultivars, 72 in leaf and 53 in embryo, respectively. The results of protein identification suggested that, regardless of leaves or embryos, none of the E. crusgalli genes were encoded in the new rice cultivar, RB207. The fact that 60% of the differentially expressed spots between R207 and RB207, however, were verified as the proteins involved in metabolism and photosynthesis makes a rather convincing argument that the DNA fragments transferred from E. crusgalli to rice are responsible for exerting the unknown influence to the expression of rice genes.

摘要

将远缘基因组DNA导入植物被提议作为一种培育新品种的新技术。例如,通过将稗草(Echinochloa crusgalli)的基因组DNA导入水稻品系R207,成功培育并稳定了一个恢复系水稻品系RB207。尽管这个变异品系的表型与受体明显不同,但其分子基础尚未阐明。在此,我们系统地研究了稗草、R207和RB207组织的差异蛋白质组,试图找到关于RB207表型变化的解释。采用双向电泳(2-DE)方法分离这些植物的叶片和胚蛋白质,随后用质谱进行蛋白质鉴定。在叶片中,稗草、R207和RB207分别检测到953±15、1084±11和1091±11个银染点,而在胚中,分别发现986±3、884±10和892±14个点。与两个水稻品系的双向电泳图像显示出许多相似性不同,稗草和水稻的双向电泳图像差异很大,无法进行比较。两个水稻品种之间有一些差异表达的双向电泳点,叶片中有72个,胚中有53个。蛋白质鉴定结果表明,无论在叶片还是胚中,新水稻品种RB207中均未编码稗草基因。然而,R207和RB207之间60%的差异表达点被证实为参与代谢和光合作用的蛋白质,这有力地证明了从稗草转移到水稻的DNA片段对水稻基因的表达产生了未知影响。

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