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通过农杆菌接种小麦幼苗基部产生的抗白粉病转基因小麦后代。

Transgenic wheat progeny resistant to powdery mildew generated by Agrobacterium inoculum to the basal portion of wheat seedling.

作者信息

Zhao Tong-Jin, Zhao Shuang-Yi, Chen Hui-Min, Zhao Qing-Zhen, Hu Zan-Min, Hou Bing-Kai, Xia Guang-Min

机构信息

School of Life Sciences, Shandong University, Jinan, 250100, PR China.

出版信息

Plant Cell Rep. 2006 Nov;25(11):1199-204. doi: 10.1007/s00299-006-0184-8. Epub 2006 Jun 14.

Abstract

To improve the transformation efficiency of wheat (Triticum aestivum L.) mediated by Agrobacterium tumefaciens, we explored the possibility of employing the basal portion of wheat seedling (shoot apical meristem) as the explants. Three genotypes of wheat were transformed by A. tumefaciens carrying beta-1, 3-glucanase gene. After vernalization, the seeds to be transformed were germinated. When these seedlings grew up to 2 approximately 5 cm, their coleoptile and half of the cotyledon were cut out, and the basal portions were infected by A. tumefaciens. A total 27 T(0) transgenic plants were obtained, and the average transformation efficiency was as high as 9.82%. Evident segregation occurred in some of the T(1) plants, as was indicated by PCR and Southern blotting analysis. Investigation of the T(2) plants revealed that some transformed plants had higher resistance to powdery mildew than the controls. Northern blotting revealed that beta-1, 3-glucanase gene was normally expressed in the T(2) plants, which showed an increased resistance to powdery mildew. The results above indicate that the exogenous gene has been successfully integrated into the genome of wheat, transmitted and expressed in the transgenic progeny. From all the results above, it can be concluded that Agrobacterium inoculum to the basal portion of wheat seedling is a highly efficient and dependable transformation method. It can be developed into a practicable method for transfer of target gene into wheat.

摘要

为提高根癌农杆菌介导的小麦(普通小麦)转化效率,我们探索了利用小麦幼苗基部(茎尖分生组织)作为外植体的可能性。携带β-1,3-葡聚糖酶基因的根癌农杆菌对三种基因型的小麦进行了转化。种子经过春化处理后萌发。当这些幼苗长到约2至5厘米时,切除其胚芽鞘和一半子叶,然后用根癌农杆菌感染基部。共获得了27株T(0)转基因植株,平均转化效率高达9.82%。PCR和Southern杂交分析表明,部分T(1)植株出现了明显的分离现象。对T(2)植株的研究发现,一些转基因植株对白粉病的抗性高于对照。Northern杂交显示,β-1,3-葡聚糖酶基因在T(2)植株中正常表达,这些植株对白粉病的抗性增强。上述结果表明,外源基因已成功整合到小麦基因组中,并在转基因后代中得以传递和表达。综合以上所有结果,可以得出结论:对小麦幼苗基部接种根癌农杆菌是一种高效且可靠的转化方法。它可发展成为一种将目标基因导入小麦的实用方法。

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