Abstract:

The vector pDM805 which harbors the selection gene bar and reporter gene gus was

transformed into embryogenic callus lines of ryegrass. In total, 52 individual

regenerated plants were obtained from bialaphos resistant callus lines. Among them, 27

were confirmed by bar PCR analysis; 15 of the 27 PCR positive plants showed strong blue

color with GUS staining. The results suggested that through optimizing callus induction

and plant regeneration conditions, an efficient Agrobacterium mediated transformation

protocol was established in ryegrass, which paves the way for its genetic improvement

through genetic engineering in the near future.

Key words: perennial ryegrass, Agrobacterium mediated transformation, GUS staining, PCR detection;Basta selection

摘要：

以胚性愈伤组织系,作为转化受体,利用农杆菌介导的遗传转化方法,将报告基因gus和抗除草剂基因bar

成功转入多年生黑麦草。共计获得52株T0代抗性植株。利用PCR方法对转基因植株的bar基因进行扩增,

确定阳性株系27株。对阳性植株叶片进行GUS染色,有15株叶片呈现较强的蓝色。对再生植株和野生型

植株叶片喷施5‰的除草剂Basta溶液,GUS阳性的植株全部表现为Basta抗性。以上结果表明,以黑麦草

成熟胚胚性愈伤组织为受体,通过优化农杆菌介导的黑麦草遗传转化体系,初步建立了多年生黑麦草的

转基因平台,这对黑麦草的遗传改良具有重要的意义。

关键词: 多年生黑麦草；农杆菌介导转化；GUS染色；PCR检测；Basta筛选