采用农杆菌介导的遗传转化方法,将来自枯草杆菌的果聚糖蔗糖转移酶基因(SacB)导入银腺杨,以提高杨树对水分胁迫的抗性。以来自无菌培养的叶片为外植体,通过大约10 0 0个叶盘与农杆菌LBA4 4 0 4共培养,将植物双元表达载体pKP中SacB基因导入银腺杨基因组,经卡那霉素筛选后,共获得10 2株卡那霉素抗性植株。经PCR特异性扩增和Southern点杂交分析,证明其中97株再生植株基因组DNA中整合了SacB基因。对其中的6 2个无性系进行RT PCR分析,结果表明SacB基因在其中的5 0个无性系中获得表达。温室生长观察表明,转基因无性系外部形态与对照相比没有稳定的显著差异,少数部分转基因无性系的生长明显受到抑制,其他转基因无性系生长正常。这些转基因无性系的获得为培育抗旱转基因杨树奠定了基础。 Agrobacterium-mediated genetic transformation was used to transfer Saccharomyces cerevisiae sucrose transferase (SacB) from Bacillus subtilis into P. gossypii to increase its resistance to water stress. The leaves from the aseptic culture were used as explants. The SacB gene was introduced into the genome of the binary vector pKP by co-cultivation with Agrobacterium tumefaciens LBA4404 by about 100 leaf discs. After screening, a total of 102 kanamycin-resistant plants were obtained. PCR-specific amplification and Southern blot analysis showed that SacB gene was integrated into the genomic DNA of 97 plants. RT PCR analysis of 6 2 of them showed that the SacB gene was expressed in 50 of them. The greenhouse growth observation showed that there was no stable and significant difference between the external morphology of transgenic clones and the control, the growth of a few transgenic clones was obviously inhibited, and the growth of other transgenic clones was normal. The acquisition of these transgenic clones laid the foundation for the cultivation of drought-resistant transgenic poplars.