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期 刊 ：植物遗传资源学报 2014年 15卷 3期

关键词：大蒜；基因枪；转化；潮霉素；GUS；

Keywords:garlic(Allium sativum L), Particle bombardment, transformation, hygromycin phosphotransferase (HPT), GUS,

摘 要 ：以大蒜无菌苗根诱导的愈伤组织为受体进行基因枪转化，根据愈伤的形态特征设计了6组基因枪转化参数进行试验。结果表明，其中4组转化试验都获得了转基因植株；轰击距离9cm、压力1100 psi 和轰击距离12.5cm、压力1300 psi各轰击1次的轰击方式获得了最高的转化效率，为2.4%。对抗性植株进行了 PCR 和 Southern blot 检测，证明pCAMBIA1301载体上的潮霉素抗性选择标记基因和gus 报告基因已整合到抗性植株的基因组中。对转化后的愈伤、胚状体、根、再生芽尖和叶片进行gus基因的表达检测，抗性材料显示出明显的蓝色反应。这些结果表明我们初步建立了基因枪介导的大蒜的遗传转化体系，将为大蒜的基因工程改良奠定基础。

Abstract:Using the calli induced from the root segments of in vitro plantlets as the recipients, six models of particle bombardment were designed to develop a biolistic transformation procedure of garlic (Allium sativum. L ).Transgenic garlic plantlets have been recovered in four models of particle bombardment. The highest transformation frequency of 28.3% had been observed when bombarding calli twice at shoot distance 9cm with pressure 1300 psi helium. But the highest regeneration efficiency of 2.4% were obtained when respectively bombarding calli once at shoot distance 9cm with pressure 1100 psi and at shoot distance 12.5cm with pressure 1300 psi helium. The resistant plantlets were analyzed by PCR and Southern blot. The results showed that hygromycin phosphotransferase (HPT) selectable marker gene and β-glucuronidase (gus) reporter gene had been integrated in the genome. GUS expression assay of bombarded calli, somatic embryos, proembryos or young embryos, embryogenic roots, regenerated shoots and leafs showed that resistant materials distinctly demonstrated blue. So we established an effective biolistic transformation procedure for garlic whicn can be used to generate transgenic garlic plants with other agronomical beneficial traits.

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