Mullins K V, Llewellyn D J, Hartney V J, Strauss S, Dennis E S
CSIRO Forestry and Forest Products, P.O. Box 4008 Queen Victoria Terrace, ACT 2600, Australia, , , , , , AU.
CSIRO Plant Industry, P.O. Box 1600, Canberra City, ACT 2601, Australia, , , , , , AU.
Plant Cell Rep. 1997 Sep;16(11):787-791. doi: 10.1007/s002990050321.
Reliable regeneration protocols for Eucalyptus camaldulensis using leaf explants from in vitro-grown plants have been developed. Out of the 24 clones tested 13 were regenerated and of these, 6 showed regeneration from more than 60% of the explants. Identical protocols were also successful in the regeneration of some clones of E. microtheca, E. ochrophloia, E. grandis and E. marginata, but at lower frequencies. Co-cultivation of E. camaldulensis leaf explants with Agrobacterium tumefaciens strains carrying a kanamycin resistance gene and the reporter gene β-glucuronidase (GUS), followed by selection on kanamycin at 9 mg l, allowed the selection of transformed shoots that could be rooted on selective media. Transformation of the plants was verified by staining for the GUS enzyme in various plant tissues, NptII assays and by Southern blotting on isolated DNA using specific probes for both the GUS and selectable marker genes. Transformed tissue was obtained with 5 clones of E. camaldulensis tested and a number of A. tumefaciens strains. However, only 1 clone regenerated transformed whole plants reliably.
已经开发出了使用体外培养植物的叶片外植体对赤桉进行可靠再生的方案。在测试的24个克隆中，有13个实现了再生，其中6个从超过60%的外植体中再生。相同的方案在微叶桉、褐皮桉、巨桉和边缘桉的一些克隆的再生中也取得了成功，但频率较低。将携带卡那霉素抗性基因和报告基因β-葡萄糖醛酸酶（GUS）的根癌农杆菌菌株与赤桉叶片外植体共培养，然后在9 mg/l的卡那霉素上进行选择，从而筛选出能够在选择培养基上生根的转化芽。通过对各种植物组织中的GUS酶进行染色、NptII检测以及使用针对GUS和选择标记基因的特异性探针在分离的DNA上进行Southern印迹分析，验证了植物的转化情况。对测试的5个赤桉克隆和多个根癌农杆菌菌株获得了转化组织。然而，只有1个克隆可靠地再生出了转化的完整植株。
