Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia.
Appl Biochem Biotechnol. 2012 Feb;166(4):1008-19. doi: 10.1007/s12010-011-9489-3. Epub 2011 Dec 20.
The possibility of controlling Fusarium wilt--caused by Fusarium oxysporum sp. cubensec (race 4)--was investigated by genetic engineering of banana plants for constitutive expression of rice thaumatin-like protein (tlp) gene. Transgene was introduced to cauliflower-like bodies' cluster, induced from meristemic parts of male inflorescences, using particle bombardment with plasmid carrying a rice tlp gene driving by the CaMV 35S promoter. Hygromycin B was used as the selection reagent. The presence and integration of rice tlp gene in genomic DNA confirmed by PCR and Southern blot analyses. RT-PCR revealed the expression of transgene in leaf and root tissues in transformants. Bioassay of transgenic banana plants challenged with Fusarium wilt pathogen showed that expression of TLP enhanced resistance to F. oxysporum sp. cubensec (race 4) compared to control plants.
通过对香蕉植株进行遗传工程改造，使其组成型表达水稻硫素蛋白（tlp）基因，来研究由尖孢镰刀菌古巴专化型（race 4）引起的枯萎病的控制可能性。利用携带 CaMV 35S 启动子驱动的水稻 tlp 基因的质粒，通过粒子轰击将转基因导入由雄性花序分生组织诱导的菜花状体簇中。潮霉素 B 被用作选择试剂。通过 PCR 和 Southern blot 分析证实了水稻 tlp 基因在基因组 DNA 中的存在和整合。RT-PCR 显示转基因在转化体的叶片和根组织中的表达。对转基因香蕉植株进行枯萎病病原菌生物测定表明，与对照植株相比，TLP 的表达增强了对尖孢镰刀菌古巴专化型（race 4）的抗性。
