Kamo Kathryn, Gera Abed, Cohen Jacob, Hammond John, Blowers Alan, Smith Franzine, Van Eck Joyce
Floral and Nursery Plants Research Unit, USDA, U.S. National Arboretum, Beltsville, MD 20705, USA.
Plant Cell Rep. 2005 Feb;23(9):654-63. doi: 10.1007/s00299-004-0888-6. Epub 2004 Oct 9.
Transgenic Gladiolus plants transformed with the bean yellow mosaic virus (BYMV) coat-protein (CP) gene in either sense or antisense (AS) orientation were developed using biolistics. Four of the plants were confirmed to carry the CP gene in the sense orientation of the gene and seven plants in the AS orientation. Two of the CP plant lines and all of the AS lines showed DNA rearrangements of the transgene in addition to an intact copy of the transgene. The copy number ranged from one to nine. Of the 11 lines, eight had only one to four copies of the transgene. Transcription of the transgene occurred for three of the CP lines and five of the AS lines as determined by Northern hybridization. All 11 plant lines were challenged with BYMV using controlled aphid transmission. One month following aphid transmission, the transgenic plants were examined by immunoelectron microscopy for presence of the virus. Several transgenic plant lines containing either antiviral transgene showed a lower incidence of infection (percentage of plants infected as detected by immunoelectron microscopy) than the non-transformed plants. Most of the CP- and AS-transgenic plants that did not contain BYMV 1 month after challenge were found to contain BYMV the next season. It appeared that BYMV infection was delayed in the CP- and AS-transgenic lines but that the transgenes did not prevent eventual infection of BYMV. This is the first report of developing a floral bulb crop with antiviral genes to BYMV.
利用生物弹道技术培育出了转有菜豆黄花叶病毒(BYMV)外壳蛋白(CP)基因的转基因唐菖蒲植株,该基因以正义或反义(AS)方向转入。其中4株植株被证实携带正义方向的CP基因,7株携带反义方向的CP基因。除了转基因的完整拷贝外,2个CP植株系和所有AS植株系均显示出转基因的DNA重排。拷贝数从1到9不等。在这11个株系中,有8个株系只有1到4个转基因拷贝。通过Northern杂交确定,3个CP株系和5个AS株系发生了转基因转录。使用控制蚜虫传播的方法,用BYMV对所有11个植株系进行了挑战。蚜虫传播1个月后,通过免疫电子显微镜检查转基因植株是否存在病毒。与未转化植株相比,几个含有抗病毒转基因的转基因植株系感染率较低(通过免疫电子显微镜检测到的感染植株百分比)。大多数在接种后1个月未感染BYMV的CP和AS转基因植株在下一季被发现感染了BYMV。似乎在CP和AS转基因株系中BYMV感染有所延迟,但转基因并不能阻止最终感染BYMV。这是关于培育带有抗BYMV病毒基因的花卉种球作物的首次报道。