Vakgroep Moleculaire Genetica, Departement Plantengenetica, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium.
Mol Plant Pathol. 2000 Nov 1;1(6):383-7. doi: 10.1046/j.1364-3703.2000.00043.x.
Summary A fast plant promoter test was developed by means of a nematode to transfer Agrobacterium tumefaciens into plant roots. Two-week-old Arabidopsis thaliana (L.) Heynh. plants were transferred to infection medium. Meloidogyne incognita or Heterodera schachtii juveniles were mixed with the Agrobacterium strain that harboured the binary vector, and this mixture was used for plant inoculation. During migration of the nematode and establishment of the feeding site inside the roots, the T-DNA was delivered into the root cells. A few days later, the infected plants could be analysed for expression of the T-DNA reporter gene in and around the nematode feeding sites (NFS), without the need to go first through the whole transformation and regeneration procedure. Depending on the construct, expression of the beta-glucuronidase gene in the NFS or along the migration path of the nematode could be seen in the roots of Arabidopsis plants. Furthermore, stably transformed plants could be regenerated from the infected roots.
摘要 通过线虫将根瘤农杆菌转移到植物根部,开发了一种快速的植物启动子检测方法。将两周大的拟南芥(L.)Heynh.植物转移到感染培养基中。将携带二元载体的根瘤农杆菌与Meloidogyne incognita 或 Heterodera schachtii 幼虫混合,并用此混合物进行植物接种。在线虫迁移和在根内建立取食部位的过程中,T-DNA 被递送到根细胞中。几天后,无需先进行整个转化和再生过程,就可以在线虫取食部位(NFS)内和周围分析感染植物 T-DNA 报告基因的表达情况。根据构建体的不同,可以在拟南芥植物的根部看到 NFS 或线虫迁移路径中β-葡聚糖酶基因的表达。此外,还可以从感染的根中再生出稳定转化的植物。