Agrobacterium-mediated transformation of grapefruit with the wild-type and mutant RNA-dependent RNA polymerase genes of Citrus tristeza virus Turunçgil tristeza virüsünün (Citrus tristeza virus) normal ve mutant RNA-baǧımlı RNA polimeraz genlerinin Agrobacterium aracılıǧıyla altıntopa aktarılması

ÇEVİK B., Lee R. F., Niblett C. L.

Turkish Journal of Agriculture and Forestry, cilt.36, sa.2, ss.195-206, 2012 (SCI-Expanded, Scopus, TRDizin) identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 36 Sayı: 2
  • Basım Tarihi: 2012
  • Doi Numarası: 10.3906/tar-1008-1192
  • Dergi Adı: Turkish Journal of Agriculture and Forestry
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.195-206
  • Anahtar Kelimeler: Citrus tristeza virus, Plant transformation, Replicase-mediated resistance, RNA-dependent RNA polymerase
  • Isparta Uygulamalı Bilimler Üniversitesi Adresli: Hayır

Özet

Citrus paradisi Macfad. 'Duncan' was transformed with constructs coding for the wild-type and mutant RNAdependent RNA polymerase (RdRp) of Citrus tristeza virus (CTV) for exploring replicase-mediated pathogen-derived resistance (RM-PDR). The RdRp gene was amplified from a CTV genome and used to generate the wild-type and 2 mutant RdRp constructs for plant transformation. One mutant had the key amino acids GDD changed to AAA (RdRpmGDD), and the second mutant had a deletion encompassing the GDD domain (RdRp-ΔGDD). Etiolated epicotyl segments of Duncan grapefruit (Citrus paradisi Macfad. 'Duncan') were transformed with each of these constructs using the Agrobacterium-mediated transformation method. From 4540 transformed epicotyl segments, 1402 kanamycinresistant shoots were regenerated. Aft er testing for expression of green fluorescent protein (GFP) and β-glucuronidase (GUS) reporter genes by fluorescence microscopy and histochemical staining, respectively, 146 GUS-positive plants were rooted and 97 surviving plants were established in soil in pots. A total of 70 plants were tested for the presence of the GUS gene and CTV RdRp transgenes by polymerase chain reaction (PCR). A total of 51 GUS and CTV transgenepositive transgenic plants (15 with RdRp, 21 with RdRp-mGDD, and 15 with RdRp-ΔGDD) were identified. © TÜBİTAK.