Genetic transformation of lime (Citrus aurantifolia Swing.): factors affecting transformation and regeneration.

Abstract:

:We have previously developed procedures for the efficient production of sweet orange (Citrus sinensis L. Osbeck) and Carrizo citrange (C. sinensis L. Osbeck×Poncirus trifoliata L. Raf.) transgenic plants using an Agrobacterium tumefaciens-mediated transformation and shoot tip grafting in vitro regeneration system. We now report on the optimization of the cocultivation, regeneration and selection conditions for efficient and reliable production of transgenic lime (C. aurantifolia Swing.) plants. Improved transformation frequencies were obtained by cocultivating the explants with Agrobacterium on feeder plates. Optimum regeneration of transgenic shoots was obtained by exposing the explants to darkness for 2 weeks and by using kanamycin at 100 mg/l as selective agent. Attempts to use geneticin as selection antibiotic were not successful. Shoot tip grafting of regenerated shoots on Troyer citrange seedlings resulted in 100% successful production of transgenic plants. The presence and expression of the transferred genes in the regenerated plants was verified by β-glucuronidase histochemical and fluorimetric assays, neomycin phosphotransferase ELISA assays, PCR and Southern analyses.

journal_name

Plant Cell Rep

journal_title

Plant cell reports

authors

Peña L,Cervera M,Juárez J,Navarro A,Pina JA,Navarro L

doi

10.1007/s002990050311

subject

Has Abstract

pub_date

1997-09-01 00:00:00

pages

731-737

issue

11

eissn

0721-7714

issn

1432-203X

pii

10.1007/s002990050311

journal_volume

16

pub_type

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