NDLI: Production of Hevea brasiliensis transgenic embryogenic callus lines by Agrobacterium tumefaciens: roles of calcium

Content Provider	Springer Nature Link
Author	Montoro, P. Rattana, W. Pujade Renaud, V. Michaux Ferrière, N. Monkolsook, Y. Kanthapura, R. Adunsadthapong, S.
Copyright Year	2003
Abstract	A procedure has been established for Agrobacterium tumefaciens-mediated genetic transformation of Hevea brasiliensis embryogenic friable calli. Precultivation of tissues on a CaCl2-free maintenance medium dramatically enhanced the transient activity of the reporter gene, gusA encoding β-glucuronidase (GUS). The increase was first noticed in highly active cells (undifferentiated or/and embryogenic), in tissues precultured for 2–8 weeks. Beyond 8 weeks of preculture, GUS activity increased again, but this time in tissues consisting of differentiated cells accumulating polyphenols. Out of five Agrobacterium strains cocultivated with CaCl2-free precultured tissues, only inoculation with EHA105pC2301 led to high transient GUS activity. Paromomycin proved more effective than kanamycin for the selection of transformed cells, as it inhibits the growth of non-transformed cells more radically. Five paromomycin-resistant callus lines were established. The presence of gusA and neomycin phosphotransferase (nptII) genes in the plant genome was confirmed by DNA amplification, and by Southern hybridization. These results confirmed that A. tumefaciens is an effective system for mediating stable transformation of rubber tree calli with a low copy number of transgenes. Transgenic callus lines constitute a useful tool for studying genes of interest on a cellular level and for regenerating transgenic rubber trees.
Starting Page	1095
Ending Page	1102
Page Count	8
File Format	PDF
ISSN	07217714
Journal	Plant Cell Reports
Volume Number	21
Issue Number	11
e-ISSN	1432203X
Language	English
Publisher	Springer-Verlag
Publisher Date	2003-04-12
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Agrobacterium tumefaciens Calcium Genetic transformation Hevea brasiliensis Paromomycin
Content Type	Text
Resource Type	Article
Subject	Plant Science Medicine Agronomy and Crop Science

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